### OECD SIDS METHANOL

1 

# SIDS Initial Assessment Report

For

SIAM 19

Berlin, Germany , 18

- 20 October, 2004

1. Chemical Name: Methanol

2. CAS Number: 67

- 56

- 1

3. Sponsor Country: United States

4. Shared Partnership with: BASF AG, Germany, Methanol Institute U. S. A., Methanor v. o

. f., Netherlands, Shell Chemicals Ltd., U. K., Statoil ASA,

Norway, TOTAL Raffinerie Mitteldeutschland GmbH,

Germany, Veba Oil Refining & Petrochemicals GmbH, Germany

5. Roles/Responsibilities of

the Partners: BASF

– Initial preparer of documents.

U.S. EPA

– reviewer and editor of documents, presenter at

SIAM

• 

### Name of industry sponsor

/consortium BASF AG, Germany

Contact person:

Dr. Hubert Lendle

GUP/CL

– Z 570

D

- 67056 Ludwigshafen

• 

### Process used The documents were written by BASF AG, Germany and edited

by U.S. EPA

6. Sponsorship History

• 

### How was the chemical or

category brought into the

OECD HPV Chemicals

Programme ? ICCA Initiative

7. Review Process Prior to

the SIAM: An iterative process was used by which BASF first prepared a

draft based on the U .S. EPA HPV Program submission from the

Methanol Institute Testing Group. U.S. EPA reviewed the draft

and provided comments to BASF. U.S. EPA also edited later

drafts of the documents and inserted relevant information into

the documents. Meetings between U.S. EPA and BASF were

also used to resolve issues.

8. Quality check process: Within the review process, data in the SIAR were compared with

data in the SIDS Dossier and for selected endpoints a

comparison with original studies was done.

U.S. EPA checke d the consistency among the documents.

9. Date of Submission: July 26, 2004

10. Date of last Update: July 26, 2004

11. Comments: In a separate evaluation, the U.S. EPA is also assessing the

hazards of methanol to obtain an Agency

- wide consensus

evalua tion for the Integrated Risk Information System (IRIS). As

### OECD SIDS METHANOL

2 

### part of the IRIS assessment, EPA will review pharmacokinetic

models to determine the best method of extrapolating animal

data to humans; this work will assist in determining the

relevance of the ef fects observed in animals to humans. As part

of the IRIS assessment, an independent Science Advisory Board,

consisting of scientific experts from several external

organizations, will review the evaluation.

### OECD SIDS METHANOL

3 

# SIDS INITIAL ASSESSMENT PROFILE

CAS No. 67

- 56

- 1

Chemical Name Methanol

Structural Formula CH 3 

### OH

SUMMARY CONCLUSIONS OF THE SIAR

Human Health

Toxicokinetics, Metabolism, and Distribution: Methanol is readily absorbed by inhalation, ingestion and

dermal contact and distributes rapidly thro ughout the body. Metabolism in humans, rodents, and monkeys

contributes up to 98 percent of the clearance, with more than 90 percent of the administered dose exhaled as

carbon dioxide. Renal and pulmonary excretion contributes only about 2

– 3 percent. The metabolism and

toxicokinetics of methanol varies by species and dose. In humans, t he half

- life is approximately 2.5

– 3 hours at

doses lower than 100 mg/kg bw. At higher doses, the half life can be 24 hours or more.

The mammalian metabolism of metha nol occurs mainly in the liver, where methanol is initially converted to

formaldehyde, which is in turn converted to formate. Formate is converted to carbon dioxide and water. In

humans and monkeys, the conversion to formaldehyde is mediated by alcohol d ehydrogenase. In rodents, the

first reaction occurs mainly via a catalase

- peroxidase pathway which is rate

- limiting. Methanol in turn

accumulates in the blood at high doses. In primates, the last reaction step, conversion of formate to carbon

dioxide by the formyl

- tetrahydrofolate synthetase, is rate

- limiting, leading to a disproportionate increase of

formate in the blood and sensitive target tissues (such as CNS and the retina).

In humans, when exposed via inhalation up to an air concentration of 0.065 mg/L, no increase of blood methanol

is expected. Up to 0.26 mg/L (single or repeated exposure) , the methanol blood level is likely to increase 2 to 4

- fold above the endogenous methanol concentration in humans, but still remains significantly below 10 mg/ L. Air

concentrations up to 1.6 mg/L resulted in similar blood methanol among rats, monkeys, and humans. However,

above 1.6 mg/L, a steep exponential increase occurs in rats, a smaller exponential increase occurs in monkeys,

and humans exhibit a linear r elationship between air concentrations and blood methanol levels.

Baseline levels of formate in blood are about 3 to 19 mg/L (0.07

– 0.4 mM) in humans. Toxic blood formate

concentrations are reported to be 220 mg/L and higher ( > 5 mM formate). Inhalation of about 1.20 mg

methanol/L for 2.5 hours contributed only insignificantly to the internal formate pool in monkeys (in the μM

range); this also held true for folate

- deficient conditions. Also a fter repeated inhalation of 2.6 mg/L (2000 ml/m 3 )

for 6 hrs/d ay, 5 days/week, for 1 or 2 weeks, monkeys showed no discernible increase in formate concentration in

blood [estimated body burden 200 to 300 mg/(kg bw

* d)], whereas formate accumulation has been observed in

primates at methanol doses greater than 500 mg/kg bw given as a bolus. The critical methanol dose that saturates

the folate pathway in humans is estimated to be ≥ 200 mg/kg bw. Based on data produced in monkeys, metabolic

saturation in humans is also less likely to happen during inhalation if the amount is divided over a prolonged time

and not incorporated as a bolus.

Acute Toxicity: Formate is considered to be the ultimate toxicant in acute methanol intoxication in humans.

Acidosis and ophthalmologic changes are typical primary effects. However, thes e effects do not occur in rodents

or rabbits., which are able to remove formate more efficiently. In these animals, CNS depression is usually the

cause of defects and finally death.

A blood level of 500 mg methanol/L in acutely poisoned patients generally is regarded as requiring hemodialysis.

This blood concentration can transiently be achieved in an adult person (70 kg) by ingestion of 0.4 ml

methanol/kg (approximately 0.32 mg/kg). Generally, in humans, transient central nervous system (CNS) effects

app ear above blood methanol levels of 200 mg/L and serious ocular symptoms appear above 500 mg/L.

The minimal acute methanol dose to humans that can result in death is considered to be 300 to 1000 mg/kg by

### OECD SIDS METHANOL

4 ingestion, and fatalities have occurred in untreate d patients with initial methanol blood levels in the range of

1500

- 2000 mg/L. However, such high blood methanol levels able to cause death are not .likely to be achieved

through inhalation exposure. For example, 2.6 or 6.5 mg/L resulted in methanol blood levels that barely exceed

100 and 200 mg/L, respectively, after an 8

- hour working shift. Exposure to 0.26 mg methanol/L for 4 hours was

without significant physiologic effects in human volunteers.

In rats, LC 50 values have been calculated to be 83.2 and 128.8 mg/L after 4 hours. In cats, the LC 50 was 85.4

mg/L after four hours. In monkeys, air concentrations of 52 mg/L after 1

- 4 hours and 13 mg/L after 18 hours led

to an unspecified level of mortality. Dermal LD 50 s in rabbits range from 15800 to 2000 0 mg/kg bw. In rats, the

dermal LD 50 is greater than 45,000 mg/kg bw. In monkeys, four daily dermal doses of 400 mg/kg bw eventually

resulted in death. Oral LD 50 s in rats range from < 790 to 13,000 mg/kg bw; in mice, the values range from 7,300

to 10,00 0 mg/kg bw; in rabbits, the LD 50 was approximately 14,200 to 14,400 mg/kg bw; and in monkeys, the

values range from 7000 to 9000 mg/kg. Although most of the references for these values provided only limited

details, the values are consistent within species and route of exposure.

Irritation/Sensitisation: Methanol exhibited no skin irritation in one study in which the chemical was

administered under occluded conditions as a 100 percent concentration. Limited information from another study

showed moderate irritation with 500 mg. Available studies show that methanol is a slight eye irritant with doses

of 0.05 to 0.1 mL. Some reversibility of effects was observed. High concentration of methanol vapours may be

irritating to mucous membranes. A guinea pig m aximization assay gave no evidence of contact sensitisation after

induction and challenge doses of 50 percent.

Repeated

- Dose Toxicity: In a whole body inhalation study in monkeys exposed to 0.013, 0.13, and 1.3 mg/L 21

hours per day 7 days per week for 7 , 19, and 29 months, several general clinical signs as well as degenerative

effects in the brain (at 0.13 and 1.3 mg/L), slight peripheral nerve damage (at 0.13 and 1.3 mg/L), very slight

degeneration of the optic nerve (concentrations not noted), increase d fat granules and slight fibrosis in the liver

(all concentrations), and Sudan positive granules in the kidney were observed (at 0.13 and 1.3 mg/L). Also, a

slight myocardial disorder (at 0.13 and 1.3 mg/L) and localized effects in the trachea and possibl e slight fibrosis

in the lungs (concentrations not noted) were observed. Although the statistical significance of the effects cannot

be verified from the study report, the number of effects and systems affected indicate a relationship with

methanol.

In another whole body inhalation study in monkeys exposed up to 20 days for 21 hours per day, coma and

lethality were observed at concentrations > 9.1 mg/(L

* d). In the brain, necrosis of the basal ganglia and cerebral

edema were observed at 6.5 mg/(L

* d) and at 3.9 mg/(L

* d), hyperplasia and fibrosis around myelin sheaths of the

basal ganglia as well as a slight to moderate increase in astroglia cells were observed. The optic nerve showed

atrophy at >3.9 mg/(L

* d), along with reduction in myelin fibers. In the liver, fibrosis was observed at 6.5

mg/(L

* d) and mild fatty degeneration was observed at 3.9 mg/(L

* d). In the kidney, partly vacuolated hyaline

degeneration was observed at 6.5 mg/(L

* d).

In rats exposed to methanol up to 6.5 mg/L for 6 hours per day, five days per week for 28 days, no adverse effects

were observed except local nasal irritation and increased relative spleen weights, which were observed only at the

middle dose. The estimated blood level of methanol was about 250 mg/L under this conditio n.

In a whole body inhalation study in mice exposed for 12 months to concentrations of 0.013, 0.13, and 1.3 mg/ L

20 hours per day, slight changes in clinical signs, body and organ weights, and some changes in histopathology

were observed. In rats exposed in the same manner, slig ht changes in body weight and organ weights were

observed at the highest dose. The NOEL is 0.13 mg/L. In rats, gavage doses of 100, 500, and 2,500 mg/(kg

bw

* d) for 90 days resulted in increased liver enzymes and reduced brain wei ghts at the highest dose resulting in a

NOAEL of 500 mg/(kg bw

* d).

Mutagenicity: Numerous in vitro assays (including seven Ames assays, four micronucleus/cytogenetic assays,

a mammalian gene mutation assay, a yeast gene mutation assay, a mouse lymphoma t est, three cell

transformation assays, and a DNA damage and repair assay) were conducted on methanol. The majority of these

assays are negative, with the exception of positive results in the mouse lymphoma test and a mitotic

recombination assay in Aspergi llus, and ambiguous results in an Ames assay for strain TA102 and in the DNA

damage and repair assay. Only limited details were available for the mouse lymphoma test. Of the eleven in

vivo assays (all micronucleus and cytogenicity assays plus a Drosophil a SLRL assay), all are negative except

one cytogenetic assay, which was positive for aneuploidy, sister chromatid exchange, and micronuclei. Limited

information was available regarding this positive result. Thus, although most studies indicate that metha nol does

not have the potential to be mutagenic, not all studies lead to that conclusion.

### OECD SIDS METHANOL

5 Carcinogenicity: Methanol was tested in two long

- term whole body inhalation studies (24 months in rats and 18

months in mice for 20 and 19 hours per day, respectiv ely). There was no evidence of a carcinogenic potential in

rats and mice exposed to air concentrations of up to 1.3 mg/L.

Reproductive/Developmental Toxicity: Monkeys. In monkeys, parents were exposed via inhalation prior to

and during breeding as wel l as during pregnancy to doses of 0.26, 0.78, and 2.34 mg/L. A late wasting syndrome

was observed at the highest dose in 2 out of 7 female descendants, associated with signs of severe malnutrition

and gastroenteritis noted at autopsy . Mild neurobehaviora l effects in offspring and some vaginal bleeding in

mothers were observed at all concentrations. However, due to the normal variance in and the low number of

animals, along with pre

- term delivery of several mothers, the observed findings are difficult to interpret.

Rats. Several inhalation studies in rats resulted in a variety of effects in offspring due to prenatal and/or postnatal

dosing. In a 2

- generation whole body inhalation reproductive study in which rats were exposed for 19

- 20

hours/day, decreas ed brain weights in the first and second generation offspring (F1, F2) resulted in a NOAEL of

0.13 mg/L. In a developmental study in which rats were exposed by whole body inhalation on gestation days 1 to

19 at the two lowest doses and days 7 to 15 at the highest dose for 7 hours/day, malformations and fetal weight

changes resulted in a NOAEL of 6.5 mg/(L

* d) (the lowest dose tested). A second whole

- body inhalation

developmental study in which rats were exposed on gestation days 7 to 17 for 23 hours/day, m alformations,

increased fetal resorptions, and decreased numbers of live fetuses were observed, resulting in a NOAEL of 1.3

mg/(L

* d).

Mice. In a study of reproductive effects, there was an insignificant increase in morphological anomalies in

spermatozoa in male mice at 1000 mg/(kg bw

* d) after oral dosing for five weeks. A developmental whole body

inhalation study in mice exposed on gestation days 6 to 15 for 7 hrs/day resulted in developmental effects

including increased exencephaly and cleft palate, ful ly resorbed fetuses, decreased numbers of live pups, and

decreased body weights; this study resulted in a NOAEL of 1.3 mg/(L

* d). Oral studies in mice resulted in various

malformations at 4000 mg/(kg bw

* d) (the LOAEL) and higher; no NOAELs could be establi shed from these

studies.

Humans. No epidemiological studies in humans have been located to demonstrate that there is a link between

methanol exposure and an increased incidence of fetal malformations or developmental impairment.

Rodent data on reproduc tive and developmental toxicity are relevant for humans despite the known differences in

methanol metabolism between rodents and humans. Rodents are adequate models for human exposure to

methanol at levels where formate does not accumulate. However, bloo d methanol concentrations associated with

serious teratogenic effects and reproductive toxicity observed in the rodent studies are in the range of 1000

– 2000 mg methanol/L blood, associated with formate accumulation in humans

, which is likely to result in

metabolic acidosis and visual and clinical effects in humans. In humans, g enerally, transient central nervous

system (CNS) effects appear above blood methanol levels of 200 mg/L, ocular symptoms appear above 500

mg/L, and fatalities have often occurred i n untreated patients with initial methanol levels in the range of 1500

- 2000 mg/L. Other effects (e.g., subtle, not yet definitive neurological effects observed in primates) may be

exhibited at lower inhalation concentrations and lower methanol blood levels

.

Environment

Methanol is a colorless, clear, highly flammable liquid with a mild alcoholic odor. Physicochemical property

values for methanol are: melting point

– 97.8 0 C, boiling point 65 0 C, density 0.79 g/m 3 , vapor pressure 128 hPa,

and log Kow

– 0 .74. Methanol is miscible with water at 20 ° C. It does not undergo hydrolysis. A value of 0.461

Pa m3/mol for the Henry’s Law constant indicates that volatilization is not a significant removal process from

the aquatic compartment.

Air is the main targ et compartment, based on a fugacity model calculation (Mackay Level III) with about 73 %

of environmental methanol distributing to air and 16 % to water. Methanol is degraded in the atmosphere by

photochemical, hydroxyl

- radical dependent reactions. The e stimated elimination half

- life is calculated to be

about 17

- 18 days with a rate constant of 0.93 x 10

- 2 cm 3 /molecule

- sec. Methanol is readily biodegradable.

Bioaccumulation in fish is expected to be low.

In several 96

- hour studies in fish in which met hanol concentrations were measured during the tests, LC50s

ranged from 15,400 to 29,400 mg/L. In Daphnia magna , 24

- hour and 48

- hour EC50s were > 10,000 mg/L

### OECD SIDS METHANOL

6 (highest dose tested). In one 24

- hour study, the actual EC50 value reported in Daphnia magna was 21,400

mg/L. In aquatic plants, an EC50 for growth inhibition of Chlorella pyrenoidosa was calculated to be 28,440

mg/L over 10

- 14 days. No information on analytical monitoring was available for the invertebrate and plant

studies.

The low toxicity of me thanol for aquatic organisms is confirmed by QSAR calculations, resulting in acute

toxicity values for fish, daphnids and algae well above 1000 mg/L.

Exposure

The world

- production volume of methanol is approximately 30 million tonnes annually in 2000, about 11

million tonnes used in countries participating in NAFTA (North American Free Trade Agreement) and about 3.3

million tonnes in Western Europe, of which 70% is being used for the manufacture of formaldehyde, MTBE and

acetic acid. Several additional compounds are synthesized from methanol.

Methanol occurs naturally in humans, animals and plants. The general population is exposed to methanol mainly

through consumption of food and beverages and through use of consumer products such as paints, sealers and

adhesives that contain methanol as a solvent. Worldwide, air concentrations of methanol have been measured at

ranges from 1.05 to 13.1 x 10

- 5 mg/L.

Releases into the environment may occur from both natural and man

- made sources. It is however released

predominantly from production and use in industrial processes. According to the U.S. Toxics Release Inventory,

about 89,225 tonnes of methanol were released on

- and offsite for facilities in all industries in the U.S. in 2000.

The largest emitter was th e pulp and paper industry. About 90 percent of the emissions were to the air and about

1.7 percent was released to surface water.

The most important routes of potential exposure are inhalation and dermal exposure at the workplace.

A mean background bod y burden of 0.5 mg methanol/kg bw has been estimated based on baseline blood levels

and elimination kinetics of methanol.



### RECOMMENDATION AND RATIONALE FOR THE RECOMMENDATION AND

NATURE OF FURTHER WORK RECOMMENDED

Human Health: This chemical is a candid ate for further work. Methanol exhibits potential hazardous properties

for human health (neurological effects, CNS depression, ocular effects, reproductive and developmental effects,

and other organ toxicity). Rapid metabolism and excretion is noted depe nding on the dose. In the US (the

Integrated Risk Information System), further work is being performed regarding the use and refinement of

pharmacokinetic models for extrapolating animal data to humans.

Environment: The chemical is currently of low prio rity for further work, due to its low hazard profile.

### OECD SIDS METHANOL

7 

# SIDS Initial Assessment Report

1 IDENTITY

1.1 Identification of the Substance

CAS Number: 67

- 56

- 1

IUPAC Name: Methanol

Molecular Formula: CH 4 

### O

Structural Formula: H 3 

### C

- OH

Molecular Weight: 32.04

Syn onyms: Methyl alcohol, Methyl hydroxide, Methylol, Monohydroxymethane

1.2 Purity/Impurities/Additives

Purity: ≥ 99.85% (w/w)

Impurities: water < 0.1% (w/w)

chloride ≤ 0.00005% (w/w)

sulphur ≤ 0.00005% (w/w)

iron ≤ 0.00001% (w/w)

ethanol ≤ 0.005% (w/w)

acetone ≤ 0.003% (w/w)

1.3 Physicochemical Properties

Table 1 : Summary of Physicochemical Properties

Property Value Reference

Physical state liquid Merck Index, 1989; BASF, 1999a

Melting point approx.

− 98 °C Merck Index, 1989; BASF, 1999a

Boiling point 65 °C Merck Index, 1989; BASF, 1999a

Relative density 0.79 g/cm 3 (20 °C) Merck Index, 1989; BASF, 1999a

Vapour pressure 127.9 hPa (20 °C) Daubert and Danner, 1989

Water solubility Miscible (at 20 °C) BASF, 1999a

Partition coefficient n

octanol/water (Log Kow) − 0.82 to

− 0.64 Hansch and Leo, 1979;

Rekker, 1977; Sangster, 1989

Henry’s law constant

(25 °C) 0.461 Pa • m 3 /mol Gaffney, 1987

Flash point 10 °C (closed cup) BASF, 1972

Auto

- ignition point 455 °C BASF, 1979



### Methanol is a colourless, clear, volatile, and highly flammable liquid. When pure, it has a mild

alcoholic odour. Methanol is miscible with water at all ratios at ambient temperature, but also

soluble in many organic solvents. Methanol is expected to have no explosive and oxidising

### OECD SIDS METHANOL

8 

### properties, as indicated by its chemical structure. Vapours may form explosive mixture with air

(BASF, 1999b).



## 2 GENERAL INFORMATION ON EXPOSURE

The methanol production process involves the catalytic steam reforming of a hydrocarbon

feedstock, typically natural gas, to give a gaseous mixture of carbon oxides and hydrog en. This

mixture is then compressed and reacted over another catalyst to give methanol and by

- products as

follows:

CO + 2H 2 

### <

- > CH 3 

### OHCO 2 

### + 3 H 2 

### <

- > CH 3 

### OH + H 2 

### O

The pure product is obtained by fractional distillation. All necessary steps are performed in closed

systems. The details of this process are outlined in the following paragraphs.

The gas is reformed over a nickel catalyst. The synthesis gas, containing a mixture of hydrogen,

carbon oxides, steam and residual methane leaves the reformer at about 8 80°C and 20 bar, and is

cooled before being compressed to synthesis pressure.

The synthesis loop comprises a circulator, converter, feed/effluent exchanger, separator, and in

some cases a heat recovery exchanger. The optimum operating pressure is typically about 80 bar for

large plants, but for smaller plants (less than 600 t/day) a pressure of 50 bar may be economical due

to the reduced material requirements. Methanol conversion is at a rate of 5% per pass hence there is

a continual recycling of the unrea cted gases in the synthesis loop.

Synthesis gas is mixed with the unreacted gas and preheated in a feed/effluent interchanger. The

converter effluent is cooled by passing through the feed/effluent interchanger and dependent on the

reactor type, a heat rec overy exchanger. Prior to the catchpot, the gas is cooled to about 40°C to

condense the product methanol. A purge is taken from the unreacted gas to remove inerts (N2,

CH4, argon, and surplus hydrogen or carbon oxides) from the loop. This purge is used as fuel in the

reformer. A purge gas expander can be used to recover power from the purge gas as it is lowered to

the fuel system pressure.

The crude methanol from the separator contains both water and low levels of by

- products, which

must be removed to a chieve required product purity.

2.1 Production Volumes and Use Pattern

In the year 2000, the world

- wide production volume was about 30 million tonnes annually

(estimated capacity approximately 35 million tonnes), which can be split as follows:

### OECD SIDS METHANOL

9 

### Table 2: Produ ction Volumes of Methanol (2000)

Region Production Volume

(in million tonnes)

North America (based on NAFTA 1 ) Production approx. 7

South America (includes Trinidad) approx. 7

Western Europe approx. 3.3

Eastern Europe approx. 0.5

Former S oviet Union approx. 2

Africa and Middle East approx. 5

Asia/Pacific approx. 5

Total approx. 30

1 NAFTA = North American Free Trade Agreement



### Production of methanol is reported to occur at 98 plants world

- wide (Methanol Institute, 2003).

Ap proximately 70 percent of the methanol produced worldwide serves as feedstock for chemical

syntheses, in particular formaldehyde, methyl tert

- butyl ether (MTBE), acetic acid, methyl

methacrylate, and dimethyl terephthalate. Only a small proportion is util ized for energy production,

although this use has great potential. The use of pure methanol as solvent is limited, although it is

often included in solvent mixtures. (Ullmann, 2000). The profile of main uses within NAFTA is

summarized below:

Table 3: Mai n Uses of Methanol in the NAFTA Region

Use Percentage

MTBE/TAME * approx. 36%

Formaldehyde approx. 23%

Acetic Acid approx. 9%

Exports approx. 7%

Solvents approx. 6%

Methyl Methacrylate approx. 3%

DMT * approx. 2%

Gasoline/Fuel approx. 0.2%

Others approx. 14%

DMT = dimethyl terephthalate

TAME = tert. Amylmethylether = 2

- methoxy

- 2

- methylbutane

MTBE = tert

- butylmethyl ether



### MTBE demand is expected to decline in 2004 in the USA because sev eral states will discontinue its

use as a gasoline additive (CMAI, 2001). Methanol is used to synthesize a large number of other

organic compounds (see Table 3 “Others”). These compounds and their uses are identified in Table

4.

### OECD SIDS METHANOL

10 

### Table 4: Additional Comp ounds Synthesized from Methanol and Accompanying Uses

Compound Uses

Formic acid Preservatives, pickling agents

Methyl esters of organic acids Solvents, monomers

Methyl esters of inorganic acids Methylation reagents, explosives, inorganic acids,

insectic ides

Methylamines Pharmaceutical precursors, auxiliaries, absorption liquids

for gas washing and scrubbing

Trimethylphosphine Pharmaceuticals, vitamins, fragrances, fine chemicals

Sodium methoxide Organic intermediate, catalyst

Methyl halides Organic i ntermediates, solvents, propellants

Ethylene Organic intermediates, polymers, auxiliaries



### Likely future uses of methanol include the synthesis of light olefins [methanol

- to

- olefin (MTO)

process], fuel for Otto engines and gas turbines of power stations, and as a fuel additive and

hydrogen carrier for fuel cell applications (CMAI, 2001; Ullmann, 2000; Kavet and Nauss, 1990).

2.2 Environmental Exposure and Fate

2.2.1 Sources of Environmental Exposure

Methanol occurs naturally in humans, animals and plants. In addit ion, methanol is released into the

environment from both natural and man

- made sources, the latter being the most significant. Natural

sources of methanol emissions include volcanic gasses, vegetation, microbes and insects

(IPCS/WHO, 1997). Of the man

- mad e sources, methanol is released predominantly from its

production and use in industrial processes. According to the US Toxics Release Inventory, 89,225

tonnes methanol were released on

- and offsite for facilities in all industries in the U.S. in 2000.

T he largest emitter in 2000 was the pulp and paper industry. About 90% of emissions (79,658

tonnes) were released into the air, and 1.7 % (1531 tonnes) into surface water (U.S. EPA, 2001).

A minor source of environmental exposure may result from use of con sumer products such as

paints, sealers and adhesives that contain methanol as a solvent.

Urban air levels of methanol world wide in the range of 1.05

– 13.1 x 10

- 5 

### mg/L have been reported.

In Sweden, methanol was detected at concentrations of 59

– 940 10

- 5 

### mg/L at dense traffic sites in

Stockholm and 0.5

– 3.0 x 10

- 5 

### mg/L at five other sites in and around Stockholm. In three US

locations (Boston, MA; Houston, TX; and Lima, OH), median methanol levels of 0.6

– 6.0 x 10

- 5



### mg/L were found in 52 samples (US EPA, 1993). Methanol was also detected at air concentrations

of 0.06

– 0.18 x 10

- 5 

### mg/L in Point Barrow, Alaska (IPCS/WHO, 1997).

2.2.2 Photodegradation

Methanol is degraded in the atmosphere by photochemical, hydroxyl

- radical dependent reactions,

with a rate c onstant of 0.932 x 10

- 12 

### cm3/molecule

. 

### sec (Atkinson, 1989). A half

- life in the

troposphere of about 17

– 18 days can be estimated for methanol

### OECD SIDS METHANOL

11 

### 2.2.3 Stability in Water

As an alcohol, methanol lacks hydrolysable groups and is chemically stable in water.

2.2.4 Transpo rt between Environmental Compartments

With a Henry’s law constant of 0.461 Pa m3/mol, methanol is not expected to significantly volatilize

from the aquatic compartment (Gaffney et al., 1987).

Methanol is the simplest of the alcohols, having only one carbon atom, and is completely miscible

in water. The dominant mechanisms of methanol loss from subsurface soil and groundwater are

expected to be biodegradation and advection (dispersion and diffusion), with little loss from

adsorption on soils due to its high solubility as well its low octanol

- water partition coefficient

(Malcolm Pirnie, 1999).

A distribution calculation performed with the Mackay Level III fugacity model (BASF, 2003) and

using emission rates derived from total releases into water, air and soi l (79658, 1531 and 658

tonnes, respectively) reported in the U.S. Toxics Release Inventory for 2001 predicts that the air is

the target environmental compartment for methanol (see Table 5).

Table 5: Environmental Distribution of Methanol (Fugacity Model )

Environmental

Compartment Emissions

Calculated from

TRI Releases

(kg/hr) Resulting Mass

Amount

(%)

Air 9093 73.3

Water 175 15.6

Soil 75 11.1

Sediment 0 0.0233



### 2.2.5 Biodegradation

Methanol is readily biodegradable based on the results of standard tests t hat show 76

– 82 percent

and 95 percent removal in standard ready tests after 5 and 20 days, respectively ( Price et al., 1974 ;

Wagner, 1976). Methanol occurs naturally in the environment due to various biological processes

in vegetation, microorganisms an d other living species. It is well established in the scientific

literature that methanol will biodegrade readily in the surface water, subsurface aquatic or soil

systems under both aerobic and anaerobic conditions (Malcolm Pirnie, 1999).

2.2.6 Bioaccumulatio n

Experimental BCFs of < 10 in fish species, including Cyprinus carpio and Leuciscus idus

, have

been measured ( Hansch and Leo, 1985, Gluth et al., 1985, Freitag et al., 1985). . These results are

expected because methanol has a high water solubility and a low octanol

- water partition coefficient

(log K

ow 

### =

- 0.82 to

– 0.64) ( Hansch and Leo, 1979; Rekker, 1977).

### OECD SIDS METHANOL

12 

### 2.3 Human Exposure

2.3.1 Occupational Exposure

The major anthropogenic sources of methanol include 1) its production, storage and use as a

solvent, 2) its us e as a chemical intermediate, and 3) as trace constituent of exhaust from vehicle

engines (IPCS/WHO, 1997). See Section 2.1 for a more complete list of uses. During the period

January 7, 1998 to November 12, 2003, a total of 950 measurements of methanol concentrations

were taken at several workplaces at BASF AG in Ludwigshafen, Germany. All concentrations are

time

- weighted

- average values over a reference period of 8 hours. The measured exposure is taken

during different activities such as production, pro cessing, quality control, pilot plant, research

laboratories, filling stations, and others. The 95 th 

### percentile value of the measured concentrations

was 0.032 mg/L (32 mg/m3).

In the European Union, the occupational exposure limit for methanol is 260 mg/m 3 (200 ppm) as an

8 hour TWA. In the US, the limit is 200 ppm as an 8 hour TWA and 250 ppm as a STEL.

The National Occupation Exposure Survey (NOES) of 1981

- 1983 estimated that about 1,700,000

employees were potentially exposed to methanol in the US (IPCS /WHO, 1997).

2.3.2 Consumer Exposure

In humans, the average background level of methanol in blood was found to be 0.73 mg/L, and

ranged from 0.32

– 2.61 mg/L (Sedivec et al., 1981). Formate, the major metabolite, ranged from 3

to 19 mg/L (0.07

– 0.4 mM). A mea n background body burden of 0.5 mg methanol/kg bw has been

estimated from the data of Sedovic et al. (1981) (Kavet and Nauss, 1990). Additional body burdens

may arise from various everyday exposures.

Methanol occurs naturally in humans, animals and plan ts. The general population may be exposed

to methanol via intake of fresh fruits, vegetables and commercial beverages like fruit juices, the

artificial sweetener aspartame, beers, wines and distilled spirits (IPCS/WHO, 1997).

Methanol is also contained as a solvent in products such as paints, sealers, and adhesives, which

may be available to consumers and used in car care, hobbies, crafts and home maintenance

(National Library of Medicine, 2003).

Additional sources of background body burdens may arise from metabolic processes such as

hydrolysis of various methyl esters, which may be contained in food and other consumer products.

For instance, L

- aspartyl

- L

- phenylalanine methyl ester (the artificial sweetener aspartame),

hydrolyzes to form methanol.

In 2002 , the American Association of Poison Control Centers found 1,049 incidences of exposure to

methanol reported to poison control centers. This represents a significant reduction from the 2,517

methanol exposure cases reported in 1997. Approximately 85 perc ent of methanol exposures (882

in 2002) resulted from “unintentional misuse” (largely ingestion) that was unplanned or not

foreseen by the patient. In 2002, over 120 of the methanol exposures were due to “intentional

abuse” relating to suspected suicide a ttempts or abuse to achieve a euphoric or psychotropic effect.

The reports for methanol represent only a very small fraction of the total “alcohol” exposures

(69,215) reported in 2002, and are much fewer than the total ethanol exposures (66,898). In many

cases, exposure to alcohols results from the ingestion of illegal or “bootleg” alcoholic beverages

where the patient has unintentionally consumed a beverage that has been “spiked” with elevated

methanol or another alcohol (Watson et al., 2003).

### OECD SIDS METHANOL

13 

### As a hydr ogen carrier fuel, methanol is likely to be contained in cartridges that provide fuel to

handheld fuel cell devices that power consumer electronic devices such as laptop computers and

cellular phones. Stringent design and performance standards are now bei ng developed for these

methanol fuel cartridges (by the Underwriters Laboratory Standard Technical Panel 2265 and

International Electrotechnical Commissions TC 105 Working Group #8) to preclude direct

consumer contact with the methanol fuel. Fuel cell

- pow ered vehicles may also use methanol as a

hydrogen carrier fuel. International standards for fueling systems for fuel cell vehicles (Society of

Automotive Engineers Standard J2579) are being developed to minimize the risk of consumer

exposure from methanol and other hydrogen fuels when refueling or operating fuel cell vehicles. A

dry

- connect, “spill

- free” methanol fueling nozzle system has been developed to prevent the

consumer from coming into direct contact with methanol during vehicle refueling.



## 3 HUMAN H EALTH HAZARDS

3.1 Effects on Human Health

3.1.1 Toxicokinetics, Metabolism and Distribution

Overview

Methanol is readily absorbed by inhalation, ingestion and dermal contact, and partitions rapidly and

equally throughout the organism in relation to the water cont ent of organs and tissues, regardless of

its exposure route. A small amount is excreted unchanged by the lungs and kidneys. Half

- lives of

methanol in the body are roughly 2.5 to 3 hours at doses less than 100 mg/kg bw. At high doses

(greater than about 1000 mg/kg bw), half

- lives can be 24 hours or more (IPCS/WHO, 1997). As

much as 90 percent of ingested low doses of methanol (e.g., 2 mg/kg bw) is exhaled as CO 2 

### in

rodents and monkeys. Even after very high doses (1000 mg/kg bw), up to almost 80 percent w as

recovered as CO 2 

### (Kavet and Nauss, 1990). However, there are pronounced differences in

metabolism and biotransformation between rodents and primates, especially at high (saturating)

dose levels.

The mammalian metabolism of methanol proceeds as follow s:

Step 1 Step 2 Step 3

Methanol  Formaldehyde  Formate  CO 2 

### + H 2 

### O

Step 1. In humans and monkeys, the initial reaction (from methanol to formaldehyde) is mediated

by alcohol dehydrogenase (ADH). In rodents, the reaction occurs mainly th rough a catalase

peroxidase system. The metabolic conversion to formaldehyde proceeds in both primates and

rodents at a similar rate under non

- saturated conditions (Kavet and Nauss, 1990; Medinsky et al.,

1997). However, in rodents, the catalase reaction is the limiting step, which means that at a critical

dose, methanol will accumulate and cause effects that may not occur in humans. In humans, ADH

levels are high enough so that accumulation of methanol does not occur except at dose levels that

are letha l or near

- lethal.

Step 2. This step occurs as a two

- reaction process. Formaldehyde is catalysed by the NAD

dependent formaldehyde dehydrogenase in the presence of glutathione to form the intermediate, S

formylglutathione. The intermediate is then hydrol ysed to formic acid by an associated thiolase.

Step 3. In this step, formate combines with tetrahydrofolic acid (THF) to form 10

- formyl

- THF

through the action of formyl

- THF synthetase. Next, 10

- formyl

- THF is converted to CO 2 

### by formyl

THF dehydrogenase . In humans and monkeys, THF levels are only 60 percent of that in rat liver

### OECD SIDS METHANOL

14 

### and 16

– 26 percent of that in mouse liver. Also, the amount of formyl

- THF dehydrogenase in

human and monkey liver preparations is only about 25 and 37 percent, respectively, of that in rat

liver (Johlin et al., 1987). Therefore, in humans and monkeys, the conversion of formate to CO 2 

### constitutes the limiting step, leading to a disproportionate increase of formate in the blood and in

sensitive target tissues such as CNS and the retina.

Formate accumulation in primates has been observed at methanol doses greater than 500 mg/kg bw

(Tephly and McMartin, 1984, as cited in NTP, 2003). Another study showed that formic acid in the

blood of rats and monkeys was similar at doses up to 600 mg/kg bw, but became substantially

higher in monkeys at 3000 mg/kg bw (Katoh, 1989 as cited in NTP, 2003). However, after repeated

inhalation of 2.6 mg/L (2000 ml/m 3 

### ) for 6 hrs/day, 5 days/week, for 1 or 2 weeks, monkeys (Rhesus

monkey, Macaca mulatta ) showed no methanol accumulation or discernible increase in formate

concentration in blood (Horton et al., 1992). The body burden can be estimated to be of the order of

200 to 300 mg/(kg bw

* 

### d), i.e. below or in the range of the critical threshold of satur ation of the

folate

- dependent formate metabolism (see also below). While this amount of methanol could

overwhelm the formate metabolism pathway if given as a bolus, the probability of saturating

metabolism with this amount divided over a prolonged time und er inhalation exposure is unlikely in

monkeys (Horton et al. 1992).

After a single exposure of 0.26 mg/L (200 ml/m 3 

### ), formate levels remained unchanged in humans,

irrespective of physical activity (Lee et al., 1992); these findings were confirmed by D ́Ale ssandro

et al. (1994) and Franzblau et al. (1993) in similar experiments on human volunteers.

The conversion of formate to CO 2 

### is also affected by the endogenous folate pool which, in monkeys

is only about 25

– 50 percent of the amount in rats (McMartin et al., 1977). Also, studies show that

formate oxidation in retinal tissue is 50 percent lower in humans than in rats, and is again thought to

be related to lower folate levels in human retinas (about 14 percent of levels in rats) (Martinasevic

et al., 1996). This difference in folate may also help explain the specific ocular and optic nerve

toxicity of methanol in humans. Furthermore, this implies that humans with a poor folate status are

supposed to be more susceptible to methanol action. On the oth er hand, in monkeys exposed

repeatedly to 900 ml/m 3 

### (approx. 1.2 mg/L) methanol for 2.5 h, the incremental increase in blood

formate was very low and remained significantly below the endogenous formate levels under folate

deficient conditions (see Appendix : Dorman et al. (1994) and Medinsky et al. (1997)). The

methanol dose that saturates the folate pathway in humans is estimated to be 210 mg/kg bw (Kavet

and Nauss, 1990).

Table 6 compares the maximal in vivo conversion rates of the above key steps of meth anol

metabolism in rats and monkeys (Kavet and Nauss, 1990; McMartin et al., 1977; Clay et al., 1975).

In monkeys, the maximum elimination rate of formate was about 34 mg/(kg

* 

### h), whereas in rats it

was about 73 mg/(kg

* 

### h) (McMartin et al., 1977). Eliminat ion half

- lives from blood were 30

– 50

minutes in the monkey and 12

– 23 minutes in rats, depending on the doses (Clay et al., 1975).

Table 6: Comparison of Reaction Rates in Rats and Monkeys

Methanol  Formate (Step 1, 2) Formate  CO 2 (Step 3)

Rat

~ 30 mg/(kg

* h) * ~73 mg/(kg

* h)

Monkey ~48 mg/(kg

* h)

~ 34 mg/(kg

* h) *

* Rate limitation of this reaction is evident from the lower maximal conversion rate.

### OECD SIDS METHANOL

15 

### Species Comparisons among Humans, Other Primates, and Rodents

The species

- specific kinetics and diffe rences in the metabolic transformation of methanol and

formate in rodents and primates allow a better under standing of the different effects encountered in

those species and their relevance for extrapolation across species. They can be illustrated with d ata

from inhalation exposures, which is a relevant exposure route for humans (see Table 7).

For doses up to 1.5 mg/L (1200 ml/m 3 

### ) for 6 h/d, a similarity in blood methanol levels (including a

linear relationship between air and blood concentrations as well as formate levels) are predicted for

rat, monkey and man. Furthermore, no increases in methanol blood levels above background occur

in rats and monkeys at 0.065 mg/L (50 ml/m 3 

### ) (Horton, 1992) or even after continuous exposure of

rats to 0.13 mg/L (100 ml /m 3 

### ) (NEDO 1987). However, above 1.5 mg/L, the increase in blood

methanol levels is non

- linear for rats (resulting in a steep exponential increase) and for monkeys

(with a flat exponential increase), whereas it remains linear for humans (Horton et al., 19 92).

Table 7: Kinetic and Blood Values after Inhalation in Rodents versus Primates

Rat 1) Mouse Monkey 1 Human 1

Exposure

conc.

[mg/L] Half

- life

[hrs] Methanol

(blood)

[mg/L]

(1x/d, 6 h) Methanol

(blood)

[mg/L]

(1x/d, 6 h) Half

- life

[hrs] Methanol (blo od)

[mg/L]

(1x/d, 6 h) Methanol (blood)

[mg/L]

(1x/d, 6 h)

0.26 0.8 ± 0.2 3.1 ± 0.4 1.1 ± 0.2 3.9 ± 1.0 3

- 8 2

0.65

- 0.780 approx. 10 approx. 10 approx. 10

1.6 1.0 ± 0.1 26.6 ± 2.0 approx. 150 3 3.2 ± 1.2 37.6 ± 8.5 approx. 25

2.6 2.1 ± 0.1 79.7 ± 6 .1 approx. 500 4 2.9 ± 0.6 64.4 ± 10.7 approx. 50

6.5 no data approx. 400 approx. 2000 4 no data approx. 240 approx. 140

1 Data taken from Horton et al. 1992, Fig. 8 and Tab. 2, unless otherwise indicated

2 From Horton et al. 1992 and other sou rces (see also above: 3.1.1 Inhalation)

3 Estimated from Perkins et al. 1995, Tab. 3

4 Mouse data are taken from Pollack and Brouwer 1996, Fig 36



### The rate of methanol accumulation in mice is two

- to three

- fold higher than in rats due to their more

rapid breathing and higher absorption, even though mice have a two

- fold higher elimination rate.

These factors possibly contribute to the greater sensitivity of mice to the teratogenic effects of

methanol, which are mediated by the parent compound (Pollack and Brouwer, 1996; Perkins et al.,

1995; 1996).

Additional metabolism data in animals and humans are discussed in Appendix A.

Conclusion

Methanol is readily absorbed by inhalation, ingestion and dermal contact, and partitions rapidly and

equally throughout the organism in relation to the water content of organs and tissues. A small

amount is excreted unchanged by the lungs and kidneys. Half

- lives of methanol in the body are

roughly 2.5 to 3 hours at doses less than 100 mg/kg bw.

At high doses disproportio nate increases of the parent compound in the blood are obtained in

rodents, but not in humans. On the other hand, in humans the metabolite formate accumulates at

high doses. This important difference mirrors the different enzymes and enzyme capacities invo lved

### OECD SIDS METHANOL

16 

### in the oxidative pathway from methanol to carbon dioxide. Specifically, two different rate limiting

processes have been identified: in rodents, high doses (after inhalation of 2.5

– 3.3 mg/L) lead to the

saturation of catalase, resulting in the accumu lation of methanol whereas formate levels remain low,

whereas in primates (especially humans), the parent compound is well oxidized and does not

accumulate, but formate increases disproportionately.

From studies in humans and monkeys exposed to concentrat ions of 0.26

– 2.6 mg/L (administered

for 6 to 8 hours), it can be concluded that methanol remains close to 50 mg/L in blood. At

inhalation exposures of 2.6 mg/L, rats also exhibit methanol blood levels that are not much higher

(at about 80 mg/L), whereas the level in mice was 400 mg/L. At a higher inhalation exposure (6.5

mg/L), humans show the lowest blood methanol level (at 140 mg/L), followed by monkeys, rats,

and mice, with the level in mice being more than 10 times higher than humans. Formate

accum ulation in primates has been observed at methanol doses greater than 500 mg/kg bw.

3.1.2 Acute Toxicity

Studies in Animals

Inhalation. LC 50



### - values in SD rats of 67,000 ml/m 3 

### (6 hours) and 98,000 ml/m 3 

### (4 hours) were

obtained corresponding to 87.5 and 128.2 mg/L , respectively (BASF, 1980a,b). Clinical signs of

toxicity were aqueous secretion of eyes and nose, labored breathing, staggering, apathy, and

narcosis.

A similar range of toxicity values is reported for the mouse: LC 50 

### (2.25 h) = approx. 79 mg/L (Von

Bu rg, 1994), narcotic doses = 40 and 55 mg/L after 2.2 and 7 h, respectively (Mashbitz et al., 1936;

Lehmann and Flury, 1943).

In cats, LC 50 

### values after 6

- hour exposures ranged from 26 to 48 mg/L (Von Burg, 1994; Witte,

1931). A shorter duration (4.5 hou rs) led to an LC 50 

### of 85.4 mg/L (Von Burg, 1994).

Studies in Rhesus monkeys indicate lethal concentrations (percent mortality not reported) at 13

mg/L (after18 hours) and 52 mg/L (after1

– 4 hours). Blindness associated with optic atrophy was

reported. Eve ntual recovery from this lesion was observed (McCord, 1931).

Dermal

According to an early report, all rats survived 35,000 mg/kg bw after occlusive skin application of

methanol, while deaths were reported at 45,000 mg/kg bw (Eulner and Gedicke, 1955). In rabbits, a

dermal LD 50 

### of about 17,000 mg/kg bw was found (Carnegie

- Mellon, 1981).

In Rhesus monkeys, four daily doses of 400 mg/kg bw (total 1600 mg/kg bw) caused sickness

within 24 hours, and eventually death. After four daily doses of 1000 mg/kg bw (t otal 4000 mg/kg

bw), severe illness took place followed by death on the second day (McCord, 1931).

Oral

In rats, mice, rabbits, and dogs, the LD 50 

### values after single oral administration range from about

5,600 to 14,400 mg/kg bw (NIOSH 1985; BASF, 1975; Kimura et al., 1971; Smyth et al., 1941;

IPCS/WHO, 1997; Munch et al. 1972; Gilger and Potts, 1955).

Rodents, rabbits and dogs suffer from ataxia, narcosis and coma after high methanol doses.

However, these animals do not exhibit the acidosis and ophthalm ologic changes typically seen in

humans at high lethal and sub

- lethal doses (Gilger and Potts, 1955).

### OECD SIDS METHANOL

17 

### In Rhesus monkeys, 1000 to 2000 mg/kg bw did not lead to mortality, while animals receiving 3000

to 8000 mg/kg bw died within two days. Treated animals showed acidosis, and some of them

exhibited semi

- coma and ophthalmologic changes (Gilger and Potts, 1955).

At higher doses, of those monkeys with severe intoxication, the retina and the optic papilla showed

extended oedema, and the pupils were wide and non

- responsive. Six of eight animals dosed with

6000 mg/kg bw exhibited cystic degeneration of the outer retinal granular layer, and in one animal

there was evidence of significant demyelinisation of the optic nerve. Histological lesions were seen

in the pu tamen and nucleus caudatus in 3 of 8 animals. All of these effects were most pronounced

at the high doses only after early compensation of acidosis using bicarbonate application, because

these monkeys generally did not survive those high doses of methanol but after early treatment with

bicarbonate (Potts, 1955; Potts et al., 1955).

Contrary to observations by Gilger and Potts (1955), there was no evidence of marked acidosis in

12 Rhesus monkeys (28 applications) after sublethal doses up to 6000 mg/kg bw. Specifically,

there was no hyperventilation, no increase in urinary excretion of organic acids, or shift in serum

bicarbonate. Blindness was seen in only one surviving monkey dosed with 9000 mg/kg bw; the

effect was transient four days after exposure. The LD 50 

### was between 7000 and 9000 mg/kg bw

(Cooper and Felig, 1961).

In a study of neurobehavioral effects, Male Long

- Evans rats were gavaged with single methanol

doses of 1000, 2000, and 3000 mg/kg bw representing about 10, 20, and 30 % of the LD 50. 

### Ten

m inutes after dosing the animals were subjected to a fixed wheel running test to assess operant

running (FR20). The rats displayed no signs of overt intoxication such as gait disturbance, but did

exhibit a significant, dose

- related reduction in FR20 respon se at all dose levels (Youssef et al.,

1993). A NOAEL for behavioural changes could not be derived.

Studies in Humans

Toxicity Syndrome

Oral ingestion dominates as the most frequent route of poisoning, but percutaneous absorption or

inhalation of vapors a re as effective as the oral route in producing methanol acute toxic syndrome.

After such acute poisoning, a blood level of ≥ 500 mg/L indicates treatment by hemodialysis is

needed. This blood concentration can be achieved in an adult person by ingestion o f 316 mg (or 0.4

ml) methanol/kg bw (Kavet and Nauss, 1990; p. 26). Generally, transient central nervous system

(CNS) effects appear above blood methanol levels of 200 mg/L, ocular symptoms appear above 500

mg/L, and fatalities have often occurred in untr eated patients with initial methanol levels in the

range of 1500

- 2000 mg/L. The minimal acute lethal dose to humans is in the range of 300 to 1,000

mg/kg bw (Roe, 1982; Kavet and Nauss, 1990; IPCS/WHO, 1997). In general, coma, seizures and

prolonged acid osis were poor prognostic signs (Naraqi et al., 1979).

Acute methanol intoxication (including acidosis and visual effects) evolves in a well

- defined

pattern. First, a mild depression of the CNS occurs and is followed by an asymptomatic latent

period comm only lasting 12 to 14 hours. Clinical symptoms include headache, dizziness, nausea,

and vomiting, abdominal pain, and labored, periodic breathing (Kussmaul breathing) and may

progress to coma and death from respiratory failure. Methanol exposure result i n ocular effects

ranging from mild photophobia, misty or blurred vision to markedly reduced visual acuity and total

blindness. Severe visual disturbances have been reported in workers who experienced methanol air

levels of about 1.5 mg/L (1200 ml/m 3 

### ) or m ore.

### OECD SIDS METHANOL

18 

### Ophthalmologic examination reveals hyperaemia of the optic disc, followed by the appearance of

oedema projecting into the surrounding of the retina from the optic disc, while the papilla itself is

not oedematous. As post

- mortem signs, focal necroses have been observed in the basal ganglia in

the brain and specifically the putamen, which participates in the control of motor activities.

Survivors of severe methanol intoxication may continue suffering from residual lesions of the

putamen and from visua l and lingual impairment (Kavet and Nauss, 1990; Erlanson et al., 1965;

Naraqi et al., 1979). The incidence of permanent ocular abnormalities was found to correlate with

the incidence of metabolic acidosis and with the amount of methanol consumed (Dethlef s and

Naraqi, 1978).

Human susceptibility to methanol toxicity may be decreased by concurrent ingestion of ethanol,

which slows the entrance of methanol into the metabolic pathway by competitive inhibition of the

ADH enzyme. Another important determinant o f susceptibility is the hepatic folate status, which

governs the rate of formate detoxification. In general, there is considerable wide interindividual

variability of the toxic dose (Kavet and Nauss, 1990; IPCS/WHO, 1997). Also blood measurements

in letha l cases revealed a high variability in formate and methanol levels (Roe, 1955; Litovitz et al.,

1988).

Human Exposure Studies

Neurophysiological effects were tested in 12 healthy subjects when exposed for 4 hours to 0.026

mg/L (control) and 0.260 mg/L in an exposure chamber in a cross

- over design. The EEG was

recorded before (as a baseline reference) and at the end of each exposure with the subject’s eyes

closed and opened and during a choice reaction test (color word stress test) (Muttray et al., 2001).

During subjects’ exposure to 0.26 mg/L, their scores for pre

- narcotic and irritating symptoms were

not different from controls. Under the open

- eye condition and during the choice reaction test, no

significant changes were found. Under the closed

- eye con dition, theta

- band and some delta

- band

brain waves were significantly less active at the end of exposure to 0.26 mg/L than in controls.

These changes suggest a slight excitatory effect of 0.26 mg/L methanol. However, the effect was

weak, as indicated by t he scores of acute symptoms, which did not change.

Osterloh et al. (1996) and Chuwers et al. (1995) studied neurobehavioral effects of methanol on 26

volunteers exposed to 0.26 mg/L methanol for 4 hours. The variables assessed were color

discrimination and contrast sensitivity, auditory evoked potentials, and neurobehavioral qualities.

Overall visual, neurophysiological, and neurobehavioral test outcomes were not significantly

affected, with the exception of some between

- subject variables. Specifically, sl ight effects on P

300 amplitude (brain waves in response to sensory stimuli) and information processing and

psychomotor skills (symbol digit testing) were apparently noted. However, these effects cannot be

adequately evaluated due to the low number of sub jects and high standard deviations for those

parameters.

Conclusion

For most experimental animals, rats, mouse, rabbits, dogs, and monkeys, the lethal dose of

methanol is high (greater than 2000 mg/kg bw after single oral administration). This also appli es

for inhalation. Lethal concentrations were found to be much higher than 10 mg/L for rats, cats and

monkeys (exposure range 4

– 7 h). This can be estimated to also correspond to body doses of

>>2000 mg/kg bw. After single dermal exposure, high lethal doses are reported in rats and rabbits,

but somewhat lower for Rhesus monkeys. Doses between 1000 and 2000 mg/kg bw proved to be

lethal. CNS suppression progressing to a narcotic state and possible fatality is the dominant effect

in non

- primate animals. The acidosis and ophthalmologic changes typical in humans do not occur

in rodents or rabbits. Also, reports of acidosis and ophthalmologic changes in Rhesus monkeys are

conflicting.

### OECD SIDS METHANOL

19 

### Generally, in humans, transient CNS effects appear above blood methanol levels of 200 mg/L.

Ocular symptoms appear above 500 mg/L. T he minimum lethal dose in humans has been shown to

be 300 to 1000 mg/kg bw, lower than generally observed in experimental animals. Fatalities have

occurred more commonly in untreated patients w ith initial methanol blood levels in the range of

1500

- 2000 mg/L. In general, coma, seizures and prolonged acidosis were poor prognostic signs.

However, exposures up to 0.26 mg methanol/L for 4 hours have been shown to be without

significant physiologic effects in human volunteers.

3.1.3 Irritation

Skin Irritation

The irritation potential of an unspecified dose of undiluted methanol in rabbits was examined under

occlusive conditions after exposure intervals of 1, 5, and 15 minutes and 20 hours. According to

Dr aize scoring, no signs of skin irritation were observed 24 hours and 8 days after treatment for any

of the exposure time periods (BASF, 1975). Another source, however, found that rabbits exposed

to 500 mg showed moderate skin irritation (NIOSH, 1982, as c ited in DGMK, 1982). However,

because these data were from a secondary source, they could not be verified.

Eye Irritation

One hour after instillation of 0.05 ml undiluted methanol into the eyes of two rabbits, slight

erythemas and cornea opacity as well as moderate oedemas associated with secretion were

observed. After 24 hours, the effects were assessed as mild, and after 8 days the animals had no

symptoms (BASF, 1975).

In six rabbits, mild to moderate conjunctivitis and edemas as well as mild iritis w ere produced after

instillation of 0.1 ml undiluted methanol into the eyes. Average scores after 24, 48, and 72 hours

were approximately 2 for conjunctivae and <1 for other effects. Primary irritation subsided after 72

hours, although redness of the conj unctivae persisted at that time (Jacobs, 1990) 1



### .

Exposure to an atmosphere saturated by methanol vapours at 20 °C produced severe irritation of

mucous membranes and milky corneal opacity in rats (time not specified) and eventually led to

mortality with all animals dead after 8 h (BASF, 1975).

Conclusion

Methanol exhibited no skin irritation in one study but moderate irritation in a study that could not

be verified. Available studies show that it is a slight to moderate eye irritant (with some

reversib ility of effects). High concentration of methanol vapours may be irritating to mucous

membranes. Based on the vapour pressure of about 130 hPa at 20 °C, the molecular weight of 32

g/mol and the molar volume of about 24 L/mol, it can be estimated that the saturation concentration

was ≥ 150 mg/L and, thus, clearly lethal.

3.1.4 Sensitisation

Studies in Animals

In one study conducted by BASF (1979a), a guinea pig maximization bioassay gave no evidence of

contact sensitisation in 10 female animals after induction a nd challenge with a 50 % aqueous

methanol solution (0.1 ml). In another BASF study using 24 female animals (two tests with 12

1 Information on effects 8 and 14 days later was not available.

### OECD SIDS METHANOL

20 

### animals each), 1 of 12 (test 1) and 2 of 12 (test 2) animals exhibited a slight skin response (score 1)

after 24 and 48 hours (BA SF, 1979a).

Conclusion

In summary, the low number (3 of 34) animals with a score of 1 show no notable sensitisation

potential of methanol.

3.1.5 Repeated

- Dose Toxicity

Studies in Animals

Several key studies are summarised in Table 8.

Table 8: Animal Studies o n Prolonged Exposure to Methanol (Inhalation, Oral)

Study Design Result Source

(Reliability)

Chronic inhalation [IUCLID 5.4, No. 1]

Macaca fascicularis monkey

0.013, 0.13, and 1.3 mg/L

(10, 100, 1000 ml/m 3 )

for ~21 h/d, 7 d/wk,

for 7, 19 and 29 months

8 animals per group, 2, 3 and 3 for

indicated intervals, respectively

Recovery group at 1.3 mg/L (1000

ml/m 3 ): 1 + 6 months General: Runny nose at 1.3 mg/L. Crouched stance at

1.3 mg/L.

Brain: Increased hyperplastic responsiveness of the

astroglia in some parts of the white substance in time

related manner (all dose levels), not considered

degenerative, absent in recovery groups. Slight

degeneration of the inside nucleus of the thalamus was

observed at 0.13 and 1.3 mg/L after 7 months or more.

Sl ight increases of responsive stellate cells were observed

at 0.13 and 1.3 mg/L. The authors conclude that the

tissues around the ventricules at the hypothalamus are

very sensitive to methanol. No clear correlation of the

effects was observed with exposur e level and duration.

No fibrosis or necrosis noted.

Peripheral Nervous System: One monkey at 0.13 mg/L

and 2 at 1.3 mg/L showed slight but clear changes in

peroneal nerves; the authors concluded that these effects

show that methanol causes damage to pe ripheral nerves.

Visual system: Some degenerative effects observed on

the optic nerve and corpus geniculatum laterale at 7 and

19 months (doses not noted).

Slight degeneration of the optic nerve suspected at 29

months (1or 2 cases at each dose level).

T hese effects were not interpreted as significant.

Liver and kidney: Increase in fat granules with dose

response from 0.013 to 0.13 mg/L. Some signs of

fibrosis at 1.3 mg/L, which were considered borderline.

In the kidney, Sudan positive granules appeared on the

renal tubule epithelium at 0.13 and 1.3 mg/L. Some

additional effects (hyalinisation of glomeruli) were

observed at 1.3 mg/L. The authors concluded that these

effects were possibly related to methanol exposure.

Cardiac: At 0.13 and 1.3 mg/L, som e effects observed

on the ECG, which were described by the authors as a

slight myocardial disorder. Histologically, a significant

increase of Sudan positive granules was noted in the 1.3

- mg group without pathological manifestations (e.g.

fibrosis) Lungs/t rachea: In the lungs, fibrosis was seen

in the interalveolar space. However, no dose response NEDO, 1987 (4)

### OECD SIDS METHANOL

21 was observed, and the effect was also seen in the controls.

In the trachea, atrophy of the epithelium of the mucous

membrane and a decrease in goblet cells wa s seen in four

cases (doses were not shown, although effects were not

observed in controls). The effect was localized.

It is not clear whether the effects observed were related to

methanol exposure.

LOAEL = 0.013 mg/L

(The LOAEL was determined based on t he qualitative

discussion of effects.)

Chronic inhalation [IUCLID 5.4, No. 2]

Fischer

- 344 rat (m; f)

0.013, 0.13, and 1.3 mg/L

(10, 100, 1000 ml/m 3 )

for ~20 h/d, 7 d/wk, for 12 mo

Examinations were comprehensive and

included optic nerve , eye, reproductive

organs No significant clinical and histopathological effects.

However, slight changes of body and organ weights were

observed within a 5 percent limit in the high

- dose groups

only.

NOEL = 0.130 mg/L NEDO, 1987 (4)

Chronic inhalatio n [IUCLID 5.4, No. 3]

B6C3F1 mice (m; f)

0.013, 0.13, and 1.3 mg/L

(10, 100, 1000 ml/m 3 )

for ~20 h/d, 7 d/wk, for 12 mo

Examinations were comprehensive and

included optic nerve, eye, reproductive

organs Significance of clinical and histopathological effects

unclear at highest dose, slight changes of body and organ

weights within 5 % limit,

slight increase in more severe (“moderate”) fatty

degeneration in livers of male mice, but considered

incidental because of a high incidence of this lesion in the

control, too. No cases that would have to be graded as

“severe” were found.

NOEL = 0.13 mg/L NEDO, 1987 (4)

Subchronic oral [IUCLID 5.4, No. 4]

SD rat (m; f)

(30 animals/sex/dose)

100, 500, 2500 mg/(kg

* d), gavage, 90 d,

interim sacrifice: wk 6

(10 animals per sex and dose) Increased liver enzyme levels; reduced brain weights

NOAEL = 500 mg/(kg

* d) U.S. EPA, 1986

[Cited in U.S. EPA,

1995] (4)

Subacute inhalation [ IUCLID 5.4, No. 5 ]

SD rat (m; f)

0.66, 2.7, and 6.6 mg/L

(500, 2000, 5000 ml/m 3 )

f or

– 6 h/d, 5 d/wk over 4 weeks

35 tissues were examined No significant histopathological effects,

but nasal irritation and ocular discharge were observed.

Relative spleen weights were increased significantly only

at 2.7 mg/L in females.

NOAEL = 6.6 mg /L (for systemic toxicity) Andrews et al., 1987

(2)

Subacute inhalation [ IUCLID 5.4, No. 6

and No. 7]

Macaca fascicularis monkey

3.9, 6.5, 9.1, and 13 mg/L

(3000, 5000, 7000, and 10000 ml/m 3 )

for up to 20 d, 21 h/d and including

recovery animals

4 ani mals per control and exposure

group were used. Blood levels at 3.9 and 6.5 mg/L:

- methanol: 80 and 5250 mg/L, respectively

- formate: 30 and 1210 mg/L, respectively

Coma and lethality: At ≥ 9.1 mg/L ( ≤ 6 d);

Brain: At 6.5 mg/L (5 and 14 d) , necrosi s in basal

ganglions of cerebrum (2/3 animals) and cerebral

oedema, recovery noted (2/2 animals).

At 3.9 mg/L (at 20 d), hyperplasia and fibrosis around

myelin sheets of basal ganglia and white substance with

slight to moderate increase in the astroglia. NEDO, 1987 (4)

### OECD SIDS METHANOL

22 Visual system: At > 3.9 mg/L: incipient atrophy of

optic nerve, reduction in myelinated fibers

Liver and kidney: At 3.9 mg/L: mild fatty degeneration

At 6.5 mg/L: fibrosis in liver, partly vacuolated, hyaline

degeneration in kidneys

LOAEL = 3.9 mg/L (This study could not be critically

reviewed because the report did not include a sufficient

amount of quantitative data, statistical results, or

histopathological information.)



### Studies in Humans

In male and female workers exposed to meth anol from 0.3 to 7.8 years, highly exposed workers

more often complained of blurred vision, headache and nasal irritation during or after work. None

of the workers reported suffering from photophobia. No retinal changes were observed. Among

three workers exposed to about 1.0

– 1.6 mg/L plus one worker exposed to 0.12 to 3.6 mg/L, two

showed retarded pupil reflex and one exhibited mild mydriosis, but no permanent eye damage.

Forgetfulness and skin sensitivity were other common complaints (Kawai et al., 19 91; IPCS/WHO,

1997).

In a study published by NIOSH (Frederick et al., 1984), teacher aides who worked at or near spirit

duplicator machines (using a 99 percent methanol duplicator fluid) and duplicating machine

operators experienced exposures between 0.4 8 and 4.0 mg/L (mean 1.38 ±0.75 mg/L). A health

survey based upon a self

- administered symptom questionnaire suggested that symptoms relevant to

methanol toxicity (headache, dizziness, and eye irritation) were significantly increased among

exposed staff as compared with a non

- exposed teacher control group at the same institute (Frederick

et al., 1984: see also IUCLID, 5.10, entry 21).

Additional studies also showed that headaches were associated with occupations that involve the

operation of duplicating m achines (NTP, 2003).

Conclusion

In a chronic exposure study in monkeys, some histological findings were observed in the cerebral

area after prolonged and almost continuous exposure to doses of up to 1.3 mg/L, although the

effects were generally of a mild n ature. Several additional effects, including dose

- related liver

effects, were observed in this study. Effects in other organs and systems were also observed. The

neurological effects were assumed to be due to methanol because several such effects were

observed, neurological effects were also seen in the subchronic monkey study, and not yet verifiable

neurobehavioral effects were observed in a reproductive/developmental study in monkeys (see

Section 3.1.7

– Burbacher et al., 1999). A LOAEL of 0.013 mg/ L (continuous exposure) was

chosen for the chronic study in monkeys. Although the statistical significance of the effects cannot

be verified from the study report, the number of effects and systems affected indicate a relationship

with methanol.

In a suba cute 20

- d study in monkeys

, 3.9 mg/L (3000 ml/m 3 

### ) was chosen as the LOAEL (subacute,

continuous exposure) where neurotoxic lesions appeared to progress in monkeys (according to

NEDO, 1987). This exposure concentration correlated with methanol blood levels 80 mg/L and

formate levels 30 mg/L.

In the NEDO (1987) studies of chronic exposure in rats and mice, the effects are difficult to

interpret from the available study and NOELs for continuous exposure could be established at the

mid

- dose of 0.13 mg/L beca use adverse effects were explicitly not reported.

### OECD SIDS METHANOL

23 

### In a 28

- day study, there was no evidence of adverse effects in rats exposed to methanol up to 6.5

mg/L (6 hours per day for 28 days), except local nasal irritation and increased relative spleen

weights, whi ch were observed only at the middle dose (Andrews et al., 1987). The estimated blood

level of methanol is expected to be about 250 mg/L under this condition, as described in Section

3.1.1, Tab. 6.

3.1.6 Mutagenicity

Studies in Animals

In vitro Studies

Methanol has been examined in numerous test systems including bacterial, mammalian and fungal

test systems. Most studies failed to demonstrate mutagenic activity, with four exceptions: 1) in

Salmonella tester strain TA 102, a questionably positive response was obs erved (DeFlora et al.,

1984); 2) in a DNA damage and repair assay with E. coli WP strains (repair proficient and deficient

types), the result was considered ambiguous (deFlora et al. 1984b); 3) in a mouse

- lymphoma assay,

a significant increase in the mutat ion rate was reported at 7.9 mg/ml methanol (McGregor et al.,

1985); and 4) in a test of mitotic chromosomal segregation in Aspergillus nidulans , a positive result

was observed (Crebelli et al., 1989). All other studies produced consistently negative resu lts (see

Table 9).

Table 9: In Vitro Genotoxicity Testing on Methanol

Bioassay Test system

(+/

- S9) Test

Concentrations Result Source (Reliability)

Bacterial Assays

Salmonella

typhimurium TA 1535, 1537, 1538, 98, 100

(+/

- S9) ≤ 2.5 mg/plate negative DeFlora, 1981 (2)

Salmonella

typhimurium TA97, 102 (+/

- S9) ≤ 7.5 mg/plate negative,

in TA 102:

ambiguous DeFlora et al., 1984a

(2)

Salmonella

typhimurium TA 1535, 1537, 1538, 98 (+S9) ≤ 3.6 mg/plate negative Gocke et al., 1981 (2)

Salmonella

typhimur ium TA 1535, 1537, 1538, 98, 100

(+/

- S9) 0.005

– 5 mg/plate negative Shimizu et al., 1985 (2)

Escherichia coli WP2uvrA (reverse mutation) (+/

- S9) 0.005

– 5 mg/plate negative Shimizu et al., 1985 (2)

Escherichia coli WP67, WP2, CM871 (increased

cytot oxicity in DNA

- repair deficient

strains as indicator of genotoxicity) 20 mg/microwell

(~ 60 mg/mL) ambiguous DeFlora et al., 1984b

(2)

Mammalian Cell Culture Systems

Micronucleus Chin. Hamster V79 (

- S9) approx. 40 mg/ml negative Lasne et al., 1984 (2)

Micronucleus Syrian hamster embryo cells No data (based on

prior range

- finding) negative Fritzenschaf et al.,

1993 (4)

Sister Chromatid

Exchange (SCE) Chin. Hamster Ovary cell (CHO)

(

- S9) ≤ 1 mg/ml,

1x/d for 8 days negative Obe and Ristow, 1977

(2)

C ell Transformation Mouse embryo fibroblasts

C3H/10T1/2Cl8 (+/

- S9), +/

- TPA 5

– 100 mg/ml negative Ragan and Boreiko,

1981 (2)

Cell Transformation Baby Syrian Hamster kidney cells

BHK21 C13 (+/

- S9) approx. 8

– 80

mg/ml negative Strobel and Greb, 1981

(2)

### OECD SIDS METHANOL

24 Gene Mutation Chin. Hamster V79, reversion to 8

- Aza, 6TG or Quabain resistance

(+/

- S9) approx. 16

– 63

mg/ml negative NEDO, 1987 (2)

Chromosomal

Aberration Chin. Hamster lung cells (+/

- S9) approx. 7

– 28

mg/ml negative NEDO, 1987 (2)

Mouse Lymphoma L5 178Y (+S9) 7.9 mg/ml positive McGregor et al., 1985

(4)

Other

Chromosomal

malsegregation Aspergillus nidulans diploid strain P1

(fungus) (

- S9) 6 % (v/v) positive Crebelli et al., 1989 (2)

Gene mutation Yeast Schizosaccharomyces pombe

(ade 6 locus)(+/

- S 9) 5 % (v/v) negative Abbondandolo et al.

(1980) (4)



### In vivo Studies

The majority of in vivo assays are negative for mutagenicity and clastogenicity. Some of these

assays were conducted under specific stress conditions using folate

- deficient mice (see: A m. Petrol.

Inst., 1991). The positive findings resulted from the same working group and represent two of ten

cases (Chang et al., 1983; Pereira et al., 1982) (see Table 10).

Table 10: In Vivo Genotoxicity Testing on Methanol

Bioassay Test system Test Dose s Result Source

Chrom.

Aberration,

Micronucleus

SCE Mouse, oral 1000 mg/kg bw positive Pereira et al., 1982 (4);

NTP, 2003

SCE C57Bl mouse,

inhal. 6 h/d, 5 d, in primary lung

- cell

culture 1.04, 5.20 mg/L

(800, 4000 ml/m 3 ) negative Campbell et al., 1991

(2)

Micronucleus ICR mouse, single dose, gavage 1050, 2110, 4210,

8410 mg/kg bw negative NEDO, 1987 (2)

Micronucleus NMRI mouse, single dose, ip. 1920, 3200, 4480

mg/kg bw negative Gocke et al., 1981 (2)

Micronucleus CD

- 1 mouse, gavage during pregn ancy

(6

- 10 gd): adult + fetus peripheral blood,

folate

- sufficient and folate

- deficient

conditions 2500 mg/(kg bw

* d) negative Fu et al., 1996 (2)

Micronucleus Swiss Webster mouse, ip., 4 d, folate

sufficient and folate

- deficient conditions 300, 600, 1200,

2500 mg/(kg bw

* d) negative Am. Petrol. Inst., 1991

(2)

Micronucleus C57Bl mouse,

inhal. 6 h/d, 5 d, in primary lung

- cell

culture and peripheral blood 1.04, 5.20 mg/L

(800, 4000 ml/m 3 ) negative Campbell et al., 1991

(2)

Chromosomal

Aberration C57Bl mouse

, inhal. 6 h/d, 5 d, in primary lung

- cell

culture 1.04, 5.20 mg/L

(800, 4000 ml/m 3 ) negative Campbell et al., 1991

(2)

Chromosomal

Aberration C57Bl mouse,

inhal. 6 h/d, 5 d, in spermatocytes of

pachytene 1.04, 5.20 mg/L

(800, 4000 ml/m 3 ) negative Cam pbell et al. ,1991

(2)

Chromosomal

Aberration Mouse, intraperitoneal, total dose split in

3 daily fractions, bone marrow 75

- 300 mg/kg bw positive Chang et al., 1983 (4)

NTP, 2003

### OECD SIDS METHANOL

25 Point or

structural

mutation Male Drosophila melanogaster SLRL

assay (S ex

- linked recessive lethal

mutation) 1000 mM in oral

feed negative Gocke et al., 1981 (2)



### Conclusion

The majority of in vitro and in vivo assays on methanol are negative for mutagenicity and

clastogenicity although a few of the in vitro and in vivo assay s were positive. The positive or

ambiguous in

- vitro findings are all associated with high methanol concentrations from 0.8 to 6 %

(v/v), but are not higher than those that proved to be negative in other tests

. Thus, although most

studies indicate that met hanol does not have the potential to be mutagenic, given positive results in

some tests, the possibility of mutagenicity cannot be ruled out.. Carcinogenicity

Inhalation

Groups of male and female Fisher rats as well as B6C3F1 mice (52 animals per sex and group) were

exposed to 0.013, 0.13, and 1.3 mg/L (10, 100 and 1000 ml/m 3 

### ) for 24 and 18 months, 20 and 19

hours per day, respectively. Clinical, ophthalmological, clinical

- chemical, hematological and

pathological/histopathological examination were done (N EDO, 1987).

No clinical signs of intoxication could be observed in treated rats and mice. No changes were

evident in hematological and biochemical parameters in either species. Body weight was slightly,

but not significantly, decreased between weeks 51 t hrough 72 in the highest female dose

- group of

rats (1.3 mg/L). No differences in survival were noted in the treatment groups compared with the

control group.

The following specific tumours appeared at a somewhat higher incidence in high

- dose groups of

bo th sexes in rats after week 79 and 104 without clear dose

- response relationship (see IUCLID, 5.7,

entry 1):

- papillary lung adenomas in males (6/52 vs. 1/52 in the control), but not in females;

- adrenal phaeochromocytomas in females (7/52 vs. 2/52), but not in males;

- 3/52 and 5/52 metastatic (transition) tumors in males and females, respectively, vs. 0/52 in

the controls.

The statistical analyses did not reveal significance. However, the Fisher ́s Exact Test was close to

the limit of applicability for testi ng the data of the phaeochromocytomas. No increase of this tumour

type was seen in male rats which showed a high spontaneous tumour rate (7/52 in the control vs.

4/51 in the high

– dose group).

In mice, there were no appreciable differences from the contr ol in either numbers of animals with

tumors or in degree of malignancy observed.

There was no evidence of an increase in liver tumours in rats or in spontaneous liver tumour rate in

mice.

After almost continuous inhalation exposure of monkeys, an increase d diffuse hyperplastic

responsiveness of the astroglia in some parts of the cerebral white substance was noted at all dose

levels of 0.013 mg/L (after 29 months), 0.13, and 1.3 mg/L (after ≥ 7 months) without histological

signs of degeneration. This effect was absent in recovery groups after 1 and 6 months (NEDO,

1987).

### OECD SIDS METHANOL

26 

### Conclusion

Overall, tumour frequencies showed no statistically significant differences between treatment

groups and controls in either rats or mice. The biological significance of the non

- s tatistically

significant increase in the incidence of the phaeochromocytomas in the aged female high

- dose rats

is considered to be low. The methanol

- related proliferative effects on the astroglia cells observed

in monkeys were of transient nature and not associated with cellular neoplastic transformation. The

weight of evidence suggests that methanol is not carcinogenic.

3.1.8 Toxicity for Reproduction

Studies in Animals

Studies of Reproduction

Key studies in rats and monkeys that address reproductive perform ance (and any resulting

developmental toxicity) are summarised in Table 11. The study by Burbacher et al. (1999) of

reproductive and developmental toxicity in monkeys is also included here because the dosing

schedule is similar to a one

- generation reprodu ctive toxicity study in rodents.

Table 11: Animal St ud ies on Reproduction and Development

Study Design Results Source

(Reliability)

Two

- generation study

Inhalation

SD rats (m, f)

0.013, 0.13, and 1.3

mg/L

(10, 100, 1000 ml/m 3 )

at 19

- 20 h/d, 7 d/wk

F0: 103

- 108 d

(pre

- mating, mating,

gestation, lactation);

F1: 61

- 62 d + 145

- 153 d;

F2: 54

- 56 d Blood methanol levels:

F1(9wks of age): 3

– 3.5 mg/L, 1

– 4.2 mg/L, and 53 (m)

– 100 (fem.) mg/L,

respectively, vs. controls (baseline) 2

– 3 mg/L.

Reproductive pe rformance: No treatment

- related alterations in F0 and

F1 for all reproductive parameters.

Development: . No pathological effects/no histological manifestation/no

post

- natal functional deficiencies (learning, behaviour). However, the

following effects were o bserved:

At 1.3 mg/L: Post

- natal differentiation apparently influenced: “descensus

testis” 0.5 to 1 d earlier than in male control groups of F1

- and F2

- pups

(statistically significant: level not defined).

(Note: There is considerable variation also bet ween the control groups of

both generations).

Absolute and relative brain weights of F1 and F2 were significantly lower

(both sexes) (affected cerebrum, cerebellum); slight but not significant

shifts were observed in organ weights: thymus and pituitary were lower,

heart, lung, and liver were higher (no quantitative data on organ weights

and statistical results included) .

At 0.13 mg/L : “Descensus testis” in male F2

- progeny about 0.5 d earlier

than in male control F2 pups. Detailed data not specified and not addressed

under “Discussion”.

NOAEL(devel) = 0.13 mg/L (NOAEL based on the absolute and relative

decreased brain weights in F1 and F2 animals. This study, however, could

not be critically reviewed because the report did not include a sufficient

am ount of quantitative data, statistical results, in particular on organ

weights, or histopathological information. NEDO, 1987;

Takeda/

Katoh, 1988 (4)

### OECD SIDS METHANOL

27 Reproductive and

developmental toxicity

Inhalation

Macaca fascicularis

monkeys

(adult f)

0.26, 0.78, an d 2.34

mg/L

(200, 600, 1800 ml/m 3

for 2.5 h/d before

breeding (120 d),

during breeding (70 d),

and during pregnancy

(about 165 d).

9

- 12 monkeys per group Mean blood methanol levels:

~5mg/L (1

- to 2

- fold), ~10 mg/L (3

- to 4

- fold), and 40

– 50 mg/L

(13

- t o 16

- fold), respectively

Blood formate levels: at baseline level at all exposures

Maternal toxicity: none

Reproductive performance (menstrual cycles, conception rate, and live

birth delivery rate): not affected, only observation: dose

- unrelated

reduction of gestation length in all methanol

- exposed animals of about 6 to

8 d, within normal range of other monkey colonies. C

- sections were

performed in 2 monkeys at 0.26 mg/L, 2 at 0.78 mg/L, and 1 at 2.34 mg/L

who experienced vaginal bleeding, probably due t o detachment of the

placenta. An additional C

- section was done in one monkey at 2.34 mg/L

following 3 nights of unproductive labor.

Developmental toxicity: Apparent behavioural effects: Delay in early

sensorimotor development was observed in the Visually Directed

Reaching Test: Ages at achievement of the test (from control to highest

dose group) were: 24, 32, 43, and 41 days (males) and 34, 33, 28, and 40

days (females). Delay in visual recognition memory (but no clear dose

response was observed).

W asting syndrome: 2/7 female offspring showing growth retardation at

12 and 17 months in the 2.34 mg/L

– exposure group. Signs of severe

malnutrition and gastroenteritis noted at autopsy.

LOAEL (devel) = 0.26 mg/L Burbacher et

al., 1999

Special study on

spermatogenesis

B6C3F1 mice (male):

10 animals, 4 mo. of age;

1000 mg/kg bw (1x/d, 5

d), oral. Preparation after

5 wk (after testicular

spermatid maturation) Sperm morphology:

No significant morphological abnormalities:

In spermatozoa isolated from dissected cauda epididymis for light

microscopy, slight, statistically insignificant increase in sperm

abnormalities (mean factor about 1.7 above neg. control, with very high

variance),

In positive control group (cyclophosphamide, 100/kg): some 6

- fold

in crease in abnormalities.

NOAEL = 1000 mg/(kg bw

* d) Ward et al., 1984

(2)



### Developmental Toxicity Studies

Methanol has been tested in numerous developmental toxicity studies using rat, mouse, rabbit and

monkey. However, the number of classic developme ntal investigations following oral

administration is limited or poorly documented. Frequently, single studies are designed to elucidate

specific questions based upon previous experimental experience. The key studies are summarized

in Table 12.

### OECD SIDS METHANOL

28 

### Table 12: Animal Studies on Pre

- Natal Development Following Exposure to Methanol

Study Design Results Source

(Reliability)

Developmental Toxicity/Teratogenicity

Developmental Toxicity

Inhalation

SD rat,

6.5, 13, and 26 mg/L

(5000, 10000, 20000 ml/m 3 )

at 7 h/d, gd 1

- 19 for the

two lowest doses or gd 7

- 15 for the highest dose Methanol blood levels:

1000

– 2170 mg/L, 1840

– 2240 mg/L, and 5250

– 8650 mg/L, resp.

Maternal toxicity: slight, unsteady gait at the highest concentration

Reproductive performance (corpera lutea, implantations, fetal

lethality, resorptions): not significantly affected.

Fetal body weight: slight decrease at ≥ 13 mg/L (p<0.05)

Malformations :

At 26 mg/L:

V isceral defects : 7/15 litters with 15/96 malformed fetuses vs. 0/15

litters and 0/107 fetuses in the untreated control

Skeletal defects : 14/15 litters with 72/92 malformed fetuses vs. 0/15

litters and 0/98 fetuses in the untreated control

At 13 mg/L (not statistically significant):

Visceral malformations were 2/15 litters with 2/1 07 fetuses , and

skeletal malformations were 2/15 with 2/115 fetuses.

At 6.5 mg/L (not statistically significant):

Visceral defects were 1/13 litters with 2/90 fetuses, and skeletal defects

were 1/13 with 1/90 fetuses.

NOAEL (maternal) = 13.0 mg/L

NOAEL (devel) = 6.5 mg/L Nelson et al.,

1985 (2)

Developmental Toxicity

Inhalation

Crl:CD SD Rats

0.26, 1.3, and 6.5 mg/L

(200, 1000, or 5000

ml/m 3 ) for 22.7 hr/d

(avg), gd 7

- 17

36 animals/group Dams: 6.5 mg/L: reduced bodyweight gain; reduced food and wat er

intake; one died; one sacrificed in extremis.

Prenatal Development: 6.5 mg/L: increased late resorptions, reduced

no. live fetuses, decreased fetal weight, increased numbers of litters

with fetuses that have malformations (ventricular septal defect),

v ariations (thymus, vertebrae, ribs), delayed ossification.

Postnatal Development: 6.5 mg/L: prolonged gestation, reduced

post

- implantation embryo survival, and number of live pups/litter,

decreased survival rate on postnatal day 4. Brain, thyroid, thymu s, and

testes weights were decreased at 8 weeks of age, but not overall body

weight. Developmental milestones not affected when parturition was

taken into account.

NOAEL (devel) = 1.3 mg/L NEDO, 1987

(4)

Developmental Toxicity

Inhalation

CD

- 1 mice

1.3 , 2.6, 6.5, 9.75, 13,

and 19.5 mg/L

(1000, 2000, 5000, 7500,

10000 or 15000 ml/m 3 )

for 7 h/d, gd 6

- 15 Mean blood methanol levels:

100 mg/l, 540 mg/L, 1600 mg/L, 3200 mg/L, 4200 mg/L, and 7300

mg/L, with the background level at about 1.6 mg/L

Maternal t oxicity: none

Fetal body weight: Statistically

- significant decrease at ≥ 13 mg/L

Malformations: at 6.5 mg/L, increase in exencephaly and cleft palate

(approx. 10 % fetuses vs. 0.3 % in the control)

Dose

- related increase in extracervical ribs or ossification sites lateral to

the 7 th cervical vertebra at ≥ 2.6 mg/L (p<0.01)

(approx. 50 %/litter vs. 26 to 34 % in the controls and approx. 34

%/litter at 1.3 mg/L)

Other Developmental Effects: Dose

- related statistically significant

increase in females with fully resorbed fetuses at 13 mg/L. Dose

related statistically significant decrease in number of live pups/litter at Rogers et al.,

1993 (2)

### OECD SIDS METHANOL

29 Study Design Results Source

(Reliability)

> 9 .75 mg/L.

NOAEL (devel) = 1.3 mg/L (based on cervical

- rib defect)

Studies to Address Specific Questions

Developm ental Toxicity

Oral

CD

- 1 mice,

4000 (= 2000 x 2) mg/kg bw or

5000 (= 2500 x 2) mg/kg bw

gd 7 only

(Special test design) Stage

- dependent methanol toxicity in organogenesis:

gd 7 considered as most sensitive

Malformations in both dose

- groups: dose

- rela ted, mainly skeletal

defects of the skull and neural tube:

- increase in cleft palate (at 19 and 14 % vs. 0 % in the control);



### - increase in

extracervical ribs to C7 vertebra: at either dose approx. 30 % of

fetuses vs. 0 % in control;

- split, fused or incomplete vertebra C2 about 41% in the high dose

fetuses vs. 7.7 in the control;

- 25 instead of 26 presacral vertebrae in 10 % vs. 2.4 % in control.

LOAEL = 4000 mg/kg bw Connelly and

Rogers, 1997

(2)

Developmental Toxicity

Inhalation

CD

- 1 mice,

6.5 , 13, 19.5 mg/L

(5000, 10000,

15000 ml/m 3 );

6 h/d on

gd 6

– 15, 7

- 9, or

9

– 11

(Special test design) Stage

- dependent methanol toxicity in organogenesis:

- neural tube and ocular defects (significant during gd 7

- 9);

- hind

- limb anomalies (sign ificant during gd 9

- 11);

- cleft palate, renal pelvic dilatation/hydronephrosis and tail anomalies

(significant during both gd period 7

- 9 and 9

- 11), evident at ≥ 13 mg/L

- Increased resorptions at 19.5 mg/L for exposures that included gd 7,

significant on ly at the top exposure level

- Decreased body weight and decreased live fetuses/litter at 19.5 mg/L

- At 6.5 mg/L, no teratogenic effects, but incipient increase in post

implantation losses (p<0.05)

LOAEL (develop) = 6.5 mg/L Bolon et al.,

1993 (2)

Dev elopmental Toxicity

Oral

CD

- 1 mice,

5000 mg/(kg bw

* d)

[= 2500 mg/(kg bw*d),

2x/d]

gd 6

– 10

(Study in combination

with folate

- deficiency

and folate

supplementation) Modulation of methanol toxicity by folate status:

Methanol and folate

- supplemented mice : 5000mg/(kg

* d) methanol

produced marginal teratogenic effect (exencephaly) under folate (5/222

vs. 1/282 cases in non

- treated, folate

- sufficient control).

Folate deficiency only: no significant increase in malformations (cleft

palate and exencephaly)

M ethanol and folate deficient mice: significant increase in cleft palate

(36/235 vs. 5/215 of folate

- poor control), no significant increase in

exencephaly (8/235 vs. 2/215 of folate

- poor control).

LOAEL = 5000 mg/(kg

* d) Fu et al.,

1996; Heid et

al. 1992 (2 )

Developmental Toxicity

Inhalation

Long

- Evans rat,

5.85 mg/L

(4500 ml/m 3 ) for 6 h/d,

gd 6 to post

- natal day 21

(Pre

- and postnatal study

focused on neurotoxic

effects) Mean blood methanol levels:

in dams: 550 ± 70 and 560 ±90 mg/L (gestation and lac tation);

in neonates: 1260 ±230 mg/mL [i.e. neonates showed about twice the

blood levels of dams persisting up to 4 weeks post

- weaning (48 d of

age)]

Neurotoxicity:

No detectable evidence of neuropathology (based on brain histo

morphology, expression of ne ural cell

- adhesion molecules (NCAM)

and behavioural testing). There was a weak transient effect on the

molecular level: reduced NCAM formation, possibly associated with

treatment. Performance on operant tests suggested subtle cognitive

effects.

NOAEL = 5.85 mg/L Weiss et al.,

1996; Stern et

al., 1996,

1997 (2)

### OECD SIDS METHANOL

30 

### Studies in Humans

The limited data available on methanol exposure during pregnancy in humans and reproductive and

developmental effects do not show an association with methanol (NTP, 2003). In an

epidemiological study, the reproductive effects of various occupations and associated exposures to

complex mixtures were examined in women who gave birth to infants with and without cleft lip or

palate (Lorente et al., 2000). No association was found b etween methanol exposure and oral clefts.

The small number of subjects exposed to methanol, lack of individual exposure data, and

confounding by other chemical exposures did not allow the authors to draw firm conclusions as to

the role of methanol on thes e outcomes.

NTP (2003) did review several studies that investigated the association between the

periconceptional use of multivitamins containing folic acid and birth defects (e.g., neural tube

defects and orafacial clefts). These studies suggest that fola te deficiency in humans may lead to

greater susceptibility to such defects. Because the association between methanol and these effects

was not directly investigated, no conclusions can be made regarding causality between methanol.

And birth defects based on the results of human studies. However, these results should be

considered along with the results of studies of increased developmental effects in folate

- deficient

rodents exposed to methanol. As noted by NTP (2003), the current evidence suggests that women

with low folate status may be more susceptible to the adverse developmental effects of methanol,

but clearly more information is needed to clarify this issue.

Conclusion

NTP (2003) concluded that rodent data on reproductive and developmental toxicity are relevant for

humans despite the known differences in methanol metabolism between rodents and humans.

Rodents are adequate models for human exposure to methanol at levels where formate does not

accumulate. However, blood methanol concentrations assoc iated with serious teratogenic effects

and reproductive toxicity are in the range associated with formate accumulation, which is likely to

result in metabolic acidosis, and visual and clinical effects in humans (NTP, 2003). Other effects

(e.g., subtle, no t yet definitive, neurological effects observed in primates) may be exhibited at lower

inhalation doses and lower methanol blood levels.

Reproduction Studies in Animals

No impairment of fertility and reproductive performance was found in male and female rats (parent

and daughter generation) exposed to continuously high doses of methanol (NEDO, 1987).

In male mice after repeated oral dosing at 1000 mg/(kg bw

* 

### d), increases in morphological

anomalies in spermatozoa occurred but the increase was insignific ant (Ward et al., 1984). The

concomitant blood level can be estimated to be in the range of >500

– 1000 mg/L in mice (see

Appendix).

There was no clear evidence of malformations or variations in the pre

- and post

- natal development

of the progeny of two rat generations (NEDO, 1987). However, slight but significant retardation in

brain growth (F1 and F2) led to the conclusion of the NOAEL (20 h/d) being 0.13 mg/L. This

corresponds to a blood level of 1

– 4 mg/L, which is not statistically different from the 0 .013 mg/L

group and the untreated control. There were also some statistically significant differences in time

to decent of the testes, although the biological significance of this difference is not clear because

the differences were so slight. Due to exp erimental dose selection, the NOAEL happened to be

very low compared with the LOAEL.

Studies in monkeys (Burbacher et al., 1999) provided some evidence of effects by prenatal

methanol exposure on the neurobehavioral development of nonhuman primate infants during the

first 9 months of life. The interpretation was somewhat limited by the normal variance and the low

number of animals However, based on these neurobehavioral results and the observance of

### OECD SIDS METHANOL

31 

### neurological effects in other studies, the LOAEL for dev elopmental effects was determined to be

0.26 mg/L.

Developmental Toxicity Studies in Animals

High concentrations of methanol may cause specific malformations in organogenesis in pre

- natal

development of rodents. In the rat, the NOAEL (inhalation for 7 hours per day) was determined to

be 6.5 mg/L, based on observance of malformations as well as slight decreased body weights. This

corresponds to a maternal methanol blood level of 1000 to 2170 mg/L (Nelson et al., 1985).

In the mouse, there is unequivoca l evidence of structural malformations in the progeny at about 6.5

mg/L (inh. 7 h/d) and some evidence at 2.6 mg/L (inh. 7 h/d) (Rogers et al., 1993). The

corresponding average blood levels of methanol were about 1600 and 550 mg/L. The NOAEL

(inh.7 h/d) is suggested to be 1.3 mg/L (the corresponding blood methanol is about 100 mg/L).

The marginally teratogenic oral dose is reported to be 5000 mg/(kg

* 

### d) in mice (Fu et al., 1996).

The estimate of the respective methanol blood concentration is of the ord er of >2000 mg/L (see

Appendix).

Folate deficiency enhanced the teratogenic effects of methanol in mice, but this was not due to an

increase in formate levels (Fu et al. 1996, see Table 12).

3.1.9 Specific Toxicity (Ocular Toxicity)

In monkeys ( Macaca mulata ), acute methanol toxicity (after a single dose) does not yield signs of

ocular toxicity (McMartin et al., 1975). On the other hand, repeated dosing succeeded in producing

ocular lesions after a high initial oral dose of 2000 mg/kg bw followed by lower dose s for up to 6

days, depending on the animal’s acidotic response in blood. The ranges of blood levels were 1000

– >2000 mg methanol/L and 350

– 550 mg formate/L (8

– 12 mEq/L) (Martin

- Amat et al., 1977).

In a subsequent study in monkeys, sodium formate in fusion (without acidosis) produced blindness

and marked optic disc oedema between 24 and 48 hours, while the retina itself including the

ganglion

- cell layer was histologically completely normal. After 10 hours, all animals had

accumulated formate in blood between about 10 to 30 mEq/L, while blood pHs had been

maintained between 7.4 and 7.6 (Martin

- Amat et al., 1978).

In a folate

- deficient state, high oral doses of methanol (1500

– 2500 mg/kg bw) induced changes in

the electroretinogram (ERG) and the potass ium

- induced Müller cell depolarisation (KIMD) in rats,

whereby formate in blood increased, with concentrations ranging from approx. 6 to 15 mM at 48

hours. No adverse ocular effects were noted at 1000 mg methanol/kg per os despite folate

deprivation (Lee et al., 1994; Garner et al., 1995a). No adverse effects were produced by oral doses

of up to 3500 mg/kg bw in rats maintained on normal diet (Garner et al., 1995b). The authors

assume that intraretinal metabolism of methanol to formate is necessary for t he initiation of

methanol

- induced retinal toxicity, based on negative results following formate infusion (Garner et

al., 1995a).

Conclusion

Rats are relatively resistant to methanol

- related effects on the visual organ in contrast to humans.

However, monk eys may exhibit clear signs of ocular damage under selective test conditions in

which formate (without associated acidosis) is the causative agent, and where effective formate

blood levels were about 450 mg/L and higher ( ≥ 10 mM). Repeated dosing in monkey s may also

lead to ocular effects.

### OECD SIDS METHANOL

32 

### 3.2 Initial Assessment for Human Health

The most prominent route of occupational exposure to methanol is inhalation. In case of an

expanded use of methanol in automotive fuels, inhalation as well as dermal exposure would bec ome

increasingly important to the general population. Methanol is rapidly absorbed by all potential

routes of exposure and readily distributed in the body.

3.2.1 Acute Toxicity, Skin and Eye Irritation, and Sensitisation

In humans methanol intoxication may lea d to ocular lesions and potential lethality. The minimal

lethal dose is considered to be 300

– 1,000 mg/kg bw. These effects are not mediated by the parent

compound but by a rise in the formate blood levels up to about 5

- >10 mM, which is associated

wi th acidosis. Acidosis may become fatal to the organism unless medical treatment is administered.

There is some evidence from studies in monkeys that the acidotic state does not substantially

contribute to visual impairment (Martin

- Amat et al., 1978: see a bove 3.1.5: Oral Studies and Ocular

Toxicity). Due to species differences in metabolism, there is at present no animal model (including

monkeys) to completely and satisfactorily reflect the toxicity syndrome in humans. In fatal human

cases after acute me thanol intoxication, post

- mortem signs of damage were observed in the basal

ganglia in the brain, specifically the putamen (which participates in the control of motor activities).

Survivors of severe methanol intoxication may continue suffering from resid ual lesions of the

putamen and visual and lingual impairment (see 3.1.2: Toxicity Syndrome).

In rats, LC 50 

### values of 128.8 mg/L (after 4 hours) and 87.5 mg/L (after 6 hours) were calculated. In

monkeys, lethal concentrations of 52 mg/L after 1

- 4 hours and 13 mg/L after 18 hours have been

observed. A dermal LD 50 

### of 17000 mg/kg bw was calculated for rabbits; four daily dermal doses of

400 mg/kg bw eventually resulted in death. Oral LD 50 

### s range from 5600 to 14400 mg/kg bw in rats,

mice, rabbits, and dogs . Monkeys that received oral doses of 3000 to 8000 mg/kg bw died within

two days.

Methanol exhibited no skin irritation in one study but moderate irritation in a study that could not

be verified. Available studies show that it is a mild to moderate eye i rritant (with some reversibility

of effects). High concentration of methanol vapours may be irritating to mucous membranes.

Results of a guinea pig maximization test show no notable sensitisation potential of methanol.

3.2.2 Repeated

- Dose Toxicity

In a chronic exposure study in monkeys, some histological findings were observed in the cerebral

area after prolonged exposure to doses of up to 1.3 mg/L, although the effects were generally of a

mild nature. Several additional effects, including dose

- related liver e ffects, were observed in this

study. Effects in other organs and systems were also observed. The neurological effects were

assumed to be due to methanol because several such effects were observed, neurological effects

were also seen in the subacute monk ey study, and not yet verifiable neurobehavioral effects were

observed in a reproductive/developmental study in monkeys (see Section 3.1.7

– Burbacher et al.,

1999). A LOAEL of 0.013 mg/L was chosen for the chronic continuous

- exposure study in

monkeys. Although the statistical significance of the effects cannot be verified from the study

report, the number of effects and systems affected indicate a relationship with methanol.

In a subacute 20

- d study in monkeys

, 3.9 mg/L (3000 ml/m 3 

### ) was chosen as the LO AEL (subacute,

continuous exposure) where neurotoxic lesions appeared to progress in monkeys (according to

NEDO, 1987). This exposure concentration correlated with methanol blood levels 80 mg/L and

formate levels 30 mg/L.

### OECD SIDS METHANOL

33 

### In the NEDO (1987) studies of c hronic exposure in rats and mice, the effects are difficult to

interpret from the available study, and NOELs (continuous exposure) could be established at the

mid

- dose of 0.13 mg/L, because adverse effects were explicitly not reported.

In a 28

- day study, t here was no evidence of adverse effects in rats exposed to methanol up to 6.5

mg/L (6 hours per day for 28 days), except local nasal irritation and increased relative spleen

weights, which were observed only at the middle dose (Andrews et al., 1987). The estimated blood

level of methanol is expected to be about 250 mg/L under this condition, as described in Section

3.1.1, Tab. 6.

3.2.3 Mutagenicity and Carcinogenicity

The majority of in vitro and in vivo assays on methanol are negative for mutagenicity and

clast ogenicity although a few of the in vitro and in vivo assays were positive or ambiguous. The

positive in

- vivo findings cannot be evaluated on the limited data base available. Although most

studies are negative for mutagenicity, the possibility that methanol can result in mutagenicity cannot

be completely ruled out.

Overall, tumour frequencies showed no statistically significant differences between treatment

groups and controls in either rats or mice. The biological significance of the non

- statistically

signi ficant increase in the incidence of the phaeochromocytomas in the aged female high

- dose rats

is considered to be low. The methanol

- related proliferative effects on the astroglia cells observed in

monkeys were of transient nature and not associated with cel lular neoplastic transformation. The

weight of evidence suggests that methanol is not carcinogenic.

3.2.4 Reproductive and Developmental Effects

No epidemiological studies in humans have been located to demonstrate that there is a link between

methanol exposure and an increased incidence of fetal malformations or developmental impairment.

In monkeys, inhalation exposure of parents prior to and during breeding as well as during

pregnancy was associated with a late wasting syndrome and not verifiable, mild neurob ehavioral

effects in offspring as well as some vaginal bleeding and unproductive labor in mothers. These

recordings resulted in the identification of 0.26 mg/L (the lowest dose tested), as a LOAEL.

However, due to the normal variance in and the low numbe r of animals, the observed findings are somewhat

difficult to interpret.



### Several inhalation studies in rats resulted in a variety of effects in offspring due to prenatal and/or

postnatal dosing. In a 2

- generation reproductive study, decreased brain weight s resulted in a

NOAEL(>19 h/d) of 0.13 mg/L. In another study on rats, malformations and fetal weight changes

resulted in a NOAEL(7 h/d) of 6.5 mg/L (the lowest dose tested). A third study resulted in

malformations, increased fetal resorptions, and decre ased numbers of live fetuses, resulting in a

NOAEL of 1.3 mg/L.

An inhalation study in mice resulted in developmental effects including increased exencephaly and

cleft palate, fully resorbed fetuses, and decreased numbers of live pups; this study resulted in a

NOAEL(7 h/d) of 1.3 mg/L. Oral studies in mice resulted in various malformations at 4000 mg/kg

bw and higher; no NOAELs could be established from these studies.

NTP (2003) concluded that rodent data on reproductive and developmental toxicity are rele vant for

humans despite the known differences in methanol metabolism between rodents and humans.

Rodents are adequate models for human exposure to methanol at levels where formate does not

accumulate. However, blood methanol concentrations associated wit h serious teratogenic effects

### OECD SIDS METHANOL

34 

### and reproductive toxicity are in the range associated with formate accumulation, which is likely to

result in metabolic acidosis, and visual and clinical effects in humans (NTP, 2003). Other effects

(e.g., subtle, not yet def initive neurological effects observed in primates) may be exhibited at lower

inhalation doses and lower methanol blood levels.

3.2.5 Ocular Toxicity

Rats are more or less resistant to methanol

- related effects on the visual organ in contrast to humans.

However , monkeys may exhibit clear signs of ocular damage under selective test conditions in

which formate (without associated acidosis) is the causative agent, and where effective formate

blood levels were about 450 mg/L and higher ( ≥ 10 mM). Repeated dosing in monkeys may also

lead to ocular effects.



## 4 HAZARDS TO THE ENVIR ONMENT

4.1 Aquatic Effects

Acute Toxicity Test Results

Several data on acute toxicity of methanol to fish, invertebrates, and algae are available. The results

of the most relevant studies are shown i n Table 13 and indicate a low toxicity for aquatic organisms

Although not all the studies have been performed in sealed vessels and with analytical control of the

exposure concentrations the low value for the Henry’s Law constant indicates that volatilisat ion is

not a significant removal process from the aquatic phase. The low acute toxicity of methanol for

aquatic organisms is confirmed by QSAR calculations, resulting in acute toxicity values for fish,

daphnids and algae well above 1000 mg/l.

### OECD SIDS METHANOL

35 

### Table 13: S hort

- term Toxicity of Methanol in Aquatic Organisms (Key Studies )

Organism Test type Species Value Rel.

code Reference

Fish flow through

flow through

flow through Lepomis

macrochirus

Salmo

gairdneri

Pimephales

promelas LC 50 (96 h) = 15,400 mg/L

(m easur.)

LC 50 (96 h) = 20,100 mg/L

(measur.)

LC 50 (96 h) = 28,100 mg/L

(measur.) 1

1

1 Poirer et al., 1986

Call et al., 1983

Call et al., 1983

Invertebrates static Daphnia

magna EC 50 (48 h) > 10,000 mg/L

(nom.) 2 Kuehn et al., 1989

Algae stati c

static Scenedesmus

quadricauda

Chlorella

pyrenoidosa Toxic Threshold (192 h) =

8,000 mg/L (nom.)

EC 50 (10

- 14 d) = 28,400

mg/L (nom.) 2

2 Bringmann and

Kuehn, 1978

Stratton and

Smith, 1988

Bacteria static Pseudomonas

putida Tox ic Threshold (16 h) =

6,600 mg/L (nom.) 2 Bringmann and

Kuehn, 1977



### Acute Toxicity to Fish

In continuous flow

- through systems mean LC50(96 h) values for freshwater fish were 15,400 mg/L

( Lepomis macrochirus ) (Poirer, 1986), 20,100 mg/L ( Salmo gairdneri ) and 28,100 mg/L

( Pimephales promelas ) (Call, 1983). These measured effect concentrations are in very good

agreement with a LC50(96 h) of 10,630 mg/L estimated with the QSAR Program ECOSAR (v0.99).

Acute Toxicity to Invertebrates

In a reliable standard st atic test with Daphnia magna no adverse effects were reported at 10,000

mg/L after 48

- h exposure (Kuehn, 1989). A QSAR calculation performed with ECOSAR (v0.99)

indicates a LC50 (48 h) for daphnids of 9375 mg/L.

Acute Toxicity to Algae

The toxicity thresho ld of methanol for the alga Scenedesmus quadricauda after an exposure of 8

days was reported to be at 8,000 mg/L (Bringmann and Kuehn, 1978). The average concentration

causing 50% inhibition in growth rate was found to be at 28,400 mg/L during an exposure period of

10

– 14 days (Stratton, 1988). A QSAR calculation conducted with ECOSAR (v0.99) estimates a

EC50 (96 h) of 4,982 mg/L for green algae.

Chronic Toxicity Test Results

No experimental data are available.

### OECD SIDS METHANOL

36 

### Toxicity to Microorganisms

The lowest toxic l imit concentrations have been reported to be at 6600 mg/L in a cell multiplication

test for 16 hours using two Pseudomonas species (according to German Standard Procedures DEV

L8) ( Bringmann, 1973; Bringmann and Kuehn, 1977).

4.2 Terrestrial Effects

No relevan t data are available.

4.3 Other Environmental Effects

No data are available.

4.4 Initial Assessment for the Environment

Air is the main target compartment for methanol when the distribution is estimated using the

releases to different environmental compartments a s reported to the Toxics Release Inventory.

However, a certain portion of methanol will distribute to the water compartment. Methanol is

readily degradable by photo

- oxidation and biologically under aerobic and anaerobic conditions. It

is of low toxicity to aquatic organisms and does not bioaccumulate in appreciable amounts in fish.

Based on the weight of evidence of the available data, methanol is not considered harmful to the

environment under normal circumstances.



## 5 RECOMMENDATIONS

For human health, th e chemical is currently a a candidate for further work. Methanol exhibits

potential hazardous properties for human health (neurological effects, CNS depression, ocular

effects, reproductive and developmental effects, and other organ toxicity). Rapid meta bolism and

excretion is noted. In the US (the Integrated Risk Information System), further work is being

performed regarding the use and refinement of pharmacokinetic models for extrapolating animal

data to humans.

.

For the environment, the chemical is cur rently of low priority for further work due to its low hazard

profile.

### OECD SIDS METHANOL

37 

## 6 REFERENCES

Abbondandolo, A., Bonatti, S., Corsi, C., et al.(1980). The use of organic solvents in mutagenicity

testing. Mutat. Res. 79, 141

- 150

American Petroleum Institute (1991). Su bmission to US EPA, TSCATS, OTS0000812, Doc. I.D.

FYI

- OTS

- 0791

- 0812, 19 July 1991 (SRI Project 1409

- C01

- 90) Andrews, J.J.C., Terrill, J.B., and

Bolte, H.F. (1987). Subchronic inhalation toxicity of methanol. J. Toxicol. Envir. Health, 20, 117

-

124

Atkinson , R. (1989). Kinetic and Mechanisms of the Gas

- Phase Reactions of the Hydroxyl Radical

with Organic Compounds. Journal of Physical and Chemical Reference Data. Monograph No. 1

BASF AG, Dept. of Toxicology (1961). Short

- term test on methanol. Unpublished re sults, (X/306),

17 Jan. 1961

BASF AG (1972). Sicherheitstechnische Kennzahlen, unpublished study, sample No. 72/0239, 17

May 1972

BASF AG, Dept. of Toxicology (1975). Short

- term studies with methanol. Unpublished data,

(XXIV 436), 03 July 1975

BASF AG (197 9). Sicherheitstechnische Kennzahlen, unpublished study, TKM/SIK 78/1255, 15

Jan. 1979

BASF AG Dept. of Toxicology (1979a). Bericht zur Prüfung auf sensibilisierende Wirkung am

Meerschweinchen

- Maximierungstest (Magnusson und Kligman). Unpublished data, (78 /333), 11

April / 11 July 1979

BASF AG (1980a). Bestimmung der akuten Inhalationstoxizität LC50 von Methanol bei

6stuendiger Exposition an Sprague

- Dawley

- Ratten. Unpublished report, (80/158), 07 Aug. 1980

BASF AG (1980b). Bestimmung der akuten Inhalationst oxizität LC50 von Methanol bei 4

- stuendiger Exposition an Sprague

- Dawley

- Ratten. Unpublished report, (80/158), 20 Nov. 1980

BASF AG Dept. of Ecology (1998). Unpublished calculation, 20 Dec. 1998

BASF AG (1999a). Safety data sheet Methanol pure, 03 Nov. 199 9

BASF AG (1999b), Sicherheitstechnik, internal communication, 02 Nov. 1999

BASF AG, Dept. of Product Safety (2003). Unpublished calculation, 14 Nov. 2003

Batterman, S.A. and Franzblau, A. (1997). Time

- resolved cutaneous absorption and permeation

rates of methanol in human volunteers. Int. Arch. Occup. Environ. Health, 70, 4341

- 351 [cited in

DFG, 1999]

Batterman, S.A. et al. (1998). Breath, urine, and blood measurements as biological exposure indices

of short

- term inhalation exposure to methanol. Int. Arch. Occup. Environ. Health, 71, 325

- 335

[cited in Bouchard et al., 1998 and NTP, 2003]

Bauman, K. and Angerer, J. (1979). Occupational chronic exposure to organic solvents. VI. Formic

acid concentration in blood and urine as an indicator of methanol exposure. Int. Arch. Occup.

Environ. Health, 42, 241

- 249

Becker C.E. (1983). Methanol poisoning. J. Emerg. Med., 1, 51

- 56.

### OECD SIDS METHANOL

38 

### Bolon, B., Dorman, D.C., Janszen, D., Morgan, K.T., and Welsch, F. (1993). Phase

- specific

developmental toxicity in mice following maternal methanol inhalation. Fund. Appl. Toxicol., 21,

508

- 516.

Bouchard, M. et al. (2001). A biologically based dynamic model for predicting the disposition of

methanol and its metabolites in animals and humans. Toxicol. Sci., 64, 169

- 184.

Bringmann, G. (1973) . Gesundheits

- Ingenieur 94(12), 366

- 369.

Bringmann, G., Kuehn, R. (1977). Zeitschrift fuer Wasser

- und Abwasser

- Forschung, 10(3/4), 87

-

98.

Bringmann, G., Kuehn, R. (1978). Vom Wasser, 50, 45

- 60.

Burbacher, T. et al. (1999). Reproductive and Offspring De velopmental Effects Following Maternal

Inhalation Exposure to Methanol in Nonhuman Primates. Part I. Methanol Disposition and

Reproductive Toxicity in Adult Females; Part II. Developmental Effects in Infants Exposed

Prenatally to Methanol; Research Report No. 89, Health Effects Institute (HEI) (October 1999)

Call, D.J. et al. (1983). Toxicity and metabolism studies with EPA priority pollutants and related

chemicals in freshwater organisms, EPA

- 600/3

- 83

- 095, PB83

- 263665

Campbell, J.A. et al. (1991). Eviden ce that methanol inhalation does not induce chromosome

damage in mice. Mutation Research 260, 257

- 264

Carnegie

- Mellon Institute of Research, unpublished data. Cited in Rowe and McCollister, 1981

Chang, LW, McMillan, L, Wynne, BR, Pereira, MA, Colley, RA, W ard, JB, and Legator, MS

(1983). The evaluation of six monitors from the exposure to formaldehyde in laboratory animals.

Environ. Mutagen., 5, 381

- 387 [cited in IPCS/WHO, 1997]

Clary, J.J. (2003). Methanol, is it a developmental risk to humans? Regul. To xicol. Pharmacol., 37,

83

- 91

Clay, K.L. et al. (1975). Experimental methanol toxicity in the primate: Analysis of metabolic

acidosis. Toxicol. Appl. Pharmacol., 34, 49

- 61

Chuwers, P., Osterloh, J., Kelly, T., D ́Alessandro, A., et al. (1995). Neurobehavior al effects of

low

- level methanol vapor exposure in healthy human volunteers. Environ. Res., 71, 141

- 150

CMAI (2001). World Methanol Analysis

Connelly, L.E. and Rogers, J.M. (1997). Methanol causes posteriorization of cervical vertebrae in

mice. Teratolog y 55, 138

- 144

Cook, M.R, Bergman, F.J., Cohen, H.D., et al. (1991). HEI Research Report Number 42: Effects

of methanol vapor on human neurobehavioral measures. Cambridge, MA. Health Effects Institute

[cited in NTP, 2003]

Cooper, J.R. and Felig, P. (19 61). The biochemistry of methanol poisoning, II. Metabolic acidosis

in the monkey. Toxicol. Appl. Pharmacol., 3, 202

- 209

Crebelli, R., Conti, G., Conti, L., and Carere, A. (1989). A comparative study on ethanol and

acetaldehyde as inducers of chromosome malsegregation in Aspergillus nidulans. Mutation

Research, 215, 187

- 195

### OECD SIDS METHANOL

39 

### D ́Alessandro, A. et al. (1994). Formate in serum and urine after controlled methanol exposure at

the threshold limit value. Environ. Health Persp., 102, 178

- 181 [cited in Medinsky an d Dorman,

1995]

Daubert T.E. and Danner R.P. (1989). Physical and Thermodynamic Properties of Pure Chemicals

Data Compilation, Washington, D. C., Taylor and Francis

Davoli, E. et al. (1986). Serum methanol concentrations in rats and in men after a single d ose of

aspartame. Food Chem. Toxicol., 24, 187

- 189 (cited in IPCS/WHO, 1997; NTP, 2003)

DeFlora, S. (1981). Study of 106 organic and inorganic compounds in the Salmonella/microsome

test. Carcinogenesis, 2, 283

- 298 DeFlora, S., Camoirano, A., Zancchi, P. an d Bennicelli, C. (1984a).

Mutagenicity testing with TA97 and TA102 of 30 DNA

- damaging compounds, negative with other

Salmonella strains. Mutat. Res., 134, 159

- 165 DeFlora, S.; Zanacchi, P.; Camoirano, A.; Bennicelli,

C.; Badolati, C. (1984b). Genotoxic act ivity and potency of 135 compounds in the Ames reversion

test and in a bacterial DNA

- repair test. Mutat. Res., 133, 161

- 198

Dethlefs, R. and Naraqi, S. (1978). Ocular manifestations and complications of acute methyl

alcohol intoxication. Med. J. Australia, 2, 483

- 485

DFG Commission for the Investigation of Health Hazards of Chemical Compounds in the Work

Area (1999). Methanol, in: Occupational Toxicants: Critical Data Evaluation for MAK Values and

Classification of Carcinogens (Greim, H., ed.), Wiley

- VCH, Weinheim

DGMK (Deutsche Gesellschaft fuer Mineraloelwissenschaft und Kohlechemie e.V.) (1982).

Wirkung von Methanol auf Mensch und Tier, DGMK

- Projekt 260

- 07, Hamburg, Dec. 1982

Dorman, D.C., Moss, O.R., Farris, G.M., Janszen, D.; Bond, J.A., Medinsky, M.A. : (1994).

Pharmocokinetics of inhaled [14C]methanol and methanol

- derived [14C]formate in normal and

folate

- deficient cynomolgus monkeys. Toxicology and Applied Pharmacology, 128, 229

- 238

Dutkiewicz B., Kónczalik J., Karwacki W. (1980). Skin absorption and per os administration of

methanol in men. Int. Arch. Occup. Environ. Health, 47, 81

- 88

Erlanson, P. et al. (1965). Severe methanol intoxication. Acta Med. Scand., 177, 393

- 408

Eulner H.H. and Gedicke K.H. (1955). Ueber die Hautresorption von Methylalkohol . Sammlung

Vergiftungsfaelle 15, 409

- 414[cited in DGMK, 1982]

Ferry, D. et al. (1980). Methanol monitoring. Int. Arch. Occup. Environ. Health 47, 155

- 163

Franzblau et al. (1993). Absence of formic aid accumulation in urine following five days of

methanol e xposure. Appl. Occup. Environ. Hyg., 8, 883

- 888 [cited from Bouchard et al., 1998]

Franzblau, A. and Batterman, S.A. et al. (1995). Breath monitoring of inhalation and dermal

exposure to methanol. Appl. Occup. Environ. Hyg., 10, 833

- 839 [cited in DFG Commi ssion, 1999]

Frederick, L.J. et al. (1984). Investigation and control of occupational hazards associated with the

use of spirit duplicators. Am. Ind. Hyg. Assoc., 45, 51

- 55 [cited in DFG, 1999 and Kavet and

Nauss, 1990]

Freitag, D., Lay, P. and Korte, F. ( 1985). Environmental Hazard Profile of Organic Chemicals: An

experimental method for the assessment of the behaviour of organic chemicals in the ecosphere by

means of simple laboratory tests with 14C labelled chemicals. Chemosphere, 14, 10, 1589

- 1616

### OECD SIDS METHANOL

4 0 

### Fri tzenschaf, H., Kohlpoth, M., Rusche, B., Schiffmannn, D. (1993). Testing of known carcinogens

in Syrian hamster embryo (SHE) micronucleus test in vitro; correlations with in vivo micronucleus

formation and cell transformation. Mutation Research 319, 47

- 53

Fu, S.S., Sakanshi, T.m., Rogers, J.M., Hong, K.H. and Keen, C.L. (1996). Influence of dietary folic

acid on the developmental toxicity of methanol and the frequency of chromosomal breakage in the

CD

- 1 mouse. Reprod. Toxicol., 10, 455

- 463

Gaffney, J.S. et al. (1987). Environ. Sci. Technol., 21, 6, 519

- 524

Garner, C.D. Lee, E.W., Louis

- Ferdinand, R.T. (1995a). Müller cell involvement in methanol

induced retinal toxicity. Toxicol. Appl. Pharmacol., 130, 101

- 107

Garner, C.D., Lee, E.W., Terzo, T.S., Louis

- Ferdinand, R.T. (1995b). Role of retinal metabolism in

methanol

- induced retinal toxicity. Toxicol. Environ. Health, 44, 43

- 56

Gilger A.P. and Potts A.M. (1955). Studies on the visual toxicity of methanol, V. The role of

acidosis in experimental methanol po isoning. Amer. J. Ophthalmol., 39, 63

- 86. [Cited in DGMK

1982.]

Gluth, G. et al. (1985). Accumulation of pollutants in fish. Comp. Biochem. Physiol., 81C, 2, 273

-

277

Gocke, E., King, M.

- T., Eckhardt, K. and Wild, D. (1981). Mutagenicity of cosmetics ingr edients

licensed by the European Community. Mutat. Res., 90, 91

- 109 Hansch, C. and Leo, A. (1979).

Substituent Constants for Correlation Analysis in Chemistry and Biology,

John Wiley & Sons, New York

Hansch, C. and Leo, A.J. (1985). Medchem. Project Iss ue No.26, Claremont CA, Pomona College

Heid, M.K. et al. (1992). Folate deficiency alone does not produce neural tube defects in mice. J.

Nutr., 122, 1198

- 1200

Heinrich, R. and Angerer, J. (1982). Occupational chronic exposure to organic solvents. X.

Biolo gical monitoring parameters for methanol exposure. Int. Arch. Occup. Environ. Health, 50,

341

- 349. [Cited in DFG, 1999.]

Horton, V.L., Higuchi, M.A. and Rickert, D.E. (1992). Physiologically based pharmacokinetic

model for methanol in rats, monkeys, and h umans. Toxicol. Appl. Pharmacol. 117, 26

- 36

Howard, P.H. (ed.) (1990). Handbook of Environmental Fate and Exposure Data for Organic

Chemicals, Volume II, Solvents, Lewis Publishers Inc., Michigan, 310

- 317

IMPCA Methanol Reference Specifications, April 1999

IPCS/WHO (1997). Methanol, Environmental Health Criteria 196, International Programme on

Chemical Safety, World Health Organization, Geneva.

Jacobs, G.A. (1990). OECD eye irritation tests on three alcohols, in: Acute Toxicity Data. J. Am.

Coll. Toxico l., 1, 56

- 57

Johlin, F.C. et al. (1987). Studies on the role of folic acid and folate

- dependent enzymes in human

methanol poisoning. Mol. Pharmcol., 31, 557

- 561

Katoh, M (1989). New Energy Development Organization data. Presented at the Methanol Vapors

a nd Health Effects Workshop: What We Know and What We Need to Know

– Summary Report.

### OECD SIDS METHANOL

41 

### Washington, DC, ILSI Risk Science Institute/U.S. Environmental Protection Agency/Health Effects

Institute/American Petroleum Institute, p. A

- 7. [cited in IPCS/WHO, 1997]

Kavet, R. and Nauss, K.M. (1990). The toxicity of inhaled methanol vapors. Critical Reviews in

Toxicology 21(1), 21

- 50

Kawai, R. et al. (1991). Methanol in urine as a biological indicator of occupational exposure to

methanol vapor. Int. Arch. Occup. Enviro n. Health, 63, 311

- 318 [cited in DFG, 1999]

Kawai, R. et al. (1991). Methanol in urine as a biological indicator of occupational exposure to

methanol vapor. Int. Arch. Occup. Environ. Health, 63, 311

- 318

Kimura E.T. et al. (1971). Acute toxicity and limit s of solvent residue for sixteen organic solvents.

Toxicol. Appl. Pharmacol., 19, 699

- 704 [cited in DGMK, 1982]

Kuehn, R. et al. (1989). Water Research, 23(4), 495

- 499

Lasne, C., Gu, Z.W., Venegas, W and Chouroulinkov, I. (1984). The in vitro micronucleus assay for

the detection of cytogenetic effects induced by mutagen

- carcinogens: Comparison with the in vitro

sister

- chromatid exchange assay. Mutat. Res., 130, 273

- 282

Leaf , G., Zatman, L.J. (1952). A study of the conditions under which methanol may exer t a toxic

hazard in industry. Br. J. Ind. Med., 9, 19

- 31[cited in IPCS/WHO, 1997]

Lee, E.W., Terzo, T.S., D ́Arcy, J.B., Gross, K.B. and Schreck, R.M. (1992). Lack of blood formate

accumulation in humans following exposure to methanol vapor at the current permissible exposure

limit of 200 ppm. Am. Ind. Hyg. Assoc. J. 53(2), 99

- 104

Lee, E.W., Garner, C.D. and Terzo, T.S. (1994). A rat model manifesting methanol

- induced visual

dysfunction suitable for both acute and long

- term exposure studies. Toxicol. Appl. Pharmacol., 128,

199

- 206

Lehmann K.B. and Flury F. (1943). Toxicology and hygiene of industrial solvents., Williams und

Wilkins, Baltimore (cited in: DGMK 1982)

Leon, A.S., Hunninghake, D.B., Bell, C., Rassin, D.K., Tephly, T.R. (1989). Safety of long

- te rm

large doses of aspartame. Arch Intern Med. 149: 2318

- 24 [cited in NTP, 2003]

Litovitz T. L., et al. (1988). Annual Report of the American Association of Poison Control Centers

National Data Collection System. Am. J. Emerg. Med., 6, 479

- 515

Lorente C. et al. (2000). Maternal occupational risk factors for oral clefts. Scan. J. Work Environ.

Health, 26, 137

- 145.

Lyman,W.J. et al. (1982). Handbook of chemical property estimation methods, McGraw

- Hill, New

York, 5

- 4

Makar, A.B. et al. (1968). Methanol pois oning in monkey. Mol. Pharmacol., 4, 471 [cited in Roe,

1982]

Malcolm Pirnie, Inc. (1999). Evaluation of the fate and transport of methanol in the environment,

prepared for the American Methanol Institute, January.

Martin

- Amat, G., Tephly, T.R., McMartin , K.E. et al. (1977). Methyl alcohol poisoning II.

Development of a model for ocular toxicity in methyl alcohol poisoning using the Rhesus monkey.

Arch. Ophthalmol., 95, 1847

- 1850

### OECD SIDS METHANOL

42 

### Martin

- Amat, G., McMartin, K.E., Hayreh, S.S., Hayreh, M.S. and Tephly, T .R. (1978). Methanol

poisoning: Ocular toxicity produced by formate. Toxicol. Appl. Pharmacol., 45, 201

- 208.

Martinasevic, M.K., Green, M.D., Baron, J. and Tephly, T.R. (1996). Folate and 10

- formyltetrahydrofolate dehydrogenase in humans and rat retina: Relation to methanol toxicity.

Toxicol. Appl. Pharmacol., 141, 373

- 381.

Mashbitz N.L. et al. (1936). J. Industr. Hyg. Toxicol., 18, 117 (cited in: DGMK 1982)

McCord, C.P. (1931). Toxicity of methyl alcohol (methanol) following skin absorption and

inhalat ion. Industrial and Engineering Chemistry, 23, 931

- 936.

McGregor, D.B., Martin, R., Riach, C.G. and Caspary, W.J. (1985). Optimisation of a metabolic

activation system for use in the mouse lymphoma L5178Y tk+tk

- mutation system. Environ.

Mutagen., 7(Supp l. 3), 10 (abstr.).

McMartin, K.E., Makar, A.B., Martin G., Palese, A. and Tephly, T.R. (1975). Methanol poisoning

I. The role of formic acid in the development of metabolic acidosis in the monkey and the reversal

by 4

- methylpyrazol. Biochem. Med., 13, 3 19

- 333.

McMartin, K.E., Martin

- Amat, G., Makar, A.B., and Tephly, T.R. (1977). Methanol poisoning V.

Role of formate metabolism in the monkey. J. Pharmacol. Exp. Ther., 201, 564

- 572.

Medinsky, M., Dorman, D.C., Bond, J.A., Moss, O.R., Janszen, D.B. and Ev eritt, J.I. (1997).

Pharmacokinetics of methanol and formate in female Cynomolgus monkeys exposed to methanol

vapors. Health Effects Institute, Res. Report No. 77

Medinsky, M.A. and Dorman, D.C. (1995). Recent development in methanol toxicity. Toxicology

Lett., 82/83, 707

- 711

Methanol Institute (2003). World Methanol Plants (through December 2001),

http://www.methanol.org

Munch J.C. (1972). Industr. Med., 41, 31

- 33, [cited in: DGMK, 1982]

Muttray A. (2001). Int. Ar ch. Occup. Environ. Health, 74, 43

- 48

Naraqi, S. et al. (1979). An outbreak of acute methyl alcohol intoxication. Aust. N.Z. J. Med., 9,

65

- 68

NIOSH, Registry of Toxic Effects of Chemical Substances, Ausgabe Juli (1982). [cited in: DGMK,

1982]

National Library of Medicine (2003). Household Products Database.

http://householdproducts.nlm.nih.gov/cgi

- bin/household/

NEDO (1987). Toxicological Research of Methanol as a Fuel for Power Station, Summary Report

on Tests with Monkeys, Rats and Mice. New Energy Development Organization, Tokyo Nelson,

B.K. and Brightwell, W.S. (1985). Teratological assessment of methanol and ethanol at high

inhalation levels in rats. Fund. Appl. Toxicol., 5, 727

- 736 NIOSH (1985). Dangerous Properties of

Industrial Materials Report

NIOSH (1982). Registry of Toxic Effects of Chemical Substances, Edition July 1982 [cited in:

DGMK 1982]

NTP (2003). NTP

- CERHR Monograph on the Potential Human Reproductive and Develo pmental

Effects of Methanol. NIH Publication No. 03

- 4478. September, 2003

### OECD SIDS METHANOL

43 

### Obe, G. and Ristow, H. (1977). Chromosomal damage by alcohol in vitro and in vivo. Mutat. Res.,

56, 211

- 213 Osterloh, J.D. et al. (1996). Serum concentrations of methanol after inha lation at 200

ppm. J. Occup. Environ. Med., 38, 571

- 576 [cited in Bouchard et al., 1998]

Pereira, M.A., Chang, L.W., McMillan, L., Ward, J.B., Legator, M.S. (1982). Battery of short

- term

tests in laboratory animals to corroborate the detection of human pop ulation exposures to genotoxic

chemicals. Environ. Mutagen., 4, 317 (abstr.)

Perkins, A.P., Ward, K.W. and Pollack, G.M. (1996). Ventilation of ambulatory rats exposed to

methanol vapor in flow

- through exposure system: measurement and input to toxicokineti c models.

Inhal. Toxicol., 8, 143

- 162

Perkins, R.A., Ward, K.W. and Pollack, G.M. (1995). Comparative toxicokinetics of inhaled

methanol in the female CD

- 1 mouse and Sprague

- Dawley rat. Fund. Appl. Toxicol. 28, 245

- 254

Poirer S.H., Knuth, L.M., Anderson

- B uchou, C.D. et al. (1986). Comparative toxicity of methanol

and N,N

- dimethylformamide to freshwater fish. Bull. Environ. Contam. Toxicol., 37, 615

- 621

Pollack, G.M. and Brouwer, K.L.R (1996). Maternal

- fetal pharmacokinetics of methanol. Health

Effects Inst itute (HEI), Res. Report No. 74

Potts, A.M. et al. (1955). Studies on the visual toxicity of methanol, VIII. Additional observations

on methanol poisoning in the primate test object. Amer. J. Ophthalmol., 40, 76

- 83 [cited in DGMK

1982]

Potts, A.M. (1955). The visual toxicity of methanol, VI. The clinical aspects of experimental

methanol poisoning treated with base. Amer. J. Ophthalmol., 39, 86

- 92 [cited in DGMK, 1982]

Price, K.S. et al. (1974). JWPCF, 46(1), 63

- 77

Ragan, D.L. and Boreiko, C.J. (1981). Init iation of C3H/10T1/2 cell transformation by

formaldhyde. Cancer Letters, 13, 325

- 331. Rekker, R.F. (1977). The Hydrophobic Fragmental

Constant, Elsevier Scientific Publishing Company,Amsterdam

Roe O. (1955). The metabolism and toxicity of methanol. Pharm col. Rev., 7, 399

- 412

Roe O. (1982). Species differences in methanol poisoning. Crit. Rev. Toxicol., 10, 275

- 286

Rogers, J.M., Mole, M.L., Chernoff, N., Barbee, B.D., Turner, C.I., Logsdon, T.R. and Kavlock,

R.J. (1993). The developmental toxicity of inha led methanol in the CD

- 1 mouse, with quantitative

dose

- response modeling for estimation of benchmark doses. Teratology, 47, 175

- 188 Rowe, V.K.

and McCollister, S.B. (1981). Alcohols in: Patty’s Industrial Hygiene and Toxicology, Vol. II C,

4527

- 4708.

Sa ngster J., J. Phys. Chem. Ref. Data, Vol. 18, No. 3, p.1150, 1989

Sedivec, V. et al. (1981). Bioogical monitoring of persons exposed to methanol vapours. Int. Arch.

Occup. Environ. Health, 48, 257

- 271

Shimizu H., Suzuki, Y., Takemura, N., Goto, S., Matsu shita, H.(1985). The results of microbial

mutation test for forty

- three industrial chemicals. Jpn. J. Ind. Health, 27, 400

- 419

Smyth H.F. et al. (1941). The single dose toxicity of some glycols and derivatives. J. Ind. Hyg.

Toxicol., 23, 259. [Cited in: Ro we V.K. and McCollister S.B.: Patty’s Industrial Hygiene and

Toxicology, Vol. II, 1981]

### OECD SIDS METHANOL

44 

### Stegink, L.D., Brummel, M.C., McMartin, K.E. et al. (1981). Blood methanol concentrations in

normal adult subjects administered abuse doses of aspartame. J. Toxicol. Environ. Health., 7, 281

- 290 [cited in NTP, 2003]

Stegink, L.D., Brummel, M.C., Filer, L.J.J., Baker, G.L. (1983). Blood methanol concentrations in

one

- year

- old infants administered graded doses of aspartame. J. Nutr., 113, 1600

- 1606 [cited in

NTP, 2003 ]

Stern, S., Reuhl, K., Soderblom, S. et al. (1996). Perinatal methanol exposure in the rat I. Blood

methanol concentration and neural cell adhesion molecules. Fund. Appl. Toxicol., 34, 36

- 46

Stern, S., Cox. C., Preston, R. et al. (1997). Perinatal metha nol exposure in the rat II. Behavioral

effects in neonates and adults. Fund. Appl. Toxicol., 36, 163

- 176.

Stratton, G.W., Smith, T.M. (1988). Interaction of organic solvents with the green alga Chlorella

pyrenoidosa, Bull. Environ. Contam. Toxicol., 40(5) , 736

- 742.

Strobel, R. and Greb, W. (1981). Reproducibility of and experiences with the BHK cell

transformation assay. Toxicology, 19, 55

- 66

Takeda, K. und Katoh, N. (1988). Long

- term effects of methanol vapor at low concentration. Proc.

of the 8th Int. Sy mp. Alcohol Fuels, Tokyo, Japan.

Tephly, T.R. and McMartin, K.E. (1984). Methanol metabolism and toxicity. In: Stegnick, L.D.,

and Filer, LJ., Jr. ed. Aspartame: Physiology and biochemistry. New York, Basel, Marcel

Dekker,. Pp. 111

- 140. [cited in I PCS/WHO, 1997]

The Merck Index, 11th edition, p. 939, 1989

Ullmann’s Encyclopedia of Industrial Chemistry, 6th Edition, 2000 Electronic Release

US EPA, Office of Solid Waste, Washington, DC. Rat oral subchronic toxicity study with methanol

(1986), [cited in US EPA/IRIS Database, 1995]

US EPA/IRIS Database (1995). Methanol, Update through October 1995

US EPA TRI Explorer Report (USCH) (2003). http://www.epa.gov/cgi

- bin/

Wagner, R. (1976). Vom Wasser 47, 241

- 2 65.

Von Burg, R. (1994). J. Appl. Toxicol. 14(4), 309

- 313.

Ward, J.B., Hokanson, J.A., Smith, E.R. et al. (1984). Sperm count, morphology, and fluorescent

body frequency in autopsy service workers exposed to formaldehyde. Mutat. Res., 130, 417

- 424.

Wats on, W., Stern, S., Soderblom, S.C. et al. (2003). 2002 Annual Report of the American

Association of Poison Control Centers Toxic Exposure Surveillance System. American Journal of

Emergency Medicine, 21, 353

- 421. http:// aapcc.org/

Weiss, B. , Stern, S., Soderblom, S.C. et al. (1996). Developmental neurotoxicity of methanol

exposure by inhalation in rats. Health Effects Institute (HEI), Res. Report No. 73.

Witte R. (1931). Dissertation Wuerzburg [cited in: DGMK, 1982]

Ya sugi, T. et al. (1992). Formic acid excretion in comparison with methanol excretion in urine of

workers occupationally exposed to methanol. Int. Arch. Occup. Environ. Health, 64, 329

- 337 [cited

in DFG, 1999].

### OECD SIDS METHANOL

45 

### Youssef, A.F., Weiss, B., Cox, C. (1993). Neuro behavioral toxicity of methanol reflected by

operant running. Neurotoxicol Teratol.,15, 223

- 227 (cited in NTP (2003)

### OECD SIDS METHANOL

46 

### APPENDIX A: TOXICOK INETIC STUDIES IN AN IMALS AND HUMANS

Studies in Animals

In Vivo Studies

Oral. After a single oral exposure to rats and mice, absorption was rapid and nearly complete. In

rats, single doses of 100 and 2500 mg methanol/kg led to maximum blood concentrations of 100

and 2000 mg methanol/L, respectively. In mice, 2500 mg/kg bw caused a maximum blood

concentration of 2500 mg/L (Pollack and Brouwer, 1996).

Inhalation

. In monkeys, m 

### ethanol air concentrations of 0.26, 0.78, and 2.24 mg/L (200, 600, 1800

ml/m 3 

### ) resulted in blood concentrations of 5 mg/L (1

- to 2

- fold), 10 mg/L (3

- to 4

- fold), and 40

– 50

mg/L (13

- to 16

- fold above baseline), respectively, after single or repeated exposure for 2.5

hours/day (Burbacher et al., 1999). After repeated inhalation of 2.6 mg/L (2000 ml/m 3 

### 6 hrs/day; 5

days/week; 1 or 2 weeks) monkeys showed no methanol or formate accumulation (Horto n et al.,

1992).

In rats, three consecutive 6

- hour exposures of about 5.85 mg/L (4500 ml/m 3 

### ) resulted in higher

initial levels of methanol up to 950 mg/L, which then levelled off to 600 mg/L in blood (Stern et al.,

1996).

The mean fractional respiratory absorption of methanol in rats and mice was 85 percent at air

concentrations from 1.3 to 6.5 mg/L (1000 to 5000 ml/m 3 

### ). At higher concentrations (13

– 26

mg/L), absorption tended to be lower in rats (60

– 70 percent), but not in mice (Pollack and

Brouwer, 1996; Perkins et al., 1995; 1996). The lower absorption efficiency from the upper

respiratory tract in rats was mainly attributed to a lower breathing rate. The rate of methanol

accumulation in mice is two

- to three

- fold higher than in rats due to their more rapid breathing and

higher absorption, even though mice have a two

- fold higher elimination rate. These factors

possibly contribute to the greater sensitivity of mice to the teratogenic effects of methanol, which

are mediated by the parent compound (P ollack and Brouwer, 1996).

Dorman et al. (1994) and Medinsky et al. (1997) investigated the influence of methanol on the

endogenous formate pool in monkeys under folate

- sufficient (FS) and folate

- deficient (FD) dietary

conditions. Radiolabelled methanol was measured during exposure of female Cynomolgus

monkeys ( Macaca fascicularis ) to 0.013, 0.059, 0.260, and 1.2 mg/L (10, 45, 200, and 900 ml/m 3 

### )

(FS) and 1.2 mg/L (900 ml/m 3 

### ) (FD) for two hours per session. All doses except the highest might

be encounter ed during use of methanol as an automotive fuel. Reported blood concentrations of

methanol 2 hours after the beginning of exposure were 0.65, 96, 672, and 3,392 mg/L for the FS

exposures, and 211 ±71 μM (6.752 μg/L) for the FD exposure. The corresponding formate levels

were 0.07 ±0.02, 0.25 ±0.09, 2.3 ±2.9, and 2.8 ± 1.7 μM (FS exposures) and 9.5 ± 4.7 μM (FD

exposure). Although there was an indication of higher formate levels under folate deficiency, these

increments were far below the endogenous blood formate level of 100

– 500 μM.

Other Routes. Intraperitoneal injection of doses of methanol did not lead to formate accumulation

in Cynomolgus monkeys ( Macaca fascicularis ) except at a high dose of 3000 mg/kg bw, which is

associated with acidosis and leth ality. No abnormality was seen in the 25

- to 600

- mg/kg dose

groups (NEDO, 1987).

In rats after intravenous administration 97 percent of the clearance of methanol occurred by

oxidative metabolism, whereas renal and pulmonary excretion contributed only to 0.6 and 2.5

percent, respectively (Horton et al., 1992).

### OECD SIDS METHANOL

47 

### Studies in Humans

In Vivo Studies

Oral

- Methanol. Volunteers receiving methanol in amounts of 0.2 ml/h for 6 hours or 0.5 ml/h for

8 hours, which correspond to total doses per person of 950 mg (14 mg/kg bw) and 2600 mg (40

mg/kg bw), exhibited urinary methanol concentrations of below 8 and 18 mg/L, respectively (Ferry

et al., 1980).

In another study, blood levels of 47 to 76 mg/L were measured 2 to 3 hours after oral uptake of 71

– 84 mg methanol/ kg bw (6.6

– 7.4 ml per person). Methanol disappearance obeyed first

- order

kinetics with a half

- life of about 3 hours. Only a very small fraction of an oral dose of 50 mg

methanol/kg (about 2 percent) was eliminated unchanged via the respiratory and urin ary routes, and

about 98 percent was metabolised (Leaf and Zatman, 1952; DFG, 1999; IPCS/WHO, 1997).

Oral

- Aspartame. In another study, fasted adults were administered 34, 100, 150, or 200 mg

aspartame/kg in 300 ml orange juice as a single dose, which c orresponded to methanol portions of

about 3.5, 10, 15 and 20 mg/kg bw. Blood methanol levels were below detection (4.0 mg/L) in the

lowest dose group. At 100 mg/kg bw, blood methanol and urinary formate were increased in a dose

related manner. However, b lood formate levels were not increased at any dose. Peak mean blood

methanol levels were 12.7, 21.4, 25.8 mg/L, at 100, 150, and 200 mg aspartame/kg. All levels

returned to normal within 24 hours after exposures ceased (Stegink et al., 1981). Studies in w hich

one

- year old infants were administered aspartame showed similar results (Stegink et al., 1983).

When aspartame was administered in a single dose (500 mg/person = approx. 50 mg methanol

equivalent) methanol blood levels were not much different from th e endogenous levels. Mean

increases close to 1 mg/L were recorded compared with pre

- exposure levels of 1.4

– 2.6 mg/L

(Davoli et al., 1986).

A longer

- term study demonstrated that ingesting aspartame in a methanol equivalent of 7.5 mg/(kg

bw

* 

### d) resulted i n blood methanol levels around 10 mg/L in adults (Leon et al., 1989), which is still

far below metabolic saturation.

Inhalation. Inhalation studies have shown a net absorption of methanol of 60 to 85 percent

(IPCS/WHO, 1997). In one study, the absorptio n half

- life in a group of 22 volunteers after

inhalation was estimated to 0.80 hours (Osterloh et al., 1996). Using data from inhalation exposure

towards 1.05 mg/L (800 ml/m 3 

### ) methanol for 30 and 120 minutes, the half

- life of methanol in blood

was 1.44 ho urs (Batterman et al., 1998).

After a single exposure of 0.26 mg/L (200 ml/m 3 

### ) methanol, blood levels in exposed humans

increased 3.5 to 4

- fold above the endogenous blood methanol concentration from 1.8 to 7.0 mg/L at

rest and from 1.9 to 8.1 mg/L under light exercise (Lee et al., 1992). Formate levels remained

unchanged, irrespective of physical activity. These findings were confirmed by D ́Alessandro et al.

(1994) and Franzblau et al. (1993) in similar experiments on human volunteers. Medinsky and

Dor man (1995) also reported similar results. Higher urinary methanol concentrations of

approximately 40 mg/L have been predicted for workers exposed to 200 ml/m 3 

### (8

- hour shift)

(Heinrich and Angerer, 1982; Kawai et al., 1991).

Twenty

- six volunteers exposed to 0.26 mg/L (200 ml/m 3 

### ) methanol for 4 hours showed peak

methanol concentrations in blood of 6.5 ± 2.7 mg/L at the end of the exposure which declined during

a 4

- hour post

- exposure period without reaching pre

- exposure levels (1.0 ± 0.6 mg/L for a water

vapor control phase and 1.8 mg/L ± 2.6 for a methanol phase); these were based on a restricted diet

without coffee, vegetables, fruit, alcohol, or aspartame (NTP, 2003). Formate levels in serum and

urine remained unchanged (Osterloh et al., 1996 and Chuwers et al., 1995).

### OECD SIDS METHANOL

48 

### In a pilot study designed to measure neurobehavioral effects, a single exposure of 0.25 mg/L for 75

minutes resulted in increased plasma and urine methanol concentrations (3.3 and 2.5 fold,

respectively). No change in plasma formate concentrat ions was observed (average 0.08 mmol/L =

3.6 mg/L) (Cook et al., 1991).

Dermal. Following dermal exposure in human volunteers, average skin absorption rates of

methanol have been reported to be at 0.192 mg/(cm 2

* 

### min) (Dutkiewicz et al., 1980) and 8.1 ± 3.7

mg/(cm 2

* 

### h) = 0.135 ± 0.062 mg/(cm 2

* 

### min) (Batterman and Franzblau, 1997). According to

Dutkiewicz et al. (1980), exposure of one hand to liquid methanol for 2 minutes would result in a

body burden of 170 mg, similar to that resulting from exposure to a n approximate air concentration

of 0.050 mg/L (38 ml/m 3 

### ) for 8 hours (Dutkiewicz et al., 1980). Full exposure of one hand for 16

minutes is equivalent to blood levels after inhalation of 400 ml/m 3 

### for one 8

- hour working shift (11

mg/L; corrected for backg round value) (Franzblau and Batterman, 1995; Batterman and Franzblau,

1997).

PBPK Modeling

PBPK model simulations show that after a five day continuous inhalation of 0.26 mg/L (200

ml/m 3 

### ), predicted blood and urine levels of methanol (5.5 mg/L and 8.1 mg/ L, respectively) were

not higher than after one exposure session, and blood and urinary formate levels (1.16 mg/L and 1.5

mg/L, respectively) also remained far below reported baseline levels (4.9

- 10.3 and 6.3

- 13 mg/L,

respectively) (Bouchard et al., 2001). These simulations were consistent with data produced in

humans (Sedivec et al., 1981; Osterloh et al., 1996; Batterman et al., 1998) and show that such

exposure levels do not achieve metabolic saturation.

### OECD SIDS METHANOL

49 

# I U C L I D

D a t a S e t

**Existing Chemical** ID: 67

- 56

- 1

**CAS No.** 67

- 56

- 1

**EINECS Name** methanol

**EC No.** 200

- 659

- 6

**Molecular Weight** 32.04 g/mol

**Molecular Formula** C H4 O

**Producer Related Part**

**Company:** BASF AG

**Creation date:** 12

- NOV

- 1992

**Substance Related Part**

**Company:** BASF AG

**Creation date:** 12

- NOV

- 1992

**Memo:** master

**Printing date:** 26

- OCT

- 2006

**Revis ion date:**  15

- DEC

- 2004

**Date of last Update: 26**

**- OCT** 

**- 2006** 

**Number of Pages: 327**

**Chapter (profile):** Chapter: 1, 2, 3, 4, 5, 6, 7, 8, 10

**Reliability (profile):** Reliability: without reliability, 1, 2, 3, 4

**Flags (profile):** Flags: wi thout flag, confidential, non confidential, WGK

(DE), TA

- Luft (DE), Material Safety Dataset, Risk

Assessment, Directive 67/548/EEC, SIDS, BPD Notification,

key study, registration BPD B

- d ocs, registration BPD

A

- docs

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

50

**1.0.1 Applicant and Company Information**

**Type:** lead organisation

**Name:** BASF AG

**Contact Person:** Dr. Rolf Sarafin **Date:**

GUP/CR

- Z570

**Street:** Carl

- Bosch

- Str

**Town:** 67056 Ludwigshafen

**Country:** Germany

**Phone:** +49 621 60 44712

**Telefax:** +49 621 60 58043

**Email:** rolf.sarafin@basf.com

**Homepage:** www.basf.com

18

- APR

- 2006

**Type:** o ther: cooperating organisation

**Name:** Methanol Institute

**Country:** United States

**Type:** cooperating company

**Name:** Methanor v.o.f.

**Country:** Netherlands

**Type:** cooperating company

**Name:** Shell Chemicals Ltd.

**Country:** United Kingdom

**Type:** cooperating company

**Name:** Statoil ASA

**Country:** Norway

**Type:** cooperating company

**Name:** TOTAL Raffinerie Mitteldeutschland GmbH

**Country :**  Germany

**Type:** cooperating company

**Name:** Veba Oil Refining & Petrochemicals GmbH

**Country:** Germany

**1.0.2 Location of Production Site, Importer or Formulator**

**-**

**1.0.3 Identity of Recipients**

**-**

**1.0.4 Details on Category/Template** 

**-**

**1.1.0 Substance Identification**

**Mol. Formula:** C H4 O

**Mol. Weight:** 32.04 g/mol

**Remark:** INDEX

- Number: 603

- 001

- 00

- X

12

- SEP

- 2005

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

51 **1.1.1 General Substance Information**

**Substance type:** organic

**Physical status:** liquid

**Purity:** >= 99.85

- % w/w

**Colour:** colourless

**Odour:** alcoholic

**Method:** GC

**Remark:** The Data of the Substance Related Part are also submitted

on behalf of the following companies:

Akzo Chemicals International B.V. (NL)

Arco Chimie France, SNC

Aragonesas, Industrias y Energia, S.A. (E)

Bayer AG (D)

BP Chemicals LTD (GB)

Brenntag AG (D)

Cepsa (E)

Cheminova Agro A/S (DK)

DEA Mineraloel AG (D)

DOW Corning Coordination Center (B)

Du Pont de Nemours GmbH (D)

Ecofuel S.P.A (I)

Elf Atochem (F)

Ertisa, S.A. (E)

Hoechst AG

Huels AG

ICI Chemicals & Polymers (GB)

Krems Chemie Aktiengesellschaft (A)

Leuna Raffine riegesellschaft mbH (D)

Methanor v.o.f (NL)

(Montefibre) (I) = Enichem Fibre (I)

Neste Resins B.V. (NL)

Neste OY (FIN)

Norsk Hydro A/S (N)

Perstorp AB (S)

Perstorp SpA, Div. Polyols (I)

Praoil (Enichem Group) (I)

Rhone

- Poulenc Chimie (F)

Rhone

- Poulenc Rhodia AG (D)

Ruetgerswerke AG (D)

Ruhr Oel GmbH (D)

Shell Nederland Chemie B.V.

Union Carbide Benelux NV (B)

VEBA Oel AG (D)

Wilhelm Grean GmbH & Co.

12

- SEP

- 2005 (1)

**Substance type:** organic

**Physical status:** liquid

**Colour:** colourless

**Odour:** alcohol

- like

12

- SEP

- 2005 (2)

**Substance type:** other: Substance classification:

Alcohols

**Physical status:** liquid

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

52 **Colour:** colourless

**Odour:** ageeable to pungent

12

- SEP

- 2005 (3)

**Purity:** >= 99

- % w/w

**Method:** GC.

12

- SEP

- 2005 (4)

**1.1.2 Spectra**

**1.2 Synonyms and Tradenames**

Methanol

Methanol (8CI, 9CI)

Methyl alcohol

Methyl hydroxide

Methylol

Monohydroxymethane

**1.3 Impurities**

**EINECS**

**- Name:**  Chloride as Cl

**Mol. Formula:** Cl

**Method:** IMPCA 002

- 92

**Remark:** <= 0.00005 % w/w (<= 0.5 mg/kg).

12

- SEP

- 2005 (5)

**CAS**

**- No:**  7704

- 34

- 9

**EC**

**- No:**  231

- 722

- 6

**EINECS**

**- Name:**  sulphur

**Mol. Formula:** S

**Method:** ASTM D 3961

- 89

**Remark:** <= 0.00005 % w/w (0.5 mg/kg).

12

- SEP

- 2005 (5)

**CAS**

**- No:**  7439

- 89

- 6

**EC**

**- No:**  231

- 096

- 4

**EINECS**

**- Name:**  iron

**Mol. Formula:** Fe

**Method:** ASTM E 394

- 94

**Remark:** Total iron.

<= 0.00001 % w/w (0.1 mg/kg).

12

- SEP

- 2005 (5)

**CAS**

**- No:**  7732

- 18

- 5

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

53 **EC**

**- No:**  231

- 791

- 2

**EINECS**

**- Name:**  water

**Mol. Formula:** H2 O

**Contents:** <= .1

- % w/w

**Method:** ASTM E 1064

- 92

(5)

**CAS**

**- No:**  64

- 17

- 5

**EC**

**- No:**  200

- 578

- 6

**EINECS**

**- Name:**  ethanol

**Mol. Formula:** C2 H6 O

**Contents:** <= .005

- % w/w

**Method:** ASTM E 346

- 94

**Remark:** <= 50 mg/kg.

12

- SEP

- 2005 (5)

**EINECS**

**- Name:**  Acidity (free acid as acetic acid)

**Contents:** <= .003

- % w/w

**Method:** ASTM D 1613

- 91

**Remark:** <= 30 mg/kg.

12

- SEP

- 2005 (5)

**CAS**

**- No:**  67

- 64

- 1

**EC**

**- No:**  200

- 662

- 2

**EINECS**

**- Name:**  acetone

**Mol. Formula:** C3 H6 O

**Contents:** <= .003

- % w/w

**Method:** ASTM E 346

- 94

**Remark:** <= 30 mg/kg.

12

- SEP

- 2005 (5)

**1.4 Additives**

**-**

**1.5 Total Quantity**

**Quantity:** 100000

- 500000 tonnes produced in 1991

**Quantity:** 100000

- 500000 tonnes imported in 1995

**Quantity:** 100000

- 500000 tonnes produced in 1995

**Qu antity:**  100000

- 500000 tonnes produced in 1999

**Quantity:** 10000

- 50000 tonnes imported in 1999

**Quantity:** 10000

- 50000 tonnes imported in 2000

**Quantity:** > 1000000 tonnes produced in 2000

**Remark:** Production volume s in the year 2000:

North America (NAFTA): Production : approx. 7 Mtonnes

Import : approx. 4 Mtonnes

- > Total supply: approx. 11 Mtonnes

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

54 use: Formaldehyde : approx. 23%

DMT * : approx. 2%

Acetic Acid : approx. 9%

MTBE/TAME * : approx. 36%

Methyl Methacrylate: approx. 3%

Gasoline/Fuel : approx. 0.2%

Solvents : approx. 6%

Others : appro x. 14%

Exports : approx. 7%

South America approx. 7 Mtonnes

incl. Trinidad

Western Europe approx. 3.3 Mtonnes

Eastern Eu rope approx. 0.5 Mtonnes

Former Soviet Union approx. 2 Mtonnes

Africa and approx. 5 Mtonnes

Middle East

Asia/Pacific approx. 5 Mtonnes

World approx. 35 Mtonnes/a capacity

* DMT = dimethyl terephthalate

TAME = tert. amylmethylether = 2

- methoxy

- 2

- methylbutane

MTBE = tert

- butyl methyl ether

**Quantity:** 5 0000

- 100000 tonnes imported in 2002

**Quantity:** > 1000000 tonnes produced in 2002

**Flag:** confidential

**Quantity:** 0 tonnes imported in 2005

18

- APR

- 2006

**Quantity:** 100000

- 500000 tonnes produced in 2005

18

- APR

- 2006

**1.6.1 Labelling**

**Labelling:** as in Directive 67/548/EEC

**Symbols:** (F) highly flammable

(T) toxic

**Specific limits:** yes

**R**

**- Phrases:**  (11) Highly flammable

(23/24/25) Toxic by inhalation, in conta ct with skin and if

swallowed

(39/23/24/25) Toxic: danger of very serious irreversible

effects through inhalation, in contact with skin and if

swallowed

**S**

**- Phrases:**  (7) Keep co ntainer tightly closed

(16) Keep away from sources of ignition

- No smoking

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

55 (36/37) Wear suitable protective clothing and gloves

(45) In case of accident or if you feel unwell, seek medical

advice immediately (show the label where possible)

12

- SEP

- 2005 (2) (6)

**1.6.2 Classification**

**Classified:** as in Directive 67/548/EEC

**Class of danger:** highly flammable

**R**

**- Phras**  **es:** (11) Highly flammable

**Specific limits:** no

12

- SEP

- 2005 (2) (6)

**Classified:** as in Directive 67/548/EEC

**Class of danger:** toxic

**R**

**- Phrases:**  (23/24/25) Toxic by inhalatio n, in contact with skin and if

swallowed

**Specific limits:** yes

**Conc./Class. 1:** C >= T; R23/24/25

- 39/23/24/25

20%

12

- SEP

- 2005 (2) (6)

**Classified:** a s in Directive 67/548/EEC

**Class of danger:** toxic

**R**

**- Phrases:**  (39/23/24/25) Toxic: danger of very serious irreversible

effects through inhalation, in contact with skin and if

swallowed

**Specific limits:** yes

**Conc./ Class.**  **1:** C >= T; R23/24/25

- 39/23/24/25

20%

**Conc./Class. 2:**  10% <= T; R20/21/22

- 39/23/24/25

C <

20%

12

- SEP

- 2005 (2) (6)

**Classified:** as in Directive 67/548/EEC

**Class of danger:** harmful

**R**

**- Phrases:**  (20/21/22) Harmful by inhalation, in contact with skin and if

swallowed

**Specific limits:** yes

**Conc./Class. 1:**  C >= T; R23/24/25

- 39/23/24/25

20%

**Conc./Class. 2:** 10% <= T; R20/21/22

- 39/23/24/25

C <

20%

**Conc./Class. 3:**  3% <= Xn; R20/21/22

- 68/20/21/22

C <

10%

12

- SEP

- 2005 (6)

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

56 **Classified:** as in Directive 67/548/EEC

**Class of danger:** harmful

**R**

**- Phrases:**  (68/20/21/22) Harmful: possible risk of irreversible effects

through inhalation, in contact with skin and if swallowed

**Specif ic limits:**  yes

**Conc./Class. 1:**  C >= T; R23/24/25

- 39/23/24/25

20%

**Conc./Class. 2:** 10% <= T; R20/21/22

- 39/23/24/25

C <

20%

**Conc./Class. 3:**  3% <= Xn; R20/21/22

- 68/20/21/22

C <

10%

12

- SEP

- 2005 (6)

**1.6.3 Packaging**

**-**

**1.7 Use Pattern**

**Type:** type

**Category:** Wide dispersive use

**Type:** industrial

**Category:** Basic industry: basic chemicals

**Remark:** Use as Feedstock for Chemical Syntheses

Approximately 70 % of the methanol produced worldwide is

used in chemical syntheses: in order of importance

formaldehyde, methyl tert

- butyl ether (MTBE), acetic acid,

methyl methacrylate, and dimethyl terephthalate. Only a

small proportion is utilized for energy production, although

this use has great potenti al.

Formaldehyde

Formaldehyde is the most important product synthesized from

methanol; in 1988, 40 % of the methanol produced worldwide

was used to synthesize this product.

The processes employed are all based on the oxidation of

methanol with atmospheric oxygen.

Methyl tert

- butyl ether is produced by reacting methanol

with isobutene on acid ion exchangers. Increa sing amounts of

methanol are used in this form in the fuel sector. The ether

is an ideal octane booster and has become extremely

important due to the introduction of unleaded grades of

gasoline and awareness of the possible harmfulness of

aromatic high

- octane components. In 1988, 20 % of worldwide

methanol production was used for MTBE synthesis; annual

increase rates of up to 12 % ar e expected.

Acetic Acid

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

57 Another 9 % of the methanol produced is used to synthesize

acetic acid, and annual growth rates of 6 % are estimated.

Acetic acid is produced by carbonylation of methanol with

carbon monoxide in the liquid phase with cobalt

- iodine,

rhodium

- iodine, or nickel

- iodine homogeneous catalysts .

Other Synthesis Products

In the intensive searc h after the oil crisis for routes to

alternative fuels, processes were developed that allowed

fuels to be produced from synthesis gas with methanol as an

intermediate. Mobil in the United States has con tributed

decisively to the development of such processes, which

involve mainly the reaction of methanol on zeolite

catalysts. The most important and, up to now, the only

industrially i mplemented process is methanol to gasoline

(MTG) synthesis.

Methanol is used to synthesize a large number of other

organic compounds:

Formic acid preservatives, pickling a gents

Methyl esters of solvents, monomers

organic acids

Methyl esters of methylation reagents, explosives,

inorganic acids insecticides

Methylam ines pharmaceutical precursors,

auxiliaries, absorption liquids

for gas washing and scrubbing

Trimethylphosphine pharmaceuticals, vitamins,

fragrances, fine chemicals

Sodium methoxide organic intermediates, catalyst

Methyl halides organic intermediates, solvents,

propellants

Ethylene organic intermediates, polymers,

auxiliaries

(7)

**Type:** industrial

**Cat egory:**  Basic industry: basic chemicals

**Remark:** Methanol to Olefins (MTO)

One of the larger potential applications for methanol is the

methanol

- to

- olefins (MTO) process.

In this proce ss,methanol is converted to light olefins,

primarily ethylene and propylene.

The quantity of methanol that in theory would be required

for a 500,000 ton per year ethylene plant is 7,000 tons per

day, or 2,450,000 tons per year.

Obviously,this is larger than any methanol unit currently

being built today.

Given the generally lower costs of the traditional cracking

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

58 tec hnology however, MTO will likely only capture niche

markets or serve as "swing" capacity for existing ethylene

producers. Nonetheless, the volume potential for methanol is

huge.

One o f the potential benefits of the MTO process is the

ability to separate the olefin production from the natural

gas location by building the methanol and the MTO units in

the most financially attractive l ocation.

(8)

**Type:** use

**Category:** other: use as Feedstock for Chemical Syntheses

**Remark:** World demand for MTBE from Methanol is expected to fall

below 8 Mio. t/a equivalent to approx. < 16% of the world

demand of methanol in 2004.

(8)

**Type:** industrial

**Category:** Fuel i ndustry

**Remark:** Use as Energy Source

Methanol is a promising substitute for petroleum products if

they become too expensive for use as fuels.

Methanol as a Fuel for Otto Engines

The use of methanol as a motor fuel has been discussed

repeatedly since the 1920s. Use has so far been restricted

to high

- performance engines for racing cars and aeroplanes.

(7)

**Type:** use

**Category:** Fuel additives

**Remark:** Biodiesel.

12

- SEP

- 2005 (4)

**Type:** use

**Category:** othe r: Other Energy Uses of Methanol

**Remark:** A use that has been discussed particularly in the United

States and implemented in pilot projects is the firing of

peak

- load gas turbines in power stations ( peak shavi ng).

Benefits include simple storage and environmentally friendly

combustion in the gas turbine.

The use of methanol as a fuel in conventionally fired boilers

obviates the need for cos tly flue gas treatment plants but is

not yet economically viable.

12

- SEP

- 2005 (7)

**Type:** use

**Category:** other: Other Energy Uses of Methanol

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

59 **Remark:** Methanol as a Hydrogen Carrier for Fuel Cell Applications

Methanol is a leading candidate to provide the hydrogen

necessary to power a range of fuel cell technology

applications. Taking an estim ate from DaimlerChrysler, by

2020, 4.2 million to 12 million new fuel cell cars will be

sold each year. DaimlerChrysler also believes that methanol

is the best hydrogen carrier fuel for personal transpo rt.

Long before the automotive fuel cell market reaches

maturity, consumers will be using cartridges of methanol to

fuel direct methanol fuel cell power packs for a range of

electronic devices such as cellular phones and laptop

computers. Methanol also will be used to power portable

fuel cell generators, and even residential fuel cell

systems.

(9)

**Type:** use

**Category:** Intermediates

**Type:** industrial

**Category:** Paints, lacquers and varnishes industry

**Type:** industrial

**Category:** Chemical industry: use d in synthesis

12

- SEP

- 2005 (2)

**Type:** use

**Category:** Laboratory chemicals

**Type:** use

**Category:** Solvents

**Type:** use

**Category:** other: RM

**Remark:** Methanol's low freezing point and its miscibility with water

allow it to be used in refrigeration systems, either in pure

form (e.g., in ethylene plants) or mixed with water and

g lycols.

It is also used as an antifreeze in heating and

cooling circuits; compared to other commonly used

antifreezes (ethylene glycol, propylene glycol, and

glycerol), it has the adva ntage of lower viscosity at low

temperature. It is, however, no longer used as an engine

antifreeze; glycol

- based products are employed instead.

Large amounts of methanol are used to protect natural gas

pipelines against the formation of gas hydrates at low

temperature. Methanol is added to natural gas at the pumping

station, conveyed in liquid form in the pipeline, and

recovered at the end of the pipeline. Methanol can be

recycled after removal of water taken up from natural gas by

distillation.

Methanol is also used as an absorption agent in gas

scrubbers. The r emoval of carbon dioxide and hydrogen

sulfide with methanol at low temperature (Rectisol process,

Linde and Lurgi) has the advantage that traces of methanol

in the purified gas do not generally interfer e with further

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

60 processing.

The use of pure methanol as a solvent is limited, although

it is often included in solvent mixtures.

12

- SEP

- 2005 (7)

**1.7.1 Detailed Use Pattern**

**-**

**1.7.2 Methods of Manufacture**

**Orig. of Subst.:** Synthesis

**Type:** Production

**Remark:** The methanol production process involves the catalytic steam

reforming of a hydrocarbon feedstock, typically natural gas,

to give a gaseous mixture of carbon oxides and hydrogen.

This mixture is then compressed and reacted over another

catalyst to give methanol and by

- products, according t o the

following reactions.

CO + 2 H2 <

- > CH3OH

CO2 + 3 H2 <

- > CH3OH + H2O.

The pure product is obtained by fractional distillation. All

necessary steps are performed in closed systems.

The gas is reformed over a nickel catalyst. The synthesis

gas, containing a mixture of hydrogen, carbon oxides, steam

and residual methane leaves the reformer at about 880oC and

20 bar, and is cooled before being compressed to synthesis

pressure.

The synthesis loop comprises a circulator, converter,

feed/effluent exchanger, separator, and in some cases a heat

recovery exchanger.

The optimum operating pressure is typically about 80 bar for

large plants, but for smaller plants (less than 600 t/day) a

pressure of 50 bar may be economic due to th e reduced material

requirements.

Synthesis gas is mixed with the recycle gas and preheated in a

feed/effluent interchanger.

The converter effluent is cooled by passing through the

feed/effluent interchanger and dependent on the reactor type,

a heat recovery exchanger.

Prior to the catchpot, the gas is cooled to about 40oC to

condense the product methanol. A purge i s taken from the

recycle gas to remove inerts (N2, CH4, Argon, and surplus

hydrogen or carbon oxides) from the loop. This is used as

fuel in the reformer. A purge gas expander may be used to

recover power from the purge gas as it is let down to the

fuel system pressure.

The crude methanol from the separator contains both water

and low levels of by

- products, which must be removed to

achieve required product purity.

12

- SEP

- 2005 (10)

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

61 **1.8 Regulatory Measures**

**-**

**1.8.1 Occupational Exposure Limit Values**

**Type of limit:** MAK (DE)

**Limit value:** 27 0 mg/m3

**Country:** Germany

**Remark:** Exceeding factor: 4.

If the MAK and BAT values are complied with, there should be

no risk for the fetus.

Cutaneous absorption.

**Source:** TRGS 900 (Germany), 08/2004

12

- SEP

- 2005 (2) (11)

**Type of limit:** MAK (DE)

**Limit value:** 200 other: ppm

**Country:** Germany

**Remark:** Exceeding factor: 4.

If th e MAK and BAT values are complied with, there should be

no risk for the fetus.

Cutaneous absorption.

**Source:** TRGS 900 (Germany), 08/2004

12

- SEP

- 2005 (2) (11)

**Type of limit:** other: EU

- TWA

**Limit value:** 260 mg/m3

**Country:** Europe

12

- SEP

- 2005 (6)

**Type of limit:** other: EU

- TWA

**Limit value:** 200 other: ppm

**Country:** Europe

12

- SEP

- 2005 (6)

**Type of limit:** BAT (DE)

**Limit value:** 30 other: mg per liter urine

**Country:** Germany

**Remark:** Time of sampling (long term exposure):

After several preceding shifts end of exposure/end of shift.

Examination material: Urine.

12

- SEP

- 2005 (3)

**1.8.2 Acceptable Residues Levels**

**-**

**1.8.3 W ater Pollution** 

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

62 **Classified by:** other: VwVwS (Germany), Annex 2

**Labelled by:** other: VwVwS (Germany), Annex 2

**Class of danger:** 1 (weakly water polluting)

**Country:** Europe

**Source:** VwVwS (Germany), 17.05.1999

12

- SEP

- 2005 (2) (12)

**1.8.4 Major Accident Hazards**

**Legislation:** Stoerfallverordnung (DE)

**Substance listed:** yes

**Country:** Germany

**Remark:** Annex I

- No.: 26

Threshold for o perating range to § 1 sec. 1:

- Record 1: 500,000 kg

- Record 2: 5,000,000 kg

12

- SEP

- 2005 (3)

**1.8.5 Air Pollution**

**Classified by:** TA

- Luft (DE)

**Label led by:**  TA

- Luft (DE)

**Number:** other: 5.2.5 (organic substances)

**Class of danger:** I

**Country:** Germany

**Remark:** Following total amounts must not be exceeded in exhaust gas:

mass flow: 0.10 kg/h or

mass concentration: 20 mg/m3

12

- SEP

- 2005 (3)

**1.8.6 Listings e.g. Chemical Inventories**

**Type:** EINECS

**Additional Info:** EINECS

- No. 200

- 659

- 6

12

- SEP

- 2005 (13)

**Type:** ENCS

**Additional Info:** ENCS No. 2

- 201

**Remark:** ENCS CLASSIFICATION:

Low Molecular Chain

- like Organic Compounds.

12

- SEP

- 2005 (13)

**Type:** ECL

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

63 **Additional Info:** ECL Serial No. KE

- 23193

ECL Toxic Chemical No. 97

- 1

- 80

**Remark:** KOREAN TCCL DESIGNATION:

ECL Toxic Chemical No. 97

- 1

- 80

12

- SE P

- 2005 (13)

**Type:** other: SWISS

**Additional Info:** SWISS No. G

- 2063

**Remark:** SWISS CLASSIFICATION:

Giftliste 1 (List of Toxcic Substances 1), 31 May 1999

. Toxic Category 3.

12

- SEP

- 2005 (13)

**Type:** TSCA

12

- SEP

- 2005 (13)

**Type:** DSL

12

- S EP

- 2005 (13)

**Type:** AICS

12

- SEP

- 2005 (13)

**Type:** PICCS

12

- SEP

- 2005 (13)

**Type:** other: ASIA

- PAC

12

- SEP

- 2005 (13)

**Type:** CHINA

**Additional Info:** Inventory of Existing Chemical Substances (IECSC), 2004

12

- SEP

- 2005 (14)

**Type:** other: New Zealand

**Additional Info:** Composite List of Single Component Substances to be considered

for Transfer (April 2003)

**Flag:** non confidential

12

- SEP

- 2005 (14)

**1.9.1 Degradation/Transformation Products**

**EINECS**

**- Name:**  Thermal decomposition:

### OECD SIDS METHANOL

1. GENERAL INFORMATI ON ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

64 Vapours may form explosive mixture with air.

12

- SEP

- 2005 (2)

**1.9.2 Components**

**-**

**1.10 Source of Exposure**

**-**

**1.11 Additional Remarks**

**Memo:** Characterisation:

Highly inflammable liquid.

Vapours form explosive mixtures with air.

Mixable with water.

High volatile.

12

- SEP

- 2005 (3)

**Memo:** Hazardous reactions:

Explosion hazard when product reacts with metals and forms

hydrogen.

Reacts with strong oxidizing agents.

Aluminium, magnesium, toxic vapours/gases.

12

- SEP

- 2005 (2)

**1.12 Last Literature Search**

**Type of Search:** Internal and External

**Chapters covered:** 1

**Date of Search:** 12

- SEP

- 2005

**Remark:** Update 2006

**Flag:** non confidential

12

- SEP

- 2005

**Type of Search:** In ternal and External

**Chapters covered:** 5.10

**Date of Search:** 06

- NOV

- 2002

**1.13 Reviews**

### OECD SIDS METHANOL

2. PHYSICAL CHEMICAL DATA ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

65 **2.1 Melting Point**

**Value:** =

- 97.8 degree C

**Reliability:** (4) not assignable

Handbook

(15)

**Value:** =

- 98 degree C

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16 )

**2.2 Boiling Point**

**Value:** = 64.7 degree C

**Reliability:** (4) not assignable

Handbook

(15)

**Value:** = 64.7 degree C

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**Value:** = 64.6 degree C

**Reliability:** (4) not assignable

Secondary quotation

(17)

**2.3 Density**

**Type:** density

**Value:** = .79 g/cm3 at 20 degree C

**Reliability:** (4) not assignable

Handboo k

(15)

**Type:** density

**Value:** = .791

- .792 g/cm3 at 20 degree C

**Reliability:** (4) not assignable

Manufacturer / producer data withou t proof

(16)

### OECD SIDS METHANOL

2. PHYSICAL CHEMICAL DATA ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

66

**2.3.1 Granulometry**

**2.4 Vapour Pressure**

**Value:** = 129 hPa at 20 degree C

**Reliability:** (4) not assignable

Manufacturer / prod ucer data without proof

(16)

**Value:** = 319 hPa at 38 degree C

**Reliability:** (4) not assignable

Secondary quotation

(18)

**Value:** = 552 hPa at 50 degree C

**Reliability:** (4) not assignable

Manufacturer/producer data

(16)

**Value:** = 128 hPa at 20 degree C

**Reliability:** (4) not assignable

Secondary quotation

(19)

**Value:** = 127.9 hPa at 20 degree C

**Me thod:**  other (calculated)

**Method:** Value calculated with the following equation:

Y= exp[A + B/T + ClnT + DT2]

and using the following equation coefficients:

A= 8.1768E+01

B=

- 6.8760E+03

C=

- 8.7078E+00

D= 7.1926E

- 06

**Remark:** Source for accurate physical property data.

Data are collected from a wide range of sources and

critically evaluated

. **Reliability:** (2) valid with restrictions

Scientifically verified data, standard reference source for

vapour pressure

19

- OCT

- 2006 (20)

**2.5 Partiti on Coefficient** 

**Partition Coeff.:** octanol

- water

**log Pow:** =

- .82

**Method:** other (measured)

**Reliability:** (2) valid with restrictions

scientifically verfied data

### OECD SIDS METHANOL

2. PHYSICAL CHEMICAL DATA ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

67 (21)

**Partition Coeff.:** octanol

- water

**log Pow:** =

- .77

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**Partition Coeff.:** octanol

- water

**log Pow:** =

- .74

**Remark:** Recommended value

**Reliability:** (2) valid with restrictions

Scientifically verified data

(22)

**Partition Coeff.:** octanol

- water

**log Pow:** =

- .64

**Method:** other (calculated)

**Remark:** Value calculated applying the hydrophobic fragmental constant

(f) for small molecules.

**Reliabili ty:**  (2) valid with restrictions

scientifically verfied data

(23)

**2.6.1 Solubility in different media**

**Solubility in:** Water

**Value:** at 20 deg ree C

**Descr.:** miscible

**Remark:** pH value: neutral

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**Solu bility in:**  Water

**Descr.:** miscible

**Reliability:** (4) not assignable

Secondary quotation

(17)

**2.6.2 Surface Tension**

**Value:** = 22.55 mN/m at 20 degree C

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(24)

### OECD SIDS METHANOL

2. PHYSICAL CHEMICAL DATA ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

68 **2.7 Flash Point**

**Value:** = 9.7 degree C

**Type:** closed cup

**Method:** other: Abel

- Pensky

**Reliability:** (2) valid with restrictions

National standard specification without detailed

documentation

(25)

**Value:** = 11 degree C

**Type:** closed cup

**Method:** other: DIN 51755

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**2.8 Auto Flammability**

**Value:** = 455 degree C

**Method:** other: DIN 51 794

**Remark:** Autoignition temperature

**Reliability:** (2) valid with restrictions

National standard specification without detailed

documentation

(26)

**Value:** = 385 degree C at 1013 hPa

**Remark:** Autoignition temp erature

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(27)

**2.9 Flammability**

**Result:** highly flammable

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**2.10 Explosive Properties**

**Result:** not explosive

**Remark:** becaus e of chemical structure

**Reliability:** (2) valid with restrictions

expert judgement

### OECD SIDS METHANOL

2. PHYSICAL CHEMICAL DATA ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

69 (28)

**2.11 Oxidizing Properties**

**Result:** no oxidizing properti es

**Remark:** because of chemical structure

**Reliability:** (2) valid with restrictions

Expert judgement

(28)

**2.12 Dissociation Constant**

**-**

**2.13 V iscosity** 

**Value:** = .59 mPa s (dynamic) at 20 degree C

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(16)

**2.14 Ad ditional Remarks** 

**Remark:** Explosion limits: 5.5

- 36.5 vol.%

Conditions to avoid, substances to avoid:

Hazardous reaction influenced by strong oxidizing agents,

(e.g. nitric acid, chromium t rioxide, perchloric acid and

salts).

Vapours may form explosive mixture with air.

Explosion hazard when product reacts with metals and forms

hydrogen: aluminium, magnesium at elevated temperature.

Hazardous reactions: vapours form explosive mixtures with

air

Hazardous decomposition products: ignitable gases / vapours

**Reliability:** (4) not assignable

Manufactu rer / producer data without proof

(16)

**Remark:** Explosion limits: 5.5

- 36.5 vol.%

**Reliability:** (4) not assignable

Handbook

(29)

**Remark:** Explosive limits: 5.5

- 31.0 vol.%

**Reliability:** (4) not assignable

Secondary quotation

(30)

**Remark:** Explosive Limits in air: 6.0

- 36.5 vol.%

**Reliability:** (4) not assignable

Manufacturer / producer data without proof

(27)

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

70 **3.1.1 Photodegradation**

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Rate constant:** = .000000000000932 cm3/(molecule * sec)

**Degradation:** = 50 % after 17.2 day(s)

**Method:** other (measured)

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Rate Constant: 0.932 (+/

- 0.233)*10^

- 12 cm^3/molecule*sec at

298 K.

Recommended value based on a unit

- weighted least squares

analysis of several published and scientific ally evaluated

experimental rate constants.

**Reliability:** (1) valid without restriction

scientifically verfied data

(31)

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Remark:** k(298K)= 7.8*10e

- 13 or 1.4*10e

- 13 cm3/molecule*sec.

Temp. dependence of k/cm3/molecule*sec: 3.3*10e

- 12 exp

(

- 380/T).

Temp. range/K: 240

- 300

**Reliability:** (2) valid with restrictions

(32)

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Conc. of sens.:** 1000000 molecule/cm3

**Rate constant:** = .000000000001 cm3/(molecule * sec)

**Degradation:** = 50 % after 8 day(s)

**Remark:** Bimolecular reaction constant

**Reliability:** (2) valid with restrictions

(33)

**Ty pe:**  air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Conc. of sens.:** 500000 molecule/cm3

**Rate constant:** = .0000000000012 cm3/(molecule * sec)

**Degradation:** = 50 % after 14.6 day(s)

**Remark:** Reaction at ca. 300 K, smog chamber tes t

**Reliability:** (2) valid with restrictions

(34)

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Conc. of sens.:** 3000000 molecule/cm3

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

71 **Remark:** Atmos pheric transformation: 1 to 5 days

**Reliability:** (4) not assignable

Secondary literature

(35)

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Rate constant:** = .000000000000944 cm3/(molecule * sec)

**Remark:** Recommended value (at 298 K)based on a unit

- weighted

least

- squares analysis of several experimental absolute rate

constants scientifically ev aluated by the author.

**Reliability:** (1) valid without restriction

scientifically verfied data

(36)

**Type:** air

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Remark:** The half

- life of methanol in the troposphere for this

reaction will obviously depend upon the concentration of

hydroxyl radicals. Using an average hydroxyl radical

concentrati on of 1*10^6 molecules/cm^3, half

- lifes for

the reaction ranged from 6.7

- 13.4 days. However, using

the global diurnal mean tropospheric concentration of

5*10^5 molecules/cm^3 quoted in some references, the

half

- lifes ranged from 13.4

- 17.6 days.

**Reliability:** (4) not assignable

secondary qoutation

(37)

**Type:** air

**INDIRECT PHOTOL YSIS** 

**Sensitizer:** OH

**Conc. of sens.:** 1000000 molecule/cm3

**Degradation:** = 50 % after 7.3 day(s)

**Remark:** Reaction at 300 K and atmospheric pressure; k=1.1*10^

- 12

cm^3/molecule*sec

**Reliability:** (4) not assignable

secondary quotation

(38)

**Type:** air

**Remark:** Methanol is listed as hazardous air pollutant under Title

III of CAAA (Clea n Air Act Amendments) with an atmospheric

lifetime of 16

- 18 days.

**Reliability:** (4) not assignable

secondary literature

(39)

**Type:** water

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

72 **Result:** Rate constant = 1*E9 l/mol*sec

(40)

**Type:** water

**INDIRECT PHOTOLYSIS**

**Sensitizer:** OH

**Method:** other (measured)

**Remark:** k(OH) = 8.46 * 10e9 liter/mol*sec

OH concentration: 5 * 10e

- 14 mol/liter

temperature: 298 K

(41)

**3.1.2 Stability in Water**

**Method:** other

**Remark:** Alcohols are generally resistant to hydrolysis.

**Reliability:** (1) valid without restriction

Handbook

(42)

**3.1.3 Stability in Soil**

**-**

**3.2.1 Monitoring Data (Environment)**

**Medium:** air

**Remark:** concentration: 6

- 60 ug/m3 (3 locations, 52 samples)

(35)

**Type of measurement:** other

**Medium:** other

**Remark:** Methanol is a naturally occurring compound, being a meta

- bolite in man, animals and plants.

(43)

**Type of measurement:** other

**Remark:** Holzer et al. (1977) identified methanol in forest air in

Alabama, 20

- 35 miles away from the closest urban activities/

habitation, and ascribed its pres ence to natural sources.

(44)

**Type of measurement:** other

**Remark:** Owens et al. (1969) identified methanol as one of the

volatile compounds emitted by a lfalfa, and Isidorov et

al. (1985) reported that methanol was given off by evergreen

cyprus in a survey of the volatile emissions from the

foliage of 17 trees characteristic of the forests of

Northern Europe and Asia.

(45) (46)

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

73 **Type of measurement:** other

**Medium:** air

**Remark:** Jonsson et al. (1985) reported the results of a 1 year study

(1982 to 1983) of air quality in Stockholm. Samples were

collected at four urban sites in central Stockholm and a

more rural site 12 km from the centre of the city. The

minimum detectable quan tity in the 2

- 3 litre samples collec

- ted was about 0.008 nl/l for a typical oxygenated compound,

i.e. about 0.01 ug/m^3 methanol. The following methanol

levels were reported:

1. Busy street, 8 samples/day collected over 7 days, 56

samples in total, March

- May 1983; average: 34.98 ug/m^3;

range: 5.31

- 94.84 ug/m^3.

2. Quiet street, 8 samples/day collected over 3 days, plus

samples taken in early morning and at night, 40 samples in

total, August

- September 1982; average: 10.96 ug/m^3; range:

2.0

- 28.17 ug/m^3.

3. Busy street, 8 samples/day collected over 12 days, 96

samples in total, September

- December 1982; average: 10.22

ug/m^3; range: 0.76

- 34.6 ug/m^3.

4. Quiet street, 8 samples/day collected over 7 days, plus

samples taken in early morning and a t night, 68 samples in

total, January

- March 1983; average: 5.02 ug/m^3; range:

0.59

- 15.07 ug/m^3.

5. Recreation area, 12 km from central Stockholm, 8 samples

/day collected over 7 days , 56 samples in total, May

- June

1983; average: 11.21 ug/m^3; range: 3.21

- 22.27 ug/m^3.

(47)

**Type of measurement:** other

**Medium:** other: water and sedi ment

**Remark:** Jungclaus et al. (1978) analysed the wastewater and

effluent

- receiving waters and sediments for a US chemical

plant manufacturing a broad range of chemicals, including

pharmaceut icals, herbicides and surfactants. The plant was

located on a small freshwater river draining into a small

brackish cove, which eventually fed into an estuary.

Methanol was found at levels of 17

- 80 mg/l in the

waste

- water but not in the effluent

- receiving waters or

associa ted sediments (no detection limits were reported).

(48)

**Type of measurem ent:**  other

**Medium:** other: condensate waters

**Remark:** Mohr and King (1985) identified methanol in one of four

samples of condensate waters taken from a fixed

- bed coal

gasification process plant. No detection limit was reported

but methanol was found in the sample at a level of 1050

mg/l.

(49)

**Type of measurement:** other

**Medium:** biota

**Remark:** Pellizzari et al. (1982) collected 42 samples of mothers'

milk from women in four urban areas of the USA. They ana

lysed all the samples by GC/MS and the mass spectra from

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

74 eig ht samples were later interpreted manually. Methanol

was found in one of these samples, however, no actual level

of occurrence was reported.

(50 )

**Type of measurement:** other

**Medium:** food

**Remark:** Methanol has been isolated and identified in the volatile

components of the following foodstuffs: baked potatoes

(Coleman et al., 1981), roasted filbert nuts (Kinlin et al.,

1972), dried legumes (Lovegren et al., 1979). Of these

occurrences, only Lovegren et al. quantified the levels of

methanol present, as follows: Lima beans (seven samples),

common beans (five samples), mung beans (one sample) and

soya beans (one sample): mean level =4.2 mg/kg; range:

1.5

- 7.9 mg/kg.

Split peas (one sample): level =3.6 mg/kg.

Lentils (two samples): mean level =4.4 mg/kg.

(51) (52) (53)

**3.2.2 Field Studies**

**-**

**3.3.1 Transport between Environmental Compartments**

**Type:** adsorption

**Media:** water

- air

**Remark:** The Henry's Law constant of 1.35*10^

- 4 atm*m^3/mole

indicates that volatilization from environmental waters may

be significant.

**Reliability:** (2) valid with restrictions

(54)

**Type:** volatility

**Media:** water

- air

**Method:** other: calculated according to HENRYWIN, V 3.10

**Result:** Henry's Law Constant at 25 °C = 4.27E

- 6 at m * m3/mol.

**Reliability:** (2) valid with restrictions

(55)

**Type:** adsorption

**Media:** water

- soil

**Remark:** Lokke (1984) studied the adsorption of methanol onto three

different soil types at 6 deg C. Only slight methanol ad

sorption occurred with the two sandy soils tested (percen

tage organic matter of 0.09% and 0.1% in the samples) and

with the clay soil (percentage organic matter was 0.22%).

Methanol solutions of concentration 0.1, 1.0, 9 and 90 mg/l

were used in 1 hour exposure adsorption studies and adsorp

tion coef ficients of between 0.13 and 0.61 were measured for

all soil types and at all concentrations. These coefficients

indicate that methanol has a low adsorptive capacity on

soils.

**Reliability:** (4) not assignable

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

75 Secondary quotation

(56)

**Type:** adsorption

**Media:** water

- soil

**Method:** QSAR calculation performed with PCKOCWIN v1. 66 (2000)

**Remark:** The very low value of the estimated Koc indicates no

significant adsorption of methanol on soils.

**Result:** Estimated Koc = 1

**Reliability:** (2) valid with restrictions

(57)

**Type:** volatility

**Media:** water

- air

**Remark:** Methanol has a estimated Henry's Law Constant of

4.4*10^

- 6 atm*m^3/mol at 25 deg C.

**Reliability:** (4) not assignable

(58)

**Type:** volatility

**Media:** water

- air

**Result:** H = 0,461 Pa*m3/mol

**Reliability:** (2) valid with restrictions

(59)

**3.3.2 Distribution**

**Media:** air

- biota

- sediment(s)

- soil

- water

**Method:** Calculation according Mackay, Level I

**Result:** water: 84.59 %

air: 15.38 %

sediment: 0.0068 %

soil: 0,0067 %

**Reliability:** (2) valid with restrictions

(60)

**Media:** air

- biota

- sediment(s)

- soil

- water

**Method:** Calculation according Mackay, Level III

**Remark:** Input parameters for fugacity model:

Molecular weight: 32.04

Vapor pressure: 129 hPa

Henr y' Law constant: 0.461 Pa*m3/mol

Log Kow:

- 0.82

Water solubility: 1E+006 mg/l

**Result:** compartment mass amount (%) emission (kg/h)

air 73,3 9093

water 15,6 175

soil 11,1 75

sediment 0,0233 0

**Reliability:** (2) valid with restrictions

(61)

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

76 **3.4 Mode of Degradation in Actual Use**

**3.5 Biodegradation**

**Type:** aerobic

**Degradation:** = 95.6 % after 5 day(s)

**Method:** other: (BOD of THOD)

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

Test methods and performance uncertain, limited

documentation available

19

- OCT

- 2006 (62)

**Type:** aerobic

**Inoculum:** domestic sewage

**Degradation:** = 53.4 % after 5 day(s)

**Method:** other: BOB

- Test (BOD of THOD)

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Concentration of test substance:2.5 p pm.

Mineralized dilution water, settled sanitary sewage as seed

and incubation at 20 °C.

Incubation time: 50 days.

**Result:** Percentage of the Theoretical Oxygen Demand (BOD of ThOD) at

day 5, 10, 15, 20, 30, 40, 50: 53.4, 62.7, 69.4, 93.4, 97.7

**Reliability:** (2) valid with restrictions

acceptable study meets basic scientific principles

19

- OCT

- 2006 (63)

**Type:** aerobic

**Inoculum:** domestic sewage, non

- adapted

**Degradation:** = 76 % after 5 day(s)

**Result:** readily biodegradable

**Kinetic:** 5 day(s) = 76 %

10 day(s) = 88 %

15 day(s) = 91 %

20 day(s) = 95 %

**Method:** other: closed bottle test (BOD of THOD)

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** A settled domestic wastewater was adde d (3 ml/bottle) as seed

material to clean BOD bottles. The bottles were half filled

with aerated dilution water containing the specified minerals

and buffer.

Small aliquots of the test chemical were added to these

bottles yielding concentrations of 3, 7 and 10 mg/l. At least

two of these concentrations were tested in duplicate.

Dissolved oxygen (DO) was monitored periodically in the

individual bottles through the use of a comm e rcial DO meter

fitted with an agitated probe.

The bottles were opened for sampling and DO measurements about

five times during the course o f the 20

- day test.

**Reliability:** (2) valid with restrictions

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

77 Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

19

- OCT

- 2006 (64)

**Type:** aerobic

**Inoculum:** domestic sewage, non

- adapted

**Degradation:** = 82.5 % after 5 day(s)

**Method:** other: Respirometric test (BOD of THOD)

**GLP:** no

**Test substance:** as prescri bed by 1.1

- 1.4

**Remark:** Determination of the reaction kinetic by means of the

respirometric dilution method.

This method involves the use of a nutritious matrix, usually

but not necessarily domestic sewage, which contains a lot of

organic substances competing as nutrient with the substrate

under test.

**Result:** ThOD: 1498, 0 mg/g

rate constant K: 1.882 1/d

BOD at day 5: 1236 mg/g

BOD/ThOD at day 5: 82.7%

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

19

- OCT

- 2 006 (65)

**Type:** aerobic

**Inoculum:** other bacteria: synthetic sewage

**Degradation:** = 71.2 % after 5 day(s)

**Method:** other: Respirometric test (BOD of THOD)

**G LP:**  no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

(65)

**Type:** aerobic

**Inoculum:** activated sludge, industrial

**Concentration:** 100 mg/l related to Test substance

**Degradation:** > 90 % after 1 day(s)

**Test substanc e:**  as prescribed by 1.1

- 1.4

**Remark:** Matsui et al. (1988) examined the biodegradability of

methanol in a batch reactor using activated sludge from

the Fukashiba industrial wastewater treatment plant,

which was acclimated to the wastewater (containing various

organic compounds) originating from the Kashima petro

- chemical complex in Japan. Methanol, at an initial concen

tration of 100 mg /l, was found to be highly biodegradable,

and 91% COD removal and 92% TOC removal was achieved after

a 1 day acclimation period and after 1 day's operation of

the batch reactor.

**Reliability:** (2) v alid with restrictions

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

78 acceptable study meets basic scientific principles

(66)

**Type:** aerobic

**Degradation:** = 99 %

**Method:** OECD Guide

- l ine 301 D "Ready Biodegradability: Closed Bottle

Test"

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

secondary quotation

(67)

**Type:** aerobic

**Degradation:** = 70.7

- 88.7 % after 5 day(s)

**Method:** other

**Test substance:** as prescribed by 1.1

- 1.4

**Method :**  standard dilution method (BOD of THOD) and sea water

dilution method

**Remark:** BOD5 value for methanol was measured by the sea water dilution

method (S.W.D.M) and the standard dilution method (S.D.M).

Concentration of test substance: 2.56 ppm.

**Result:**

S.D.M. S.W.D.M.

BOD5 (g/g) 1.33 1.06

ThOD (g/g) 1.50 1.50

BOD/ThOD (%) 88.7 70.7

**Reliability:** (2) valid with restr ictions

19

- OCT

- 2006 (68)

**Type:** aerobic

**Inoculum:** activated sludge, domestic, non

- adapted

**Degradation:** = 80 %

**Method:** other: Respirometric test (Sapromat)

**Year:** 1973

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

Secondary quotation (IUCLID Data Sheet), Data to EU

(69)

**Type:** aerobic

**Inoculum:** other bacteria: marine inoculum

**Degradation:** = 69 % after 5 day(s)

**Method:** other: closed bottle test (BOD of THOD)

**GLP:** no

**Test substance:** as prescribed by 1. 1

- 1.4

**Remark:** BOD 5/10/15/20 of THOD = 69/84/85/97 %.

concentration: 3

- 10 mg/l of test substance

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

79 (64)

**Type:** aerobic

**Inoculum:** other: contaminated soil

**Contact time:** 30 day(s)

**Degradation:** ca. 19 % after 30 day(s)

**Method:** other: see freetext

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Microcosm studies were done in 0.5l

- glass bottles with caps

containing 150 g soil impacte d with relatively high

concentrations of solvents inclusive Methanol (MeOH) and

approximately 0.4 l groundwater. H2O2 wasadded as an oxygen

source to maintain 1 to 2 mg/l oxygen in solution. In the

Microcosms complemented with NH4Cl and Na2HPO4 in ratio

COD:N:P= 100:10:2 the result was 19% degradation of MeOH in

30 days.

**Reliability:** (2) valid with restrictions

Special study de sign, sufficiently documented, meets

generally accepted scientific principles, acceptable for

assessment

(70)

**Type:** aerobic

**Inoculu m:**  other: contaminated soil

**Degradation:** ca. 100 % after 32 day(s)

**Method:** other: see freetext

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** The continuous

- flow reactor consisted of a 30

- gal gl ass

reactor and the center portion was filled with site soil

and groundwater. Suction and discharge tubing was enveloped

in pea stone, perforated baffles were used to separate the

zon es. Aerobiosis were achieved by addition of H2O2.

**Result:** The half live time for MeOH was : t1/2 = 16 days

**Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

gen erally accepted scientific principles, acceptable for

assessment

(70)

**Type:** aerobic

**Inoculum:** activated sludge, adapted

**Degradation:** = 5 0

- 80 %

**Remark:** Swain and Somerville (1978) monitored the biodegradation

of methanol in a model activated sludge system. Methanol

was not broken down when added transiently (at levels of

0. 23% by volume) to the system operating with a retention

time of 11 hours. However, adaption of the sludge in the

system to 0.1% by volume methanol occurred over a period

of several days, and after 2 day s acclimation, about 50%

of the methanol was utilised. After 6 days acclimation,

at least 80% of the methanol had been degraded.

**Reliability:** (4) not assignable

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

80 secondary quotation

(71)

**Type:** aerobic

**Remark:** Goldsmith assessed the potential for methanol degradation

in subsurface soil and groundwater from sites in Penn

- sylvania, New York and Virginia under aerobic conditions.

Substantial bacterial populations were present in all

samples and methanol was readily degraded. A range of

methanol concentrations from 1 to 1000 mg/l were used and

rates of degradation from 0.8 to 20.4 mg/l/day were recorded

in these aerobic studies and in parallel anaerobic studies

using material from the same sites. The rates of degra dation

were generally greater in the anaerobic samples than in the

aerobic.

**Reliability:** (4) not assignable

secondary quotation

(72)

**Type:** aerobic

**Remark:** Further details of the study of Goldsmith were reported

by Novak et al. They stated that methanol at concentrations

up to 1000 mg/l was degraded at all three si tes to such an

extent that contamination would be reduced to non

- measurable

amounts within a year. The soils and overlying water used

in these tests were not thought to have been previously

contaminated. In one aerobic sample of soil from a Penn

- sylvania site, complete degradation of 100 mg/l methanol

occurred in less than 30 days.

**Reliability:** (4) not assignable

secondary quo tation

(73)

**Type:** aerobic

**Remark:** Biodegradation of methanol in not heavily polluted aerobic

aquifer material: half

- live 58

- 260 days (loc ation: Clay loam

Virginia)

**Reliability:** (4) not assignable

Secondary literature

(74)

**Type:** anaerobic

**Inoculum:** other: bacterial population from a sewage sludge

**Degradation:** = 75

- 80 %

**Remark:** Bekes et al. (1975) studied the degradation of methanol by

the bacterial population from a sewage sludge. Under an

-

aerobic cond itions rapid degradation occurred, leading to

75

- 80% disappearance of the methanol. Rapid liberation of

methane and carbon dioxide was observed.

**Reliability:** (4) not assignable

secondary quotation

(75)

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

81 **Type:** anaerobic

**Concentration:** 500 mg/l related to Test substance

**Degradation:** = 66 %

**Remark:** Chou et al. (1978) studied the acclimation and degradation

of several petrochemical wastewater components (including

methanol) in an anaerobic fermentation system. Methanol, at

a concentration of 500 mg/l, was found to be metabolised by

enriched bacterial cultures (developed on acetate substrate)

after a period of acclimation. After a 6 day lag period, 66%

of the methanol was removed rapidly at a rate of 182 mg/l/

day from serum bottle inocula.

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(76)

**Type:** anaerobic

**Degradat ion:**  = 83

- 91 % after 3 day(s)

**GLP:** no

**Remark:** Oremland et al. (1982) studied the degradation of methanol

in anaerobic sediments collected from the upper 15 cm of a

salt marsh in San Franci sco Bay. The sediments were highly

reduced and contained methane and hydrogen sulphide. The

sediments were homogenised anaerobically with San Francisco

Bay water and 310

- 340 umol methanol/flask was adde d to make

up the inocula. After 3 days incubation, 83

- 91% conversion

of the methanol had occurred. The products of methanol

degradation were methane, CO2 and water.

**Reliability:** (2) valid with res trictions

2.1; accpetable study, meets basic scientific principles

(77)

**Inoculum:** other: methylotrophic organisms

**Remark:** In a survey of meth ylotrophic organisms, i.e. those micro

- organisms recognised by their ability to grow on compounds

that contain no carbon

- carbon bonds and to assimilate car

- bon as formaldehyde or as a mixture of forma ldehyde and

carbon dioxide, Hanson (1980) identified the following

strains/genera of microorganism capable of using methanol

as a growth substrate: Pseudomonas sp. (including P. AM1,

P . C, P. 8 and P. oleovorans), Methylobacterium organophi

- lum XX, Hyphomicrobium sp., Streptomyces sp., Rhodopseudo

monas acidophila, Paracoccus denitrificans, Microcyclus

aquaticus, Thiobacillus novel lus, Micrococcus denitrificans,

Achromobacter 1L and Mycobacterium (isolated from activated

sewage sludge). Hansen also identified one strain of

methanol

- oxidising fungus, Trichoderma lignorium, and the

following genera of methanol

- oxidising yeasts: Pichia,

Saccharomyces, Hansenula, Rhodotorula, Kloeckera, Candida,

Torulopsis.

**Reliability:** (4) not assignable

secondary quotation

(78)

**Inoculum:** other

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

82 **Remark:** Other organisms identified as being capable of utilising

methanol as a growth substrate in other reports included :

Methanosarcina sp. (anaerobe, transformed methanol to

methane), Candida sp. (a yeast isolated from Niger Delta

sediment), Mycobacterium sp. (isolated from various samples

of Japanese soil) and a yeast isolated from palm wine.

**Reliability:** (4) not assignable

secondary quotation

(79) (80) (81) (82)

**3.6 BOD5, COD or BOD5/COD Ratio**

**3.7 Bioaccumulation**

**Species:** Cyprinus carpio (Fish, fresh water)

**Remark:** Gluth et al. studied the accumulation of methanol by carp,

Cyprinus carpio, exposed to the chemical for up to 72 hours.

The amount of radioacti vity derived from the 14

- C

- labelled

chemical was determined in the liver, kidneys, intestine,

muscle, blood and gills of carp exposed to methanol at 5

mg/l. The initial uptake of methanol into the diffe rent

tissue types was the same after 24 hours, and levels re

mained constant over 72 hours in the liver, kidneys, gills

and intestine but dropped slightly in the blood and muscle.

Bio accumulation factors of about 1 were proposed for the

fish tissues and organs.

**Reliability:** (2) valid with restrictions

(83)

**Species:** Leuciscus i dus (Fish, fresh water)

**BCF:** < 10

**Remark:** The bioconcentration factor of methanol experimentally

measured in fish (golden ide) was less than 10. Based on

a log Kow of

- 0.77 the BCF value for meth anol can be

estimated to be 0.2 from a recommended regression

- derived

equation.

**Reliability:** (2) valid with restrictions

(84) (85) (86)

**Species:** other: Chlorella fusca (green alga)

**Exposure period:** 24 hour(s) at 25 degree C

**Concentration:** 50 μg/l

**BCF:** = 28400

**Method:** other

**GLP:** no data

**Method:** Algae with a dry weight of 20 mg/200 ml were exposed in a

500 ml Erlenmeyer flask with glass stoppers to 14C

- labelled

methanol in nutrient solution for at least 24 hours at room

temperature (19

- 25°C) under illumination (16 hr/d) and

agitation.

After 24 hours the algae were removed by centrifugation at

at 4000 rpm.

The 14C activity of the algae was determined by combustion

### OECD SIDS METHANOL

3. ENVIRONMENTAL FAT E AND PATHWAYS ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

83 in a Packard Tri Carb Sample Oxidize r and counting in a

Packard Tri Carb liquid scintillation counter Model 3385.

The bioaccumulation factor on the basis of dry weight is

calculated by dividing the chemical concentration of the

algae by the final concentration in the water after 24

h ours.

**Reliability:** (3) invalid

The derived high bioconcentration factor is anomalous compared

to those for other aquatic organisms. It is likely due to the

fact that methanol is taken up very rapidly and metabolized by

the algae. The 14C

- label, which is measured to calculate the

BCF value, is therefore incorporated into the algae i n

metabolic forms other than methanol.

In addition the test was conducted in a wide temperature range

and with an unappropriate exposure time.

(87)

**3.8 Additional Remarks**

**Memo:** more information see IPCS report no. 196

(88)

**Result:** Methanol biodegradation has been shown to take p lace also

under anaerobic conditions (all electron acceptors besides

oxygen).

Microbial growth on methanol with nitrate as the electron

acceptor is the second most energetically favore d mode of

methanol metabolism. There are over 100 wastewater treatment

plants in the United States that currently use methanol as

a carbon source to remove nitrate (NO3

- ) from water by

anaerobic denitrification.

**Reliability:** (1) valid without restriction

(89)

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

84 **AQUATIC ORGANISMS**

**4.1 Acute/Prolonged Toxicity to Fish**

**Type:** flow through

**Spec ies:**  Lepomis macrochirus (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** yes

**LC50:** = 15400

**Method:** other

**Year:** 1986

**GLP:** no data

**Rem ark:**  Method: Test procedures followed the basic flow

- through test

method described by the U. S. Environmental Protection

Agency (EPA

- 660/3

- 75

- 009, 1975).

The exposure system for fish consisted of several modified

proportional diluters with dilution factors ranging from 0.6

to 0.8. Each diluter cycle delivered 1.0 l of water and

toxicant, where appropriate, to each of five test

concentrations and a control. Each test concentration and

control was duplicated by dividing the original volume

equally using flow

- splitting tanks.

The glass exposure tanks contained 6.3 l water. D iluters

were cycled at a rate to yield 5 to 8 tank volumes/day.

Exposure samples containing methanol were analyzed by direct

aqueous injection gas

- liquid cromatography. The percent

rec overy of methanol

- spiked exposure water for bluegill test

was 101.5 +/

- 2.7% (n = 5).

Bluegills were held in 530

- l fiberglass holding tanks at

their respective test temperatures and fed a commercial

formulation of dry food.

Groups of 10 bluegills were randomly assigned to the

exposure tanks at the start of each test.

Fish were not fed 24 h before or during a test.

A verage weights of exposed fish were obtained by weighing

control fish from each test. Average weight of blugills was

3.07 g +/

- 1.38

Death was the primary endpoint, although effects on

equilibrium, behavior and coloration were noted

Median effect concentration, EC50 was 12700 mg/l (11800

-

13700).

Dead fish were counted and removed at 1, 3, 6, 12 and 24 h

after init ial exposure and at 24

- h periods thereafter.

Determination of LC50 and EC50 and their 95% confidence

intervals were made using the Trimmed Spearman Karber method

(Hamilton et al., 1977). Mortality data from duplicate

exposure tanks were combined before LC50 and EC50

determinations were made.

**Test condition:** Water for all tests was untreated and obtained directly

from Lake Superior and heated to me an test temperature.

Water temperature of the test: 19.8 +/

- 2.3 °C

Mean hardness: 46.6 mg/l CaCO3 (40.4

- 56.3)

Mean alkalinity: 41.7 mg/l CaCO3 (30.0

- 46.0)

pH range: 7.04

- 7.97

Mean dissolved oxygen: 78.8 % (54.3

- 88.9)

**Test substance:** Methanol, no further data.

**Reliability:** (1) valid without restriction

1c: Test procedures in accordance with accepted standard

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

85 methods, sufficiently documented, acceptable for assessment

(90)

**Type:** flow through

**Species:** Pimephales promelas (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Uni t:**  mg/l **Analytical monitoring:** yes

**LC50:** 28100

**Method:** other

**Year:** 1983

**GLP:** no data

**Remark:** The test was conducted with Lake Superior water. The flow

- through test used a proportional diluter system, in which

there were five toxicant concentrations plus a control all

in duplicate.

Individual exposure chambers measured either 26x17x15 cm, or

20x35x15 cm and contained 3.1 l and 6.3 l of water. The

diluter system delivered 0.5 l of fresh water alone to the

controls or 0.5 l of fresh water plus toxicant in the case

of the exposure g roups every 15

- 19 min.

Water temperature was mantained at 25 °C and a 16

- h light

photoperiod was used. Ten to 20 fish of age 30

- 32 days were

placed into each chamber. fish were not fed during the acute

test.

Observations for mortalities and other gross behavioral

effects were made at regular intervals through 96 hrs.

Several water quality parameters (temperature, dissolved

oxygen, hardness, alkalinity, acidity and pH) were

monitored throughout the tests.

Toxicant concentrations were measured daily in acute tests.

All twelve chambers were analyzed at 0 and 96 h, with

alternating replicates analyzed at 24, 48 and 72 h.

Methanol concentrations were determined by using Rhodamine B

dye as a tracer. The stock solution contained 25 mg/l which

was de livered to the exposure tanks. The concentration of

alcohol was calculated from the analysis of Rhodamine B,

which was measured a spectrofluorometer.

The excitation and emission wavelengths were 554 nm and 578

nm respectively. Standard were prepared in Lake Superior

water and lake water was used as the reference blank.

The exposure tanks contained from 0.1 to 1.3 μg/l Rhodamine

B.

**R esult:**  LC50 (24h) = 28400 mg/l

LC50 (48h) = 28400 mg/l

LC50 (72h) = 28400 mg/l

**Test substance:** methanol, purity > 99 %

**Reliability:** (1) valid without restriction

1c: Test procedures i n accordance with accepted standard

methods, sufficiently documented, acceptable for assessment

(91)

**Type:** flow through

**Species:** Pimephales promelas (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** yes

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

86 **LC50:** = 28200

**Method:** other: 5 concentrations and 4 fish/concentration were used

**Year:** 1932

**GLP:** no data

**Test substance:** other TS

**Remark:** 11 alcohols with increasing chain length were tested to

create a structure activity relationship with LC50 and

octanol

- water partition coefficient.

**R esult:**  A bilinear relationship was found between log LC50 and log P

with increasing toxicity with increasing log P.

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

(92)

**Type:** flow through

**Species:** Salmo gairdneri (Fish, estuary, fresh water)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:**  yes

**LC50:** = 20100

**Method:** other

**Year:** 1983

**GLP:** no data

**Remark:** The test was conducted with Lake Superior water. The flow

- through test used a proportional diluter sys tem, in which

there were five toxicant concentrations plus a control all

in duplicate.

Individual exposure chambers measured either 26x17x15 cm, or

20x35x15 cm and contained 3.1 l and 6.3 l of water. The

diluter cycled every 1o

- 16 min, providing from 6.8 to 10.6

volume additions per day.

Water temperature was mantained at approximately 12 °C and a

16

- h light photope riod was used. Ten fish were tested per

chamber. Fish were not fed during the acute test.

Observations for mortalities and other gross behavioral

effects were made at regular intervals through 96 hrs.

Several water quality parameters (temperature, dissolved

oxygen, hardness, alkalinity, acidity and pH) were

monitored throughout the tests.

Toxicant concentrations were measured daily i n acute tests.

All twelve chambers were analyzed at 0 and 96 h, with

alternating replicates analyzed at 24, 48 and 72 h.

Methanol concentrations in the exposure water were

determined by GLC using direct aqueous injection on a Tenax

GC packed column and flame

- ionization detection

**Result:** LC50 (24h) = 20300 mg/l

LC50 (48h) = 20100 mg/l

LC50 (72h) = 20100 mg/l

**Test substanc e:**  methanol, purity > 99 %

**Reliability:** (1) valid without restriction

1c: Test procedures in accordance with accepted standard

methods, sufficiently documented, acceptable for assessment

(91)

**Type:** flow through

**Species:** Salmo gairdneri (Fish, estuary, fresh water)

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

87 **Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** yes

**LC50:** = 20100

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** Method: Flow

- through acute toxicity test

EC50: median effect concentration, EC50=13000 mg/kg

**Test substan ce:**  Methanol, no further data.

**Reliability:** (4) not assignable

(90)

**Type:** other: flow through or static

**Species:** Pimephales promelas (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Unit:** g/l **Analytical monitoring:** yes

**LC50:** 29.4

**EC50 :**  28.9

**Method:** other

**GLP:** no data

**Test substance:** other TS

**Remark:** Test organism: fathead minnow, age 28

- 29 days, overall mean

length 21 +/

- 2 mm

Test conditions:

Test temperature: 25 degrees centigrade

Dissolved oxygen: 7.3 mg/l

Hardness: 43.5 mg/l CaC03

Alkalinity: 46.6 mg/l CaCO3

Tank volume: 6.3 l

pH: 7.66

Test concentrations nominal:

4000; 6700; 11200; 18600; 31000 mg/l

EC50: 28900 mg/l/96 h

**Test substance:** methanol, purity > 99 %

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(93)

**Type:** semistatic

**Species:** Poecilia reticulata (Fish, fresh water)

**Exposure period:** 7 day(s)

**Unit:** mg/l **Analytical monitoring:**

**LC50:** = 10860

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary quotation

(94) (95)

**Type:** semistatic

**Species:** other: Agonus cataphractus

**Exposure p eriod:**  96 hour(s)

**Unit:** mg/l **Analytical monitoring:**

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

88 **LC50:** >= 7900

- 26070

**Remark:** Seawater

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary quotation

(96)

**Type:** static

**Species:** Alburnus alburnus (Fish, estuary)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monit oring:**  no data

**LC50:** = 28000

**Method:** other: not indicated

**Year:** 1984

**GLP:** no data

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(97) (98)

**Type:** static

**Species:** Cyprinus carpio (Fish, fresh water)

**Exposure period:** 72 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**EC :**  >= 790

**Method:** other: effect on clinical chemical parameters

**Year:** 1985

**GLP:** no data

**Test substance:** other TS

**Method:** 6 fish were used per group. Exposure time was 6, 24 or 72

hours. The following parameters were examined: serum

glucose, serum cortisol, serum protein, serum cholesterol,

liver glycogen, muscle glycogen.

**Result:** Methanol treatment did not affect serum glucose but led to

increased se rum cortisol, decreased serum protein,

decreasedserum cholesterol, decreased liver glycogen,

increased muscle glycogen.

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(99)

**Type:** static

**Species:** Cyprinus carpio (Fish, fresh water)

**Exposure period:** 72 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**EC :**  = 790

**Method:** other: effects on clinical chemical parameters were measured

**Year:** 1984

**GLP:** no data

**Test substance:** other TS

**Me thod:**  Exposure time was 72 hours. Exposure temperature was 12, 17

or 22 øC. The following parameters were measured: serum

cortisol, serum glucose, liver glycogen, muscle glycogen,

serum protei n, serum cholesterol.

**Result:** The treatment led to increased serum cortisol and serum

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

89 glucose and to decreased liver glycogen, muscle glycogen,

serum protein and serum cholesterol.

**Test substance:** Methanol, n o further data.

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(100)

**Type:** static

**Species:** Lebistes reticulatus (Fish, fresh water)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**SG :**  = 500

**Method:** other: DIN 38412 T.15

**Year:** 1973

**GLP:** no

**Test substance:**  as prescribed by 1.1

- 1.4

**Remark:** SG = Schaedlichkeitsgrenze (limit of toxicity)

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary quotation

(101)

**Type:** static

**Species:** Leuciscus idus melanotus (Fish, fresh water)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**LC0:** = 7900

**LC50:** > 10000

**LC100:** > 10000

**Method:** other: not indicated

**Year:** 1978

**GLP:** no data

**Test substance:** other TS

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assi gnable

(102)

**Type:** static

**Species:** Oncorhynchus mykiss (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical mo nitoring:**  no data

**LC50:** = 19000

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary quotati on

(103)

**Type:** static

**Species:** Semolitus atromaculatus (Fish, fresh water)

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical mo nitoring:**  no data

**NOEC:** < 8000

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

90 **EC :**  >= 8000

- 17000

**Method:** other: the "critical range of toxicity" was measured

**GLP:** no data

**Test substance:** other TS

**Remark:** The test result is described as the critical range of

toxicity, but the signs of toxicity examined are not

reported in detail.

**Test substance:** Methanol, no further data.

**Reliability:** (3) invalid

(104)

**Species:** Cyprinus auratus

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**TLm :**  = 1659

**Method:** other: no details indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** The test substance concentration is given as 0.21 %, the

test result is described as median tolerance limit without

further details.

**Test substance:** Methanol, no fu rther data.

**Reliability:** (4) not assignable

(105)

**Species:** Cyprinus carpio (Fish, fresh water)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**TLm :**  = 28440

**Method:** other: no details indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** The test substance concentration is given as 3.6 % and the

te st result is described as median tolerance limit (no

detailed definition).

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(105)

**Species:** Leuciscus idus (Fish, fresh water)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**LC0:** > 1000

**Method:** other: DIN 38412 Teil 15

**GLP:** no data

**Test substance:**  other TS

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(106)

**Species:** Oncorhynchus mykiss (Fish, fresh water)

**Exposure period:**  96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

91 **LC50:** = 10800

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** 13680 ppm (original data)

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(107)

**Species:** Oncorhynchus mykiss (Fish, fresh water)

**Exposure period:** 96 hour(s)

**Uni t:**  mg/l **Analytical monitoring:** no data

**NOEC:** = 8530

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** 10800 ppm (original data)

**Test substance:** Methan ol, no further data.

**Reliability:** (4) not assignable

(107)

**Species:** Oryzias latipes (Fish, fresh water)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**TLm :**  = 41080

**Method:** other: no details indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** The test substance concentration is given as 5.2 %, the

result is described as median tolerance limit without

further details.

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(105)

**Species:** Semolitus atromaculatus (Fish, fresh water)

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:** no data

**:** = 6320

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** River (water) is indicated as test environment. The

concentration indicated is given as toxic threshold.8000 ppm

(original data).

**Test substance:** Methanol, no further data

. **Reliability:** (4) not assignable

(107)

**Species:** other: Armed Bullhead

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** n o data

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

92 **LC50:** 7900

- 27700

**Method:** other: not indicated

**GLP:** no data

**Test substance:** other TS

**Remark:** 10000

- 35000 ppm (original data)

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

(107)

**Type:** other

**Species:** other

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50:** 1 0630

**Remark:** QSAR Estimation of acute toxicity of methanol to fish

conducted with ECOSAR v.099.

ECOSAR Class: Neutral Organics

**Reliability:** (2) valid with restrictions

(61)

**4.2 Acute Toxicity to Aquatic Invertebrates**

**Type:** static

**Species:** Daphnia magna (Crustacea)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**EC0:** > 10000

**EC50:** > 10000

**Method:** other: acute Toxicity to daphnia magna

**Year:** 1989

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** DIN 38412 Teil 11, Bestimmung der W irkung von

Wasserinhaltsstoffen auf Kleinkrebse

**Remark:** Test procedure:

From the solutiom with the pollutant, dilution series were

prepared using dilution water of the specified composition.

The concentration steps of the test solution were selected

so as to give 3

- 4 EC values in a range between EC0 and

EC100, of which at least one value was below and one above

EC50.

The test vessel were 50 ml beakers.

Two parallel preparations were made for each concentration

step. Loading amounted to one test animal per 2 ml test

medium as ten 6

- 24 h old daphnids were pleaced in each test

and control vessel, i.e. 20 animal per concentration test.

After 24 h and again after 48 h the number of animals in the

control and test solutions that could swim was counted.

After 48 h the oxygen content were determined in order to

ensure that it had not fallen below a minimum oxygen

concentration of 2 mg/l. Furthermore the ph value was

determined at the end of the test.

The test was considered as valid when fewer than 10% of the

animals in the control solutions were unable to swim, when

the pH value was not below 7.0 and the oxygen concentration

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

93 was not below 4.0 mg/l.

The EC0 and EC100 values were taken from the results

obtained for the test solution, the 24 and 48 h EC 50s

calculated arithmetically from the concentration/effect

ratio.

**Result:** EC0 (24 h) > 10000 mg/l

EC50 (24 h) > 10000 mg/l

**Test condition:** 6

- 24 h old daphnids were used as test animals.

Specific composition of dilution water used:

acid c apacity: 0.8 mmol/l

total hardness: 2.4 mmol/l

Calcium to magnesium ratio: 4:1

Sodium to potassium ratio: 10:1

initial pH value: 8.0 +/

- 0.2

The test solutions were k ept at 20 °C in an incubator. The

test period lasted 24 and 48 h. No feed was given during the

test period.

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally acce pted

scientific principles, acceptable for assessment.

(108)

**Species:** Daphnia magna (Crustacea)

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC50:** > 10000

**Method:** other: inhibition of swimming ability

**Year:** 1977

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

(109)

**Species:** Daphnia magna (Crustacea)

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC50:** > 10000

**EC100:** > 10000

**Method:** other: DIN 38412 Teil 11, Bestimmung der Wirkung von

Wasserinhaltsstoffen auf Kleinkrebse

**Year:** 1982

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

(110)

**Species:** Nitocra spinipes (Crustacea)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**LC(I)50 :**  = 12000

**Method:** o ther: acute toxicity

**Year:** 1979

**Test substance:** as prescribed by 1.1

- 1.4

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

94 **Reliability:** (4)not assisnable

(98)

**Type:** static

**Species:** Daphni a magna (Crustacea)

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** nominal concentration

**Reliability:** (4) not assignable

(111)

**Species:** Artemia salina (Crustacea)

**Exposure period:** 24 hour(s)

**Unit:** g/ l **Analytical monitoring:**

**EC50:** = 43.6

**Method:** other

**Year:** 1994

**Result:** = 1360 mM

**Reliability:** (4) not assignable

(112)

**Species:** Artemia salina (Crustacea)

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**LC50 :**  > 10000

**Method:** Endpoint evaluated was death.

**Remark:** Concentration s tested: 100, 1000 and 10000 mg/l

**Reliability:** (4) not assignable

(64)

**Type:** static

**Species:** Asellus intermedius (Crustacea)

**Exposure period:** 96 hour( s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Remark:** nominal concentration

**Reliability:** (4) not assignable

(111)

**Species:** Daphnia magna (Crustacea)

**Exposure period:** 24 hour(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 21.4

**Year:** 1994

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

95 **Result:** = 668 mM

**Test substance:** methanol, purity > 97 %

**Reliability:** (4) not assignable

(112)

**Type:** other

**Species:** Daphnia sp. (Crustacea)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**EC50:** 9374

**Remark:** QSAR Estimation of acute toxicity of methanol to daphnid

conducted with ECOSAR v.099.

ECOSAR Class: Neutral Organics

**Reliability:** (2) valid with restrictions

(61)

**Type:** static

**Species:** Gammarus fasciatus (Crustacea)

**Exposure period :**  96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** nominal concentrat ion

**Reliability:** (4) not assignable

(111)

**Type:** static

**Species:** other aquatic mollusc: Helisoma trivolvis

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Remark:** nominal concentration

**Reliability:** (4) not assignable

(111)

**Type:** static

**Species:** other aquatic worm: Dugesia tigrina

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Remark:** nominal concentration

**Reliability:** (4)not assignable

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

96 (111)

**Type:** static

**Species:** other aquatic worm: Lumbriculus variegatus

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**LC50 :**  > 100

**Method:** other

**Year:** 1985

**GLP:** no

**Remark:** n ominal concentration

**Reliability:** (4) not assignable

(111)

**Species:** other: Anodonta imbecilis

**Exposure period:** 48 hour(s)

**Unit:** mg/l **A nalytical monitoring:** 

**LC50 :**  = 37.02

**Reliability:** (4) not assignable

(113)

**Species:** other: Brachionus clyciflurus

**Exposure period:** 24 hour(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 35.9

**Method:** other

**Year:** 1994

**Result:** = 1120 mM

**Reliability:** (4) not assignable

(112)

**Species:** other: Ceriodaphnia dubia

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**LC50 :**  = 11

**Reliability:** (4) not assignable

(113)

**4.3 Toxicity to Aquatic Plants e.g. Algae**

**Species:** Scenedesmus quadricauda (Algae)

**Endpoint:** biomass

**Exposure period:** 8 day(s)

**Unit:** mg/l **Analytical monitorin g:**  no

**TGK :**  = 8000

**Method:** other: growth inhibition

**Year:** 1978

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The toxicity threshold of methanol for the green alga

S cenedesmus quadricauda was determined in the cell

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

97 multiplication inhibition test.

Culture media and conditions of culturing were standardized

already for stock and preliminary cultures because a

standardized history of the test organisms is essential

insofar as history and resistance to pollutants show a close

physiological relationship. Composition of culture media and

conditions of culturing are described in the paper.

Prior of preparations of test cultures, the pollutant

solution of known content was neutralized.

Dilutions at different volume ratios with the factor 2 were

prepared in flasks from the pollutant solution and from

bidistillated water under sterile conditions.

In the dilution series to be inoculated an addition of 5 ml

of each of the stock nutrien t solution and of the algal

suspension from the preliminary culture having a known

adjusted extinction value was made to obtain the rated value

of 50 ml. The quantity of the cell material to be used for

inoculation was determined turbidimetrically and was

standardized.

The flasks of the non

- inoculated dilution series were

completed to the rated value of 50 ml by addition of 5 ml

each of the stock nutrient solution and bidistilled water.

While shaking the contents, 10 ml each were taken from each

flask of the inoculated dilution series and transferred to 3

culture tubes each and also from each flask of the

non

- inoculated dilution series, however transferred to one

culture tube only.

Test cultures and control cultures were kept then under

standar dized conditions for a period of 8 days, exposed to

constant lighting at 27 °C and a relative umidity of 50%.

At the end of the test the concentration of the algal

suspension was measured turbidimetric ally and expressed as

the extinction of the primary light of the monocromatic

radiation at 578 nm for a 10 mm layer.

For graphical evaluation of the toxicological findings after

termin ation of the test period, the median value (A) of the

extinction of all test cultures not under toxic influence

nor stimulated (provided that their extinction values were

within the range of a <3% stand ard deviation) and the median

value (B) of the extinction of the three test cultures

having the lowest toxic pollutant concentration were

determined.

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(114)

**Species:** other algae: Plancton algae

**Endpoint:** other: photosynthetic oxygen production

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC10:** = 1600

**EC50:** = 12000

**Method:** other: assimilation test

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** gra phic evaluation (probability distribution)

nominal concentration

**Reliability:** (2) valid with restrictions

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

98 (115)

**Species:** Chlorella pyrenoidosa (Al gae)

**Endpoint:** growth rate

**Unit:** g/l **Analytical monitoring:** no data

**EC50:** = 28.44

**Method:** other

**Year:** 1988

**GLP:** no

**Remark:** The geen alga Chlorella pyrenoidosa was obtained from the

National Research Council of Canada, Atlantic Research

Laboratory , Halifax, Nova Scotia, Canada.

Stock cultures were grown in 250

- ml erlenmeyer flasks

containin g 150 ml of an inorganic, nitrogen

- free medium

supplemented with 1.5 mg/l of NaNO3. Flasks were incubated

at 25 +/

- 1 °C and a light intensity of 7000 lux on a 12

hour light

- dark cycle.

During solvent bioassay experiment test organisms were

cultured in test tubes (150 x 25 mm O.D) in both presence

and absence of solvent.

Growth was monitored by following the increase in optical

density over time for 10 to 14 days by means of a

spectrophotometer. The most suitable wavelength was 660 nm.

Cell concentrations were determined microscopically using a

haemocytometer.

A total of 5

- 10 concentrations between 2% and 6% methanol

by volume were tested (15.8

- 47.4 g/l). Each concentration

was replicated five to ten times. Appropriate control

systems containin g no solvent were included in each

experiment.

Bioassay systems contained 9.5 ml of growth medium, an

appropriate volume of solvent and 0.5 ml of inoculum

standardized to yield an init ial cell concentration of 6.5

to 7.8 E+04 cells/ml.

In each experiment percent inhibition values, relative to

growth in control systems, were calculated daily using

spectrophotometric data.

The EC50 values (the concentrations causing 50% reduction in

algal growth) were calculated using linear regression

analysis of percentage inhibition values against solvent

concen tration data. All correlations coefficients were

>0.960.

The EC50 value calculated was 3.6% v/v methanol (28.44 g/l).

**Test condition:** Exposure period: 10

- 14 days

**Reliability:** (2) valid with restrictions

2.1; accpetable study, meets basic scientific principles

(116)

**Species:** other algae: green algae

**Endpoint:** other

**Exposure period:** 96 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC10:** calculated

**EC50:** = 4982

**Remark:** QSAR Estimation of acute toxicity methanol to green algae

conducted with ECOSAR v.099

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

99 ECOSAR Class: Neutral Organics

**Reliability:** (2) valid with restrictions

(61)

**4.4 Toxicity to Microorganisms e.g. Bacteria**

**Type:** aquatic

**Species:** Pseudomona s fluorescens (Bacteria)

**Exposure period:** 16 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**TGK :**  = 6600

**Method:** other: Vereinfachte Modifikation des DEV L8 (1960); Messung

der beginnen den Hemmung der Glucose

- Assimilation;

turbidometrische Zellzahlbestimmung

**Year:** 1973

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The toxicity threshold of methanol for Pseudomonas

fluorescens (bacteria) was determined in the

glucose

- assimilation inhibition test.

Pure cultures of single

- cell model organism were used in the

test. Culture media and conditions for the pr epations of

stock cultures, preliminary cultures and test cultures of

the model organism were standardized and described in the

paper.

Prior of preparations of test cultures, the pollu tant

solution of known content was neutralized (pH: 7.0).

From the pollutant solution four parallel dilution series

were prepared in 300 ml Erlenmeyer

- flasks stoppered with

plastic cap s.

The dilution series were preapared as follows: the first

flask of each series contained 160 ml of pollutant solution.

Starting from this flask the subsequent dilution steps were

pre pared at a constant dilution ratio by consistently mixing

80 ml of preliminary pollutant dilution and 80 ml

bidistilled water.

Consequently each flask contained 80 ml of culture liquid at

the start. Each flask of the three dilution series to be

inoculated was completed to 100 ml by adding 5 ml of each of

the stock solutions and of 10 ml each of the prepared

bacterial suspension from the preliminary culture having a

known adjusted extinction.

The quantity of the cell material to be used for inoculation

was determined turbidimetrically and was standardized.

The fla sks of the non

- inoculated dilution series were

completed to 100 ml by adding 5 ml each of the stock

solutions and 10 ml saline.

Both inoculated and non

- inoculated dilution series were

kept at 25 °C for 16 h.

After termination of the test period the concentration of

the bacterial suspension was measured turbidimetrically: it

is expressed by the extinction of the monochromatic

radiation Hg 436 nm.

The concentration at which the inhibitory action of a

pollutant starts will be present in that step of a dilution

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

100 series of the pollutant having an extinction value at t he

end of the test that is >3% below the mean value of the

extinction for non

- toxic dilutions of the test cultures.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and curre nt standards,

sufficiently documented, acceptable for assessment

(117)

**Type:** aquatic

**Species:** Pseudomonas putida (Bacteria)

**Exposure period :**  16 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**TGK :**  = 6600

**Method:** other: growth inhibition test

**Year:** 1977

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The toxicity threshold of methanol for Pseudomonas putida

(bacteria) was determined in the cell multiplication

inhibition test.

Pure cultures of single

- cell model organism were used in the

test. Culture media and conditions for the prepations of

stock cultures, preliminary cultures and test cultures of

the model organism were standardized and described in the

paper.

Prior of preparations of test cultures, the pollutant

solution of known content was neutralized.

From the pollutant solution four parallel dilution series

were prepared in 300 ml Erlenmeyer

- flas ks stoppered with

plastic caps.

The dilution series were preapared as follows: the first

flask of each series contained 160 ml of pollutant solution.

Starting from this flask the subse quent dilution steps were

prepared at a constant dilution ratio by consistently mixing

80 ml of preliminary pollutant dilution and 80 ml

bidistilled water.

Consequently each flask cont ained 80 ml of culture liquid at

the start. Each flask of the three dilution series to be

inoculated was completed to 100 ml by adding 5 ml of each of

the stock solutions and of 10 ml each of the prepar ed

bacterial suspension from the preliminary culture having a

known adjusted extinction. The quantity of the cell material

to be used for inoculation was determined turbidimetrically

and was standardized.

The flasks of the non

- inoculated dilution series were

completed to 100 ml by adding 5 ml each of the stock

solutions and 10 ml saline.

Both inoculated and non

- ino culated dilution series were

kept at 25 °C for 16 h.

After termination of the test period the concentration of

the bacterial suspension was measured turbidimetrically: it

is expressed by the extinction of the primary light of the

monochromatic radiation at 436 nm for a layer of 10 mm

thickness.

The concentration at which the inhibitory action of a

pollutant starts will be present in that step of a dilution

series of the pollutant having an extinction value at the

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

101 end of the test that is >= 3% below the mean value of the

extinction for non

- toxic dilutions of the t est cultures.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

(118)

**Type:** aquatic

**Species:** activated sludge of a predominantly domestic sewage

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**SG :**  = 5000

**Method:** ETAD Fermentation tube method "Determination of damage to

effluent bacteria by the Fermentation Tube Method"

**Year:** 1973

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** SG = toxicity l imit

**Reliability:** (4) not assignable

secondary quotation

(69)

**Type:** aquatic

**Species:** Entosiphon sulcatum (Protozoa)

**Exposure period:** 72 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**TGK :**  > 10000

**Method:** other: growth inhibition test

**Year:** 1978

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

(119)

**Type:** aquatic

**Species:** Microcystis aeruginosa (Bacteria)

**Exposure period:** 192 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**TGK :**  = 530

**Method:** other: growth inhibition test

**Year:** 1978

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

(114)

**Type:** aquatic

**Species:** Tetrahymena pyriformis (Protozoa)

**Exposure period:** 48 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**IC50 :**  = 18756.3 4

**Method:** other: Population Growth Inhibition Test

**Year:** 1990

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

102 **GLP:** no data

**Test substance:** other TS

**Test substance:** purchased from Aldrich Chemical Co., Milwaukee, Wisconsin,

USA with a purity of 9 5%

**Reliability:** (4) not assignable

(120)

**Type:** aquatic

**Species:** Uronema parduzci (Protozoa)

**Exposure period:** 20 hour(s)

**Unit:** mg/l **Analytical monitoring:** no

**TGK :**  > 10000

**Method:** other: growth inhibition test

**Year:** 1980

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Reliability:** (4) not assignable

(121)

**Type:** aquatic

**Species:** Vibrio fisheri (Bacteria)

**Exposure period:** 5 minute(s)

**Unit:** mg/l **Analytical monitoring:** no

**EC50:** = 4440 0

- 88500

**Method:** other: ToxAlert 10, Microtox and LumiStox standard assays

using the appropriate manufacturers's luminometer (see

respective manual)

**GLP:** no

**Test substance:** no data

**Result:** EC50 (5 min): 83600 mg/l (ToxAlert10), 44400 mg/l

(Microtox), and 88500 mg/l (Lumistox);

data related to nominal concentrations;

**Test condition:** temperature: 15 degree C, pH 7

reference s tandard: 440 mg/l Zinc sulphate in 2% NaCl

The same stock chemical solution was used to prepare the

dilution series for each assay (ToxAlert 10, Microtox and

Lumistox, respectively) and all tests were carried out by

the same person on the same day.

test criterion: inhibition of light emitted by the

bioluminescent bacteria Vibrio fischeri after 5 min

compared with zero dos e expressed as EC (effect

concentration);

**Reliability:** (2) valid with restrictions

scientifically acceptable, commercially used assay systems

(122)

**Type:** aquatic

**Species:** other bacteria: Bakterienmischpopulationen

**Exposure period:** 24 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC10:** = 13000

**EC50:** = 30000

**Method:** other: oxygen consumption

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

103 **Test substance:** as prescribed by 1.1

- 1.4

**Remark:** graphicevaluation

nominal concentration

**Reliability:** (4) not assignable

(115)

**Type:** aquatic

**Species:** other bacteria: Nitrosomonas sp./ Nitrobacter sp.

**Exposure period:** 2 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**EC50:** ca. 160

**Method:** othe r: inhibition

**Year:** 1973

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Result:** Hooper and terry showed methanol to be completely inhibitory

to ammonia oxidation by Nitrosomonas bacteria at a

concentration of 5*E

- 3 M (about 160 mg/l).

**Reliability:** (4) not assignable

(123)

**Type:** aquatic

**Species:** other bacteria: activated sludge fr om a municipal sewage

treatment plant

**Unit:** g/l **Analytical monitoring:**

**EC50:** ca. 71.21

**Method:** other: Activated Sludge Respiration Inhibition Test

**Reliability:** (4) not assignabl e

secondary quotation

(124)

**Type:** aquatic

**Species:** other bacteria: cyanobacteria

**Unit:** g/l **Analytical monitori ng:** 

**EC50:** = 20.3

- 43.29

**Method:** other

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** A minimum of 10 concentrations were tested for each species,

ra nging from 0.6% to 10% methanol by volume (4.74

- 79.00

g/l). The EC50 values were calculated from percentage in

hibition values measured daily relative to growth in the

controls.

Anaba ena cylindrica: EC50=2.57% v/v (20.3 g/l); Anabaena

inaequalis: EC50=2.68% v/v (21.17 g/l); Anabaena sp.:

EC50=3.13% v/v (24.73 g/l); Nostoc sp.: EC50=5.48% v/v

(43.29 g/l).

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

(125)

**Species:** Nitrosomonas sp. (Bacteria)

**Exposure period:** 24 hour(s)

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

104 **Unit:** mg/l **Analytical monitoring:**

**IC50 :**  = 880

**Remark:** Inhibition in ammonia consumption

**Reliability:** (4) not assignable

(126)

**Species:** Paramaecium caudatum (Protozoa)

**Exposure period:** 4 hour(s)

**Unit:** mg/l **Analytical monitoring:**

**LC50 :**  = 7690

**Remark:** Raji ni et al. (1989) exposed the ciliated protozoan

Paramaecium caudatum to methanol. The 4 hour LC50 value

was quoted as 1.0% (v/v) methanol or 0.24 M, which corres

- ponds to a concentration of 7690 mg/l ( calculated from the

molar concentration reported). The 10 minute LC50 value was

quoted as 6.0% (v/v) methanol or 1.4 M, i.e. 44860 mg/l

calculated from the molar concentration quoted.

**Reliability:** (4) not assignable

secondary quotation

(127)

**Species:** Photobacterium phosphoreum (Bacteria)

**Exposure period:** 5 minute(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 157

**Remark:** EC50 (5/15/30 minutes) = 157.00/285.56/320.40 g/l.

**Reliability:** (4) not assignable

Secondary quotation

(128)

**Species:** Photobacterium phosphoreum (Bacteria)

**Exposure period:** 15 minute(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 14.7

**Reliability:** (4) not assignable

(129)

**Species:** Photobacterium phosphoreum (Bacteria)

**Exposure period:** 24 hour(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 29.3

**Resu lt:**  = 916 mM

**Reliability:** (4) not assignable

(112)

**Species:** other bacteria: Steptocephalus proboscideus

**Exposure period:** 24 hour(s)

**Unit:** g/l **Analytical monitoring:**

**EC50:** = 1020

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

105 **Result:** = 1020 mM

**Reliability:** (4) not assignable

(112)

**Species:** other bacteria: aerob ic heterotrophs

**Exposure period:** 15 hour(s)

**Unit:** g/l **Analytical monitoring:**

**IC50 :**  = 20

**Remark:** Inhibition of oxygen uptake

**Reliability:** (4) not assignable

(126)

**Species:** other bacteria: methanogenes

**Exposure period:** 48 hour(s)

**Unit:** g/l **Analytical monitoring:**

**IC50 :**  = 22

**Remark:** Inhibition in gas production

**R eliability:**  (4) not assignable

(126)

**4.5 Chronic Toxicity to Aquatic Organisms**

**4.5.1 Chronic Toxicity to Fish**

**4.5.2 Chronic Toxicity to Aquatic Invertebrates**

**TERRESTRIA L ORGANISMS** 

**4.6.1 Toxicity to Sediment Dwelling Organisms**

**4.6.2 Toxicity to Terrestrial Plants**

**Species:** other terrestrial plant

**Expos. period:** 14 day(s)

**Remark:** Plants such as Phaseolus vulgaris, Raphanus sativus

radicula, Lepidum sativum, Trifolium pratense, Saintpaulia

and Petunia hybrids were exposed to air containing methanol

at concentrations between 0.4 and 2.5 mg/m^3 for 14 days.

Retarded growth, mea sured as the effect on both wet and dry

weight of the plants, was reported for each species.

**Reliability:** (4) not assignable

secondary quotation

(130)

**Species:** other terrestrial plant

**Remark:** US EPA (1976) reported that methanol has not been implicated

in vegetation damage as other pollutants, eg. ethylene,

nitrogene oxides, sulphur dio xide and ozone, have. However,

a Russian study has indicated that plants may be sensitive

to methanol vapour at concentrations >0.15 ppm (>0.2

mg/m^3). Branches from eight different species of tree

were studied and the permissible standard (0.15 ppm or

0.2 mg/m^3) was taken as the concentration which did not

### OECD SIDS METHANOL

4. ECOTOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

106 produce a decrease in photosynthesis after 5 minutes ex

posure.

**Reliability :**  (4) not assignable

secondary quotation

(131) (132)

**4.6.3 Toxicity to Soil Dwelling Organisms**

**-**

**4.6.4 Toxicity to other Non**

**- Mamm. Terrestrial Species** 

**-**

**4.7 B iological Effects Monitoring** 

**-**

**4.8 Biotransformation and Kinetics**

**-**

**4.9 Additional Remarks**

**Memo:** more information see IPCS report No. 196

(88)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

107 **5.0 Toxicokinetic s, Metabolism and Distribution** 

**-**

[See Section 5.11]

**5.1 Acute Toxicity**

**5.1.1 Acute Oral Toxicity**

**Type:** LD50

**Species:** rat

**Value:** = 5628 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test subst ance:**  Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (107)

**Type:** LD50

**Species:** rat

**Strain:** Sprague

- Dawley

**Sex:** male/female

**No. of Animals:** 10

**Vehicle:** other: as 15

- 35

- % aqueous solution

**Doses:** 7914; 5390 and 3672 mg/kg

**Value:** > 7914 mg/kg bw

**Method:** other: BASF

- Test

**Year:** 1 975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** No mortality was observed in any dose group.

No pathological findings after post

- observation period of 7

days.

ke y study

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study, limited documentation,

acceptable for assessment, key study

25

- OCT

- 2006 (133)

**Type:** LD50

**Species:** rat

**Strain:** Sprague

- Dawley

**Value:** = 7000 mg/kg bw

**Year:** 1971

**GLP:** no

**Test substance:** no data

**Remark:** T he LD50 was determined in adult male, old rats (300

- 470 g).

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

108 25

- OCT

- 2006 (134)

**Type:** LD50

**Species:** rat

**Strain:** Sprague

- Dawley

**Value:** = 10300 mg/kg bw

**Year:** 1971

**GLP:** no

**Test substance:** no data

**Remark:** The LD50 was det ermined in young adult male rats (80

- 160 g).

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (134)

**Type:** LD50

**Species:** rat

**Strain:** Sprague

- Dawley

**Value:** = 5800 mg/kg bw

**Year:** 1971

**GLP:** no

**Test substance:** no data

**Remark:** The LD50 was determined in rats in 14

- d old rats, weight

16

- 50 g, not yet sexually mature.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (134)

**Type:** LD50

**Species:** rat

**Strain:** Wistar

**Sex:** male/female

**No. of Animals:** 10

**Vehicle:** water

**Doses:** applied in log scale

**Value:** = 13000 mg/kg bw

**Method:** other

**Year:** 1941

**GLP:** no

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study, however, documentation

limited.

25

- OCT

- 2006 (135) (136)

**Type:** LD50

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

109 **Species:** mouse

**Strain:** C57BL

**Value:** = 7300

- 10000 mg/kg bw

**Method:** other: no details reported

**Year:** 1982

**GLP:** no data

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not as signable

Secondary literature

25

- OCT

- 2006 (88) (137)

**Type:** LD50

**Species:** rabbit

**Value:** = 14400 mg/kg bw

**Method:** other: no details inidi cated

**Year:** 1972

**GLP:** no data

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (138)

**Type:** other: LD values

**Species:** rat

**Strain:** no data

**Vehicle:** water

**Doses:** no data

**Year:** 1955

**GLP:** no

**Test substance :**  as prescribed by 1.1

- 1.4

**Remark:** Methanol was administered as 50

- % aqueous solution by

gavage.

non

- BASF study

**Result:** Severe narcosis was produced without latency, however, no

acidosis and no changes of the eye fundus.

Mean LD10 was at 9000 mg/kg

mean LD70 was at 9500 mg/kg

mean LD100 was at 10000 mg/kg.

**Test substance:** Methanol, 99.5 % pure from Merck, no further data

. **Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (139)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

110 **Type:** other: lethal dose

**Species:** rabbit

**Value:** = 7000 mg/kg bw

**Method:** other: no details indicated

**Year:** 1955

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** 7 g Methanol/kg bw per gavage led to mortality within 1

- 3

days. All animals showe d ataxia, nystagmus and coma but no

acidosis and no ophthalmological changes.

non

- BASF study

**Test substance:** Methanol, 99.5 % pure from Merck, no further data.

**Reliability:** (4) not assignable

Se condary source

25

- OCT

- 2006 (139)

**Type:** other: lethal dose

**Species:** dog

**Value:** >= 4000

- 7000 mg/kg bw

**Method:** other: only single animals were used

**Year:** 1955

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Methanol was applied orally together with each 20 g

Saccharose to prevent irritant effects. The animals

receiving 9000 mg/kg body weight died within 46

hours, one after 4000 mg/kg, while 7 other animals receiving

between 2500 and 8000 mg/kg bodyweight survived.

All animals showed signs of an influence on the centr al

nervous system like ataxia, nystagmus, coma, narcosis,

euphoria and vomiting. No ophthalmological changes occurred

and mostly no acidosis.

non

- BASF study

**Test substance:** Methanol, 99.5 % pure from Merck, Germany, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (139)

**Type:** LD50

**Species:** monkey

**S train:**  other: Rhesus macaca

**Sex:** no data

**No. of Animals:** 12

**Vehicle:** water

**Doses:** 500

- 4000, 5000, 6000, 7000, 8000, and 9000 mg/kg

**Value:** ca. 7000

- 9000 mg/kg bw

**Year:** 1961

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** There was no evidence of marked acidosis after high doses

(no hyperventilation, no increase in urinary excretion of

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

111 organic acids, or shift in serum bicarbonate). Signs of

acidosis in only one monkey receiving 6 g/kg were not

accompanied by the characteristic "Kussmaul respiration"

(Cooper and Felig, 1961).

Blindness was not generally noted, but only 4 days after

exposure in one very high

- dosed monkey (9 g/kg) who

survived. This effect apparently was transient, because the

animal showed again no rmal responses to visual stimuli and

normal behaviour by the following day (Cooper and Felig,

1961).

Characterisation of typical symptomatology:

- Ataxia, weakness, and lethargy: 1 to 2 h after exposure

and disappeard with 24 h.

- Transient coma: Usually 0.5 h after exposure intoxication

evident: progressive ataxia, weakness, and lethargy

resulting in coma 1 to 4 h later for 5 to 12 h, but appeared

to fully recover after 24 h.

- Death: previous narcosis then death within 20 to 30 h.

(Cooper and Felig, 1961).

-------------------

**Test condition:** Methanol was administered orally by catheder, diluted to 20

- 30 % in water.

12 animals received doses from 0.5 to 9 g/kg, with 7 and 8

monkeys at sublethal doses and with 6 to 2 monkey at lethal

doses. Some animals were re

- used after a time of recovery,

on the whole 28 applications with 12 animals.

----------------

**Test substance:** Methanol, 99.9 % pure, Fisher Scientific Comp.

**Conclusion:** The lethal dose in this study (Cooper and Felig, 1961) was

significantly higher than published elsewhere for monkeys

(minimal lethal dose at approx. 3000 mg/kg from Potts, 1955;

Gilger et al., 1956) a nd also above reported fatal doses in

humans (approx. 3000 mg/kg). In this study 6000 mg/kg were

sublethal, while Gilger et al.(1959) found a mortality of

78%.

The clinical picture was always one of intoxication

progressing to a narcotic state which in fatal cases was

irreversible. The characteristic features of human

poisoning, such as a latent period, rapid and deep

breathing, vomiting, and blindness, were almost never be

found, but may temporarily occur. Therefore, acute methanol

intoxication in monkeys appears to resemble that in rodents

and other lower animal s (Cooper and Felig, 1961).

------------------ **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, limited, but sufficiently documented

25

- OCT

- 2006 (140) (141) (142) (143)

**Type:** other: intoxication and lethality

**Species:** monkey

**Strain:** other: Rhesus

**No. of Animals:** 8

**Doses:** 6000 mg/kg

**Year:** 1955

**GLP:** no

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

112 **Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** Eight young adult rhesus monkeys received 6000 mg/kg

methanol by gavage.

After 1 h: narcosis at maximum after 4 h.

after 15 h: normal appearance without apparent anomalities;

after 20

- 24 h: marked decrease in alkaline blood capacity.

Following bicarbonate supplementation, 4/8 animals survived.

Clinical observations:

Eye fundus: In case of mild intoxication symptoms, no or

only transitory alterations of the eye fundus were observed.

In severe cases, an extended edema of the r etina and the

optic papilla as well as wide, non

- responsive pupils.

Histological changes: 6/8 animals exhibited cystic

degeneration of the outer retinal granular layer, in 1

animal th ere was evidence of significant demyelinisation of

the optic nerve.

In the elektroretinogramme, a

- wave signal was negative,

while the b

- wave was extinguished.

Neurological symptoms were clinically expressed as rigor and

tremor, histological lesions were seen in the putamen and

nucleus caudatus (3/8 animals).

-----------------------

**Test substance:** Methanol, 99.5 % pure from Merck, no further data.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

113 **Reliability:** (2) valid with restrictions

25

- OCT

- 2006 (144) (143)

**Type:** other: lethal dose

**Species:** monkey

**Value:** >= 3000

- 8000 mg/kg bw

**Method:** other: only single animals were used

**Year:** 1955

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Methanol was applied orally to male rhesus monkeys. 1 to 2

g/kg bodywe ight did not lead to mortality while animals

receiving 3 to 8 g/kg bodyweight died within two days.

Treated animals showed acidosis and partly semicoma and

ophthalmological changes. 20 g Saccharose was applied

concurrently to prevent irritant effects (Gilger and Potts,

1955).

[Compare also Cooper and Felig, 1961 (other entry this

Chapter) who found no mortality in this dose range.]

non

- BASF study

**Test substance:** Methanol, 99.5 % pure from Merck, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (140) (139)

**Type:** LD50

**Species:** rat

**Strain:** Sprague

- Dawley

**Value:** < 790 mg/kg bw

**Year:** 1971

**GLP:** no

**Test substance:** no data

**Remark:** Neonatal rats, 24

- 48 h old, weight 5

- 8 g:

The LD50 could not be exactly determined, but was below 790

mg/kg.

non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (134)

**Type:** LD50

**Species:** rat

**Value:** = 6200 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (107)

**Type:** LD50

**Species:** rat

**Value:** = 9100 mg/kg bw

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

114 **Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (145)

**Type:** LD50

**Species:** rat

**Value:** = 12900 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (146)

**Type:** LD0

**Species:** rat

**No. of Animals:** 10

**Vehicle:** water

**Doses:** limit test: 6.4 ml/kg

**Value:** >= 5065 mg/kg bw

**Method:** other: BASF

- Test

**Year:** 1961

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** Oral application of 5065 mg/kg (6.4 ml/kg) did not lead to

mortality in any of 10 animals used. Only clinical symptoms

including staggering, narcotic effects.

Postobservation period: 7 days

**Test condition:** Methanol was administered as 50

- % aqueous solution.

**Test substance:** Methanol, containing 1.5 % water, no other contamination

**Rel iability:**  (2) valid with restrictions

Comparable to guideline study, limited documentation,

acceptable for assessment

25

- OCT

- 2006 (147)

**Type:** LDLo

**Species:** mouse

**Value:** = 420 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (107)

**Ty pe:**  LD50

**Species:** rabbit

**Value:** = 14200 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (148)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

115 **Type:** LDLo

**Species:** rabbit

**Value:** = 7500 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (107)

**Type:** other: ND50

**Species:** rabbit

**Value:** = 6000 mg/kg bw

**Test substance:** no data

**Remark:** ND50: narcotic dose in 50 % of the animals used.

non

- BASF study

**Test substan ce:**  Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (138)

**Type:** LDLo

**Species:** dog

**Value:**

= 7500 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (107)

**Type:** other: acute intoxication

**Species:** miniature swine

**Strain:** other: minipig YU, CR

**Sex:** female

**No. of Animals:** 3

**Vehicle:** water

**Doses:** 1000, 2500, or 5000 mg/kg (20 % solution)

**Year:** 1993

**GLP:** no data

**Test substan ce:**  no data

**Method:** Three animals were used per dose group and treated by

gavage.

Control tests were conducted with formate administration.

------------

**Remark:** non

- BASF study

**Result:** Dose

- dependent signs of acute intoxication including mild

CNS depression, tremor, ataxia, and recumbency, developed

within 1.5 to 2 h, and resolved by 52 h. Methanol

- and

formate

- dosed animals did not develop optic nerve lesions,

toxicologically relevant formate accumulation, and metabolic

acidosis.

Average maximum methanol levels in blood:

3100 +

- 700, 6200 +

- 2300, and 15200 +

- 900 ug/ml,

respectively, within 0.5 to 4 h. T/2 ranged from about 9

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

116 (low dose) to about 19 and 22 h for the higher doses.

Formate plasma levels: transiently 1.74

- 3.4 mEq/L vs. 0.5

+

- 0.3 mEq/L (control) within 4 to 30 h post

- treatment.

------------

**Test substance:** Methanol, no further data.

**Conclusion:** The minipig, maintained on a normal folate

- diet, does not

appear to be a satisfactory model for human methanol

poisoning.

**Reliability:** (2) valid with restrictions

25

- OCT

- 2006 (149)

**5.1.2 Acute Inhalation Toxicity**

**Type:** LC50

**Sp ecies:**  rat

**Sex:** male/female

**No. of Animals:** 100

**Exposure time:** 4 hour(s)

**Value:** 128.2 mg/l

**Method:** other: BASF

- Test

**Year:** 1980

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**R emark:**  10 male and 10 female Sprague

- Dawley rats (185 +

- 15 g bw)

were tested per dose group. Following analytical / nominal

methanol vapor concentrations were tested in a head

- nose

inhalation system:

87.6 / 127 mg/l

115.9 / 159 mg/l

139.0 / 168 mg/l

150.9 / 193 mg/l

151.1 / 242 mg/l.

Mainly in the high concentrations, where also mortali ty

occurred following clinical symptoms were described: aqueous

secretion of eyes and nose, closed eyes, laboring breathing,

stagger, apathy, narcosis, prone position, opacity of the

cornea. Animals died within 5 days after substance

administration. Necropsy showed cardiac dilatation and

hyperaemic congestion as well as pulmonary edema with

extended or scattered hyperaemia.

Some animals, surviving, still showed some

effect after the 14

- day post observation period.

key study

**Test substance:** Methanol, purity 99.8 % (Merck)

**Reliability:** (2) valid with restrictions

Comparable to guideline study, meeting current standards,

acceptable for assessment, key study

25

- OCT

- 2006 (150)

**Type:** LC50

**Species:** rat

**Sex:** male/female

**No. of Animals:** 160

**Exposure time:** 6 hour(s)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

117 **Value:** 87.5 mg/l

**Method:** other: BASF

- Test

**Year:** 1980

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** 10 male and 10 female Sprague

- Dawley rats were tested per

dose group. Following analytical / nominal methanol vapor

concentrations were tested in a head

- nose inhalation system:

31.1 / 36.1 mg/l

64 .2 / 93.3 mg/l

65.22 / 81.3 mg/l

66.5 / 103.0 mg/l

83.2 / 104.0 mg/l

88.36 / 121.9 mg/l

94.4 / 113.3 mg/l

160.0 / 197.6 mg/l.

Mai nly in the high concentrations following clinical

symptoms were described: aqueous secretion of eyes and nose,

closed eyes, laboring breathing, stagger, apathy, narcosis,

prone position, opacity of the cornea. Animals were dying

within 7 days after substance administration. Necropsy

showed cardiac dilatation and hyperaemic congestion as well

as pulmonary edema and scattered hyperaemia.

Some animals of high concentration groups, surving, still

showed some effect after the 14 day post observation period.

No effect was observed in the lowest concentration group.

key study

**Test substan ce:**  methanol, purity 99.8 %

**Reliability:** (2) valid with restrictions

Comparable to guideline study, meeting current standards,

acceptable for assessment, key study

25

- OCT

- 2006 (151)

**Type:** LC50

**Species:** mouse

**Exposure time:** 134 minute(s)

**Value:** = 79.43 mg/l

**Method:** other: no details reported

**Year:** 1994

**GLP:** no data

**Test substance:** no data

**Remark:** The substance concentration is given as 61100 ppm.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Acceptable study, meets basic scie ntific principles

25

- OCT

- 2006 (152)

**Type:** other: narcotic dose

**Species:** mouse

**Exposure time:** 3.2 hour(s)

**Value:** = 40 mg/l

**Year:** 1936

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

118 **GLP:** no

**Test substance:** other TS

**Remark:** 40 mg/l for 3.2 hours led to narcosis.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- O CT

- 2006 (153)

**Type:** LC50

**Species:** cat

**Exposure time:** 6 hour(s)

**Value:** = 43.68 mg/l

**Method:** other: no details reported

**Year:** 1994

**GL P:**  no data

**Test substance:** no data

**Remark:** The substance concentration is given as 33600 ppm.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Accpetable study, meets basic scientific principles

25

- OCT

- 2006 (152)

**Type:** other: narcotic dose

**Species:** mouse

**Exposure time:** 7 hour(s)

**Value:** = 55 mg/ l

**Year:** 1943

**Test substance:** no data

**Remark:** Exposure to 55 mg/l for 7 hours is reported as narcotic

dose.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (154)

**Type:** LC50

**Species:** cat

**Exposure time:** 4.5 hour(s)

**Value:** = 85.41 mg/l

**Method:** other: no details reported

**Year:** 1994

**GLP:** no data

**Test substance:** no data

**Remark:** The substance concentration is given as 65700 ppm.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reli ability:**  (2) valid with restrictions

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

119 Accpetable study, meets basic scientific principles

25

- OCT

- 2006 (152)

**Type:** LC50

**Species:** cat

**Exposure time:** 6 hour(s)

**Value:** = 26

- 48 mg/l

**Method:** other

**Year:** 1931

**Test substance:** no data

**Remark:** Cats were exposed for 6 hours to concentrations of 26; 48;

97; 210 mg/l or for 3.5 hours to 380 mg/ l.

Concentrations of 26 and 48 mg/l led to 50 % mortality

whichincreased with higher concentrations. The number of

animals used is not indicated.

non

- BASF study

**Test substance:** Metha nol, no further data.

**Reliability:** (4) not assignable

Secondary literature

25

- OCT

- 2006 (155)

**Type:** other: lethal dose

**Species:** monkey

**Strain:** other: Rhesus

**Sex:** male/female

**No. of Animals:** 11

**Exposure time:** 18 hour(s)

**Value:** >= 13 mg/l

**Method:** other

**Year:** 1931

**GLP:** no

**Test substance:** no data

**Remark:** Exposure to 1 .3 mg/l (1000 ppm) for 41 hours, to 10000 ppm

(13 mg/l) for 18 h or to 52 mg/l (40000 ppm) for 1

- 4 hours

led to mortality.

Blindness based on ophthalmology associated with optic

atrop hy is reported (conditions not specified). Recovery

from this lesion is observed.

non

- BASF study

**Test substance:** Methanol, "synthetic", >95 % pure, no further data.

**Reliability:** (2) valid with restrictions

Test procedures based on scientific principles, limited

documentation, acceptable for assessment

25

- OCT

- 2006 (156)

**Type:** LC50

**Species:** rat

**E xposure time:**  4 hour(s)

**Value:** = 83.2 mg/l

**Method:** other: no details reported

**GLP:** no data

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

120 **Test substance:** no data

**Remark:** Substance concentration is given as 64000 ppm.

non

- BASF study

**Te st substance:**  Methanol, no further data.

25

- OCT

- 2006 (152)

**Type:** other: Hormone status

**Species:** rat

**Strain:** Long

- Evans

**Sex:** male

**No. of Animals:** 10

**Doses:** 200, 5000 or 10000 ppm (0.26, 6.5 or 13 mg/l)

**Exposure time:** 6 hour(s)

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Two experiments were conducted to evaluate th e acute effects

of inhaled methanol on serum hormones associated with

reproductive function in the male rat:

1. 10 rats per group (60 d old) exposed to methanol (0,

200, 5000 and 10, 000 ppm) for 6 hours were killed at the end

of the exposure period or the following morning. The effect

of the handling associated with placing the rat in the

exposure chamber was also evaluated by comp aring hormonal

changes in sham

- and methanol

- exposed groups acclimated for

two weeks with groups that were not acclimated. [note: Prior

handling resulted in an increase in serum lutenizing hormone

greater in the non

- acclimated groups than in the acclimated

group.]

2. Groups of acclimated and non

- acclimated rats were

exposed to 0 or 5000 ppm methanol for 1, 2 and 6 hours and

killed immediately after removal from the chamber.

--------------------

**Remark:** non

- BASF study

**Result:** Serum luteinizing hormone, testosterone and follicle

stimulating hormone values were not different in sham

- vs

methanol

- exposed rats at any time point. An effect of prior

handling was noted: In general, the concentrations of these

hormones and serum prolactin in the non

- acclimat ed rats were

greater than those observed for acclimated rats. However,

methanol exposure appeared to result in increased prolactin

concentrations under both handling conditions.

-----------------------

**Conclusion:** Effects on serum hormone levels (luteinizing hormone,

follicle stimulating hormone, testosterone, prolactine) were

inconsistent with partly no effects and partly contradictory

results. According to the author methanol treatment led to

increased prolactin levels in serum.

------------------------ **Reliability:** (2) valid with restrictions

Test procedure s based on scientific principles, sufficiently

documented, acceptable for assessment

25

- OCT

- 2006 (157)

**Type:** other: IRT

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

121 **Species:** rat

**Exposure time:** 7 hour(s)

**Method:** other: based on H.F. Smyth et al. Am. Ind. Hyg. Ass. J.

23,95

- 107, (1962)

**Year:** 1980

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** No mortality was observed wh en rats were exposed for 7

hours to an atmosphere that has been saturated at 20 degrees

celsius with the volatile part of the compound. Irritant

effects on the respiratory tract were observed.

**Test subs tance:**  Methanol, no further data.

25

- OCT

- 2006 (158)

**Type:** other: IRT

**Species:** rat

**No. of Animals:** 6

**Doses:** approx. 150 ml/l (estimated from vapour saturatio n properties

and measured decrease in substance during vapour generation)

**Exposure time:** 4 hour(s)

**Method:** other: BASF

- Test

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Mortality was o bserved when rats were exposed for more than

1 hour to an atmosphere that had been saturated at room

temperature with the volatile part of the compound (11

- 13

Vol%): 3/6 dead after 2 h; 6/6 dead after 4 h.

No mortality occurred up to 1 hour.

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Screening test

(159)

**Type:** other: Inhalation Risk Test

**Species:** rat

**Sex:** male/female

**Exposure time:** 8 hour(s)

**Method:** other: BASF

- Test

**Year:** 1975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** No mortaility was observed when 12 rats were exposed for 3

hours to an atmosphere that had been saturated with the

volatile part of the compound. The inhalation of this

atmosphe re over 8 hours caused mortality of all 6 rats

tested.

Clinical symptoms were strong irritation of mucosa, opacity

of the cornea and narcosis.

-------

**Test substance:** methanol

(160)

**Type:** other: lethal dose

**Species:** rat

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

122 **Exposure time:** 18 hour(s)

**Value:** >= 41 mg/l

**Method:** other

**GLP:** no

**Test substance :**  no data

**Remark:** Six concentrations were tested at different exposure times:

Rats were exposed to Methanol concentrations of 2.6; 11.5;

30 mg/l 8 hours, 17 mg/l 0.33 hours, 65.5 mg/l 2.5 hours and

41 mg/l 18

- 20 hours. Only 41 mg/l for 18

- 20 hours was

lethal.

non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (161)

**Type:** other: lethal dose

**Species:** rat

**Exposure time:** 6 hour(s)

**Value:** >= 41

- 91 mg/l

**Method:** other: various concentrations and exposure times were used

**Test substance:** no data

**Remark:** Exposure to 65

- 100 mg/l for 1.5 hours and 51

- 70 mg/l for 3

hours did not lead to mortality. Exposure to 70

- 111 mg/l for

3 hours or 30

- 120 mg/l for 6 hours led to a dose

related increase in mortality. Clinical signs indica ting

effects on the central nervous system occurred in treated

animals.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary sour ce

25

- OCT

- 2006 (162)

**Type:** LCLo

**Species:** cat

**Exposure time:** 6 hour(s)

**Value:** = 44 mg/l

**Test substance:** no data

**Remark:** non

- BASF study

**Test subs tance:**  Methanol, no further data.

25

- OCT

- 2006 (107)

**Type:** LC0

**Species:** dog

**Exposure time:** 1 hour(s)

**Value:** = 65.5 mg/l

**Test substance:** no data

**Remark:** Dogs were exposed for 24 hours to 2.6 mg/l, for 4 hours to

18 mg/l, for 8 hours to 48 mg/l or for 1 hour to 65.5 mg/l.

At 48 or 65.5 mg/l signs of effects on the central nervous

system occurre d.

non

- BASF study

**Test substance:** Methanol, no further data.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

123 25

- OCT

- 2006 (163)

**5.1.3 Acute Dermal Toxicity**

**Type:** other: lethal dose

**Species:** rat

**Val ue:**  >= 45000 mg/kg bw

**Method:** other

**Year:** 1955

**GLP:** no

**Test substance:** no data

**Remark:** Occlusive application of up to 35000 mg/kg body weight to rats

did not lead to mortality, w hile 45000 mg/kg was lethal.

key study

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

, key study

25

- OCT

- 2006 (164)

**Type:** LD50

**Species:** rabbit

**Value:** = 17100 mg/kg bw

**Year:** 1981

**GLP:** no data

**Test substance:** no data

**Remark:** non

- BASF study

**Result:** No detailed data: LD50 given as 20 ml/kg.

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (165)

**Type:** other: lethal dose

**Species:** monkey

**Strain:** other: Rhesus

**Sex:** male/female

**No. of Animals:** 8

**Doses:** 4x 0.5 and 1.3 ml/(kg*d) under occlusive condition

**Value:** 1600

- 4000 mg/kg bw

**Method:** other: occlusive patch test

**Year:** 1931

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** Four doses of 400 mg/kg daily caused sickness within 24 h,

and ev entually death. At 1000 mg/kg (4 doses daily), severe

illness takes place, followed by death the second day.

**Test substance:** Methanol, "synthetic", >95 % pure, no further data.

**Reliability:** (2) valid with restrictions

Test procedures based on scientific principles, limited

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

124 documentation, acceptable for assessment

25

- OCT

- 2006 (156)

**Type:** LD50

**Species:** rabbit

**Valu e:**  = 20000 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (107)

**Type:** LD50

**Species:** rabbit

**Value:** = 15800 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (148)

**Type:** LDLo

**Species:** monkey

**Value:** = 393 mg/kg bw

**Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (148)

**5.1.4 Acu te Toxicity, other Routes** 

**Type:** LD50

**Species:** rat

**Route of admin.:** i.p.

**Value:** = 9540 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(107)

**Type:** LD50

**Species:** rat

**Route of admin.:** i.p.

**Value:** = 7529 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.p.

**Value:** 10500

- 11500 mg/kg bw

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

125 **Test substance:** other TS

**Test substance:** Methanol, no further data.

(166)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.p.

**Value:** = 10765 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.p.

**Value:** 7914 mg/kg bw

**Method:** other: BASF

- Test

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Test substance:**  Methanol, no further data.

(160)

**Type:** LD50

**Species:** rabbit

**Route of admin.:** i.p.

**Value:** = 1826 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD50

**Species:** guinea pig

**Route of admin.:** i.p.

**Value:** = 3556 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD50

**Species:** hamster

**Route of admin.:** i.p.

**Value:** = 8555 mg/kg bw

**Test substan ce:**  other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD0

**Species:** rat

**Sex:** male

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

126 **No. of Animals:** 25

**Vehicle:** physiol . saline

**Route of admin.:** i.p.

**Value:** >= 3000 mg/kg bw

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Methanol was applied to male F344 rats i. p. at dose levels

of 25; 125; 600 und 3000 mg/kg. After 1; 3; 10; 24; 30 und

48 hours a range of clinical chemical parameters were

determined (formate level, organic acids, folic acid, pH,

pCO2, glucose, urea, Na, K, Cl). The highest dose level led

to a slight increased formate level in blood, but no

decrease in the blood pH. No other parameters were changed.

**Test substance:** Methanol, reagent special grade of Junsei Chemicals Co., no

further data.

**Reliability:** (2) valid with restrictions

(167)

**Type:** LDLo

**Species:** mouse

**Route of admin.:** i.p.

**Value:** = 1 20 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(107)

**Type:** other: effective dose

**Species:** rat

**Route of admin.:** i.p.

**Val ue:**  >= 6000 mg/kg bw

**Year:** 1975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Rats were given 6 g/kg bw Methanol i. p.. The treatment led

to depressionen of the central nervou s system for 4

- 6 hours.

**Test substance:** Methanol, reagent grade, no further data.

(168)

**Type:** other: lethal dose

**Species:** rat

**Route of admin.:** i.p.

**Value:** = 5065 mg/kg bw

**Method:** other: BASF

- Test

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The application of 2530 mg/kg did not lead to mortality in 5

animals but 5065 mg/kg led to mortality in 10 of 10 rats.

**Test substance:** Methanol, no further data.

(169)

**Type:** other: lethal dose

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

127 **Species:** mouse

**Route of admin.:** i.p.

**Value:** >= 5065 mg/kg bw

**Method:** other: BASF

- Test

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The application of 1270 mg/kg did not lead to mortality in 5

animals but 5065 mg/kg led to mortalit y in 1 of 5 animals

and 7910 mg/kg in 6 of 10 animals.

**Test substance:** Methanol, no further data.

(169)

**Type:** other: lethal dose

**Species:** m onkey

**Strain:** other: Macaca fascicularis

**Sex:** male

**No. of Animals:** 5

**Vehicle:** physiol. saline

**Doses:** 0; 25; 125; 600 and 3000 mg/kg.

**Route of admin.:** i.p.

**Value:** = 3000 mg/kg bw

**Method:** ot her

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** After 1; 3; 10; 24; 30 and 48 hours the following clinical

chemical parameters were examined: pH; pCO2; formate,

organ. acids, folate, glucose, urea, K, Na, Cl.

No abnormalities were seen up to 600 mg/kg bw. All monkeys

exposed to 3000 mg/kg fell into a coma immediately after

administration and remained recumbent for 3 h to 2 d.

Additionally, anorexia was observed. Autopsy and

pathological examination revealed no lesions in the eye,

optic nerve or central nervous system except for slight

effects in the liv er (p. 65). During another part of the

study (p. 66), 3 animal died after injection of 3000 mg/kg

within 24 to 48 h.

The highest dose level led to increased formate level,

decreased p H in blood as well as delayed excretion of

methanol and secondary products.

------------------------

**Test condition:** Results created within a metabolism study (NEDO, 1987, p.

61): Methanol was dilut ed in water and applied to male

monkeys (Macaca fascicularis) i.p. A total of 5 monkeys were

repeatedly used.

**Test substance:** Methanol, reagent special grade of Junsei Chemicals Co., no

further data.

**Reliability:** (2) valid with restrictions

(167)

**Type:** other: lethal dose / acidosis

**Species:** monkey

**Strain:** other: adult rhesus Macaca mulatta ( females) and Macaca

nemestrina (pigtail monkey) (females + 1 male)

**Sex:** male/female

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

128 **No. of Animals:** 10

**Vehicle:** physiol. saline

**Doses:** 2000, 3000, and 4000 mg/kg bw (as 20

- or 25 % solution)

**Route of adm in.:**  i.p.

**Value:** = 4000 mg/kg bw

**Year:** 1975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Objective of the study was

1. to find the suitable model animal to represent methan ol

intoxication in humans (acidose model) and

2. to identify those organic acids which are responsible

for development of acidosis.

Blood buffer status was measured: pH, pCO2, CO2,

bicarbonate.

Further blood parameters: ion status, glucose, urea.

Blood formate was measured by colorimetric analysis. Formate

and formate

- borne metabolites were analysed in blood, urin e

and exhaled air by using 14C

- formate. Blood controls were

from the same test animals 12 h prior to methanol exposure.

Organic acids were measured using GC

- MS.

------------------ --------

**Remark:** : The pigtail monkey was tested for effects of

toxic methanol

exposure to establish an animal model for

human methanol

toxicity. Blood acidosis was measured as

major toxic effect

of methanol and the responsible acid was

identified.

In pigtail monkeys but not in rhesus

monkeys significant

blood acidosis was produced by a single

i.p. dose of 4 g/kg

bodyweight methanol. Blood concentration

of formate was

correlated with acidosis, no other organic

acid was found

atincreased levels in acidotic monkeys.

Test substance: Methanol, reagen t grade, no further data.

Mortality occurred at the highest dose

level in one monkey, all treated animals showed depression

of

the central nervous system and acidosis

**Result:** 1. Symptomatology and changes in various blood parameters

After 4000 mg/kg bw, i.p., 3/4 rhesus macacques exhibited no

apparent signs of toxicity within 24 h and later, although a

mild acidosis was produced: pH at 7.22

- 7.24, pCO2 at 30

-

34 mmHg vs pH 7.40 and 37 mmHg for the control.

1/4 rhesus macacque became progressively weaker after

treatment and developed a severe metabolic acidosis which

led to coma and death: His blood values were pH = 7.03 and

pCO2 = 14.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

129 After 4000 mg/kg bw, i.p., 4/4 pigtail monkeys exhibited

distinct signs of methanol intoxication: depression of the

CNS within 30 min, thereafter apparently normal apperance,

followed by weakness and apathy at 15

- 18 h later and

eventually coma and death.

All displayed a sharp decrease in bloo d pH (7.03

- 7.10) and

in pCO2 (15 to 20, and 33 mmHg): 1/4 died after 4

- h coma and

22

- h post treatment, 2/4 recovered from coma, 1/4 in coma

after 32 h and sacrificed after 36 h. The acidosis developed

within about 10

- 12 h accompanied and followed by a marked

decrease in blood bicarbonate, total CO2 and blood pH over

about 20 h post

- treatent.

The two pigtail monkeys given 2000 an d 3000 mg/kg showed no

apparent signs of toxicity, but displayed a mild acidosis

(pH 7.21 and 7.29; pCO2 30 and 32 mmHg).

Inhibition of ADH prior to methanol treatment (4

-

methylpyraz ole, 50 mg/kg i.v.) prevented the occurrence of

acidosis in the pigtail monkey at a dose which otherwise

routinely produced acidosis.

----------------------------- Blood analysis ident ified only bicarbonate as the sole ion

to change during development of acidosis. Additionally, an

time

- dependent increase in blood glucose was evident (<=106

mg/100 ml vs. 73 mg/100 ml of the control).

------------------------------

2. Blood organic acids

Bood formate increased steadily in pigtail monkeys given

4000 mg/kg methanol and reached >800 mg/l (note: In SD rats

given 6000 mg/kg, no increase occured. Concomitantly,

bicarbonate declined.

Other metabolic organic acids measured (lactate,

alpha

- OH

- butyrate, beta

- OH

- butyrate, alpha

- ketobutyrate,

etc.) showed a significant increase versus control values,

but the excess sum in these anions accounted for only about

0.5 mEq./L anf therefore, for not more than 2

- 3% of the

observed decrease i n bicarbonate (p. 54).

3. Formate elimination

The calcualted half

- lifes for NaFormate (50 mg

- 470 mg/kg

bw, i.v.) ranged from 31 min (50 mg/kg) to 51 min (470

mg/kg) in monkey (nor specfied which, and no comparisao

between both strains).

[note in rat: half

- lifes were significant shorter from 12

min (100 mg/kg) to 23 min (670 mg/kg) (p.57)]

------------- ---------------

**Test substance:** Methanol, reagent grade, no further data.

**Conclusion:** The pigtail macacque appeared to serve as a suitable

experimental animal in which to study human methanol

acidosis: Biochemica l and clinical parameters as well as the

parallelism of the time course for the development of

symptoms and acidosis observed in human cases of acute

intoxication are comparable to those found in this m onkey.

Results support the view that formic acid is the major agent

to produce acidosis and methanol intoxication in monkeys and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

130 humans: I appears that formate, a fixed base, replaces

bicarbonate, a volatile base, in the blood and that the

resultant decrease in blood buffer capacity renders the

organismen incapable of mantaining blood pH within normal

limits (p. 58).

---------------------- **Reliability:** (2) valid with restrictions

1c: Test procedures in accordance with accepted standard

methods, sufficiently documented, acceptable for assessment

(168)

**Type:** LD50

**Species:** mouse

**Route of admin.:** s.c.

**Value:** ca. 4160 mg/kg bw

**Test substance:** other TS

**Remark:** LD50 in newborn animals.

**Test substance:** Methan ol, no further data.

(170)

**Type:** LD50

**Species:** mouse

**Route of admin.:** s.c.

**Value:** = 4100 mg/kg bw

**Test substance:** other TS

**Test substance:** Me thanol, no further data.

(107)

**Type:** LD50

**Species:** mouse

**Route of admin.:** s.c.

**Value:** = 9800 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(107)

**Type:** LD50

**Species:** rat

**Route of admin.:** i.v.

**Value:** = 2131 mg/kg bw

**Test substance:** other TS

**Test substance :**  Methanol, no further data.

(148)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.v.

**Value:** = 5673 mg/kg bw

**Test substance:** other TS

**Test subst ance:**  Methanol, no further data.

(107)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

131 **Type:** LD50

**Species:** mouse

**Route of admin.:** i.v.

**Value:** = 5660 mg/kg bw

**Test substance:** other TS

**Test su bstance:**  Methanol, no further data.

(171)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.v.

**Value:** = 4710 mg/kg bw

**Test substance:** other TS

**Tes t substance:**  Methanol, no further data.

(148)

**Type:** LD50

**Species:** mouse

**Route of admin.:** i.v.

**Value:** = 4710 mg/kg bw

**Method:** other: no details reported

**GLP:** no data

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(172)

**Type:** LD50

**Species:** rabbit

**Route of admin.:**  i.v.

**Value:** = 8907 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** LD0

**Species:** dog

**Route of admin.:**  i.v.

**Value:** = 4700 mg/kg bw

**Test substance:** other TS

**Remark:** Methanol was applied i. v. to 4 female dogs at dose levels

of 2.5

- 4.7 mg/kg bodyweight. No mortality occurred but

ataxia a nd severe inflammation of the eyes with blindness

for some days.

**Test substance:** Methanol, no further data.

(173)

**Type:** LDLo

**Species:** cat

**R oute of admin.:**  i.v.

**Value:** = 118 mg/kg bw

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

132 **Test substance:** other TS

**Test substance:** Methanol, no further data.

(107)

**Type:** LDLo

**Species:** cat

**Route of admin.:** i.v.

**Value:** = 4641 mg/kg bw

**Test substance:** other TS

**Test substance:** Methanol, no further data.

(148)

**Type:** other: lethal dose

**Specie s:**  rabbit

**Route of admin.:** i.v.

**Value:** = 4200 mg/kg bw

**Test substance:** other TS

**Remark:** 4200 mg/kg Methanol applied as 50 % solution led to

mortality after 25 min..

**Test substance:** Methanol, no further data.

(139)

**5.2 Corrosiveness and Irritation**

**5.2.1 Skin Irritation**

**Species:** rabbit

**Concentration:** 100 %

**Exposure:** Occlusive

**Exposure Time:** 20 hour(s)

**N o. of Animals:**  5

**Result:** not irritating

**EC classificat.:** not irritating

**Method:** other: BASF

- Test

**Year:** 1975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** The irritation potential of methanol was examined after

several exposure periods of 1, 5, and 15 min and 20 h.

Effects were assessed 24 h and 8 days after treatment

according to Draize scoring.

A piece of gauze was soaked with an unspecified amount

methanol, placed on the shaved skin and covered by an

impervious bandage.

20

- h exposure was conducted on the skin of the back as well

as the ear

.

--------------

**Remark:** key study

**Result:** After 1, 5, 15 min and 20 h of exposure, no signs of

irritation were apparent after 24 h and 8 d.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

133 **Test substance:** Methanol, not further specified

**Relia bility:**  (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented, acceptable for assessment

, key study

25

- OCT

- 2006 (133)

**Species:** rabbit

**Concentration:** 500 mg

**Result:** moderately irritating

**Method:** other: no data

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Tes t substance:**  Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (174)

**5.2.2 Eye Irritation**

**Species:** rabbit

**Concentrat ion:**  100 %

**Dose:** .05 ml

**Comment:** not rinsed

**No. of Animals:** 2

**Result:** irritating

**Method:** other: BASF

- Test

**Year:** 1975

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** After 1 h, mild erythema and cornea opacity as well as

moderate edema associated with secretion were observed.

After 24 h, the effects were assessed as mild.

After 8 days the animals were without s ymptoms.

key study

**Test substance:** methanol, not further specified

**Conclusion:** This screening test gives evidence of mild to moderate

irritation of the mucous membrane on contact with pure

metha nol, but which is reversible and, therefore, gives no

rise to classification as eye irritating.

----------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study and curr ent standards, limited

documentation

, key study

25

- OCT

- 2006 (133)

**Species:** rabbit

**Concentration:** undiluted

**Dose:** .1 ml

**Comment:** not rinsed

**N o. of Animals:**  6

**Result:** slightly irritating

**Method:** OECD Guide

- line 405 "Acute Eye Irritation/Corrosion"

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

134 **Year:** 1990

**GLP:** no data

**Test substance:** no data

**Remark:** non

- BASF study

**Result:** New Zealand White rabbits treated according to OECD

guidelines and assessed based on Draize scoring criteria

exhibited the following mean scores for irritation effects:

Time after application [h]

1 4 24 48 72

----------------------------------------------------

Mean scores

conjunctivitis 0.89 2.00 1.67 2.28 2.22

chemosis 2.00 2.00 0.67 1.00 0.50

iritis 0.33 1.00 1.00 1.00 0.33

corneal opacity 0.00 0.00 0.50 0.50 0.67

-----------------------------------------------------

Eight

- and 14

- d findings not shown.

**Test substance:** Methanol, no further data.

**Conclusion:** The mean scores of the single symptoms did not exceed the

respective limits for classification: Therefore, this result

can be interpreted in terms of a mild irritation potential

not requiring classification as eye irritating.

--------------------------- **Reliability:** (2) valid with restrictions

Guideline study, limited documentation, only abstract

, key study

25

- OCT

- 2006 (175)

**Species:** rat

**Dose:** ot her: saturated air (20 °C)

**Exposure Time:** 3 hour(s)

**Comment:** other: vapour

**No. of Animals:** 12

**Result:** irritating

**Method:** other: inhalation risk test

**Year:** 1975

**GLP:** no

**Test substance:** as prescri bed by 1.1

- 1.4

**Result:** Clinical symptoms were strong irritation of mucosa, opacity of

the cornea.

No mortaility was observed when 12 rats (male/female) were

exposed for 3 hours to an atmosp here that had been saturated

with the volatile part of the compound.

------- **Reliability:** (2) valid with restrictions

Acceptable study, meets basic scientific principles

28

- DEC

- 2004 (160)

**Species:** rabbit

**Result:** irritating

**EC classificat.:** irritating

**Method:** other: Smyth Carpenter

**Year:** 1946

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

135 **Test substance:** no data

**Remark:** non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (176)

**5.3 Sensitization**

**Type:** Guin ea pig maximization test

**Species:** guinea pig

**Concentration 1st:** Induction 50 other: % active substance with and without

Freund ́s Adjuvans (see Method) intracutaneous

**2nd:** Induction 50 % a ctive substance open epicutaneous

**3rd:** Challenge 50 % active substance occlusive epicutaneous

**No. of Animals:** 34

**Vehicle:** water

**Result:** not sensitizing

**Method:** other: modified Magnussen

- Kligman test

**Year:** 1979

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Two studies were conducted using 10 test animals (1st study)

and 24 test animals (2nd study) with each 5 control animals.

During challenge in the first study, a formaldehyde solution

(25 %) was applied in order to identify possible

sensitisation effect that might be caused by

metabolism

- related transformation of methanol to

formaldehyde.

The induction scheme was modified as follows (1 wk between

each treatment): 0.1 ml volume, 50 % test substance

- 1st intradermal induction (including 6 pa rallel

injections with Freund ́s Adjuvans solely, with methanol

solely and both combined)

- 1st epidermal induction

- 2nd intradermal induction (including 4 parallel

injections with methanol solely and combined with

Freund ́s Adjuvans)

- 2nd epidermal induction.

---------------

**Remark:** key study

**Result:** In the first study, no contact sensi tisation was observed,

in the parallel test with formaldehyde, also a negative

result was seen.

In the second study using 24 female animals (two tests à 12

animals), 1/12 (test 1) and 2/12 (test 2) exhibited a slight

skin response (score 1) after 24 and 48 h which can be

interpreted as a weak sensitising potential.

The intracutane induction produced necroses and some open

ulcerations.

----------------

**Conclusion:** In summary, the low number of 3/24 animals with score 1

gives no evidence of a notable sensitisation potential of

methanol.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

136 **Reliability:** (2) valid with restrictions

, key study

25

- OCT

- 2006 (177) (178)

**5.4 Repeated Dose Toxicity**

**Type:** Chronic

**Species:** monkey **Sex:** no data

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Exposure period:** a) 7 months; b) 1 year + 7 months; c) 2 years + 5

months

**Frequency of treatment:** 21 h/d

**Post exposure period:** none

**Doses:** 1000; 100; 10 ppm (1.3; 0.13; 0.013 mg/l)

**Control Group:** yes, concurrent no treatment

**NOAEL:** 10 ppm

**LOAEL:** 100 ppm

**Method:** other: see Method

**Year:** 1985

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term studies, repr oductive assays and carcinogenicity

studies:

Whole

- body exposure: 4 monkeys were housed in one inhalation

chamber.

8 animals per exposure group were used. Sacrifices were

conducted 7 months (2/group), 19 months (3/group), and at 29

months (3/group) (Tab. 3, p. 29). A further group at 1000

ppm was included for recovery (30 d and 6 months) (p. 31 f).

Clinical, clini cal

- chemical, hematological,

ophthalmological, and pathological/histopathological

examiniation were done. Furthermore, endocrine status was

tested after 1 year and 7 months as well as after 2 years

and 5 months.

Analytical control of chamber concentration: Analytical

values close to nominal ones. (p. 20 ff)

-------------------------

**Remark:** The blood levels of methanol a nd formate were not measured.

Note: Exposure per day was 20 h, which implies that

prolonged steady

- state blood levels were reached which even

may have been higher than in studies using the same exposur e

concentration, but shorter exposure times.

Relevance of observations limited by the low number of

animals available at each time interval.

--------------------

key study

non

- BASF study

**Result:** The animals showed normal body

- weight gain, food and and

water consumption throughout. The monkeys of the 1000

- ppm

group showed abnormal scratching of the skin and clinical

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

137 symptoms such as frequent yawning and nasal discharge (see

also recovery test, p. 31 f).

In both upper dose groups, the ECG of 1/8 of the 100

- ppm

exposed monkey and 3/8 of the 1000

- ppm exposed monkeys

showed abnormalities (negative T

- or Q

- wave changes and

flattening of T

- wave) which indicated slight myocardial

disorder (p. 21).

Examinations of the e ye fundus revealed no changes in any

group.

Haematological examinations revealed no deviations from

normal in Hb, Ht, red and white blood

- cell count, or

percentage of white blood cell s in any group.

No significant differences in serum values were seen, but

one monkey in the 100

- ppm group for most liver parameters

(total protein, GOT, GPT, total cholesterol, free

c holesterol, thymol and glucose). This was not considered

treatment

- related.

BRAIN:

In both upper exposure groups, the number of responsive

stellate cells containing increased endoplas mic reticulum

(minimal hypertrophy of astroglia) appeared in the basal

ganglions after 1 year and 7 months of exposure. In the

cerebral white substance, slight hyperplasia of the

astro glia was noted in the 100 and 1000

- ppm groups, in

particular in the 100

- ppm group after 7 months (NEDO, 1987,

p.53). Yet, this was not characteristic of a degenerative

process and was unrelated to dosin g, because no such

phenomena were noted after 2 years and 5 months.

In the cerebral white substance, diffuse increases of

responsive stellate cells centered in the subcortical white

s ubstance and the semioval center of the frontal and

parietal lobes was noted at the higher dose groups, but also

emerged in 3/8 monkeys of the 10

- ppm group after 29 months

(p. 23). This was apparently a reversible effect, as shown

from monkeys after recovery.

There were a few cases of degeneration of the optic nerve

and the corpus geniculatum laterale after 7 months and 19

months (o phthalmencephalon). In one or two cases in the groups

of 8 animals exposed for 2 years and 5 months, including the

10

- ppm group, slight degeneration of the optic nerve was

suspected, but not considered as significant (p. 23).

Slight degeneration processes including increased

responsiveness of the astroglia were noted in the thalamus

after exposure to 100 and 1000 mg for 7 months and longer (p.

24).

Peripheral Nervous System:

One monkey at 0.13 mg/L and 2 at 1.3 mg/L showed slight but

clear changes in peroneal nerves; the authors concluded that

these effects show that methanol causes damage to peripheral

nerves (p. 25/26).

There were mild degenerations of the livers and kidneys

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

138 which showed no clear correlation with exposure levels and

exposur e time. There was some evidence of an increase in

fatty granules in liver parenchyma at 1000 ppm along with

signs of fibrosis in the hepatic cord more pronounced than

at the lower concentration. Sudan

- p ositive granules were

observed at 100 and 1000 ppm in the renal tubular epithelium.

(NEDO, 1987, p. 26/27).

There was an increase in the Sudan

- positive granules in

heart tissue at 100 0 ppm, but not manifested in any

morphological lesion of the heart muscle (e.g. fibrosis).

LUNG and TRACHEA:

There were no cases of pneumonia in any of the exposed

monkeys. In the lun gs, fibrosis was seen in the interalveolar

space. However, no dose response was observed, and the effect

was also seen in the controls. In the trachea, atrophy of

the epithelium of the

mucous membranes and reduction of goblet cells were observed

in a total of 4 cases of exposed animals, but not correlated

with the amount of methanol inhaled. This was not seen in

the controls (p. 2 8).

---------------------------

**Test condition:** Exposure time 21 hours per day, significantly longer than

according to guideline (6 hrs) (see p. 5). This does not

represent normal industrial and con sumer use.

---------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride, <3 ppm formaldehyde

------------

**Conclusion:** Taking into account slight myocardial efects and slight

hyperplasia of the astroglia in the cerebral white substance

already noted at 100 ppm, then already 100 ppm (130 mg/m3)

have to be defined as LOAEL with a NOAEL of 10 ppm (13

mg/m3).

But no clear adverse effects are reported at 1000 ppm:

no necrotic effects occurred in the nervous

tissue. Hyperplasia of astroglia, not considered as

degenerative, might be a transient methanol

- dependent effect

which appears to subside upon cessation of long

- term

methanol exposure (p. 52/53). But there was no clear

correlation to the exposure c oncentration and time.

Therefore, the biological relevance is questionable.

There was evidence of an increase in mild fatty changes in

the liver and kidney upon exposure to 1000 ppm, which were

associated with signs of fibrosis. This effect was still

present after recovery (see other entry). Given the small

histological effect (p. 27), the pathological relevance

appears to be low, b ut indicates that long

- term exposure to

1000 ppm methanol is on the borderline to toxicologically

relevant, histological manifestations (authors statement, p.

27).

There were histolog ical signs of diffuse responsiveness of

the astroglia in some parts of the cerebral white substance

at all exposure levels, at high exposure after shorter time

period, after 10 ppm after 29 months in a few animals. The

relevance was not clearly commented by the authors.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

139 However, under test conditions (continuous exposure), there

was no evidence of irreversible effects arising from

lo ng

- term exposure to up to 1000 ppm methanol.

Therefore, the NOAEL for continuous exposure may be

established at 1000 ppm (1300 mg/m3)m, but the low number of

animals and limited description do not allo w to draw firm

conclusions on the apparent neural effect.

-------------------------- Note: This study served as basis for risk assessment by

Vyskocil and Viau (2000): A 1

- h reference concentration

(RfC) was derived to be at about 100 mg/m3, taking into

account sensitive people. A very conservative chronic RfC

was obtained and proposed at 0.38 mg/m3, based on the

a ssumption that 10 ppm has to be used as NOAEL and 100 ppm

as the LOAEL for "neurotoxic effects". This appears to be

not in compliance with the authors ́ observations and is not

suggested in the report.

--------------------------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, however, documentation and number of anaimals

limited.

, key study

25

- OCT

- 2006 (167) (179)

**Type:** Chronic

**Species:** rat **Sex:** male/female

**Strain:** Fis cher 344/DuCrj

**Route of administration:** inhalation

**Exposure period:** 12 months

**Frequency of treatment:** continuous, average about 20 h/d, total exposure time

7318

- 7341 h (males); 7474

- 7496 h (females)

**Post exposure perio d:**  none

**Doses:** 10; 100; 1000 ppm ( 0.013; 0.13; 1.3 mg/l)

**Control Group:** yes, concurrent vehicle

**LOAEL:** 1000 ppm

**NOEL :**  100 ppm

**Method:** other: in accordance with OECD Guideline O ct. 1980

**Year:** 1983

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studies and carcinogenicity

studies (NEDO, 1987):

Whole

- body exposure: The animals were housed individually in

wire

- mash cages attached to the inhalation ch amber.

20 animals per sex and dose group were tested. Clinical,

ophthalmological, clinical

- chemical, hematological and

pathological/histopathological examiniation were done. The

latte r included among others the optic nerve, the eye, and

reproductive organs.

Analytical control of chamber concentration: Analytical

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

140 values close to nominal ones.

-------------------- -----

**Remark:** The blood levels of methanol and formate were not measured.

key study

non

- BASF study

**Result:** One female rat of the 1000 ppm dose group and one male rat

of the 10 ppm do se group died or were sacrificed in

extremis.

This was considered spontaneous on the basis of the absence

of a consistent dose relationship.

A very slight transient suppression of bod y

- weight gain was

observed for males and females of the 1000 ppm dose group

from week 27 through 44 (less than 5 %). This has to be

attributed to temporary occurrence of slight diarrhea in

these groups during this period. The otherwise addressed

significant decreases in body weight at the end of the study

are not noticable in Fig. 3 representing the time

- course of

body weight develop ment. The high

- dose males showed a

minimal, but significant decrease in food consumption from

week 30 to the end. However, this was not more than about

- 5

% from the control.

Data of the organ/body weight ratio revealed a dose

- related

upward tendency in the liver and spleen for females which

remained within a 5

- % range.

Urinalysis showed no changes suggesting effects from

exposure to methanol.

Hematologic examinations revealed no clear changes which

could be attributed to exposure to methanol.

Serum biochemical examination showed a small tendency to

decrease for alkaline phosphatase, the enzymatic activities

of GOT, GPT, LDH and gamma

- GTP did not show any differences.

Free fatty acid showed lower values in all exposure groups

without dose

- res ponse relationship. Cholesterol and

triglyceride remained unchanged.

Changes of other clinical

- chemical parameters showed no

correlation with the methanol exposure.

Histopathological examinations showed a variety of

non

- tumoral changes in various organs, many of them were

infrequent findings which were possibly accidental. For

tumoral changes, similarly, almost all the findings obse rved

were considered spontaneous ones due to aging.

-------------------------------

**Test condition:** Exposure time 20 hours per day, significantly longer than

according to guideline (6 hrs). This d oes not represent

normal industrial and consumer use.

---------------

**Test substance:** Methanol of first grade by Junsei Chemicals, <1 ppm vinyl

chloride, <3 ppm formaldehyde

--- --------------

**Conclusion:** According to the authors, only in the 1000 ppm dose

- group

changes could be considered treatment

- related. But based on

the minor degree and severity, these changes have no

pathological meaning and may be considered as toxicologically

irrelevant.

Levels of 100 ppm or less (at 20 h/d) did not produce any

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

141 effect (= NOEL). 1000 ppm is adopted as LOAEL, although it may

be considered as a NOAEL.

(see also IPCS/WHO, 1997)

----------------------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting cu rrent

standards, documentation limited.

, key study

25

- OCT

- 2006 (88) (167)

**Type:** Chronic

**Species:** mouse **Sex:** male/fe male

**Strain:** B6C3F1

**Route of administration:** inhalation

**Exposure period:** 12 months

**Frequency of treatment:** continous, average ca. 20 h, total 7202

- 7225 h

(males); 7352

- 7373 h (females)

**Post exposure pe riod:**  none

**Doses:** 10; 100; 1000 ppm (0.013; 0.13; 1.3 mg/l)

**Control Group:** yes, concurrent no treatment

**NOAEL:** 1000 ppm

**NOEL :**  100 ppm

**Method:** other: in accordance with OECD Guid eline Oct. 1980

**Year:** 1983

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studies and carcinogenicity

studies (NEDO, 1987):

Whole

- body exposure: The animals were housed individually in

wire

- mash cages attached to the inhala tion chamber.

30 animals per sex and dose group were tested. Clinical,

ophthalmological, clinical

- chemical, hematological and

pathological/histopathological examiniation were done. The

latter included among others the optic nerve, the eye, and

reproductive organs.

For interim examination, 10 animals each were sacrificed

after 6 months.

Analytical control of chamb er concentration: Analytical

values close to nominal ones.

-------------------------

**Remark:** The blood levels of methanol and formate were not measured.

Note: Exposure per day was 20 h, whic h implies that

prolonged steady

- state blood levels were reached which even

may have been higher than in studies using the same exposure

concentration, but shorter exposure times.

-------------

key study

non

- BASF study

**Result:** One female animal of the 100

- ppm group died on day 174,

another one had to be killed in moribund state on day 178.

During the whole study, there were no changes in the general

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

142 appearance that could suggest any relation to treatment.

The body

- weights of both sexes of the treated groups tended

to be higher than the controls in the second half of the

study (NEDO, Fig. 3, p. 168).

There was a decreased food consumption in high

- dose females

during the first 2 months and after 7 months without effect

on body

- we ight gain.

All clinical and blood test failed to reveal any

methanol

- related alteration in parameters from normal.

In the group of high

- dose females, an increase in organ

weight with significant differences was noted for the liver

after 6 months and for the kidney and spleen after 12

months, however, the latter without statistical significance

and a clear dose

- response.

After 12 months, the incidence of more severe fatty

degeneration of hepatocytes was pronounced in high

- dose

males: This was graded as moderate in 8/20, mild in 8/20 and

negative in 4/20 survivin g males vs. 1/20 (moderate), 9/20

(mild) and 10 negative in the male control group. On the

other hand, this effect was common and found to similar

extent in untreated/external male mice. The female mice

showed no difference among the groups.

--------------------------

**Test condition:** Exposure time 20 hours per day, significantly longer than

according to guideline (6 hrs). This does not represen t

normal industrial and consumer use.

---------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride, <3 ppm formaldehyde

----- --------------------------

**Conclusion:** According to the authors, only in the 1000 ppm dose

- group

changes could be considered treatment

- related. But based on

the minor degree and severity, these changes have no

pathological meaning and are considered as toxicologically

irrelevant.

Levels of 100 ppm or less did not produce any effect (=

NOEL).

1000 ppm can be established as NOAE L, while it is the LOEL

for non

- pathological, apparently treatment

- related effects.

(see also IPCS/WHO, 1997)

----------------------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, however, documentation limited.

, key study

25

- OCT

- 2006 (88) (167) (180)

**Type:** Sub

- chro nic

**Species:** rat **Sex:** male/female

**Strain:** Sprague

- Dawley

**Route of administration:** gavage

**Exposure period:** 90 days

**Frequency of treatment:** daily

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

143 **Doses:** 0, 100, 500 or 2500 mg/kg body weight

**Control Group:** yes, concurrent vehicle

**NOAEL:** = 500 ml/kg bw

**LOAEL:** = 2500 mg/kg bw

**Method:** other: EPA Test protocol

**GLP:** yes

**Test substance:** no data

**Method:** 30 animals per sex and group were used. After 6 weeks, 10

rats per sex and dose group were subjected to interim

sacrifice.

The following parameters were examined: bodyweight, food

consumption, clinical signs, ophthalmological evaluations,

mortality, blood and urine chemistry, gross and microscopic

evaluations.

------------------

**Remark:** non

- BASF study

**Result:** Elevated levels of SGPT, SAP and increased (not

statistically significant) liver weights in males and

females of the high dose group. Brain weights of both high

dose males and females were signific antly less than those

of controls. There were no histopathological effects.

The NOAEL was determined at 500 mg/kg/day, based on

increased liver enzyme levels and reduced brain weights.

---------------------

**Test substance:** Methanol, no further data.

**Conclusion:** The subchronic NOAEL was determined at 500 mg/kg/day, based

on increased liver enzyme levels and reduced brain weights.

Due to th e low pathological relevance of effects, this value

is considered to be a NOEL.

---------- **Reliability:** (4) not assignable

Guideline study, only abstract available

25

- OCT

- 2006 (181) (182)

**Type:** Sub

- acute

**Species:** rat **Sex:** male/female

**Strain:** Sprague

- Dawley

**Route of administration:** inhalation

**Exposure period:** 4 weeks

**Frequency of treatment:** 6 h/d, 5 d/wk

**Doses:** 0, 520, 1980 and 5010 ppm (0.663; 2.65; 6.63 mg/l)

**Control Group:** yes, concurrent no treatment

**NOAEL:** 6.63 mg/l

**GLP:** no data

**Test sub stance:**  as prescribed by 1.1

- 1.4

**Method:** The study used 5 rats/sex/group. Parameters evaluated

included, ophthalmoscopic exam, body weight, clinical signs,

organ weights, histopathology and survival.

--------------

**Remark:** A good screening study. Daily exposure was 6 hours per day,

the normal daily exposure length for inhalation studies

evaluating potential workplace exposure.

--------------

key study

non

- BASF study

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

144 **Result:** All animals survived. In rats nasal and eye discharge

(mucoid, nasal, red nasal, lacrimation) was noted in the

tre atment groups.

Only mucoid nasal discharge appeared to be dose related.

There were no treatment

- related effects on body weight.

No ocular abnormalities were noted at the terminal

ophth almoscopic exam. No treatment related histopathological

effects (35 tissues) were noted, but spleen weights were

increased in female rats exposed at 2,000 ppm (not at 5,000

ppm). No other organ weight e ffects were noted.

----------------

**Test substance:** Methanol, 99.85 % pure from Celanese Corp. NY, no further

data.

**Conclusion:** Rats were exposed to up to 5,000 ppm (6 hr/d, 5d/wk for 4

week s) showed no treatment related histopathological

effects.

Inhalation exposure had resulted in some slight

treatment

- related signs.

**Reliability:** (2) valid with restrictions

Test p rocedures in accordance with accepted standard

methods, well documented, acceptable for assessment

, key study

25

- OCT

- 2006 (183)

**Type:** Sub

- acute

**Species:** monkey **Sex:**

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Exposure period:** >= 15 d

**Frequency of treatment:** 21 h/d

**Post exposure period:** none

**Doses:** 1 0000; 7000; 5000; 3000 ppm (about 13; 9.1; 6.5; 3.9

mg/l)

**Control Group:** yes, concurrent no treatment

**LOAEL:** 3000 ppm

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studies and carcinogenicity

studies: Four animals were used per test and control group.

Exposure period was variable, but was expected to be at

least 15 days (p. 12 ff).

Test procedure with respect to exposure times not well

documented.

---------------

**Remark:** key study

non

- BASF study

**Result:** Blood levels (p. 51):

3000 ml/m3 5000 ml/m3

============= ================================

Methanol: 80 mg/l 5250 mg/l

Formate: 30 mg/l 1210 mg/l

=============================================

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

145 The animals at the top dose showed lethargy, and after 3

treatment fell into coma and died.

The animals of the 7000

- ppm group had to be killed because

of suffering.

The animals at 5000 and 3000 ppm exhi bited weakness and

uncoordinated movement. Food consumption was reduced at 5000

ppm. The body weight was normal.

Haematological findings displayed no changes, except for a

somewhat hi gher leukocyte count at the two highest exposure

levels.

Blood pH in one animal of the 5000

- ppm and at the highest

dosed groups were significantly decreased showing acidoses.

No change s were found in other clinical

- chemical blood and

liver values, but in the 7000

- ppm group alkaline phosphatase

increased.

At 5000 ppm, necrosis of the basal ganglion of the cerebrum

i s reported, associated with marked cerebral edemas.

These effects were not seen at 3000 ppm.

Effects (mild atrophy) on the visual system (retina, optic

nerve, state of myelinated fibres) were hardly n oticeable

and considered as insignificant at 3000 ppm and below, but

pronounced at 5000 ppm.

Fatty degeneration in the liver occurred at 3000 ppm and

more pronounced at the higher exp osures, with necrosis after

5000 ppm. Also at this exposure level, kidneys were partly

affected, showing vacuolated degeneration.

--------------------

**Test condition:** Exposure time 21 hours per day , significantly longer than

according to guideline (6 hrs). This does not represent

normal industrial and consumer use.

---------------

**Test substance:** Methanol of first grade by Junsei Chemicals, < 1 ppm vinyl

chloride, <3 ppm formaldehyde

----------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, limited docu mented

, key study

25

- OCT

- 2006 (167)

**Species:** monkey **Sex:** no data

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Ex posure period:**  1000 ppm: 7 months; 2000 ppm + 3000 ppm 20 d; 5000 ppm

6 d + 4000 ppm 6 d

**Frequency of treatment:** 21 h/d

**Post exposure period:** 30 d; 6 months; 10 or 4 months (see Method)

**Doses:** 1000; 20 00; 3000; 5000+4000 ppm (1.3; 2.6; 3.9; 6.5+5.2

mg/l)

**Control Group:** yes, concurrent no treatment

**Method:** other: Special acute to chronic inhalation test programme

including recovery phases

**Year:** 1985

**GLP:** no data

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

146 **Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term studies, reproductive assays and carcinogenicity

studies:

Whole

- body exposure: 4 monkeys were housed in one inhalation

chamber.

Here recovery study with specific foc us on neurological

effects apart from organ toxicology:

1000 ppm: 7 months exposure + 30 d or 6 months recovery

2000 and 3000 ppm: 20 d exposure + 30 d, 6 or 10 months

recovery

5000 ppm: 6 d exposure + 6 d 4000 ppm exposure with 30 d or

4 months recovery.

Clinical, clinical

- chemical, hematological,

ophthalmological, and pathological/histopathological

examiniation were done.

Analytical control of chamber concentration: Analytical

values close to nominal ones.

-------------------------

**Remark:** key study

non

- BASF stu dy

**Result:** At the concentration of 4000 and 5000 ppm, irreversible

lesions/degenerations appeared to be likely in the basal

ganglions of the cerebrum, because they did not subside

within the r ecovery period, while after 3000 ppm (20 d

exposure) the slight necrotic degenerations were not

progressive as evidenced after recovery.

The increase of responsive astroglia seen in the cerebral

white substance was still present after recovery from

3000

- ppm exposure.

Degenerative changes in the visual system were evident after

exposure to 3000 ppm and higher: atrophy of the optic n erve

and reduction in myelinated fibers. Recovery was unequivocal

at exposures below 1000 and 2000 ppm, but at 3000 ppm,

there appeared to be a trend of these lesions to progress,

alth ough

- due to the low number of animals

- this could also

reflect individual variability acc. to authors.

Lesions of the liver occurred in all dose groups with

round

- cell infiltration and fibrosis noted in dose

- related

manner. They were still present histologically after the

recovery phase.

Changes of the kidneys were observed in all groups:

hyalinisation of glomeruli, cell infiltrati ons into the

renal tube stroma. These effects were no longer noted after

recovery.

-----------------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride; <3 ppm formaldehyde

------------

**Conclusion:** At 1000 ppm (7 months exposure and higher; see chronic

exposure other entry), some irreversibility of degenerative

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

14 7 changes (mild fibrosis presumably following fatty

degeneration) unrelated to the recovery period was noted in

the liver (p.54)[see also p. 27: chronic study].

These effects were histologically small and, theref ore, of

little pathological relevance.

All other effects

- if there were any at all

- were not

persistent and not considered pathologically relevant at

that level, but were significantl y more pronounced at 3000

ppm and higher after shorter exposure (20 d), associated

with eventual lack of recovery.

Between 2000 and 3000 ppm appears to be a threshold

concentration whe re methanol

- induced neurotoxic effects may

become biological relevant (p. 35).

-------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting curren t

standards, however, documentation and number of animals

limited.

, key study

25

- OCT

- 2006 (167)

**Type:** Sub

- acute

**Species:** monkey **Sex:** male

**Strain:** other: rhesus Macaca mulatta

**Route of administration:** gavage

**Exposure period:** 1.5 to 6 d

**Frequency of treatment:** variable

**Doses:** initial 2000 mg/kg, thereafte r 500 or 1000 and 2000

mg/kg (variable frequency)

**Control Group:** other: internal / same animal prior to treatment

**Method:** other: (see Method)

**Year:** 1978

**GLP:** no

**Test substance:** as pre scribed by 1.1

- 1.4

**Method:** Test model in monkeys for methanol

- induced ocular toxicity

to characterise the toxicity syndrome and histological

manifestations. Methanol was applied as 20% solution by

nasogastric tube (see: McMartin et al., 1975).

Ophthalmoscopic examinations and ultrastructural

histopathology (electronmicroscopy) were performed.

-----------------------

**Remark:** non

- BASF study

**Result:** 1. Under methanol treatment acc. to this test design,

formate levels were between min. 7.2 and max. 14.4 mEq/L in

blood and cerebrospinal fluid, blood bicarbonate min. 4.0

and max. 10.2, and blood pH min. 7.13 and max. 7.28.

Methanol levels ranged from 1540 to 2840 mg/L (Martin

- Amat

et al., 1977).

2. All six animals developed fundus changes at the head of

the optic nerve (optic disc) within 43 to 171 h after

methanol ingestion, expressed as intraaxonal swellings

(Hayreh et al, 1977). Electronmicroscopic studies revealed

swelling of the nerve fibers with an accumulation/clustering

of swollen mitochondria in the optic nerve head being

maximally in the lamina cribrosa region. Furthermore, in the

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

148 retrolaminar and intraorbital optic nerve, swel ling of

astrocytes was prominent as well as swelling of the

cytoplasm of the oligodendroglial cytoplasm in contact with

the axons (Baumbach et al., 1977).

Alterations were not observed in the retina itself: The

ganglion cells of the retina were intact with only minimal

swellings of the mitochondria and loss of cristae. But these

findings were also present in the control tissue (Baumb ach

et al., 1977).

----------------------

**Test condition:** A total of six monkeys was treated such as to maintain a

state of metabolic acidosis at attenuated toxicity (arterial

bl ood pH 7.1 to 7.3 and bicarbonate at 10 mEq/L or above)

for a prolonged time: A high initial dose was followed by

lower doses depending on the animal ́s acidotic response in

blood.

[not e: Experience had told (McMartin et al., 1975) that

after a single dose of 3000 mg/kg bw., in general, the

animals died within 20 to 30 h without demonstrating ocular

abnormalities.] (Martin

- Amat et al ., 1977).

----------------------

**Test substance:** Methanol, highest purity available, no further data (see:

McMartin etal., 1975)

**Conclusion:** 1. While acute methanol toxicity in monkeys (after a

single dose) does not yield ocular signs, repeated dosing

succeeded in producing ocular lesions (Martin

- Amat et al,

1977).

The only detectable ocular change was optic disc edema (of

the optic papilla) which was similar to that seen in raised

intracranial pressure in humans, but without this pressure

after methanol (Hayreh et al, 1977).

The primary sites of ocular injury were th e optic nerve

heads and the anterior segment of the optic nerve rather

than the retinal ganglion cells themselves. It appears that

interference with oxidative phosphorylation causes

mi tochondrial damage, thus disruption of active axoplasmic

flow in the retrolaminar optic nerve (Baumbach et al., 1977;

Hayreh et al., 1977).

[note: In humans it has been hypothised that optic atrophy,

which often follows acute methanol intoxication, is

secondary to injury of the retinal ganglion cells.]

2. Mechanistically, there is a close causal relationship

between the prolonged increase in formic acid from methanol

and the development of optic edema. Similar effects can be

produced by i.p. administration of formate without acidosis

(see other entry 5.11: Martin

- Amat et al., 19 78).

--------------------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

25

- OCT

- 2006 (184) (185) (186) (187) (188) (189)

**Type:** Sub

- acute

**Species:** rat **Sex:** male

**Strain:** Sprague

- Dawley

**Route of administration:** inhalation

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

149 **Exposure period:** 1; 2; 4; or 6 Wk

**Frequency of treatment:** 6 h/d, 5 d/Wk

**Post exposure period:** 16 h after the last exposure. One group exposed for 6

wks: recovery 2 wks

**Doses:** 200, 2,000 or 10,000 ppm (0.265; 2.65; 13.3 mg/l)

**C ontrol Group:**  yes, concurrent no treatment

**NOAEL:** 13.3 mg/l

**Year:** 1982

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Limited study: Only biochemical/physiological and

cytological parameters of the lung were examined

(tissue

- specific: lung weight, tissue DNA and protein

contents, RNase, Protease; in the filtered lavage fluid:

surfactant, number of cells, PMN, protein, surface LDH and

N

- acetyl

- beta

- glucosaminidase).

Histologic analyses of lung tissue were not conducted.

Four rats per group were used.

Chamber concentration was analysed by GC.

--------

**Remark:** non

- BASF study

**Result:** Even the highest concentration caused no signs of lung

inflammation or irritation.

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Special study design, meets generally accepted scientific

principles, restrictions in assessment

25

- OCT

- 2006 (190)

**Type:** Sub

- chronic

**Species:** rat **Sex:**

**Strain:** no data

**Route of administration:** inhalation

**Exposure period:** 3 months

**Frequency of treatment:** 12 h/d, 5 d/wk

**Doses:** a) 1.77; 49 mg/m3; b) 0.57; 5.3 mg/m3

**Control Group:** yes, concurrent no treatment

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Special neurophysiological study design: chronaximetry =

ratio of the minimum time necessary for a stimulus of twice

the absolute threshold intensity to evoke a response

measured as muscle contractions in response to an electric

current applied to an animal ́s hind leg. Normally, the

flexor chronaxia is shorter than the extensor chronaxia, and

their ratio is stated to be relatively stable (NTP

- CERHR,

2001, p. 80).

------------

**Remark:** non

- BASF study

**Result:** Low concentrations of methanol inhaled for 3 months by rats

are reported to exhibit changes in the average motor

chronaxia ratio between antagonistic muscles. No significant

effects were noted at 1.77 mg/m3 (Chao, 1959) and 0.57 mg/m3

(Ubaydullayev, 1966). At the other doses the chronaxia

returned to normal during the recovery period.

Unspecified histopatholog ical changes in relation to the

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

150 mucous membranes of the respiratory tract were seen at 450

mg/m3, but not at the lowest doses. Some shifts in liver

parameters in blood were also mentioned.

-------------------

**Test substance:** Methanol, no further data.

**Conclusion:** The analysis by Kavet and Nauss (1990) indicated that these

studies do not provide adequate evidence of an association

between neurobehavioral effects and low

- level exposure to

methanol: limited animal number, insufficient reporting of

methodss, and unclear statistical analysis. The biological

significance of changes in the chr onaxia ratio is considered

as uncertain. (see also NTP

- CERHR, 2001, p. 80/81)

------------------- **Reliability:** (4) not assignable

Test methods and performance uncertain, limited

documentation available

25

- OCT

- 2006 (191) (192) (193)

**Type:** Sub

- acute

**Species:** rat **Sex:** male

**Route of administration:** inhalation

**Expos ure period:**  1; 2; 4 or 6 weeks

**Frequency of treatment:** 8 h/d 5 d/wk

**Doses:** 200, 2000, and 10000 ppm (0.265; 2.65; 13.3 mg/l)

**Control Group:** yes, concurrent no treatment

**Method:** other

**GLP:** no dat a

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Study design was to elucidate sexual hormone status in

mature male rats (Sprague

- Dawley): determination in blood

after exposure of circulating free testos terone, luteinizing

hormone and follicle stimulating hormone.

------

**Remark:** Conflicting results as compared to findings of others (see

also other entry 5.1.2: Cooper et al., 1992; and 5.4: L ee et

al., 1991).

non

- BASF study

**Result:** Significantly decreased levels of circulating free

testosterone were observed among rats exposed at 200 ppm for

2 and 6 weeks (32 % o f control) and 6 weeks at 2,000 ppm.

The high dose group (10,000

- ppm) showed no change. The

authors interpreted this as evidence that methanol exposure

had lowered testicular production of testosterone

. In addition, significant increases in circulating LH were

observed after six weeks of exposure to 10,000 ppm. No

changes in follicle stimulating hormone levels were

observed.

-----------------

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

generally accepted scientific principles, restrictions in

assessment (see Remark)

25

- OCT

- 2006 (194)

**Species:** rat **Sex:** male

**Strain:** Sprague

- Dawley

**Route o f administration:**  inhalation

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

151 **Exposure period:** 1 d or 1 wk

**Frequency of treatment:** 6 h/d

**Post exposure period:** none or 18 h

**Doses:** 200 ppm (0.265 mg/l)

**Control Group:** yes, concurrent no treatment

**Method:** o ther

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Study design was to elucidate sexual hormone status in

mature male rats (Sprague

- Dawley): determination in blood

after exposu re of circulating free testosterone, luteinizing

hormone and follicle stimulating hormone.

**Remark:** Conflicting results as compared to findings of others (see

also other entry 5.1.2: Cooper et al., 1992; and 5.4 : Lee et

al., 1991).

non

- BASF study

**Result:** SD rats exposed to 200 ppm methanol for 6 h for 1 day or

1x/d for one week showed a significant depression of serum

testosterone ( 59 %) immediately after the exposure, but not

after one

- week daily 6

- h exposure or after 18

- h recovery.

There was no evidence of an effect on LH.

----------------

**Test substance:** Methanol, no fu rther data.

**Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

generally accepted scientific principles, restrictions in

assessment (see Remark)

25

- OCT

- 2 006 (195)

**Type:** Sub

- acute

**Species:** rat **Sex:** male

**Strain:** Sprague

- Dawley

**Route of administration:** inhalation

**Ex posure period:**  a) 1, 2, 4, and 6 wk; b) 13 wk

**Frequency of treatment:** a) 8 hours/day, 5 days/week; b) 20 hours/day, 7

days/week

**Doses:** a) 200 ppm (260 mg/m3); b) 50, 200, 800 ppm (1040

mg/m3)

**NOAEL:** = 800 ppm

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Special study design: The potential toxic effects of

methanol vapors on testicular producti on of testosterone (a)

and the morphology of testes (b) were investigated using

normal or methanol

- sensitive folate

- reduced rats.

(a) Nine rats per group were used.

(b) 11 to 12 anima ls (10 months old) for three exposure

levels, and 8 animals (18 months old) for a 800

- ppm group

were used.

-----------

**Remark:** non

- BASF study

**Result:** Methanol inhalation did not r educe serum testosterone levels

in normal rats. Testes isolated from methanol

- exposed (200

ppm) rats had the same capability as those from air

- exposed

rats in synthesizing testosterone, whether testes were

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

152 incubated in the absence or presence of hCG.

The testes

- to

- body weight ratio and testicular morphology of

rats exposed up to 800 ppm methanol for up to 13 weeks (20

hours/day, 7 days/week) were not different from control

animals, irrespective of the nutritional state, normally fed

or folate deficient.

A greater incidence of testicular degeneration was seen only

in 18 month old, folate

- deficient rats exposed to 800 ppm

methanol for 13 weeks (20 hours/day, 7 days/week). The

results are equivocal.

800 ppm, the top level, can be established as subchronic

NOAEL based on testicular histopathology.

--------------

**Test substance:** Methanol, no further data

**Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented, acceptable for assessment

25

- OCT

- 2006 (196)

**Type:** Sub

- acute

**Species:** rat **Sex:** male

**Strain:** Long

- Evans

**Route of administration:** gavage

**Exposure period:** 21 d

**Frequency of treatment:** 2 x/d

**Post exposure period:** none

**Doses:** 800; 1600 mg/kg/d

**Control Group:** yes, concurrent no treat ment

**Method:** other: special test on endocrin function and male reproduction

parameters

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** Treatment with th e highest dose produced a transient, but

significant decrease in serum testosterone. On day 21 of

treatment, the testosterone level was normal again, but a

significant increase in the luteinising hormon e was

analysed. The testis weights were reduced as well as the

number of morphologically normal sperms in the epididymis.

The acute treatment with high doses of 800, 1600, and 3200

mg /kg (2x 12 h apart) had no effect on testis weight, and on

sperm size. At day 1 after treatment, a reduced testosterone

value was found in serum and in the interstitial fluid.

After 7, 14, and 21 d post

- application, testosterone in both

compartments were normal without significant difference from

the control. LH was distinctly higher on day 21.

(see also other entry 5.1.2: Cooper et al., 1992)

-------------------------

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Only abstract

25

- OCT

- 2006 (197)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

153 **Type:** Sub

- acute

**Species:** rabbit **Sex:** male

**Route of administration:** gavage

**Exposure period:** 5

- 7 d

**Frequency of treatment:** daily

**Doses:** 4.68 g/kg as 30

- % solution

**Remark:** non

- BASF study

**Result:** Lethal dose for all 14 rabbits within the following 5 to 7

days.

**Test substance:** Methanol, no further data.

25

- OCT

- 2006 (198)

**Type:** Chronic

**Species:** dog **Sex:**

**Route of administration:** inhalation

**Exposure period:** 379 d

**Frequency of treatment:** 8 h/d, 7 d/wk

**Doses:** 0.585

- 0.65 mg/l

**GLP:** no

**Rem ark:**  non

- BASF study

**Result:** Four dogs were tested for a broad spectrum of haematological

parameters and ophthalological alterations. There was no

evidence of a treatment

- related effect.

**Test substance :**  Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (199)

**Type:** Sub

- acute

**Species:** monkey **Sex:** male/female

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Exposure period:** 4 wk

**Frequency of treatment:** 6 h/d, 5 d/wk

**Post exposure period:** none

**Doses:** 500, 2000 , and 5000 ppm (0.663; 2.65; 6.63 mg/l)

**Control Group:** yes, concurrent no treatment

**NOAEL:** 6.63 mg/l

**Year:** 1987

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Three monkeys per sex and dose were use d. Chamber methanol

concentration was measured regularly by IR.

Histopathological examinations included ophthalmoscopy at

the end of exposure, but apparently no microscopical

inspectio n of brain and neural tissue.

-----------

**Remark:** non

- BASF study

**Result:** There were no clinical signds of intoxication. There were no

methanol

- related abnormalities.

**Test substance:** Methanol, pur ity 99,85 % (from Celanese Corp., N.Y.)

**Reliability:** (4) not assignable

Test procedures in accordance with accepted standard

methods, insufficiently documented, limitations in tissue

examination ( see Method).

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

154 25

- OCT

- 2006 (200)

**Species:** monkey **Sex:** male

**Route of administration:** gavage

**Exposure period:** 3 d

**Frequency of treatment:** daily

**Doses:** 2340 mg/kg as 30

- % solution

**Test substance:** no data

**Remark:** non

- BASF study

**Result:** Lethal dose for all 7 animals under test after 3 days.

**Test substance:** Methanol, no further data.

**Reliability:** (2) v alid with restrictions

Meets generally accepted scientific standards, sufficiently

documented

25

- OCT

- 2006 (198)

**5.5 Genetic Toxicity 'in Vitro'**

**Type:** Ames test

**System of testing:** Salmonella typhimurium TA1535, TA1537, TA1538, TA98,

TA100 (plate incorporation standard test)

**Concentration:** <= 2.5 mg/plate

**Cytotoxic Concentration:** non

- toxic within t he range of testing

**Metabolic activation:** with and without

**Result:** negative

**Method:** other: acc. to Ames et al., 1975

**Year:** 1981

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Test condition:** S9 mix was prepared from Arochlor

- induced SD rat livers.

**Test substance:** Methanol, from Carlo Erba, no further data.

**Reliability:** (2) valid with restrictions

Comprehensive test programme using test procedures in

accordance with accepted standard methods, but limited

documentation

--------------

25

- OCT

- 2006 (201)

**Type:** Ames test

**System of testing:** Salmonella typhimurium TA97, TA102 (plate incorporation

standard system)

**Concentration:** <= 7.5 mg/plate

**Metabolic activation:** with

**Result:** negative

**Meth od:**  other: acc. to Maron and Ames, 1983

**Year:** 1984

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** key study

non

- BASF study

**Result:** In TA102, a slightly positive t rend was indicated [(+

- )

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

155 ambiguous], reproducible in 5 parallel independent

experiments, but never exceeding the revertant ratio of 2:

Spontaneous mutation rate 200

- 300/plate, while in the

presence of high methanol doses, an increase in the mutation

frequency above background of 140 revertants was found.

**Test condition:** S9 mix was prepared from Arochlor

- induced SD rat livers.

**Test substance:** Methanol, from Carlo Erba, no further data.

**Conclusion:** The methanol

- related increase in the mutation frequency in

TA102 did not fulfil the accepted criteria for mutagenic

activity. Along with the high methanol doses required to

induce such a weak effect and the negative results observed in

all other Ames tests, the overall evidence clearly

demonstrates that methanol is not mutagenic in bacterial

reverse

- mutation sy stems.

------------- **Reliability:** (2) valid with restrictions

Comprehensive test programme using test procedures in

accordance with accepted standard methods, but limited

docu mentation

--------------

, key study

25

- OCT

- 2006 (202) (203)

**Type:** Ames test

**System of testing:** Salmonella typhimurium TA1538, TA98, TA1537, TA1535

**Conc entration:**  <= 3.6 mg/plate

**Metabolic activation:** with and without

**Result:** negative

**Method:** other: acc. to Ames

**Year:** 1981

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Metabolic activation system was derived from liver S

- 9

fraction of Aroclor

- 1254 pretreated rats.

non

- BASF study

**Test substance:** Methanol, from Merck, Germany, no further data.

**Reliability:** (2) valid with rest rictions

Comprehensive test programme using test procedures in

accordance with accepted standard methods, limited

documented

25

- OCT

- 2006 ( 204)

**Type:** Ames test

**System of testing:** Salmonella typhimurium TA100, TA98, TA1537, TA1538,

TA1535; Escherichia coli WP2uvrA

**Concentration:** 5, 10, 50, 100, 500, 1000, and 5000 μg/plate

**Metabolic activation:**  with and without

**Result:** negative

**Year:** 1985

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Metabolic activation system was derived from liver S

- 9

fra ction of KC500(polychlorinated biphenyl)

- pretreated rats.

non

- BASF study

**Test substance:** Methanol, 99.6 % pure, no further data.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

156 **Reliability:** (2) valid with restrictions

Comprehensive test programme using test pr ocedures in

accordance with accepted standard methods, sufficiently

documented

25

- OCT

- 2006 (205)

**Type:** DNA damage and repair assay

**Syste m of testing:**  Escherichia coli WP67, WP2, CM871

**Concentration:** no details

**Cytotoxic Concentration:** > 40000 μg/well (no details, derived from the MIC)

**Metabolic activation:** with and without

**Result:** ambiguous

**Method:** other: no Guideline indicated

**Year:** 1984

**GLP:** no data

**Test substance:** no data

**Method:** E. coli strains used were all trp

- dependent (trp

- ), carrying

an ochre nonsense mutation blocking an intermed iate process in

the synthesis of tryptophan:

E. coli WP2 (wild

- type, repair proficient)

E. coli WP67 (repair deficient: uvrA

- , polA

- )

E. coli CM8761 (repair deficient: uvrA

- , recA

- , l exA

- ).

For each tester strain, the Minimal Inhibitory Concentration

(MIC) was determined. The endpoint is cytotoxicity. The ratio

the MIC of the repair proficient to the MIC of the repair

deficient strains was taken as indicator for genotoxicity. A

ratio of greater than 2 was accepted as significant only if

reproducible in 5 parallel independent experiments.

------------------

Trials were conducted in miniature form (liquid micromethod

procedure: 350 μl) using microwell plates that contained

nutrient broth, and as 2

- h preincubation assay

("treat

- and

- plate met hod" on nutrient broth agar).

--------------------

The initial concentration was governed by the solubility or by

the toxicity of the TS as inferred from preliminary testing.

Starti ng from this, eight 2

- fold dilution steps followed in

general.

-------------------

Bacterial inocula were obtained from 16

- h cultures in nutrient

broth with about 10^3 cells per wel l. The exact bacterial

density was determined by seeding the same volume of cell

culture (0.1 ml) onto a nutrient agar plate after 16

- h

incubation at 37 °C.

--------------------

Bacterial growth in the test wells was visually evaluated

after 16 h at 37°C by observing the increase in turbidity of

the medium and/or formation of pellets of settled cells.

----------- -----

**Remark:** non

- BASF study

**Result:** The microwell method resulted in a negative result in the

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

157 presence of S9 and in a positive result in the absence of S9

at 20 mg/well (Flora et al., 1990, 1984). Bu t the

preincubation procedure was negative without S9, but ambiguous

with S9 (Flora et al. 1984).

Note: The MIC concentrations were very high (40 and 20 mg/well

= about 120 g/L and 60 g/l).

-------------------

**Test substance:** Methanol, no further data.

**Conclusion:** Given the high concentrations and the conflicting findings, it

is concluded that observations made at the margin of

significance (ratio = 2) are of low reliability and biological

relevance. The weak relative increase of toxicity in

repair

- deficient strains may be an increase in unspecific

cytotoxicity rather t han solely "genotoxicity".

(Note: A similar result was obtained with ethanol at somewhat

lower concentrations).

-------------- **Reliability:** (2) valid with restrictions

2c: Mee ts generally accepted scientific standards, well

documented with methodological restrictions (see Conclusions)

, key study

25

- OCT

- 2006 (203) (206)

**Type:** other: M icronucleus Test

**System of testing:** V79 Chinese hamster cells

**Concentration:** 50 μl/ml (approx. 40 mg/ml)

**Metabolic activation:** without

**Result:** negative

**Year:** 1984

**GLP:** no data

**Remark:** Comparative study including alcohols, acetone, and various

alkylating agents. Exposure was 48 h for alcohols. 4000

- 7000 interphase cells were scored per treatment.

-----------------

key study

non

- BASF study

**Result:** No MN increases induced by any alcohol or acetone, whereas

the alkylating agents produced significant MN frequences

above medium controls.

**Test substan ce:**  Methanol [CAS 67

- 56

- 1], 95 % pure from Prolabo, no further

data.

**Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documente d, acceptable for assessment

, key study

25

- OCT

- 2006 (207)

**Type:** other: Micronucleustest in vitro

**System of testing:** Syrian hamster embryo cells

**Concentration:** not indicated: derived from range finding

**Metabolic activation:** without

**Result:** negative

**Method:** other: according to Schmuck et al. (1988)

**Year:** 1988

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

158 **Method:** Comparative study including a broad spectrum of chemicals.

Exposure time was 18 hours, no further details of method are

reported.

Report refers to:

Schmu ck, G. et al. (1988): Characterisation of an in vitro

micronucleus assay with Syrian hamster embryo fibroblasts.

Mut. Res., 203, 397

- 404:

Cell cultures established from 13

- d old embryos. Tests

performed with tertiary cultures in a humidified atmosphere

with 12 % CO2 in air at 37°C.

Tertiary cell cultures were incubated for 24 h (1.5 x10exp5

cells on cover slips in 35

- mm petri dishe s).

Then various concentrations of the TS were added in DMSO

(final concentration of the solvent =<0.1%) (Dose ranges had

been determined prior to the main test, but no details are

giv en concerning the concentrations.)

Incubation time 5 h, followed by washing.

Further incubation for 18 h.

Thereafter, the cells were fixed, stained and scored for

micronuclei.

Monitoring of micronuclei: The number of cells containing

single or multiple micronuclei was counted among 2000 cells

for each dose.

STATISTICAL ANALYSIS: Student ́s t

- test

-------------------

----------------------

**Remark:** non

- BASF study

**Test substance:** Methanol, analytical grade

**Reliability:** (4) not assignable

4e: Documentation insufficient for assessment, test pro cedures

in accordance with accepted standard methods, but limited

documentation.

25

- OCT

- 2006 (208) (209)

**Type:** Sister chromatid exchange assay

**System of testing:** CHO

- Zellen

**Concentration:** up to 0.1%, 1x/d for 8 days

**Metabolic activation:** without

**Result:** negative

**Method:** other

**Year:** 1977

**GLP:** no

**Test substance:** no data

**M ethod:**  Repeated exposure design: ethanol, propanol, butanol, and

acetalaldhyde were also tested. No standard positive control

substance was included. Test medium was change every second

day, b ut test substances appeared to be added every day "up

to a final concentration of 1 %".

-----------

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

159 **Remark:** non

- BASF study

**Result:** The alcohols produced no increases in the SCE rate compared

to the control, but acetaldehyd induced a dose

- related,

significant increase.

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Only short communication. Test proced ure largely accordance

with current standards and based on scientific principles.

Positive result of acetaldehyde raises the level of

reliability for the proper functionality of the test system.

25

- OCT

- 2006 (210)

**Type:** other: Cell transformation assay

**System of testing:** C3H/10T1/2 Cl 8 cells (mouse embryo fibroblasts cells)

**Concentration:** 5, 10, 20, 50, 10 0 mg/ml

**Cytotoxic Concentration:** 50 mg/ml (> 30 % inhibition of growth)

**Metabolic activation:** with and without

**Result:** negative

**Year:** 1981

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** 12

- O

- Tetradecanoyl

- phorbol

- 13

- acetat TP

non

- BASF study

**Result:** Methanol and formic acid induced no foci type I or III either

in the presence or absence of TPA

(12

- O

- Tetradecanoyl

- phorbol

- 13

- acetate) as promoting agent.

[note: Formaldehyde, on the other hand, caused a high number

of transformed cells in the presence of TPA, but failed to in

its absence.]

**Test substance:** Methanol, HPL C grade from Fisher, no further data.

----------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

25

- OCT

- 2006 (211)

**Type:** other: Cell transformation assay

**System of testing:** BHK 21 C 13 (Baby Syrian Hamster Kidney Cells)

**Concentration:** 10, 50, and 100 ul/ml (approx. 8, 40, and 80 mg/ml)

**Cytotoxic Concentration:** LC50 approx. 60 mg/ml

**Metabolic activation:** with and without

**Result:** negative

**Year:** 1981

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Metabolic activation system was derived from liver S

- 9

fraction of Aroclor

- 1254 pretreated male NMRI mice (500

mg/kg, i.p.).

non

- BASF study

**Result:** No increase in transformants, while mitomyci n, MMN,

cyclophosphamide and dichromate were highly active.

---------------

**Test substance:** Methanol from Merck, Germany, p.a., no further data.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

160 **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

25

- OCT

- 2006 (212)

**Type:** Mammalian cell gene mutation assay

**System of te sting:**  Chinese Hamster V79 Zellen 8

- Azaguanin

- , 6

- Thioguanin

- , Ouabain

- Resistenz

**Concentration:** 15.8, 31.7, 47.4, and 63.3 mg/ml

**Cytotoxic Concentration:** 63.3 mg/ml (approx. 70% inhibition of colony formation)

**Metab olic activation:**  with and without

**Result:** negative

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** key study

non

- BASF study

**Result:** No increas es in mutant frequency in gene mutation to drug

resistance vs. neg. control, whereas the pos. control DMN

produced increases in dose

- related manner.

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride, <3 ppm formaldehyde

------------------ **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

stan dards, sufficient documentation, acceptable for

assessment

, key study

25

- OCT

- 2006 (167)

**Type:** Chromosomal aberration test

**System of testing:** Chinese hamster lung cell

**Concentration:** 7.1, 14.3, and 28.5 mg/ml

**Cytotoxic Concentration:** 28.5 mg/ml (approx. 50% inhibition)

**Metabolic activation:** with and without

**Result:** negative

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** MNNG was used as pos. control (0.5 and 1.0 ug/ml) in the

absence of S9 and 2

- aminoanthracene (2 and 4 ug/ml) in the

presence of S), along with two negative control (untreated

and solvent = medium). 200 metaphases were scored after 6,

24, and 48

- h exposure (p. 242).

------------

**Remark:** key study

non

- BASF study

**Result:** No (dose

- dependent) increases in any type of chromosome and

chromatid breaks and gaps were noted at any time interval,

while MNNG and 2

- AA produced clear dose

- related increases in

breaks and gaps ( Tables, p. 250

- 255).

----------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride, <3 ppm formaldehyde

------------------

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

161 **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, sufficient documentation, acceptable for

assessment

, key study

25

- OCT

- 2006 (167)

**Type:** Mouse lymphoma assay

**System of testing:** mouse lymphoma cells L5178Y

**Concentration:** 7.9 mg/ml

**Metabolic activation:** with

**Result:** positive

**Method:** other: acc. to Clive and Spector. Mut. Res., 31, 17

- 29, 1975

**Year:** 1985

**GLP:** no data

**Test substance:** no data

**Method:** Objective: Optimisation of the amount of the S9

- mix to be

used to obtain m aximum yield of mutations. Comparative study

including typical mutagens.

Exposure time 4 h.

---------------------

**Remark:** key study

non

- BASF study

**Result:** In the presence of 10

- 15 ul/ml S9 and methanol (7.9

mg/ml), there was a significant increase in mutation

frequency. No detailed results presented.

---------------------

**Test substance:** Methanol, no fur ther data.

**Reliability:** (4) not assignable

Only abstract, no detailed data.

, key study

25

- OCT

- 2006 (213)

**Type:** other: Mitotic recombination in ascom ycetes

**System of testing:** Aspergillus nidulans diploid strain P1

**Concentration:** 5.2, 5.6, 6.0, and 7.0 % (v/v)

**Cytotoxic Concentration:** 7.0 % (lowest concentration to arrest conidial

germination)

**Metabolic activatio n:**  without

**Result:** positive

**Method:** other: no data

**Year:** 1989

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Mitotic segregants were detected as homozygous (yAA2/yA2) or

hemizygous (yA2/0) yellow sectors or patches in heterozygous

(yA2/yA2+) pale

- green colonies growing on complete medium.

Yellow segregants were isolated, analyzed for their

nutritional r equirements and classified as mitotic cross

- over,

non

- disjunctional diploids and haploids.

----------------------

Treatment of conidia (10 exp^5/ml) with the TS at early

germinatio n, i.e during the first mitotic divisions, in liquid

complete medium at 37 °C under agitation until emergence of

germ tubes (check by light microscopy).

Plating onto complete agar for determination of s urvival and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

162 detection of mitotic segregants.

---------------------

**Remark:** Approx. 3 % chromosomal malsegregation was induced.

The result was statistically significant at two concentration s

and a dose

- response relationship was evident (IPSC/WHO 1997).

non

- BASF study

**Result:** There was a concentration

- related increase in non

- disjunctions

with a maximum of about 3 %. No aneuploidi es were noted at the

lowest test concentration. No other forms of malsegregation

were statistically significant (1/265 cross

- over seen at 6.0

%).

The survival was reduced in dose

- rela ted manner.

Conc. Survival Scored Aneuploids(yellow segregants)

[%] [%] colonies Counts %

=======================================================

5.2 26 268 0 0

5.6 19 407 6 1.47*

6.0 10 265 8 3.02**

7.0 5 176 0 0

--------------------------------------------------------

Spontaneous control

0 100 2673 7 0.26

========================================================

* = P<0.01; ** = P<0.001

Note: Similar results were obtained with ethanol and other

pri mary aliphatic alcohols. The aneuploidy rate of ethanol

reached a maximum of about 10 %.

-----------------------

**Test substance:** Methanol [CAS 67

- 56

- 1], purity >99.9%

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

25

- OCT

- 2006 (214)

**Type:** Yeast gene m utation assay

**System of testing:** Schizosaccharomyces pombe (ade 6 locus)

**Concentration:** <= 5 %

**Cytotoxic Concentration:** 10 %

**Metabolic activation:** with and without

**Result:** negative

**Year:** 1980

**GLP:** no

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comparative study including various solvents:

As indicator for mutation served the phenotypic change from

the red to the white colour of the colonies. The P1 strain

produces red colonies, owing to a mutation at the ade6 locus

which blocks the adenine biosynthesis pathway, associated with

the accumulation of a red pigment, 5

- amino

- 4

- imidazole

ribotide. Any mutation at each 5 different loci blocking the

pathway to the pigment will lead to the development of white

colonies.

---------------------

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

163 **Remark:** There was no incre ase in mutant frequency as compared to the

negative controls (buffer and DMSO) at a the maximum conc.

of 5 % with or without post

- mitchondrial fraction from

phenobarbital

- induced mouse liver.

--------------

non

- BASF study

**Test substance:** Methanol, analytical grade, from Carlo Erba, Milano, no

further data.

**Reliability:** (4) not assignable

4e: Documentation insufficient fo r assessment (see Method)

25

- OCT

- 2006 (215)

**Type:** Ames test

**System of testing:** Salmonella typhimurium TA98, TA100, TA1535, TA1537

(spot test)

**Concentration:** 3 μmol/plate

**Metabolic activation:** with and without

**Result:** negative

**Year:** 1979

**GLP:** no

**Test substance:** no data

**Remark:** Metabolic activation system was derived from liver S

- 9

fraction of Aroclor

- 1254 and 3

- methylcholanthrene

- pretreated

rats.

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (3) invalid

Significant methodolog ical deficiency: Test concentration

may have been too low, no dose

- response relationship given

25

- OCT

- 2006 (216)

**Type:** Ames test

**System of testing:** Salmonella typimurium TA1535, TA1537, TA1538, TA98,

TA100

**Concentration:** <= 5 mg/plate

**Metabolic activation:** with and without

**Result:** negative

**Method:** other: acc. to Ames

**Year:** 1 977

**GLP:** no

**Test substance:** no data

**Remark:** Metabolic activation system S9 mix was prepared and applied

according to Ames

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Comprehensive test programme using test procedures in

accordance with accepted standard methods, limited

documented

25

- OCT

- 2006 (217)

**Type:** other: Cell transformation assay

**System of testing:** Syrian Hamster Embryo Cells

**Result:** negative

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

164 **Year:** 1983

**GLP:** no data

**Test substance:** no data

**Rema rk:**  non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Secondary source

25

- OCT

- 2006 (218) (219)

**Type:** Ba cterial reverse mutation assay

**System of testing:** Salmonella typhimurium TA1535, TA 1537, TA1538, TA100,

TA98 + E. coli WP2 uvvA (trp dependent)

**Concentration:** 10, 50, 100, 500, 1000, and 5000 μg/plate

**Metabolic act ivation:**  with and without

**Result:** negative

**Method:** other: largely in compliance with OECD Guide

- lines

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Metabolic activation system was derived from liver S

- 9

fraction of PCB

- pretreated rats (500 mg/kg, single i.p.) (p.

241).

---------------

non

- BASF study

**Test substance:** Methanol, reagent sp ecial grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride, <3 ppm formaldehyde

------------------ **Reliability:** (4) not assignable

Comprehensive test programme using test procedures in

accordance with accepted standard methods, only short

communication.

25

- OCT

- 2006 (167)

**Type:** other: mutagenic inactivation of a bacteriophage

repressor

**System of testing:** E. coli RK+ (lysogenic)

**Concentration:** up to 23 % (0.23 ml/ml = 0.18 g/ml)

**Metabolic activation:** without

**Result:** positive

**Method:** other

**GLP:** no data

**Test substance:** no data

**Method:** The mutagenic inactivation of a bacteriophage repressor

gene leading to subsequent replicative killing of the

bacteria was used as mutagenicity screening model. Bacte ria

were exposed for 10 min. at 30 øC.

**Remark:** non

- BASF study

**Result:** Methanol was toxic and mutagenic at concentrations of 23 %

(v/v) and more (extreme high concentrations necessary to

produce the "mutagenic" effect).

**Test substance:** Methanol, no further data.

**Reliability:** (3) invalid

Questionable, not validated test system, extreme high

concentrations necessary to produce the "mutagenic" e ffect.

25

- OCT

- 2006 (220)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

165 **5.6 Genetic Toxicity 'in Vivo'**

**Type:** other: Cytogenetic assays including chromosomal aberration,

SCE, and micronuclei

**Species:** mouse **Sex:** no data

**Strain:** no data

**Route of admin.:** oral unspecified

**Exposure period:** no data

**Doses:** 1000 mg/kg

**Result:** positive

**Method:** other: no data

**Year:** 1982

**GLP:** no data

**Test substance:** no data

**Remark:** non

- BASF study

**Result:** The incidences of chromosomal aberrations, particularly

aneuploidy and SCE in bone marrow cells as well as

micronuclei in pol ychromatic erythrocytes are reported to be

increased. No detailed results presented.

--------------------------

**Test substance:** Methanol, no further data

**Reliability:** (4) not assignable

Only abstract, no detailed data.

25

- OCT

- 2006 (221)

**Type:** Sister chromatid exchange assay

**Species:** mouse **Sex:** male

**Strain:** C57BL

**Route of admin.:**  inhalation

**Exposure period:** 6 h/day, 5 days

**Doses:** 800 or 4000 ppm (1.04 or 5.2 mg/l)

**Result:** negative

**Method:** other: see Method

**Year:** 1991

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive test programme (see also other entries this

chapter): Primary cultures of lung cells of treated mice and

controls were established and 50 second

- division metaphases

were scored for SCE induction. Replication indices were

determined of 200 metaphases and mitotic indices of 1000

cells.

**Remark:** non

- BASF study

**Result:** No treatment related SCE induction was seen : SCEs ranged

from about 11

- 12 per metaphase irrespective of exposure

and control.

The replicative and mitotic indices were not significantly

different from the control.

-----------------------

**Test substance:** Methanol, 99.9 % pure from Fisher Scientific Corporation,

no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current sta ndards,

sufficient documentation, acceptable for assessment,

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

166 however, missing positive control

25

- OCT

- 2006 (222)

**Type:** Micronucleus assay

**Speci es:**  mouse **Sex:** male

**Strain:** ICR

**Route of admin.:** gavage

**Exposure period:** single, 24 h

**Doses:** 1050; 2110; 4210; 8410 mg/kg

**Result:** negative

**Method:** other: see Method

**Year:** 1987

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Six animals were used per group. A positive control group

was enclosed (TEM, 0.5 mg/kg). Bone

- marrow was sampled 24 h

post

- treatment; 1000 polychromatic erys were scored per

mouse for the frequency of micronuclei.

---------------

**Remark:** non

- BASF study

**Result:** There was no methanol

- related increase in the induct ion of

micronuclei: In treated groups, the mean MN rate varied from

0.5 to 2.2/1000 vs. 1.5/1000 in the neg. control and 45/1000

in the pos. control group.

---------------------

**Te st substance:**  Methanol, reagent special grade from Junsei Chemicals Co.,

no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards, limited

do cumentation, restriction: only one sampling time

25

- OCT

- 2006 (167)

**Type:** Micronucleus assay

**Species:** mouse **Sex:** male/female

**Strain:** NMRI

**Route of admin.:** i.p.

**Exposure period:** single

**Doses:** 1920; 3200; 4480 mg/kg

**Result:** negative

**Method:** other: see Method

**Year:** 1981

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Met hod:**  Comparative study including a broad spectrum of 30 h after a

single dose, bone

- marrow cells were isolated from 2 male and

2 female mice per dose, and 1000 polychromatic erythrocytes

per m ouse scored.

-----------------

**Remark:** non

- BASF study

**Result:** There was no significant effect on the frequency of

micronuclei in treated as compared to control animals:

1.3/1000 ( control and low dose), 2.0/1000 (2nd dose), and

3.7/1000 (top dose).

----------------

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

167 **Test substance:** Methanol, from Merck, Germany, no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards, limited

documentation, restriction: only one sampling time

25

- OCT

- 2006 (223)

**Type:** Micronucleus assay

**Speci es:**  mouse **Sex:** female

**Strain:** CD

- 1

**Route of admin.:** gavage

**Exposure period:** gd 6 through 10 during pregnancy

**Doses:** 2500 mg/kg/d

**Result:** negative

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Study performed during a developmental investigation under

folate

- deficient and

- sufficient conditions (see other entry

5.8.2).

Maternal peripheral blood was collected 48 h after the final

exposure to methanol (still during pregnancy), and fetal

blood was taken on gd 18. The frequency of MN was scored in

1000 reticulocytes.

----------------------

**Remark:** key study

non

- BASF study

**Result:** The MN rate in maternal and fetal reticulocytes was similar

among the groups. There was no association between either

maternal or fetal hepatic folate concentrations.

Furthermore, the comparison of MN frequencies in malformed

and normal litters gave no evidence of an increase of MN.

-------- ----------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

, key study

25

- OCT

- 2006 (224)

**Type:** Micronucleus assay

**Species:** mouse **Sex:** male

**Strain:** Swiss Webster

**Route of admin.:** i.p.

**Exposure period:** 4 days

**Doses:** 300, 600, 1200, 2500 mg/kg bw

**Resu lt:**  negative

**Method:** other: ASTM 1988

**Year:** 1990

**GLP:** yes

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** Objective of the study: possible effects of methanol were to

be evalua ted as function of folate status, because folate

deficiency is known to increase the spontaneous incidence of

micronucleated erythrocytes and strongly enhance the

incidence of micronuclei induced by cer tain chemicals, and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

168 because methanol oxidation via formate to CO2 depends on

tetrahydrofolate.

---------------------

key study

non

- BASF study

**Result:** 4/1 0 folate

- deficient animals receiving 2500 mg/kg died

between days 2 and 3.

No difference in micronucleus frequency (MN in PCEs) and in

RNA

- positive blood erythrocytes was seen between treated

groups and controls while animals treated with the positive

control substances responded as expected:

+folate: MN 0.17

- 0.23 % vs. 0.23 % in saline control

- folate: MN 0.31

- 0.37 % Vs. 0. 38 % in saline control.

Caffeine (+folate): MN 0.34 %

Caffeine (

- folate): MN 2.42 %

Urethane (+folate): MN 2.52 %.

----------

**Test condition:** Mice were made folate

- deficient by maintaining on a diet

with no folic acid and 1 % succinyl sulfathiazole, inhibitor

of the catalase, and another group received

folate

- supplemented diet containing 5 mg/kg folic acid.

Blood samples were taken at 24 hours after the last dosing.

1

10 mice per dose group were used.

Folate limitation resulted in an about 15

- 20 times lower

folate blood level than in the high

- folate groups.

---------------

**Test substance:** Methanol 99.9 % pure, no further data.

**Conclusion:** The results indicate that methanol is nonclastogenic to the

developing erythroblast in bone marrow and does not inhibit

red blood cell production in either normal or

folate

- deficient mice.

---------- **Reliability:** (2) valid with restrictions

Comparable to guideline study and cu rrent standards,

sufficiently documented, acceptable for assessment

, key study

25

- OCT

- 2006 (225)

**Type:** Micronucleus assay

**Species:** mouse **Sex:** male

**Strain:** C57BL

**Route of admin.:** inhalation

**Exposure period:** 6 h/day, 5 days

**Doses:** 800 or 4000 ppm (1.04 or 5.2 mg/l)

**Result:** negative

**Method:** other: see Method

**Year:** 1991

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive test programme (see also other entries this

chapter): Five animals per dose were used.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

169 1. Peripheral blood MN: Blood sme ars from tail vein blood

were used to score for 2000 polychromatic and 2000

normochromatic erythrocytes per animal.

2. Primary cultures of lung cells: Lung MN were analyzed

in 1000 binucleated cells per animal after cell isolation

from exsanguinated and perfused lung.

------------------

**Remark:** key study

non

- BASF study

**Result:** No treatment related effect o n micronuclei frequency or

cell kinetics and no toxicity was seen at any dose level:

Blood erys: Mean MN rates were 3.1/1000 and 5.2/1000 in PCE,

and 3.4/1000 and 3.6/1000 in NCE at either dose,

respectively, vs. 5.0/1000 and 3.7/1000 of respective

controls.

Lung cells: Mean MN rates were about 20/1000 to 24/1000

irrespective of a dose or control group.

The ratio of bi

- to mononucleated cells was not influenced

by the treatment, neither was there evidence of a

treatment

- related increase in the incidence of

multi

- nucleated cells.

----------- ------------

**Test substance:** Methanol, 99.9 % pure from Fisher Scientific Corporation,

no further data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment, however,

missing positive control

, key study

25

- OCT

- 2006 (222)

**Type:** Cytogenetic assay

**Species:** mouse **Sex:** male

**Strain:** C57BL

**Route of admin.:** inhalation

**Exposure period:** 6 h/day; 5 days

**Doses:** 800 or 4000 ppm (1.04 or 5.2 mg/l)

**Result:** negative

**Method:** other: lung

- cell analysis

**Year:** 1991

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive test programme (see also other entries this

chapter): Primary cultures of lung cells): Chromosome

aberrations were scored in primary culture lung cells of

treated mice. Primary cultures of lung cells were

established from 5 mice per group and 100 first

- division

metaphases per

animal were examined for chromosome aberrations.

No positive control group was concomitantly examined.

--------------------

**Remark:** key study

non

- BASF study

**Result:** No increase i n the frequency of aberrations per cell and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

170 percent aberrant cells and no toxic effects were induced by

the treatment:

Aberration rate per cell (including chromatid and chromosome

brea kage as well as rearrangement events) was from 0.04 to

0.07 in treated animals vs. 0.09 in the negative control.

The percentage of aberrant cells ranged from 3.5 to 6.2 %

vs. 6.9 in the neg. control.

The replicative and mitotic indices were not significantly

different from the control.

------------

**Test substance:** Methanol, 99.9 % pure from Fisher Scientific Corporation,

no furt her data.

**Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment, however,

missing positive control

, key study

25

- OCT

- 2006 (222)

**Type:** Cytogenetic assay

**Species:** mouse **Sex:** male

**Strain:** C57BL

**Route of admin.:** inhalation

**Exposure period:** 6 h/d; 5 d

**Doses:** 1.06; 5.3 mg/l (800; 4000 ppm)

**Result:** negative

**Method:** other: spermatocyte analysis

**Year:** 1991

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Compreh ensive test programme (see also other entries this

chapter): Spermatocytes of the pachytene substage of the

meiotic prophase after 5 days of last exposure were

isolated, and isolates fixed for electronm icroscopy: Several

morphological parameters concerning the meiotic chromosome

structure were examined (synaptonemal complex analysis).

The so

- called "synaptonemal complex" is the attachment site

of homologues chromosomes during meiotic prophase. An

increase in the frequency of synaptic lesions may be an

indicator for exposure to mutagens.

No positive control group was concomitantly examined.

--------------------

**Remark:** key study

non

- BASF study

**Result:** Methanol exposure gave no evidence of a dose

- related

increase in structural abnormalities of the synaptonem al

complex.

------------- Spermatocytes of the pachytene substage of the meiotic

prophase after 5 days of last exposure were isolated, and

isolates fixed for electro nmicroscopy: Several morphological

parameters concerning the meiotic chromosome structure were

examined (synaptonemal complex analysis).

The so

- called "synaptonemal complex" is the attachment site

of homologues chromosomes during meiotic prophase. An

increase in the frequency of synaptic lesions may be an

indicator for exposure to mutagens.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

171 --------------------

**Test substance:** Methanol, purity 99 %

**Reliability:** (2) valid with restrictions

Test procedures based on scientific principles and

standards, sufficiently documented, acceptable for

assessment, however missing po s. control.

, key study

25

- OCT

- 2006 (222)

**Type:** Cytogenetic assay

**Species:** mouse **Sex:**

**Strain:** no data

**Route of admin.:** i.p.

**Doses:** 75

- 300 mg/kg (3x/d)

**Result:** positive

**Method:** other: no data

**Year:** 1983

**GLP:** no data

**Test substance:** no data

**Remark:** There is some evidence that bone marrow cytogenetic analysis

indicated a dose

- related response for structural aberrations,

especially centric fusions in mice treated with three daily

doses of methanol (IPCS/WHO 1997).

non

- BASF study

**Test substance:** Me thanol, no further data

**Reliability:** (4) not assignable

Only abstract, no detailed data

25

- OCT

- 2006 (226) (88)

**Type:** Drosophila SLRL test

**Species:** Dros ophila melanogaster **Sex:** male

**Strain:** other: Berlin K (wild type, male) and Basc (female)

**Route of admin.:** oral feed

**Doses:** LD50

**Result:** negative

**Method:** other: probably largely according to OECD Guide

- l ine 477

**Year:** 1981

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** non

- BASF study

**Result:** Methanol was not mutagenic in the "Basc

- Test in Drosophila".

**Test condition:** One dose (1000 mM) clos e to the LD50 was administered to adult

male wild

- type flies in 5 % saccharose and mated to virgin

females. About 1200 X

- chromosomes were tested per experiment

in each of 3 successive broods (3, 3, and 4 days).

**Test substance:** Methanol from Merck, Germany, no further data.

**Reliability:** (2) valid with restrictions

Acceptable study, meets basic scientific principles

25

- OCT

- 2006 (227)

**Type:** Micronucleus assay

**Species:** mouse **Sex:** no data

**Strain:** Swiss

**Route of admin.:** unspecified

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

172 **Exposure period:** 4 days

**Doses:** 300, 600, 1200, 2500 mg/kg bw

**Result:** negative

**Method:** other

**Year:** 1991

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Groups of folate deficient and folate supplemented mice

were compared to clarify t he role of tetrahydrofolate

mediated formate metabolism in the genotoxic activity of

methanol.

------------- Ten mice per dose level, no further details indicated.

**Remark:** non

- BASF study

**Result:** No genotoxic effects were observed in any treated group.

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Only short communication, abstract: But see Amer. Petrol.

Inst.: probably the same study.

25

- OCT

- 2006 (228)

**5.7 Carcinogenicity**

**Species:** rat **Sex:** male/female

**Strain:** Fische r 344/DuCrj

**Route of administration:** inhalation

**Exposure period:** 24 months

**Frequency of treatment:** continous, 20 h/d

**Post exposure period:** none

**Doses:** 10; 100; 1000 ppm (0.013; 0.13; 1.3 mg/l): 14255

-

14323 h (males); 14407

- 14468 h (females)

**Result:** negative

**Control Group:** yes, concurrent no treatment

**Method:** other: acc. to OECD Guide

- line for Chronic toxicity, Oct. 1980

**Year:** 1985

**GLP:** n o data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studi es and carcinogenicity

studies:

Whole

- body exposure: The animals were housed individually in

wire

- mash cages attached to the inhalation chamber.

52 animals per sex and dose group were tested (initial age 6

weeks). Clinical, ophthalmological, clinical

- chemical,

hematological and pathological/histopathological

examiniation were done.

Analytical control of chamber con centration: Analytical

values close to nominal ones.

STATISTICS:

All data obtained were analysed by t

- test, Fischer ́s exact

test or Armitage ́s chi2

- test.

--------- ----------------

**Remark:** The blood levels of methanol were measured but not documented

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

173 in detail (reported to be similar to those found in the

two

- generation study: see there). Formate was not measured.

"Blood dose levels in the low

- dose (10 ppm) and mid

- dose (100

ppm) group were almost comparable to those in the control

group. In the high

- dose group (1000 ppm), however, they were

54 ppm fo r males and 88 ppm for females." (Discussion part p.

153).

Note: Exposure per day was 20 h, which implies that prolonged

steady

- state blood levels were reached which even may have

bee n higher than in studies using the same exposure

concentration, but shorter exposure times.

--------------------

key study

non

- BASF study

**Result:** 19.2 to 34.6 % of males a nd 32.7 to 40.4 % of females died

or had to sacrificed in extremis. There were no significant

differences from the control groups.

No abnormalities of the general state could be observed in

treated animals. Body weight was somewhat but not

significantly retarded between weeks 51 through 72 in the

highest female dose

- group (1000 ppm).

Urinanalysis revealed a positive trend in glucose and a

decrease in pH in males of the 1000

- ppm group, but values

were within normal range. In 1000

- ppm females, higher

bilirubin values were found, yet within normal range.

No changes were evident in haematological and biochemical

parameters.

Non

- neoplastic findings were similar to those commonly found

in the control animals of 24 months (p. 282).

NEOPLASTIC CHAN GES:

Incidences (number of tumor

- bearing animals/total animals

(Tab. 6, p. 146, and Discussion, p. 154/155):

MALES 0 10 100 1000 ml/m3

================================ =========

A 1/52 5/50 2/52 6/52

B 4/52 1/50 5/52 4/52

-----------------------------------------

A+B 5/52 6/50 7/52 10/52

-------- ---------------------------------

C 7/52

-- -- 4/51

=========================================

-- = not examined

A) Papillary adenomas (males)

B) Ad ematoids = hyperplastic changes (pre

- neoplastic) (males)

C) Adrenal chromaffinoma (males)

FEMALES 0 10 100 1000 ml/m3

=========================================

A 2 /52

-- -- 2/52

B 3/52

-- -- 1/52

-----------------------------------------

A+B 5/52

-- -- 3/52

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

174 -----------------------------------------

C 2/50 3/51 2/49 7/51

=========================================

-- = not examined

A) Papillary adenomas (males)

B) Adematoids = hyperplastic changes (pre

- n eoplastic) (males)

C) Adrenal chromaffinoma (females)

There was no dose

- dependence of tumour development for types

A, B and the sum of A and B in males. The spontaneous tumor

rate of the Fischer strain in this laboratory and in the

published literature was in the range of 14 %. Therefore, the

tumours are unlikely to be treatment

- related.

No clear dose

- dependence was noted for C (ch romaffinomas) in

females, but at the highest dose the number of animals with

this kind of tumors was significantly increased and was higher

than the spontaneous rate published for the Fischer strain

(3.2

- 3.5 %) (p. 155) [Note: not clear whether the given data

relate only to females or to both sexes. It should be noted

that the spontaneous rate of male rats was high in this test

com pared with that of females.]

Overall, tumor frequencies showed no significant differences

between all groups. There was no evidence of an increase in

liver tumors. Specific tumours appeared at a somew hat higher

incidence in high

- dose groups of both sexes:

- papillary lung adenomas in males (without dose

relationship and not statistically significant: 6/52 vs. 1/52

in the control ), but not in females (Tab. 8, p. 149, see also

Takeda and Katoh 1988);

- adrenal phaeochromocytomas in females (based on Fisher ́s

Exact Test: not statistically significant: 7/51 vs. 2/50), but

not in males;

- 3/52 and 5/52 metastatic (transition) tumors of

common origin in males and females, respectively (originated

from tumours prone to metatasis (see p. 155/156).

These events having emerged after week 79 and 104 were

considered incidental and not treatment

- related.

------------------------------------

**Test condition:** Exposure time 20 hours per day, significantly longe r than

according to guideline (6 hrs). This does not represent

normal industrial and consumer use.

---------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride; <3 ppm formaldehyde

**Conclusion:** The results gave no evidence of a clearly cancerogenic

potential of methanol in rats. The biological significance of

the increase in the incidence of the phaeochromocytomas in the

aged female high

- dose rats is considered to be low.

------------------------ **Reliability:** (2) valid with restrictions

Comparable to guideline study, also large ly meeting current

standards, documentation limited, acceptable for assessment.

, key study

26

- OCT

- 2006 (88) (167) (180)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

175 **Species:** mouse **Sex:** male/female

**Strain:** B6C3F1

**Route of administration:** inhalation

**Exposure period:** 18 months

**Frequency of treatment:** continuous, ca. 19 h/d

**Post exposure period:** none

**Doses:** 10; 100; 1000 ppm (0.013; 0.13; 1.3 m g/l: 10436

- 10550

h (males); 10573

- 10642 h (females)

**Control Group:** yes, concurrent no treatment

**Method:** other: according to OECD Guide

- line for chronic toxicity, Oct.

1980

**Year:** 1985

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- t erm and reproduction studies and carcinogenicity

studies:

Whole

- body exposure: The animals were housed individually in

wire

- mash cages installed in the inhalation chamber.

52 animals per sex and dose group were tested (initial age 6

weeks). Clinical, ophthalmological, clinical

- chemical,

hematological and pathological/histopathological

examiniation were done.

Analy tical control of chamber concentration: Analytical

values close to nominal ones.

-------------------------

**Remark:** The blood levels of methanol and formate were not measured.

Note: Exposure per day was 20 h, which implies that

prolonged steady

- state blood levels were reached which even

may have been higher than in studies using the same exposure

concentration, but shorter exposure times.

--------------------

key study

non

- BASF study

**Result:** Survival was high without significant diffferences as

compared to the control, mortality ranging between no deaths

and 7.5% deaths by the end of the study: 0% (1000 ppm males,

10 ppm females), 2% (controls), 6% (10 and 100 ppm males)

and 7.5% (100 and 1000 pm females).

No abnormalities of the genera l state could be observed in

treated animals. Body weight was somewhat increased in

treated group, significantly in 1000 ppm males, between

months 6 through 12 in males and months 9 through 12 in

females.

There was no evidence of a treatment

- related effect in

clinical/blood parameters.

Non

- neoplastic organ findings were similar to those commonly

found in the contro l animals of 12 months.

Individual tumor frequencies showed no significant

differences between all groups; there was no difference in

the severity of tumors between the groups.

The tu mors exclusively observed in the 1000

- ppm group were

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

176 all within the spontaneous range and included fatty cell

tumor at the submaxillary lymph node (1/52 males),

chromophobe adenoma of the pituary gland (3/52 females),

adrenal pheochromocytoma (1/52 females), dermal

fibrosarcoma (1/52 males), and meningeal sarcoma (1/52

females) vs. 0/52 in all other groups.

The spontaneous liver

- tum ors rate naturally high in this

species was not increased by methanol treatment (p. 171;

Tab. 5, p. 172).

------------------------

**Test condition:** Exposure time 19 hours per day, significantly longer than

according to guideline (6 hrs). This does not represent

normal industrial and consumer use.

---------------

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride; <3 ppm formaldehyde

**Conclusion:** The result gave no evidence of a cancerogenic potential of

methanol in mice.

--------------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, documentation limited, acceptable for assessment.

, key study

26

- OCT

- 2006 (167) (180)

**Specie s:**  rat **Sex:** no data

**Strain:** no data

**Route of administration:** oral unspecified

**Exposure period:** no data

**Post exposure period:** no data

**Doses:** no data

**Result:** negative

**Method:** other: no data

**GLP:** no data

**Test substance:** no data

**Remark:** Tabular compilation of BT Institute data without details:

The author reports that Methanol was tested in a

carcinogenicity study with oral application in the rat. The

substance was clearly not carcinogenic according to the

author. No further details are reported.

-------------

non

- BASF study

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Only short communication

26

- OCT

- 2006 (229)

**5.8.1 Toxicity to Fertility**

**T ype:**  Two generation study

**Species:** rat

**Sex:** male/female

**Strain:** other: Sprague Dawley (Crj:CD)

**Route of administration:** inhalation

**Exposure Period:** F0: 103

- 108 d; F1: 61

- 62 d and 145

- 153 d; F2: 54

- 56 d (see Method)

**Frequency of treatment:** continuous (19

- 20 h/d)

**Premating Exposure Period**

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

177 **male:** 60 d (approx. 1200 h)

**female:** 60 d (approx. 120 0 h)

**No. of generation studies:** 2

**Doses:** 0.013; 0.13; 1.3 mg/l (10; 100; 1000 ppm)

**Control Group:** yes, concurrent no treatment

**NOAEL Parental:** = 1000 ppm

**NOAEL F1 Offspring:** = 100 ppm

**NOAEL F2 Offspring:** = 100 ppm

**other: LOAEL F1/F2 offspring :** 

= 1000 ppm

**Method:** other: in accordance with OECD Guidelines, Oct. 1980

**Year:** 1985

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Metho d:**  Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studies and carcinogenicity

studies:

Whole

- body exposure: One multi

- stage inhalation chamber was

used for each group.

The duration of exposure was as follows (p. 184):

F0

- Males from age of 8 wk through end of mating (16 wk or

after);

F0

- females from age of 8 wk through mating period (16 wk or

after) to end of lactation.

F1

- males from birth through end of mating (at 14 wk or

after);

F1

- females from birth through end of mating (at 14 wk or

after) to weaning of F2 pups (21 d after delivery).

F2

- Males and

- females from birth to 21 d (but to age of 8 wk

for 1/sex/litter).

F0 generation consisted of 30 animals per sex and group.

Additionally, 15 animals were reared for a second mating (p.

185).

Mating period was maximally 21 d. The pairs without eviden ce

of insemination within 21 d were again cohabited with

untreated animals (2nd mating) to determine the fertility of

each animal, in this case without exposure (p.186).

Pathohistolog y:

After mating all males were necropsied, and testes as well

as all accessory reproductive organs preserved. Females

either after second mating or not inseminated ones were

necropsied and examined for reproductive organs. Likewise,

all other females having delivered or not were sacrificed

for histopathology at due time.

Litters: Part of the pups of each generation were examined

for intra

- uterine, another one for neo

- and postnatal

development/morphological differentiation including

behavioral/function testing and histopathology until one

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

178 week after weaning (p. 1 87/188).

Gametogenesis: Histological examination of morphology of

ovaries and sperms was not included.

In 9

- week old F1 pups, blood methanol was measured, but not

formate (p. 191).

Analytical exposure control: Analytical concentration values

of methanol were close to nominal ones.

-------------------------

Statistical ANALYSIS:

All data obtain ed were analysed by t

- test, Fischer ́s exact

test, U

- test of Mann

- Whitney or Armitage ́s chi2

- test.

------------------------

**Remark:** Blood levels of methanol measured in the F1 offsprings (age

9 weeks) (NEDO, 1987, p. 191):

controls (baseline): approx. 2

- 3 ug/ml

10 ppm methanol: approx. 3

- 3.5 ug/ml

100 ppm " : approx. 1

- 4.2 ug/ml

1000 ppm " : a pprox. 53 (males)

- 100 (females) ug/ml.

There are no data on formate.

Note: Exposure per day was 20 h, which implies that

prolonged steady

- state blood levels were reached which even

ma y have been higher than in studies using the same exposure

concentration, but shorter exposure times.

--------------------

key study

non

- BASF study

**Result:** F0

- Generation: No treatment

- related alterations in all

reproductive parameters.

F1

- Generation: None of the fertility indices including

sexual cycle, mating time, fertility and pregnancy rate

showed a significant difference from untreated F1 controls.

In male pups of the 1000

- ppm group, post

- natal morphological

differentiation appeared to be influenced with respect to

the descensus testis occurrin g 0.5 to 1 d earlier (see same

effect in F2 generation) (p. 199/200) [not mentioned by Takeda

and Katoh, 1988]: This time

- dependent parameter was evaluated

by relating the completion of downward migrati on

of the testes (final length of the gubernaculum reached) to

the post

- natal body

- weight gain (The more reliable body length

was not available):

F1

- generation:

In t he F1 pups derived from the 1000

- ppm group(108 males),

this process was completed

within 16 through 20 post

- natal days with the climax at day 17

and 18 (32 and 39 %, respectively), while in the respecti ve

control (113 males), descent was complete from 16

through 21 days with the maximum at day 19 (32 %), but also

relatively high percentages on the days before and after: day

18 (22%), day 17 (19%), day 20 (18%) (p. 199).

Absolute and relative brain weights were significantly lowered

in the high

- dose groups of either sex at an age of 8 and 16

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

179 weeks. This was still found in females n ecropsied

after 24 weeks. Also other organs showed slight shifts in

weights: thymus, pituitary (lower), heart, lung, liver

(higher).

However, there were no histological manifestations.

No effects on testes or ovaries reported.

Likewise, there were no significant differences in functional

tests (movement, emotion, learning) as compared with the

control or the other groups.

F2 generation:

As in F1 males, an apparently dose

- related earlier descensus

testis was noted after 1000

- ppm exposure: F2 (94 males) on day

16 (42%), day 17 (40%), day 18 (15%) vs. control (91 males) on

day 16 (10%), day 17 (39%), day 18 (31%), day 19 (14%) (p.

200).

After 100

- ppm, "descensus testis" in male F2

- progeny was about

0.5 d earlier than in male co ntrol F2 pups (p. 195). Detailed

data not specified and not addressed under "Discussion".

Organ weights showed similar tendencies as found in the

F1

- generation without histological changes. No effects on

testes or ovaries reported.

----------------------

**Test condition:** Exposure time 19

- 20 hours per day, significantly longer than

according to guideline (6 hrs).

------------- --

**Test substance:** Methanol, reagent special grade from Junsei Chemicals Co.,

<1 ppm vinyl chloride; <3 ppm formaldehyde

**Conclusion:** No firm conclusions can be drawn about fertility of either

sex, as the copulati on time of 21 d was comfortably long for

successful insemination (but in accordance with guidelines

at that time) and gametogenesis was not considered.

In the F1 and F2 progeny (both sexes), a decrease in brain

weights was evident at 1000 ppm methanol, but without

noticeable histological changes and functional impairments.

This phenomenon is believed to represent a change occurring

during the perinatal period (Takeda and Katoh, 1988).

But no quantitative data and the statistical level are

documented for organ weights.

The information about organ weights is lacking, with much

weight placed on the "descensus testis".

The meaning of an apparent shift of testis descent in male

offspring in relation to body

- weight development of the pups

in the two following gener ations is unclear and was not

directly addressed by Takeda and Katoh (1988), but detailed

by NEDO (1987) and considered a significant difference from

untreated controls (p. 285). Furthermore, it is obvi ous that

this parameter showed considerable variation also between

the control groups of both generations (see p. 199/200).

Although there was no obvious pathological effect in the

pr ogeny of 1000

- ppm exposed groups, the effects observed may

be considered as biologically relevant under these test

conditions and, therefore, 1000 ppm (1.3 mg/L) is established

as LOAEL

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

180 and 100 ppm (0.13 mg/L) as NOAEL for post

- natal development

while for

parental effects, the NOAEL is 1000 ppm.

The NOAEL(develop) = 0.13 mg/L is also compatible with a NOEL.

---- -------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, documentation limited, acceptable for assessment

, key study

26

- OCT

- 2006 (167) (180)

**Type:** Fertility

**Species:** monkey

**Sex:** female

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Exposure P eriod:**  prebreeding about 120 days; breeding 70 days;

pregnancy about 165 days

**Frequency of treatment:** 2.5 h/d

**Premating Exposure Period**

**male:** none

**female:** prebreeding about 120 days

**Duration of test:** birth, first 9 months of life of the offsprings

**Doses:** 200; 600; 1800 ppm (0.262; 0.786; 2.358 mg/l)

**Control Group:** yes

**NOAEL Parental:** = 1800 ppm

**other: NOAEL(Repro) (mother) :** 

= 1800 ppm

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Study design limited to exposure of female animals

(pre

- breeding, breeding, and pregnancy) and

qualitative/clinical signs of toxicity. It comprised

- methanol pharmacokinetics (see other entry: 5.11)

- maternal toxicity

- reproductive toxicity (see other entry: 5.8.2)

- developmental neurotoxicity (see other entry: 5.8.2).

Specific aims of the study were to determine whether

- repeated exposure to methanol alters methanol metabolism

in adult female monkeys,

- pregnancy alters methanol metabolism,

- long

- term exposure to methanol produces overt adult

toxicity, or reproductive toxicity, or both,

- long

- term exposure to methanol in ut ero affects

offspring development, especially neurobehavioral

development.

Groups of 11

- 12 adult female monkeys were exposed to

methanol vapor in the concentrations of 0; 200; 600 or 1800

ppm, daily for 2.5 hour before breeding, during breeding

and during pregnancy. The study was performed in 2 cohorts:

24 females/cohort and 2 males/cohort.

For postnatal devel opmental evaluation, 8 to 9 infants were

available per group, in total 34, among them 26 in

- utero

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

181 treated offsprings (see other entry: 5.8.2).

------------------------------------

**Remark:** k ey study

non

- BASF study

**Result:** Reproductive performance:

Methanol exposure had no effect on most measures of

reproductive performance, including menstrual cycles,

conception rate, and live

- birth delivery rate. However, all

methanol

- exposed animals had a decrease of about six to

eight days in duration of pregnancy compared to control

animals (Tab. 17, p. 45).

It is not clear whether this decrease was related to

methanol exposure, as there was no dose response and no

differences among offspring groups in body weight or other

physical parameters.

Prenatal exposure to methanol had no effect on infant growth

and physical development for the first 9 months.

-------------------

**Test substance:** Methanol, no further data

**Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, well documented, limitation due to low number of

animals.

, key study

26

- OCT

- 2006 (230)

**Type:** Fertility

**Species:** mouse

**Sex:** male

**Strain:** B6C3F1

**Route of administration:** gavage

**Exposure Period:** 5 d

**Frequency of treatment:** 1x/ d, 5 consecutive days

**Premating Exposure Period**

**male:** sperm morphology, no mating

**female:** no females

**Doses:** 5 mal 1000 mg/kg

**Method:** other: sperm morphology

**Year:** 1984

**GL P:**  no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Ten male mice (4 months of age) were treated and maintained

without exposure for 5 weeks (the time needed for testicular

spermatogenesi s and migration of spermatides to the

epididymidis), spermatozoa were isolated from dissected

cauda epididymidis and fixed and mounted on slides, and

stained for light microscopy. 500 sperms were evalua ted per

animal.

-----------

**Remark:** non

- BASF study

**Result:** In the methanol

- treated animals, a slight, but statistically

insignificant increase in sperms abnormalities (mean facto r

about 1.7 above neg. control, with very high variance: 1.86

+

- 0.91 vs. 1.12 +

- 0.39 in the vehicle control) was observed,

while the treatment with cyclophoshamide (100 mg/kg)

resulted in an about 6

- fold increase (5.84 +

- 1.94).

-----------------

**Test substance:** Methanol, no further data.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

182 **Reliability:** (2) valid with restrictions

Test procedures based on scientific principles and

standards, limited documention, acceptable for assessment

26

- OCT

- 2006 (231)

**5.8.2 Developmental Toxicity/Teratogenicity**

**Species:** rat **S ex:**  female

**Strain:** Sprague

- Dawley

**Route of administration:** inhalation

**Exposure period:** gd 1

- 19, at the top dose: gd 7

- 15

**Frequency of treatment:** 7h/d

**Duration of test:** 21 d

**Doses:** 6.63; 13.3; 26.6 mg/l (5000; 10000; 20000 ppm)

**Control Group:** yes, concurrent no treatment

**NOAEL Maternal Toxity:** = 10000 ppm

**NOAEL Teratogenicity:** = 5000 ppm

**Method:** other: acc. to standards

**Year:** 1985

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Whole

- body exposure: Mean analytical chamber concentrations

corresponded well to the nominal ones.

15 dams per group were used. Methanol blood levels w ere

measured in concurrently exposed non

- pregnant groups by GC

on day 1, 10, and 19.

Dams were sacrificed on gd 20, one half of the fetuses was

subjected to examination for visceral, the other half for

skeletal abnormalities.

----------------

**Remark:** The biological relevance of this study for humans has to be

questioned for the following caveats:

1. The exposure concentrations of methanol relate to doses

which significantly exceed lethal doses in man: lowest dose

(5000 ppm, 7 h) in rats about 1800 mg/(kg bw*d) based upon

calculations (NTP

- CERHR, 2 001, Tab. 2

- 10) [(human lethal

dose: 300

- 1000 mg/(kg*d)].

2. This high dose which would be lethal to humans proved to

be ineffective in rats.

3. The study was performed in a range where methanol

metabolism is totally saturated in rats, not considered as

relevant for humans.

4. Consequently, the blood methanol levels in rats exposed

to 5000 ppm are estimated to be at a factor of 20

- 50 higher

than in humans exposed e.g. to 1000 ppm (7

- 8 h).

5. The differences in metabolism (predominance of catalase

in rodents vs. alcohol dehydrogenase (ADH) in humans) has to

be regarded: rate limiting steps are catalase given rise to

methanol, but not formate, in blood of rodents, while in

humans rate

- limiting is formate degradation giving rise to

increased formate levels in human blood upon kinetic

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

183 saturation.

---------------------------------

key study

non

- BASF study

**Result:** Blood levels (dams):

1.0

- 2.17 mg/ml (5000 ppm); 1.84

- 2.24 mg/ml (10000 ppm),

5.25

- 8.65 mg/ml (20000 ppm).

No background levels available.

Maternal toxicity:

Only in the high

- dose group, slightly unsteady gait and

significantly reduced food consumption during the first week

without adverse effect on body

- weight gain. No signs of

maternal toxicity in the other groups.

Effects on maternal reprodu ction:

No influence on the number of corpera lutea and of

implantations. No effect on fetal lethality and resorption.

Effects on litters/fetuses:

Statistically significant and dose

- re lated decrease in fetal

body weight in the two high

- dose groups (p<0.05) [note: The

significance is poor for the male/female offsprings of the

10000

- ppm group when compared to the control group of other

studies (see: ethanol, Nelson et al., Tab. 2, p. 731].

At the highest concentration, an increased number of litters

with skeletal and visceral malformations was noted, these

included in particular rudimentary and extra cervical ribs

and exencephaly and encephalocele, and, to minor extent,

cardiovascular and urinary

- tract defects:

Visceral defects: 7/15 litters with 15/96 malformed fetuses

vs. 0/15 litters and 0/107 fetuses in the untreated control.

Skeletal defects: 14/15 litters with 72/92 malformed fetuses

vs. 0/15 litters and 0/98 fetuses in the untreated control.

Similar abnormalities also appeared in the progeny from the

10000

- ppm group, but not statistically significant: visceral

and skeletal defects 2/15 litters each with 2/107 and 2/115

fetuses, re spectively).

In the 5000

- ppm group, visceral defects were 1/13 litters with

2/90 fetuses, and skeletal defects were 1/13 with 1/90

fetuses.

-------------------------

There was no evidence of embryotoxic/teratogenic activity of

methanol at 5000 ppm.

------------------------

**Test substance:** Methanol, 99.1 % purity

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

184 **Conclusion:** (For biological relevance for humans: see Remarks.)

1. The CERHR Expert Panel placed high weight on the

reliability of results of this prenatal study (NTP

- CERHR,

2001).

2. Based on a logistic quantitative model of Risk

Assessment for Developmental Neurotoxicants (Slikker and

Gaylor, 1997, 1998) using the inhalation data in SD rats

from Nelson et al., 1995, the following conclusions were

drawn and est imations made:

The use of continuous foetal weight data was as sensitive as

foetal brain malformation data in estimating excess risk to

methanol inhalation exposure in developing rat. Excess risk

(assumed to be the additional risk that foetal weight is

below the first percentile of control animals) was estimated

by using a logistic model: 0.1 at 16000 ppm, 0.01 at 5400

ppm, and 0.0 01 at 980 ppm.

------------------------ **Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

, key st udy

26

- OCT

- 2006 (232) (233) (234) (235)

**Species:** rat **Sex:** female

**Strain:** Sprague

- Dawley

**Route of administration:** inhalation

**Exposure period:** 7

- 17 gd

**Frequency of treatment:** continuous, ca. 22.7 h/d (NEDO, 1987, p. 215)

**Doses:** 200; 1000; 5000 ppm (0.26; 1.3; 6.64 mg/l)

**Control Group:** yes, concurrent no treatment

**NOAEL Maternal Toxity:** > 1000 ppm

**NOAEL Teratogenicity:** > 1000 ppm

**LOAEL Maternal Toxicity :**  = 5000 ppm

**LOAEL Teratogenicity :**  = 5000 ppm

**other: NOEL maternal/fetal :** 

= 1000 ppm

**Method:** other: acc. to national standards (see Method)

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Comprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reprodu ction studies and carcinogenicity

studies (analog Segment II)

Whole

- body exposure: Pregnant females were housed

individually in a cage placed in the inhalation chamber.

36 dams were u sed per test and control group, including 12

dams allowed for natural delivery (p. 211).

The study comprised of three segments:

- intra

- uterine development/teratogenicity,

- post

- natal development F1 (lactation, 21 d, and >=5 d

after weaning) (without exposure) (p. 212), and

- reproductive performance of F1 to produce F2.

Analytical exposure control: Analytical concentra tion values

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

185 of methanol were close to nominal ones.

-------------------------

**Remark:** Note: Exposure per day was >20 h, which implies that

prolonged steady

- state blood levels were reached w hich even

may have been higher than in studies using the same exposure

concentration, but shorter exposure times.

--------------------

key study

non

- BASF study

**Res ult:**  The exposure to 200 and 1000 ppm methanol failed to produce

any maternal and embryonal/fetal toxicity or fetal

malformations.

At 5000 ppm, maternal toxicity was noted as a decrease in

body

- weight gain. Food and drinking water consumption were

reduced during gd 7 through 12.

One dam died, another one had to be killed before delivery.

After Cesarian section:

Embryolethality was increased (late resorptions: 10.4 % vs.

0.6 % in control, p<0.05), but the variance between single

litters was high (Tab. 2, p. 219). No differences were

evident in the numb er of corpora lutea, of implantations,

and preimplantation resorptions (p. 219). Mean body weight

of live fetuses after Cesarian section was reduced (about

- 20 %, p<0.001) (p. 220).

M orphological abnormalities in fetuses:

Only after intra

- uterine exposure to 5000 ppm, the

incidences of various malformations and variations were

increased:

About 50 % of the fetuses w ith ventricular septal defects

(visceral malformation in 16/20 litters or 64/131 fetuses)

vs. 0% or near 0% in all other groups, and residual thymus

(variation in all 20 litter or 70/131 fetuses) vs. ab out 2.4

to 2.9 % in 4 litters each of all other groups (p. 22/223).

Other changes included increased incidence of skeletal

variations: "atresia of cervical arch/vertebra foramen

costo transversarium" (45 %), " bifurcated vertebral center"

(14 %) and "cervical rib" (65 %) as well as "excessive

sublingual neuropore" (50 %), all of which malformations

having no or little relevance in th e other group (p.

224/225) except of "atresia foramen" with about 25 % in the

control and about 4 to 8 % in the other exposure groups.

After delivery, neo

- /postnatal findings:

After 50 00 ppm, mean gestation time was prolonged by 0.7

days (p. 221). Food and drinking water consumption of dams

was reduced also after birth during lactation.

Among descendants from the high

- dose group, mo rtality was

prominent during the first 4 d after birth with live fetuses

showing poor vitality in this time (ca. 10% = 2/12 pups per

litter died as compared with 1 to 2% fatal cases in the

other groups). Survivors showed normal appearance after day

4. Slight retardation of growth was still significant at

weaning. Water consumption was slightly reduced, in

particular for females.

Early indicators for post

- natal development:

In pups from the 5000

- ppm group, eruption of upper incisor

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

186 and opening of eyelid for both sexes and descensus testis

for males were significa ntly earlier than in the controls in

relation to term of delivery, but not in relation to the

whole gestation time which was prolonged for this group.

There were no differences in behavioral and functi onal tests

as compared to control and other test groups.

At the age of 8 weeks, brain, thyroid (males), thymus and

testis (males) weights were lower (p<0.01), and

pituary

- gland weight of males was higher (p<0.05). But

histological examination revealed no treatment

- related

changes.

16.5 % of the offsprings (15/91 in 8/12 litters) had

hemilateral thyroprivia (missin g thyroid lobe, mostly left).

There was no histopathological lesion in the tissue. The

defect was attributed to an impairment of organogenesis (p.

233).

Reproductive performances of F 1 (from 5000 ppm):

No significant effects on sexual cycle, genital function and

reproductive performance of the F1 progeny were noted.

--------------------

**Test condition:** Exposure time about 22 ho urs per day, significantly longer

than according to guideline (6 hrs). This does not represent

normal industrial and consumer use.

---------------

**Test substance:** Methanol, reagent special grade fro m Junsei Chemicals Co.,

<1 ppm vinyl chloride; <3 ppm formaldehyde

**Conclusion:** The exposure to 5000 ppm of pregnant rats during gestation

(>20 h/d) produces maternal toxicity, fetal malformation,

increased perinatal mortality and developmental delay in

surviving progeny. Teratogenic effects occurred only at

maternally toxic exposure concentration.

Exposure levels of 1000 ppm or less did not indu ce toxic

symptoms in maternal animals, structural abnormalities or

delay in growth or functional development in the

F1

- generation.

Therefore, the NOEL for maternal and developmental to xicity

is considered to be 1000 ppm.

-------------------- **Reliability:** (2) valid with restrictions

Comparable to guideline study and current standards, also

largely meeting c urrent standards, documentation limited,

acceptable for assessment

, key study

26

- OCT

- 2006 (167) (180)

**Species:** mouse **Sex:** female

**Stra in:**  CD

- 1

**Route of administration:** inhalation

**Exposure period:** day 6

- 15 of gestation

**Frequency of treatment:** daily, 7 hours/day

**Duration of test:** up to day 17 of gestation

**Doses:** 1000, 2000, 5000, 7500, 10000 or 15000 ppm (1.3, 2.6,

6.5, 9.75, 13 or 19.5 mg/l)

**Control Group:** yes, concurrent vehicle

**NOAEL Maternal Toxity:** = 15000 ppm

**NOAEL Teratogenicity:** = 1000 ppm

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

187 **Method:** other

**GLP :**  no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Whole

- body eposure: Mean measured chamber concentrations

were close to the nominal ones. An air

- exposed and an

unhandled control group was included. Furthermore, for

comparison, an orally treated group receiving 2x 2000 mg

methanol/kg bw/d by gavage (as 12.5

- % solution in water) was

studied.

Blood concentrations were analysed (gd 6, 10, 15).

60 to 6 litters were produced depending on toxicity of the

exposure levels.

The following parameters were examined: maternal body

weight and clinical signs, fetal weight, number of

implantation sites, live and dead fetuses, resorptions,

gross external anomalies, skeletal and visceral

malformations, blood methanol concentrations in dams.

The 7500

- and 10000

- ppm group as well the oral group were

only examined for resorption, exencephaly and cleft palate

because earlier studies have demonstrated that these were

the major effects.

---------------------

**Remark:** The biological relevance of this study for humans has to be

questioned for the following caveats:

1. The exposure concentrations of methanol relate to doses

which significantly exceed lethal doses in man: lowest dose

(1000 ppm, 7 h) in mice about 800 mg/(kg bw*d) based upon

calculations (NTP

- CERHR, 2001, Tab. 2

- 10) [(human lethal

dose: 300

- 1000 mg/(kg*d)].

2. The minimum dose in mice which is likely to be lethal to

humans proved to be ineffective in mice. Those

concentrations which unequivocally showed malformations were

500 0 ppm and above, clearly lethal to humans.

3. The study was performed in a concentration range where

methanol metabolism becomes saturated in mice, not

considered as relevant for humans: 2000 ppm which may be

considered as marginally effective (LOAEL) caused already

saturation (see: increase in blood level of methanol).

4. Consequently, the blood methanol levels in mice exposed

to 1000 ppm are estimated to be at a factor of approx. 2 to

4, those of mice exposed to 2000 ppm at a factor of 10 to 20

higher, and those of mice exposed to distinctly teratogenic

5000 ppm at a factor of 30 to 60 higher than in humans

exposed e.g. to 1000 ppm (7

- 8 h).

5. The differences in metabolism (predominance of catalase

in rodents vs. alcohol dehydrogenase (ADH) in humans) has to

be regarded: rate limiting steps are catalase given rise to

methanol, but not formate, in blood of rodents, while in

humans rate

- limiting is formate degradation, giving rise to

increased formate level s in human blood upon kinetic

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

188 saturation.

----------------

key study

non

- BASF study

**Result:** Mean blood levels:

approx. 100 ug/ml (1000 ppm), 540 (2000 pp m), 1650 (5000

ppm), 3180 (7500 ppm), 4200 (10000 ppm), 7300 (15000 ppm) in

ug/ml. In the oral group (4000 mg/d), about 4000 ug/ml were

found, corresponding about to inhalation exposure of 10000

ppm. The controls had a background level of about 1.6 ug/ml.

Maternal and fetal effects.

1. There were no signs of maternal toxicity.

2. Significant increases in the incidence of exencephal y and

cleft palate were observed at 5000 ppm (approx. 10 % per

litter: 14 fetuses/11 litters vs. 0.3 % in the control) and

above, increased embryo/fetal death at 7500 ppm and above:

At the highest dose, almost complete resorption of embryos

in most litters occurred.

Reduced fetal weight was not noted, but at 10000 ppm and

above.

A dose related increase in cervical r ibs or ossification

sites lateral to the seventh cervical vertebra was

significant at 2000 ppm (approx. 50 %/litter vs. 26 to 34 %

in the controls; at 1000 ppm: approx. 34 %/litter).

T he NOAEL for developmental toxicity was determined at 1000

ppm with relation to the most sensitive defect, the

cervical

- rib event.

The calculation of benchmark doses for a 5

- % added risk from

the lower 95

- % confidence limit on the maximum likelihood

estimates were generally consistent with the NOAELs for

various developmental defects: the 5

- % added risk above

background level was at 3667 ppm for combined cleft palate,

exencephaly, and resorption, for cervical

- rib defect at 305

ppm.

(see also: NTP

- CERHR, 2001)

--------------------------------

**Test substance:** Methanol, high purity from Fisher Scientific

**Reliability:** (2) valid with restrictions

Comparable to guideline study, also largely meeting current

standards, with acceptable restrictions (see Method),

sufficient documentation, acceptable for assessment

, key study

26

- OCT

- 2006 (233) (236) (237)

**Species:** mouse **Sex:** female

**Strain:** no dat a

**Route of administration:** gavage

**Exposure period:** single treatment on day 7 of gestation (split into

two equal doses)

**Frequency of treatment:** single

**Duration of test:** up to day 18 of gestation

**Doses:** 0, 4 or 5 g/kg bodyweight

**Control Group:** yes, concurrent vehicle

**Method:** other: Special prenatal exposure study

**GLP:** no data

**Test substance:** no data

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

189 **Method:** Study was to elucidate time

- relate d manifestations and

mechanisms of methanol

- induced skeletal malformations in

mouse. Therefore, one single dose prior to gd 9, the

sensitive period of skeletal development. Six and 7 dams

were used, representing about 70 to 80 fetuses.

-------------------

**Remark:** non

- BASF study

**Result:** The treatment led to dose

- related increased incidences in

several skeletal malformations such as additional rib on C7

(approx. 30 % of fetuses in both dose groups vs. 0 % in

control) or split, fused or incomplete vertebra C2 (also

about 41 % in the high dose fetuses vs. 7.7 in the control),

25 instead of 26 presacral vertebrae (10 % vs. 2.4 % in

control).

These effects suggest posterior transformation of cervical

vertebrae by altering segment patterning in the developing

mouse embryo (posteriorization of cervical vertebrae).

It is assumed that interference of methanol with retinol

conversion to retinoic acid may play a crucial role in these

events.

----------------------

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

generally accepted scientific prin ciples, acceptable for

assessment

26

- OCT

- 2006 (238) (239)

**Species:** mouse **Sex:** female

**Strain:** CD

- 1

**Route of adminis tration:**  inhalation

**Exposure period:** day 6

- 15, 7

- 9 or 9

- 11 of gestation

**Frequency of treatment:** daily, 6 hours/day

**Duration of test:** up to day 17 of gestation

**Doses:** 5000, 10000 or 15000 ppm (6.5, 13 or 19. 5 mg/l)

**Control Group:** yes, concurrent vehicle

**NOAEL Maternal Toxity:** = 5000 ppm

**NOAEL Teratogenicity:** ca. 5000 ppm

**LOAEL Embryotoxicity :**  = 5000 ppm

**Method:** other

**GLP:** no data

**Test substance:** as prescribe d by 1.1

- 1.4

**Method:** Whole

- body exposure: The analytical chamber concentration

was close to the nominal one.

17

- 22 litters per group were examined.

The following parameters were examined : maternal toxicity as

clinical signs and bodyweight changes, embryo/fetal toxicity

as % resorptions, live fetuses and fetal bodyweight, gross,

skeletal and visceral malformations and histopathology (ne ural

tube defects).

-----------------

**Remark:** non

- BASF study

**Result:** Maternal toxicity was seen at 13000 and 19500 mg/m3 as reduced

body weight and clinical signs, fetal toxicity occurred as

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

190 increased resorptions dose

- related at all dose levels.

Teratogenic activity was observed in dose

- related manner at

the two upper dose levels as increased incidences of neural

tube defects and ocular defects (significant during gd 7

- 9),

hind limb anomalies (during gd 9

- 11), cleft palate, renal

pelvic dilatation/hydronephrosis and tail anomalies

(significant during either gd period).

(gd 7

- 9) 10000 ppm 15000 ppm

% litters (% fetuses)

-------------------------------------------------------

Neural tube d efect 30 (3.6) 65 (14.7)

Cleft palate 50 (14.6) 88 (50.4)

------------------------------------------------------- At 5000 ppm no difference from control: no neural tub e

defect, cleft palate 4

- 9 % litters and 0.3

- 0.7 %

fetuses affected.

At 5000 ppm, the incidence of pelvic dilatation (not

hydronephrosis) appeared to be increased although high i n

the untreated control, too: The increase was statistically

significant with p<0.05 in the Shirley ́s test.

Furthermore, post

- implantation resorptions tended to be

increased and were s ignificant with p<0.05 for the number of

litters with >=1 resorption.

But no increases in structural malformations were observed.

----------------

**Test substance:** Methanol, HPLC

- grade from J.T. Baker, no further data.

**Conclusion:** The data suggest that the spectrum of teratogenic effects is

dependent on the number of methanol exposures and the stage

of embryogenesis.

Major methanol

- induced events included malformations of the

brain, eyes, and the urinary tract as well as decreased

fetal weight. External abnormalities of the calvaria, hard

palate, jaw, and tail also suggest a detri mental

methanol

- related effect on the axial skeleton.

No clear NOAEL can be derived for developmental effects due

to increased resorptions and renal pelvic cavitation after

5000 ppm m ethanol. Therefore, the LOAEL(embryo

- tox) is 5000

ppm.

---------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods , sufficiently documented, acceptable for assessment

26

- OCT

- 2006 (240)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:**  inhalation

**Exposure period:** GD 6 through postnatal day 21

**Frequency of treatment:** 6 hours daily

**Duration of test:** until adult phase

**Doses:** 4500 ppm

**Control Group:** yes

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

191 **Method:** other: Neurotoxicological assay after high

- dose exposure

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Whole

- body exposure design: Four cohorts of pregnant rats (n

= 12), each consisting of an exposure and a control group,

were exposed to 4500 ppm methanol vapor for 6 hr daily

beginning on GD 6 through post

- natal day (PND) 21.

Average analytical chamber values were close to nominal ones

from about 4350 to 4580 ppm.

1. Methanol blood levels were determined in both dams: blood

samples immediately collected from dams on gd 7, 13, and 19

after 6

- h exposure, and from both dam s and pups on PND 7,

14, and 21 during lactation. In two cohorts,

neuro

- histopathological examinations were carried out in

neonates on PND 1 and 21. In selected adults from one cohort

(following long

- term behavorial tests, see below), brain

histomorphology and molecular examination on NCAM (neural

cell adhesion molecule, by immunoblotting) were performed.

In one cohort, post

- weaning blood concentrations in relation

to increasing age were measured, extended to 52 d of age,

including blood sampling (Stern et al., 1996).

Several behavioral procedures were used to assess exposure

effects in the offspring (Stern et al., 1997).

**Remark:** key study

non

- BASF study

**Result:** 1. Reproductive Performance, Blood Levels, Neurological

Tests:

Control and ex posed cohort with similar litter sizes, sex

distribution, and dam and pup weight gains. Any variations

were not related to exposure.

Blood levels in dams fairly constant with averages of 0.55

+

- 0.07 and 0.56 +

- 0.09 mg/ml during gestation and laction.

Blood levels in pups about twice as high as in dams, mean

1.26 +

- 0.23 mg/ml. In an extra test, it was conirmed that

the dam

- pup diff erence persisted until 48 days of age, i.e.

about 4 wk post

- weaning (fig. 4, p. 42).

Morphology/light microscopy of brain tissue: There was no

detectable evidence of neuropathology. However, NCAM indic es

revealed a significant treatment difference in the

cerebellum of neonates on PND 4, while after 15 months of

age previously exposed offsprings, no effects were seen

(Stern et al., 1 997).

2. Behavioral Tests:

Male

- female littermates were studied whenever possible to

examine sex differences, with one pair from a litter for

each procedure. Exposure to methanol did not affect suckling

latency and nipple attachment on PND 5 or perfomance on a

aversive olfactory condition procedure on PND 10. Exposure

to methanol did alter performances in motor activity

procedure.

Methanol

- exposed neonates wereless active on PND 18, but

more active on PND 25 than the equivalent control pups. In a

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

192 conditioning procedure (rotate a running wheel a specified

number of revolutions to obtain food

- pellet ) males showed

decreases, females increases in the rate of running. In a

stochastic spatial discrimination procedure methanol

- exposed

rats respo nded less efficienty at asymptotic levels of

performance than controls.

--------------------

**Test substance:** methanol, hplc grade

**Conclusion:** Blood methanol levels in adult rats was similar to those

found by other. The distinctly higher levels in offsprings

may be explained by the still immature states of the alcohol

dehydrogenase system. This may have implications for

post

- natal neuronal d evelopment/synaptogenesis and learning

profile and behavior.

However, no histopathological evidence was provided,

although on the molecular level, a certain deficiency of

cell

- adhesion molecule pattern (NCAM 180 and 140

- 10% to

- 20

%) appears to reflect a delay in the cell

- cell contact

formation associated with synaptogenesis. But this could no

longer be demonstrated in adult, previo usly post

- natally

exposed rats any more. The biological meaning remains to be

elucidated.

According to the authors, methanol

- exposure was associated

with subtle behavioral changes in both neonates and adults.

---------------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, well documented

, key study

26

- OCT

- 2006 (241) (242) (243)

**Species:** monkey **Sex:** female

**Strain:** Macaca Fascicularis

**Route of administration:** inhalation

**Exposure period:** pr ebreeding about 120 days; breeding 70 days;

pregnancy about 165 days

**Frequency of treatment:** 2.5 hours daily

**Duration of test:** birth, first 9 months of life of the offsprings

**Doses:** 200; 600; 1800 ppm (0.262; 0.786; 2.358 mg/l)

**Control Group:** yes

**NOAEL Maternal Toxity:** = 1800 ppm

**NOAEL Teratogenicity:** = 1800 ppm

**NOAEL Fetotoxicity :**  = 1800 ppm

**Method:** other

**Year:** 1996

**GLP:** yes

**Test subst ance:**  as prescribed by 1.1

- 1.4

**Method:** Study design limited to exposure of female animals

(pre

- breeding, breeding, and pregnancy) and

qualitative/clinical signs of toxicity. It comprised

- methanol pharmacokinetics (see other entry: 5.11)

- maternal toxicity (see also entry: 5.8.1)

- reproductive toxicity

- developmental neurotoxicity

Specific aims of the stu dy were to determine whether

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

193 - repeated exposure to methanol alters methanol metabolism

in adult female monkeys,

- pregnancy alters methanol metabolism,

- long

- term exposure to methanol produces overt adult

toxicity, or reproductive toxicity, or both,

- long

- term exposure to methanol in utero affects

offspring development, especially neurobehavioral

development.

Groups of 11

- 12 adult female monkeys were exposed to

methanol vapor in the concentrations of 0; 200; 600 or 1800

ppm, daily for 2.5 hour before breeding, during breeding

and during pregnancy. The study was performed in 2 cohorts:

24 females/cohort and 2 males/cohort.

For postnatal developmental evaluation, 8 to 9 infants were

available per group, in total 34, among t hem 26 in

- utero

treated offsprings.

------------------------------------

**Remark:** Limitation of the study: low number of animals, which

precludes weak effects from detection with certainty ( see

also: NTP

- CERHR, 2001).

non

- BASF study

**Result:** A. Blood levels (see details below):

Exposure to methanol vapors did not affect the health of the

adult monkeys prior to or during pregnancy. Single 2.5

- hour

exposures to methanol vapors caused short term elevations in

blood methanol concentrations of approx. 1

- to 2

- fold in the

200 ppm exposure group, 3

- to 4

- fold in the 6 00 ppm group

and 13

- to 16

- fold in the 1800 ppm group.

After long

- term exposure, peak blood methanol concentrations

declined slight over the first month and remained constant

thereaft er.

The concentration of plasma formate remained at baseline

levels during the entire course of the study in all exposure

groups.

Pregnancy had no effect on methanol disposition.

Serum folate concentrations were not affected by pregnancy

and methanol exposure.

B. Reproductive performance:

Methanol exposure had no effect on most measures of

repr oductive performance, including menstrual cycles,

conception rate, and live

- birth delivery rate. However, all

methanol

- exposed animals had a decrease of about six to

eight days in duration of pregnancy compared to control

animals.

It is not clear whether this decrease was related to

methanol exposure, as there was no dose response and no

differences among offspring groups in body wei ght or other

physical parameters.

Prenatal exposure to methanol had no effect on infant growth

and physical development for the first 9 months.

The results or behavioral assessments o f offspring did not

indicate methanol

- exposure effects on most domains of early

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

194 behavioral development. No consistent effects due to

methanol exposure were observed on early reflex responses,

gross motor development, spatial and concept learning and

memory, and social behavior.

Methanol exposure was associated with ratings of "low

arousal" on the Neonatal Behavioral Scale. The effec ts were

observed when all of the methanol

- exposed infants were

compared with controls. Further comparisons, however did not

indicate that the effect was dose

- dependent. In addition,

ma ny of the methanol

- exposed infants that were the last to

receive optimal scores for the arousal items had been

delivered via C

- section. Thus, this effect may not be

directly related to methanol exposure independent of mode of

delivery.

Methanol exposure was also associated with a delay in early

sensorimotor development of male infants only (Visually

Directed Reaching Test). The eff ect was observed after

controlling for the shortened gestation length observed for

the 3 methanol

- exposed groups. The delay was dose

- dependent

and ranged from 9 days for the 200 ppm exposure group to ov er

2 weeks for the 600 ppm

- and 1800

ppm

- exposure groups Part II, p. 96). This observation is based

on results from group sizes of 8 to 9 infant animals, with 2

to 5 males and 4 to 7 f emales per group (II, p. 86).

The statistical significances (linear contrast test based on

ANOVA) of p = or <0.04:

This test indicated a significant difference between control

infants compared with all methanol

- exposed offsprings

combined, as well as with the 600

- and 800

- ppm groups for

males (p<0.04). The comparison between the control group and

the 200

- ppm was nerly significant (p= 0.09) (Part II, p. 87).

The results of the Fagan

- Test of Infant Intelligence indicated

a possible effect of methanol exposure on visual recognition

memory when complex stimuli (social problems) were us ed in

testing. Although there were no mean group differences in the

novelty scores across the 4 exposure groups, only the control

group exhibited a significant novelty preferences for social

stimuli.

---------------------------

C. Other observations:

Finally, prenatal methanol exposure was associated with the

occurrence of a wasting syndrome in 2/7 female offsp rings in

the 1800 ppm

- exposure group after approximately 1 year of

age, both living in the same cohort. Those females began to

show growth retardation with 12 resp. 17 months of age. They

became very weak and had to be euthanized at the age of 20

resp. 36 months. Assay results for viral infection, blood

chemistry, complete blood count, and liver, kidney, thyroid,

and pancreatic funct ion were within the normal range.

Necropsies showed signs of severe malnutrition and

gastroenteritis. This symptom was not observed in any of

the offsprings of the other cohort.

-- -----------

D. Blood levels:

Methanol: No significant differences in the blood

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

195 concentrations between the 4 phases (measured 0.5 h

post

- exposure, n = 11 or 12 (non

- pregnant); n = 9 or 10

(pregnancy) (Fig 7, Tab. 11, Fig. 14, Part I):

Baseline range approx. 2 ug/ml [approx. 0.06 mM]

200 ppm: range approx. 5 ug/ml [approx. 0.15 mM]

600 ppm: range approx. 10 ug/ ml [approx. 0.30 mM]

1800 ppm: range approx. 35

- 40 ug/ml [1mM

- range]

At 1800 ppm, after 5 h elimination, the residual methanol

level was near baseline (max. 2

- fold higher).

The mean estimated elimination half

- lives (for 600 and 1800

ppm) ranged between about 60 to 90 min.

Formate: Irrespective of concentration levels or exposure

intervals, there was no evidence of a significant i ncrease

above the background range of about 0.15

- 0.30 mM (approx.

7 to 14 ug/ml) (Fig. 8, Fig. 15, Tab. 12, Part I).

Folate in serum: There was no significant shift in the

folate le vels during pregnancy, independent of exposure

(Tab. 13, Part I).

-------------------

**Test substance:** methanol, no further data

**Conclusion:** Peer

- reviewed conclusion: "The investigators findings suggest

that repeated inhalation exposure to methanol vapors as high

as 1800 ppm would not result in accumulation of

blood formate above baseline levels. With the exception of an

unexpected shortening of gestation, methanol exposure had no

effect on reproductive performance. The most significant

result to emerge from this study was the wasting syndrome

observed in two monkeys exposed in utero to 180 0 ppm.

Overall, the results provide no evidence of a robust effect of

prenatal methanol exposure on the neurobehavioral development

of nonhuman primate infants during the first nine months of

life..." (HEI Statement, in Burbacher 1999).

The conclusions are not unanimous about wasting syndrome,

gestation length, and sensomotoric and intelligence testing

(see NTP

- CERHR, 2003, Appendi x II

- 77 to II

- 81), mainly because

interpretation is limited by the normal variance of and by the

low number of animals under test, the lack of robust findings,

and no control for multiple comparisons in statistical

analyses. The neurobehavioral findings are considered

important from a qualitative perspective. But the wasting

syndrome did not draw much attention of the Expert Panel (p.

II

- 79).

--------------

---------------------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, well documented, limitation for develomental

effects (see Remark)

26

- OCT

- 2006 (230) (233)

**Species:** mouse **Sex:** female

**Strain:** CD

- 1

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

196 **R oute of administration:**  gavage

**Exposure period:** day 6

- 10 of gestation

**Frequency of treatment:** twice daily

**Duration of test:** up to day 18 of gestation

**Doses:** 2500 mg/kg bodyweight in water (2x/d)

**Control Group:** yes, concurrent vehicle

**other: LOAEL :**  5000 mg/kg bw

**Method:** other: see Method

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Study designed to clarify the role of folate in the

developmental toxicity of methanol in folate

- deficient and

folate

- sufficient mice. Additionally to the developmental

toxicity parameters, the micronuclei formation in maternal

and feta l blood was determined (see other entry 5.6).

The female animals were fed folate

- free diet supplemented

either with 400 or 1200 nmol folic acid as well as 1 %

succinylsulfathiazole (as inhibitor of int estinal folate

synthesis) 5 weeks prior to mating and throughtout breeding

and gestation.

The short dosing interval of gd 6 through 10 was selected

because this was the sensitive peri od for the induction of

the observed malformations (Fu et al. 1996, p. 461).

21

- 24 litters were examined per group, representing 215 to

282 fetuses per test and control group.

Fola te was determined in maternal and fetal liver and blood

plasma and red cells.

-------------------

**Remark:** key study

non

- BASF study

**Result:** A. Folate levels (measured on gd 18, 8 d after the final

methanol treatment):

Low

- folate diet produced a decline in folate in the maternal

liver (total folate approx.

- 30%), in maternal plasma

(approx.

- 30%), in maternal ery s (approx.

- 30%) vs.

normal

- folate diet and in the fetal liver (approx.

- 60 to

- 70%) (Fu et al., 1996, Tab. 2).

Methanol had no marked influence on maternal and folate

levels irrespec tive of folate supplementation, except in

maternal plasma where there was some evidence of a reduction

of about 20 % .

Methanol treatment was slighty fetotoxic (reduced mean fetal

wei ght and reduced mean crown

- rump length), but had no impact

other reproductive parameters. It showed some evidence of a

teratogenic effect (increased incidences of cleft palate and

exencephaly) under fol ate

- adequate supply, but this was hardly

statistically significant:

cleft palate (2/222 vs. 0/282) and execephaly (5/222 vs.

1/282 in the respective high

- folate control).

Likewise, fo late deficiency failed to produce significant

malformations (in accordance with previous reports: e.g.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

197 Heid et al., 1992):

cleft palate (5/215 vs. 0/282) and execephaly (2/215 vs.

1/28 2 in the respective high

- folate control).

However, cleft palate, but not exencephaly was significantly

increased in the presence of methanol: 39/235 vs. 5/215 and

8/235 vs. 2/215 of folate

- poor control , respectively.

Methanol did not induce micronuclei in maternal or fetal

blood.

--------------------------

B. In pregnant CD

- 1 mice given methanol (5 g/kg/d) from

gestation day 6

- 15, one group receiving folate deficient,

the other folate supplemented diet, there were no

differences in the formate

- blood levels between both groups:

5.13 +

- 0.68 mmol/L (folate

- d ef.) and 3.90 +

- 0.94 mmol/L

(folate

- suppl.) vs. 0.36 +

- 0.13 mmol/L (untreated control).

But developmental toxicity was significantly higher in

folate

- deficient dams (Hong et al. 1997).

The results indicate that increased plasma formate levels in

dams do not underlie the increased developmental toxicity of

methanol in mice fed low dietary folate (Hong et al. 1997).

-----------

**Tes t substance:**  Methanol, highest HPLC grade, from Fisher Scientific Comp.

**Conclusion:** Folate deficiency enhanced the teratogenic effects of

methanol in mice.

**Reliability:** (2) valid with restrictions

Test proc edures in accordance with accepted standard

methods, sufficiently documented

, key study

26

- OCT

- 2006 (244) (224) (245) (246)

**Species:** rat **Sex:** f emale

**Strain:** Long

- Evans

**Route of administration:** inhalation

**Exposure period:** day 7

- 19 of gestation

**Frequency of treatment:** daily for 7 hours/day

**Duration of test:** up to postnatal day 160

**Doses:** 19.5 mg/l (15000 ppm) corresponding to ca. 6100

mg/kg/day

**Control Group:** yes, concurrent vehicle

**Method:** other: postnatal offspring development and behaviour

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Whole

- body exposure: The analytical chamber concentrations

varied by less than 5 % and were fairely close to the

nominal ones. Six and 7 dams were used per group.

The following parameters were recorded: maternal

bodyweight, maternal blood methanol levels, offspring

bodyweight, offspring mortality, motor activity (day31

- 21,

30, 60), olfactory learning (da y18), behavioural

thermoregulation (day 20

- 21), T

- maze learning (day 23

- 24),

acoustic startle response (day 24, 60), reflex modification

audiometry (day 60), pubertal landmarks (day 31

- 56), passive

avoidance (day 72), visual

- evoked potentials (day 160).

5 or 6 dams were used per group with 10

- 11 offspring.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

198 Behavioural tests were performed on 4

- 5 offspring animals.

----------- ---

**Remark:** The number of pups available and tested (mostly,

1/sex/litter) was probably too small to draw firm

conclusions about postnatal developmental effects on

behavior.

**Result:** Maternal blood methanol levels were between 3.8 and 3.1

mg/ml. The treatment led to reduced maternal body weight,

the only effect noted in dams. No increase in

post

- implantation loss. Postnatal bodyweig ht were modestly

but statistically lower after treatment. No increase in

postnatal mortality.

No other test revealed significant effects of methanol

treatment.

**Test substance:** Methan ol, 99.9 % purity, Fisher Scientific

**Reliability:** (3) invalid

Test procedures based on scientific principles and

standards, well documented, limited value for

neurological/behavorial assessment

(247)

**Species:** mouse **Sex:** female

**Strain:** CD

- 1

**Route of administration:** inhalation

**Exposure period:** day 7

- 9 of gestation

**Frequency of treatment:** daily, 6 hours/day

**Duration of test:** up to day 8.5, 9.5 or 10.5

**Doses:** 15000 ppm (19.5 mg/l)

**Control Group:** yes, concurrent vehicle

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Whole

- body exposure: The analytical chamber concentration

was close to the nominal one.

4

- 9 litters and 26

- 99 live fetuses were examined p er

group. The following parameters were examined: maternal body

weightand clinical signs, gross, skeletal and visceral

malformations in fetuses, histopathology and scanning

electon mic roscopy in fetuses.

--------------

**Remark:** non

- BASF study

**Result:** Maternal toxicity was seen as clinical signs and reduced

bodyweight, in fetuses the treatment induced reduced

bo dyweight and a range of gross neural defects

(anencephaly,encephalocele, holoprosencephaly, ocular

anomalies) in up to 41 % of live embryos.

---------

**Test substance:** Methanol, HPLC gradde, from Ba ker

**Conclusion:** The result suggest that exposure to high concentrations of

methanol injures multiple stem cell populations in the

neurolating mouse embryo.

**Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, well documented, acceptable for assessment

26

- OCT

- 2006 (248)

**Species:** mouse **Sex:** female

**Strain:** CD

- 1

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

199 **Route of administration:** inhalation

**Exposure period:** 6 hours

**Frequency of treatment:** single treatment on day 8 of gestation

**Duration of test:** up to day 10 of gestation

**Dose s:**  10000 or 15000 ppm (13 mg/l)

**Control Group:** yes, concurrent vehicle

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** 12

- 14 litters per group were evaluat ed.

The effects of methanol and its major metabolite formate as

well as the influence of a pretreatment with the

alcohol

- dehydrogenase inhibitor 4

- methylpyrazole were

compared. The fo llowing parameters were examined:

crown

- rump length, head length, gross malformations and

neural tube defects, methanol concentrations in fetal and

maternal tissue. The folate status in RBC and decidual

tissue was monitored at 15000 ppm methanol.

----------------------

**Remark:** Valuable scientific information about mechanisms. However,

the biological relevance for humans is poor, becau se the

estimated systemic dose im mice [>=4000 mg/(kg*d)] is

distinctly above doses lethal to humans (300

- 1000 mg/kg

bw).

------------------

non

- BASF study

**Resul t:**  Methanol treatment (10000 ppm) led to a significant increase

in the incidence of open neural tubes (9.65 +

- 3.13% in 12

litters vs. 2.02 to 2.69 +

- 1.09% in controls, p<0.05). After

4

- methylp yrazole pretreatment (4

- MP), the percentage of this

effect was 7.21 +

- 2.65%, but not significant. No such effect

was induced by formate given orally (750 mg/kg). The other

parameters examined were not a ffected by the treatment

(Dorman et al., 1995).

Methanol concentrations were equal in fetal and maternal

tissues with maximum maternal plasma level at 65 to 75 mM,

and 62 to 85 mM in decidual swelling, irresp. of

pretreatment with 4

- MP. Likewise, mean formate blood and

decidual peaks were at 0.5 to 0.6 mM and 1.8 to 2.1 mM,

respectively. Furthermore, after 15000 ppm, too, no

significant changes of formate were noted in either

compartment.

[That means, 4

- MP failed to influence methanol and formate

levels significantly as well as embryonal dysmorhogenesis.]

(Dorman et al., 1995).

Folate status:

15000 ppm methanol resulted in transient, albeit

statistically insignificant decreases in total maternal red

blood cell and decidua fo late levels (Dorman et al., 1995).

-----------------------------

**Test substance:** Methanol HPLC

- grade from Sigma Chemical Corporation, no

further data.

**Conclusion:** Formate seems to play no apparent role in the development of

methanol

- induced exencephaly, and methanol itself is the

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

200 proximate teratogen in pregnant CD

- 1 mice exposed to high

concentration of methanol vapor, because

1. no format e accumulation in methanol

- treated mice,

2. exencephaly (failure of anterior neuropre to close)

occurred in the absence of increased formate levels,

3. no exencephaly after oral treatment with high do ses of

formate,

4. methanol, but not formate, caused neural tube defects

in whole

- embryo culture.

The results (although only from single dosing) did not

provide e vidence that folate deficiency gave rise to

methanol

- related dysmorphogenesis and appears to be

irrelevant at least at low methanol burdens (compare also

test results in monkeys: Dorman et al., 1994). B ut based on

observations by others (e.g. Sakanashi et al., 1994), an

interaction of folate deficiency and methanol cannot be

ruled out under certain conditions. (Dorman et al., 1995;

D orman and Welsch, 1996; see also: Medinsky and Dorman,

1995)

--------------------------- **Reliability:** (2) valid with restrictions

26

- OCT

- 2006 (249) (250) (251) (252) (253)

**Species:** mouse **Sex:** female

**Strain:** CD

- 1

**Route of administration:** inhalation

**Exposure period:** see method

**Frequency of treatment:** see method

**Duration of test:** unt il GD 17

**Doses:** 10000 ppm (13 mg/l)

**Control Group:** yes

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Critical periods for the developmental toxicity of met hanol

were assessed in pregnant CD

- 1 mice exposed to 10000 ppm for

7 hours/day on 2 consecutive days during GD 6

- 13 (GD 6 and

7; GD 7 and 8; GD 8 and 9, GD 9 and 10; GD 10 and 11; GD 11

and 12; GD 12 and 13) or to single day (7 hour) exposures

during GD 5 and 9. Controls were exposed in the same way to

filtered air. Mice received water but not food during

exposure. Maternal blood met hanol was determind at times

during, at the end of and subsequent to a single 7 hr

exposure on GD 7.

On GD 17 mice were killed and gravid uteri removed. Live,

dead and resorbed fetuses were counted, and live fetuses

were examined, weighed and preserved in 70 % ethanol. All

fetuses were examined externally and for cleft palate,

eviscerated, and stained with Alizarin red for skeletal

examination.

Test and control groups consisted out of 12 to 17 pregnant

mice.

-----------

**Remark:** Valuable scientific information about mechanisms. However,

the biological relevance for humans is poor, because the

estimated systemic dose im mice [some 4000 mg/(kg*d)] is

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

201 distinctly above doses lethal to humans (300

- 1000 mg/kg

bw).

-- ----------------

non

- BASF study

**Result:** Peak maternal blood methanol concentration at the end of the

exposure was about 4 mg/ml, methanol was cleared from

maternal blood within 24 hr.

Some fully resorbed litters were observed with 2

- day

methanol exposure on gd 6

- 7 (3/12) and gd 7

- 8 (1/16) or

1

- day exposure on gd 7 (3/17). With 1

- day methanol exposure

on gd 7, the number of live fetuses (4.4 per litter) was

lower than with exposure on anyother day.

Cleft palate, exencephaly and skeletal defects were the

fetal anomalies observed:

cleft palate:

occurred with 2

- day exposures on gd 6

- 7 through gd 11

- 12

(peak on gd 7

- 8) and with 1

- day exposures on gd 5 through 9

(peak on gd 7)

Exencephaly: occurred with 2

- day exposures on gd 6

- 7 through

gd 8

- 9 (peak on gd 6

- 7) and with 1

- day exposure on gd 5

through 8 (peak on gd 7).

Skeletal elements malformed included the

- exoccipital (peak on gd 6

- 7 (22.5 %); gd 5 (9.9%))

- atlas (peak on gd 6

- 7 (72.3 %); gd 5, 6 (55.5 %, 55.3 %))

- axis (peak on gd 6

- 7(22.3 %); gd 7 (28.8 %))

- cervical vertebra 7 with a rib (peak on gd 6

- 7 (73.7 %);

gd 7 (45.4 %))

- lumbar vertebra 1 with a rib (peak on gd 7

- 8 (68.3 %);

gd 7 (39.4 %).

An increase incidence of fetuses with 25 presacral vertebrae

(normal 26) was observed wiht methanol exposure on gd 5;

whereas an increased incidence of fetuses with 27 presacral

vertebrae was observed with methanol exposure on gd 7.

According to the authors the results of this study indicate

tha t gastrulation and early organogenesis represent a period

of increased embryonal sensitivity to methanol.

**Test substance:** Methanol, high purity from Fisher Scientific

**Reliability:** (2) valid with restrictions

Tes t procedures in accordance with accepted standard

methods, well documented, acceptable for assessment

26

- OCT

- 2006 (254)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:** drinking water

**Exposure period:** gd 15

- 17 or gd 17

- 19

**Frequency of treatment:** continuous

**Doses:** 2% Methanol in drinking water (ca. 2500 m g/kg/d)

**Control Group:** yes, concurrent no treatment

**Method:** The study design was to elucidate postnatal neurological

effects in rat pups after intrauterin exposure to methanol:

Two behavioral test were selected,

1) the suckling behavior test which measures the latency

time to nipple attachment (postnatal day 1) and

2) the nest

- seeking/homing behavior test (postnatal day 10)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

202 30 primigravid dams were used, 10 per group.

-------------

**Remark:** non

- BASF study

**Result:** Maternal toxicity was not observed. In both test groups no

differences were found in litter sizes, neonatal b irth

weight as well as lethality of offsprings as compared with

the progeny of the untreated control.

Both behavioral tests on post

- natal disorders showed

significant retardations in learning: Pubs from treated rats

required a longer time to begin suckling on post

- natal day 1

(80 to 90 sec. vs. 60 sec of controls) and to locate nesting

material on post

- natal day 10 than control pups (3 time

- and

distance

- related indicators evaluated).

-------------

**Test substance:** Methanol, no further data.

**Conclusion:** The data suggest that prenatal methanol exposure may induce

neuroto xic effects in the offsprings which are not

associated with evident or overt toxicity.

--------------- **Reliability:** (2) valid with restrictions

Test procedures based on scientific principles, sufficiently

documented, acceptable for assessment

26

- OCT

- 2006 (255)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:** drinking water

**Exposure period:** day 6

- 15 of gestation

**Frequency of treatment:** daily

**Duration of test:** not indicated, presumably to the end of gestation

**Doses:** 350, 700 or 1400 mg/k g bodyweight (0.5, 1 or 2% in

water)

**Control Group:** yes, concurrent vehicle

**Method:** other: teratogenicity study with folate deficient animals

**GLP:** no data

**Method:** Pilot study/Screeni ng: Rats were rendered folate deficient

by supplying a folate

- deficient diet. 7 to 9 dams were used

per group.

**Remark:** non

- BASF study

**Result:** The average blood levels were given as 0.21, 0.26, and 0.6 7

mg/L, respectively.

At 1 and 2 % methanol in water fetal resorptions/deaths as

well as skeletal malformations were significantly

increased. In the 0.5 % group minimal respective cha nges

indicate a NOEL for maternal and developmental below 0.5 %

which corresponds to a blood level of 0.21 mg/ml.

No further details are reported.

**Test substance:** Methanol, no further data.

**Reliabili ty:**  (4) not assignable

Only short communication, abstract

26

- OCT

- 2006 (256)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:** drinking water

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

203 **Exposure period:** days 6

- 15 of gestation

**Frequency of treatment:** daily

**Duration of test:** up to day 20 of gestation

**Doses:** 357.1, 714.3 or 1428.6 mg/kg b odyweight (0.5, 1 or

2%)

**Control Group:** yes, concurrent vehicle

**NOAEL Maternal Toxity:** = 357.1 mg/kg bw

**Method:** other: comparison with folate deficient dams

**GLP:** no data

**Test substance:** o ther TS

**Method:** 5 dams or litters were used per group, all treated groups

and one control group were made folate deficient before

treatment. The examination for teratogenic effects was not

pe rformed at the date of publication.

**Remark:** non

- BASF study

**Result:** The highest dose level was highly embryotoxic, of 5

expected litters only one survived. In the mid dose group

resorptions were also s ignificantly increased.

Maternal toxicity as reduced bodyweight or bodyweight gain

was seen in the mid and high dose group.

Teratogenic effects were not yet examined at the date of

pub lication.

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Comparable to guideline study, lacking important results,

documentation limited

26

- OCT

- 2006 (257)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:** gavage

**Exposure period:** single application on day 10 of gestation

**Duratio n of test:**  not indicated, presumably to the end of gestation

**Doses:** 1027, 2054 or 4108 mg/kg bodyweight (1.3, 2.6 or 5.2

ml/kg)

**Control Group:** yes, concurrent vehicle

**NOAEL Maternal Toxity :**  = 2054 mg/kg bw

**Method:** other: no details indicated

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** 10 to 13 dams per group were treated. Wilson sectioning,

gross necropsy and ali zarin red staining of fetuses was

performed. No further details are reported.

**Remark:** non

- BASF study

**Result:** Maternal toxicity was seen only as body weight loss in the

highest dose group (> 20 % decre ase).

Methanol at all doses failed to produce any significant

change in standard reproductive indices (e.g.

postimplantation loss). A significant decrease in fetal body

weight (11 to 1 9.5 %), however, was associated with a

dose

- dependent increase in anomalies (0.6 % in control,

3.7%; 7% and 16.5% in the dose groups). The dose

- related

anomalies were undescended testes, exophthalmia an d

anophthalmia.

------------

**Test substance:** Methanol, HPLC grade

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

2 04 **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, test design limited, sufficiently documented

26

- OCT

- 2006 (258) (259) (260)

**Species:** rat **Sex:** female

**Strain:** Wistar

**Route of administration:** gavage

**Exposure period:** day 6

- 15 of gestation

**Frequency of treatment:** daily

**Duration of test:** up to day 21 of gestation

**Doses:** 2.5 g/kg bodyweight

**Control Group:** yes, concurrent vehicle

**Method:** other

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Well nourished and malnourished groups of rats were

compared. The following parameters were examined: number of

live f etuses, resorptions, corpora lutea, implantation

sites, fetal weight, gross and skeletal malformations.

12

- 17 dams and 110

- 177 live fetuses per group were

examined.

---------- ------

**Remark:** non

- BASF study

**Result:** The treatment induced maternal toxicity.

The treatment led to increased incidences of skeletal

malformations in well nourished (45.4 % compared to 5.6 %

in controls) and malnourished rats (38.8 % compared to 3.8 %

in controls). Malformations were primarily found on ribs

(extra ribs) and vertebra (irregular shape).

Malnutrition did not increase the incidence of

malformations, but fetal growth was retarded.

An

increase in skeletal malformation,

primarily cerivical extra

ri bs was noted in the methanol treated

rats when compared to

controls. This is only a

single dose study and the 2.5 g/kg/day

dose is above the

lethal dose in humans.

**Test substance:** Methanol, no further data.

**Conclusion:** This is only a single dose study and the 2.5 g/kg/day dose

is above the lethal dose in humans.

**Reliability:** (3) inv alid

Unsuitable test design (see Conclusions)

26

- OCT

- 2006 (261)

**Species:** rat **Sex:** female

**Strain:** other: Hol tzman

**Route of administration:** gavage

**Exposure period:** GD 1

- 8

**Frequency of treatment:** daily

**Duration of test:** up to day 9, 11 or day 20 of gestation

**Doses:** 1600; 2400 or 3200 mg/kg bw

**Control Group:** y es, concurrent vehicle

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

205 **Method:** other: early pregnancy study

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Early pregnancy study:

Eight dams per group with 11

- 13 fetuses per dam were used.

Animals were killed on days 9, 11, or 20 of pregnancy, and

maternal, embryonic, or fetal parameters were assessed:

Effects on early pregnancy were determined on day 9 as

bodywei ght, uterine weight, number of implantation sites,

number of atypical sites, mean implantation site weight,

ovarian weight and number of corpora lutea per ovary.

Decidual cell response was determined on day 9 as uterine

weight, uterine function and normal implantation processes.

Effects on mid pregnancy was determined on day 11 as number

of live fetuses, presence of heartbeat, yolk sac

circulation, axial rotation, closure of neural tube,

presence of limb buds and sensory organs, yolk sac

diameter, crown

- rump length, head length and somite number.

Effects on late pregnancy were de termined on day 20 of

gestation as number of live fetuses, number of resorptions,

gross external abnormalities and fetal weight.

Additionally, decidualisation was assessed in pseudopregnant

rats using a DCR(decidual cell response) technique.

-------------------

-----------------

**Remark:** non

- BASF study

**Result:** The only effect of methanol treatment was seen on early

pregnancy (day 9) as reduced uterine and implantation site

weight and increased number of atypical, small implantation

sites (with extravasated blood) in the high

- dose group.

All other para meters examined showed no statistical

significant differences between treated and control groups

at any gestation day. There was no treatment

- related effect

on serum estradiol, progesterone, lutenizing hormone and

prolactin on day 9 (1 day after after cessation of

exposure).

No effects on embryo/fetal survival or development including

external anomalities were noted at any gestation day.

The 3200 mg/kg/day dose of methanol produced a reduction in

body weight gain by day 9, but not day 20, which may be

considered an indication of non

- specific maternal toxicity.

Decidualisation

After methanol treatment of pseudopregnant rats (all 3

doses), a dose

- related inhibition of uterine growth was

observed (day 9), but not significant at 1600 mg/kg/d.

------------------

**Test substance:** methanol, high purity grade, no furthr data

**Conclusion:** Results indicated that effects on uterine weight and

implatantion sites on gd 9 may have resulted from

methanol

- induced i nhibition of the DCR. However, this

apparent potential impact on decidua development was not

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

206 expressed by a depression of day

- 20 embryo/fetal survival or

development, when treatment ended on gd 8.

The biological relevance of these results is unclear, in

particular, failing a comprehensive data base on hormone

status and DCR (see also: NTP

- CERHR, 2001).

------------- **Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

generally accepted scientific principles, but technical

limitations (see Conclusion).

26

- OCT

- 2006 (262)

**Species:** rat **Sex:** female

**Strain:** Long

- Evans

**Route of administration:** oral feed

**Exposure period:** day 8 of gestation up to pa rturition

**Frequency of treatment:** daily

**Duration of test:** up to some days after parturition

**Doses:** 0.6, 0.9 or 1.6 % (v/v) (278.6, 417.9 or 742.9 mg/kg

bodyweight)

**Control Group:** yes, c oncurrent vehicle

**Method:** other: no details reported

**GLP:** no data

**Test substance:** other TS

**Method:** Dams were fed a liquid diet containing methanol. Number of

animals not indicated, no detailed descript ion of method.

**Remark:** non

- BASF study

**Result:** Methanol did not affect fecundity but reduced maternal

weight gain, decreased litter sizes, increased perinatal

and postnatal mortality and decreased weig hts at weaning.

Teratogenic effects are not reported.

**Test substance:** Methanol, no further data.

**Reliability:** (4) not assignable

Only short communication, abstract

26

- OCT

- 2006 (263)

**Species:** mouse **Sex:** female

**Strain:** C57BL

**Route of administration:** i.p.

**Exposure period:** 2 injections 4 hours apart on gestational day 7

**Frequency of treatment:**  2 treatments

**Duration of test:** up to day 17 of gestation

**Doses:** 1.7 or 2.45 g/kg bodyweight

**Control Group:** no data specified

**Method:** other: no details reported

**GLP:** no data

**Test substance:** other TS

**Method:** Study design was to elucidate pathogenic mechanisms in

methanol intoxication: Ethanol and methanol were compared

with respect to their teratogenic activity on the brain.

Ethanol was given at a dose level of 2.45 g/kg bodyweight

(Rogers, 1995).

In a further series, so

- called confocal laser scanning

microscopy was used to elucidate the pathogenesis of crani o

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

207 facial defects (Rogers et al., 1996).

-----------------------

**Remark:** non

- BASF study

**Result:** 1.7 g/kg methanol induced 30 % resorptions and midfacial

deficiencies and micro/anoph thalmia in over 50 % of live

fetuses. 2.45 g/kg resulted in 55 % resorptions and 100 %

of live fetuses having malformations of the

holoprosencephaly

spectrum (micro/anophthalmia, ceboc ephaly, absence of

forebrain, premaxillary agenesis).

Cell death of cells were observed in embryos on gd 8 at the

edge s of the neural plate and on gd 9 in the midbrain and

visceral a rches. On gd 8, sparse mesenchyme and edema were

apparent beneath the cranial neuroepithelium. Prosencephalic

deficiences observed included small or telencephalic and

optic vesicles (Rogers et al., 1996 ).

-----------------

**Test substance:** Methanol, no further data.

**Conclusion:** The pathogenesis of methanol

- induced craniofacial

malformations is similar to that previously observed for

ethanol (Rogers et al., 1996)(for the ethanol effect: see

also review Rogers and Daston, 1997).

---------------- **Reliability:** (2) valid with restrictions

Special study design, sufficiently documented, meets

generally accepted scientific principles, acceptable for

assessment

26

- OCT

- 2006 (264) (265) (266)

**Species:** mouse **Se x:**  female

**Strain:** C57BL

**Route of administration:** i.p.

**Exposure period:** gestation day 6, 7, 8, 9 or 10

**Frequency of treatment:** 2x/d with 4 h apart

**Doses:** 4900 mg/kg bw., split in 2 equal doses on resp.

gestation day

**Control Group:** yes, concurrent vehicle

**Method:** other: Screening test for sensitive stages

**Year:** 1998

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Remark:** The results included external and soft tissue defects, still

excluded skeletal malformations.

non

- BASF study

**Result:** The total incidence of malformations was highest with

gestation d ay 7, indicating that gastrulation is sensitive

to methanol similar to ethanol.

The defects included the eye (an

- and microphthalmia,

coloboma), the brain and face (holoprosencephaly, facial

clefts) as wel as cardiovascular system (vetricuar septal

and great vessel defects).

Contrary to ethanol, no external digit defects were

observed.

**Reliability:** (4) not assignable

Only abstract.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

208 26

- OCT

- 2006 (267)

**Species:** other: rat and mouse **Sex:** female

**Strain:** other: Sprague

- Dawley rat and CD

- 1 mouse

**Route of administration:**  other: in vitro whole embryo culture

**Exposure period:** 24 or 48 hours

**Frequency of treatment:** single

**Duration of test:** 48 hours

**Doses:** 0, 8, 12 or 16 mg/ml (rat), 0, 2, 4 or 8 mg/ml

(mouse)

**Control Group:** yes, concurrent vehicle

**Method:** other: in vitro whole embry culture

**GLP:** no data

**Method:** Screening test including developmental stages of

embryogenesis:

The following parameters were examined: viability, yolk sac

circulation, heart beat, crown

- rump length, head length,

somite number, Feulgen staining to localize cell death and

proliferat ion, scanning electon microscopy and histology

(only rat embryos).

**Remark:** non

- BASF study

**Result:** In rat embryos methanol affected adversely yolk sac

diameter, head length, developmental score, somit e number

and mortality. At the highest concentration 100 % of

embryos were abnormal and cell death was increased in

various

parts of the nervous system.

In mouse embr yos methanol affected adversely crown rump

length, head length, developmental score and somite number.

At the highest concentration the percentage of abnormal

embryos reached 94 %. Cell death was not in duced in treated

cultures.

------------------------

**Test substance:** Methanol, no further data.

26

- OCT

- 2006 (268)

**Species:** rat **Sex:** no data

**Strain:** no data

**Route of administration:** other: whole embryo culture

**Exposure period:** 24 hours

**Frequency of treatment:** single

**Duration of test:** 48 hours

**Doses:** 0, 4, 6, 8, 12 or 16 mg/ml

**Control Group:** yes, concurrent vehicle

**Method:** other: whole embryo culture, no details indicated

**GLP:** no data

**Method:** Screening test including developmental stages of

embryogenesis:

Embryos were explanted on day 9 of gestation, cultured for

24 hours in methanol supplemented serum and cultured for

further 24 hours in pure serum. The following parameters

were measured: embryolethality, somite number, crown rump

length, developmental score, head length. Number of embryos

used not indicated.

**Remark:** non

- BASF study

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

209 **Result:** 4 and 6 mg/ml was not toxic and did not affect embryonic

development. Higher concentrations led to a dose related

decrease in somite number and overall development. 12 and

16mg/ml led to increased lethality. Head length and crown

rumplength were not affected by the treatment.

**Test substance:** Methanol, no further data.

26

- OCT

- 2006 (269)

**Species:** rat **Sex:** female

**Strain:** Sprague

- Dawley

**Route of administration:** other: whole embryo culture

**Exposure period:** 24 hours

**Frequency of treatment:** single

**Duration of test:** 48 hours

**Doses:** 0, 2, 4, 8 , 12 or 16 mg/ml

**Control Group:** yes, concurrent vehicle

**Method:** other: in vitro culture of whole embryos

**GLP:** no data

**Method:** Screening test including developmental stages of

embryogenesis:

The following parameters were examined: somite number,

crownrump length, developmental score, yolk sac diameter,

head length, incidence of anomalies.

**Remark:** non

- BASF study

**Result:** Ra t embryos were explanted and cultured in 0, 2, 4, 8, 12 or

16 mg Methanol/ml rat serum for 24 h and then transferred to

rat serum alone for 24 h. Embryonic development of the 2 and

4 mg groups was not s ignificantly different from the

controls whereas the higher concentrations resulted in a

concentration related decrease in somite number, head length

and developmental score. The 12 mg/l dose resulted i n some

embryolethality as well as dysmorphogenesis, while the

highest dose was embryolethal. Methanol was dysmorphogenic

in vitro in rat embryos at a methanol concentration

comparable to that reported in maternal serum following

teratogenic in vivo exposures.

**Test substance:** Methanol, no further data.

26

- OCT

- 2006 (270)

**Species:** rat **Sex:** no data

**Strain:** Sprague

- Dawley

**Route of administration:** other: whole embryo culture

**Exposure period:** 48 hours

**Frequency of treatment:** single

**Duration of test:** 48 hours

**Doses:** 2.08

- 16.67 μl/ml (1.64

- 13.17 mg/ml)

**Control Group:** yes, concurrent vehicle

**Method:** other: in vitro culture of whole embryos

**GLP:** no data

**Method:** Screening test including developmental stages o f

embryogenesis:

9

- 17 embryos were used per treatment scheme, the effect

ofmethanol and its major metabolite, formic acid, was

compared. The following parameters were recorded: yolk sac

circulation, dorsally bending, heart beat, crown

- rump

length, somite number and protein content.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

210 **Remark:** non

- BASF study

**Result:** Crown

- rump length, somite number and protein content was

dose related decreased firstly at 11.6 ul/ml (9.16 mg/ml),

at 8.75 ug/ml (6.91 mg/ml) no treatment related effects

wereobserved.

**Test substance:** Methanol, no further data.

26

- OCT

- 2006 (271)

**Species:** mouse **Sex:**

**Strain:** CD

- 1

**Route of administration:** other: whole embryo culture

**Exposure period:** 24 hours

**Frequency of treatment:** single

**Duration of test:** 24 hours

**Doses:** 0, 2, 4, 6, 8 mg/ml

**Control Group:** yes, concurrent vehicle

**Method:** other

**GLP:** no data

**Method:** Screening test including developmental sta ges of

embryogenesis:

Day

- 8 mouse embryos were explanted and cultured in 0, 2, 4,

6 or 8 mg Methanol/ml culture medium (75 % rat serum, 25 %

Tyrode's salt solution) for 24 h.

**Remark:** non

- BASF study

**Result:** CD

- 1 mice: Embryonic development in the 2

- mg/ml group was

not significantly different from the controls but all higher

concentration groups had a significant decrease in

developmental score and crown

- rump length. The high

concentration group als suffered 80 % embryolethality.

C57/BL/6J mice: Somite number was not affected. 2 mg/ml and

higher produced a concen tration

- dependent increase in

dysmorphology in the absence of embry

- lethality, including

open neural tubes, branchial arch hypoplasia, and

prosencephalic hypoplasia. Additionally, 4 and 6 mg/ml

caused mes

- and rhombencephalic hypoplasia.

These results indicate that C57 mice are more sensitive than

CD

- 1 mice, suggesting that differences in strain sensitivity

to in

- vitro methanol exp osure are more likely due to

intrinsic differences in the developing embryo rather than

maternal factors.

-----------------------------

**Test substance:** Methanol, no further data.

**Reliability:** ( 2) valid with restrictions

26

- OCT

- 2006 (270) (272)

**Species:** mouse **Sex:** no data

**Strain:** CD

- 1

**Route of administration:** other: whole embryo culture

**Exposure period:** 12 hours

**Frequency of treatment:** single

**Duration of test:** 45 hours

**Doses:** 62, 125, 187, 250 or 375 mM (ca. 2, 4, 6, 8 or 12

mg/ml)

**Control Group:** yes, concurrent vehicle

**Method:** other: in vitro culture of whole embryos

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

211 **GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Method:** Screening test including developmental stages of

embryogenesi s:

The following parameters were examined: percentage of live

embryos, cephalic dysraphism, prosencenphalic lesions,

branchial arch hypoplasia.

**Remark:** non

- BASF study

**Result:** Methano l treatment led to dose related increased incidences

of all anomalies examined for.

**Test substance:** Methanol, HPLC

- grade from Sigma Chemical Corporation, no

further data.

26

- OCT

- 2006 (250)

**Species:** other: rat and mouse **Sex:**

**Strain:** other: Sprague

- Dawley rat, CD

- 1 mouse

**Route of administration:** other: whole embryo culture

**Exposure period:** 48 h

**Fre quency of treatment:**  single

**Duration of test:** 48 h

**Doses:** about 2 to 9 mg/ml (also in combination with formate)

**Control Group:** yes, concurrent vehicle

**Method:** other: in vitro culture of whole embryos

**GLP:** no data

**Method:** Screening test including developmental stages of

embryogenesis:

Follow

- up studies based on previous ones: Methanol and

formate as well as combination of both substances were

examined.

The following parameters were examined: somite number,

crownrump length, developmental score, yolk sac diameter,

head length, incidence of anomalies.

The developmental scoring system took into account embryonic

groth parameters as well as stages of development.

**Remark:** In a further preliminary study, there was some evidence that

methanol may inter fere with ADH and thus prevent oxidative

conversion of retinol into retinoic acid, a presumed

mediator in embryonic development, which may explain part of

methanol

- induced disorders (Andrews et al., 199 8b).

non

- BASF study

**Result:** 1. In rats, a less than additive

- dose effect was found:

the combination of methanol and formate proved to be less

toxic than would be predicted from their individ ual

toxicities, suggesting antagonistic interaction (Andrews et

al., 1998a).

A standard methanol dose

- response line has been constructed,

based on the developmental scoring system. Co mbinations with

low formate concentrations (<1 mg/ml), along with various

methanol concentrations, generally fell within the 95

- %

confidence interval of the standard methanol

- curve (Andrews

et al., 1998a).

2. Formate alone in whole

- embry culture, given to the

medium of 9

- d rat embryos as well as to 8

- d mice embryos at

0.2

- 3.0 mg/ml (for 24 or 48 h) exhibited trends towards

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

212 reduction of growth and development and increases in the

number of dead and abnormal embryos. Formate is embryotoxic

at concentrations >= 4fold lower than those for methanol

(Ebron

- McCoy et al., 1994)

**Test substance:** Methanol, no further data

26

- OCT

- 2006 (273) (274) (275)

**5.8.3 Toxicity to Reproduction, Other Studies**

**-**

**5.9 Specific Investigations**

**Endpoint:** Behavioural Effects

**Species:** rat

**Strain:** Long

- Evans **Sex:** male

**Route of administration:** oral, gavage

**No. of animals:** 11

**Vehicle:** water

**Frequency of treatment:** single do se

**Doses:** 1000, 2000, and 3000 mg/kg (50

- % aqueous solution)

**Control Group:** other: test animals = control group (water) prior to

treatment

**Method:** other: fixed wheel running ratio tes t

**Year:** 1993

**GLP:** no data

**Test substance:** as prescribed by 1.1

- 1.4

**Result:** The rats displayed no signs of overt intoxication such as gait

disturbance, but a significant, dose

- related reduction in FR 20

response was observed at all dose

- levels.

A NOAEL for behavioural changes cannot be derived.

--------------------------

**Test condition:** DOSE selection: 10, 20, and 30 % of the LD50.

TEST ANIMALs: The same animals were re

- used for each dose on

different days.

BEHAVORIAL TESTs:

Ten minutes after treatment, the animals were subjected to the

fixed wheel runnin g ratio test to assess operant running. The

test required the animal to run inside a wheel and rotate it

under a fixed ratio of 20 times (FR20) in order to receive a

food reward.

STAT ISTICS:

Data were evaluated by conducting repeated measures analysis

of variance (ANOVA), determining linear trend, correcting for

degrees of freedom, and performing analysis of residuals to

outliers and skewed distribution.

--------------

**Test substance:** Methanol, HPLC grade, purity 99.93 %, Aldrich

(233) (276)

**5.10 Exposure Experience**

**Type o f experience:**  other: review, general

**Remark:** Humans (and non

- human primates) are uniquely sensitive to

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

213 methanol poisoning and the toxic effects in these species

is

characterized by formic acidemia, metabolic acidosis,

ocular

toxicity, nervous system depression, blindness, coma and

death. Nearly all of the available information on methanol

toxicity in humans relates to the consequences of acute

rather than chronic exposures. A vast majority of

poisonings

involving methanol have occurred from drinking adulterated

bever ages and from methanol

- containing products. Although

ingestion dominates as the most frequent route of

poisoning,

inhalation of high concentrations of methanol vapor and

pe rcutaneous absorption of methanolic liquids are as

effective as the oral route in producing acute toxic

effects. The most noted health consequence of longer

- term

exposure to lower levels of met hanol is a broad range of

ocular effects.

The toxic properties of methanol are based on factors that

govern both the conversion of methanol to formic acid and

the subseque nt metabolism of formate to carbon dioxide in

the folate pathway. The toxicity is manifest if formate

generation continues at a rate that exceeds its rate of

metabolism.

T he lethal dose of methanol for humans is not known for

certain. The minimum lethal dose of methanol in the absence

of medical treatment is between 0.3 and 1 g/kg. The minimum

dose causing perma nent visual defects is unknown.

The severity of the metabolic acidosis is variable and may

not correlate well with the amount of methanol ingested.

The

wide interindividua l variability of the toxic dose is a

prominent feature in acute methanol poisoning.

Two important determinants of human susceptibility to

methanol toxicity appear to be (1) concurrent ingestio n of

ethanol, which slows the entrance of methanol into the

metabolic pathway, and (2) hepatic folate status, which

governs the rate of formate detoxicification.

The sympt oms and signs of methanol poisoning, which may not

appear until after an asymptomatic period of about 12 to 24

hours, include visual disturbances, nausea, abdominal and

muscle pain, dizziness, weakness and disturbances of

consciousness ranging from coma to clonic seizures. Visual

disturbances generally develop between 12 and 48 h after

methanol ingestion and range from mild photopho bia and

misty

or blurred vision to markedly reduced visual acuity and

complete blindness. In extreme cases death results. The

principal clinical feature is severe metabolic acidosis of

anion

- gap type. The acidosis is largely attributed to the

formic acid produced when methanol is metabolized.

The normal blood concentration of methanol from endogenous

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

214 sources is less than 0.5 mg/litre (0.02 mmol/litre), but

dietary sources may increase blood methanol levels.

Generally, CNS effects appear above blood methanol levels

of

200 mg/litre (6 mmol/litre), and fatalities have occurred

in

untreated patients with initial methanol levels in the

range

of 1500

- 2000 mg/litre (47

- 62 mmol/litre).

Visual disturbances of several types (blurring,

constriction

of the visible field, changes in colour perception, and

temporary or permanent blindness) have been reported i n

workers who experienced methanol air levels of about 1500

mg/m3 (1200 ppm) or more.

A widely used occupational exposure limit for methanol is

260 mg/m3 (200 ppm), which is designed to protect workers

from any of the effects of methanol

- induced formic acid

metabolic acidosis and ocular and nervous system toxicity.

No other adverse effects of methanol have been reported in

humans except minor skin and eye irritation at exposures

well above 260 mg/m3 (200 ppm).

**Reliability:** (2) valid with restrictions

see remark on selection of literature

24

- DEC

- 2004 (88)

**Type of experience:** other: kinetics, inhalation (Osterloh)

**Remark:** During a randomized double

- blind study of the potential

neurobehavioral e ffects of inhaled methanol at 200 ppm for

4

hours methanol analysis was performed. Methanol was rapidly

absorbed by inhalation. Serum methanol conc. were increased

by more than fourfold at the end of exposure period, as

were

urinary methanol excretion rates, although formate conc.

were not increased over background conc. The overall

- 205/327

-

elimation half

- life was 3.2 + 2.3 h.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

24

- DEC

- 2004 (277)

**Type of experience:** other: acute toxicity, CNS (Chuwers)

**Remark:** This study assessed whether acute exposure to methanol

vapor

at the current industrial threshold limit value of 20 0 ppm

for 4 hr has adverse effects on human neurobehavioral

performance. Twenty

- six healthy subjects (15 men, 11 women;

ages 26

- 51 years) were exposed to methanol or water vapor

for 4 hr while seated in a chamber. The subjects served as

their own controls in a randomized, double

- blind study

design. The variables assessed were serum and urine

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

215 methanol

and formate levels; visual qualities (color discrimination

and contrast sensitivity); and neurophysiological (auditory

evoked potentials) and neurobehavioral qualities.

Exposure to met hanol increased serum concentrations and

urinary excretions of methanol, but did not affect formate

levels. Overall visual, neurophysiological, and

neurobehavioral test outcomes were not signif icantly

affected, unless certain between

- subject variables are

considered. Slight effects on P

- 300 amplitude and Symbol

Digit testing were noted.

**Reliability:** (2) valid with restrictio ns

basic data given, acceptable restrictions

24

- DEC

- 2004 (278)

**Type of experience:** other: review, general (Becker)

**Remark:** Methanol poisoning is an uncommo n but an extremely

hazardous

intoxication. Since methanol is a versatile fuel and is

having increasing usage in an energy

- conscious society, a

high index of suspicion and s wift laboratory confirmation

is

essential in managing this poisoning. Methanol poisoning

may

occur in sporadic or epidemic circumstances. Chronic

expos ure may occur in the occupational setting. Man is

uniquely susceptible to methanol toxicity, perhaps

dependent

upon folate metabolism. Classic symptoms of methanol

toxicity can only occur in laboratory animals who are

rendered folate deficient. Folate may be useful in humans

enhancing removal of the toxic products of methanol

poisoning. The enzyme responsible for metabolism of

methanol

is alcohol dehydrogenase. Ethanol has a higher affinity for

this enzyme and is preferentially metabolized. Simultaneous

ethanol and methanol adminis tration may confuse the onset

of

the intoxication. Pyrazoles may also be used to inhibit

alcohol dehydrogenase thus preventing the intoxication. The

most important initial symptom of methanol poisoning is

visual disturbance. The symptoms may be delayed up to 24

hours after ingestion due to simultaneous alcohol

administration and metabolic processes. Laboratory ev idence

of severe metabolic acidosis with increased anion and

osmolar gaps strongly suggest the clinical diagnosis. There

may be an important association between mean corpuscular

volume which is significantly higher in cases of severe

methanol poisoning than in mild cases. Once the diagnosis

is

suspected, a blood level from methanol should be returned

rapidly. Treatment of methanol toxicity after good

supportive care is to diminish the metabolic degradation of

methanol with simultaneous ethanol and then to perform

hemodialysis and a lkalinization to counteract metabolic

acidosis. Folate should be administered to enhance

metabolic

breakdown of formate. Alcoholic patients may especially

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

216 susceptible to me thanol poisoning due to relative folate

defiency.

**Reliability:** (2) valid with restrictions

see remark on selectionof literature

28

- DEC

- 2004 ( 279)

**Type of experience:** other: review, general (Roe 1955)

**Remark:** During methanol poisoning in man the concentration of

formic

acid in the blood is quite variable. In 5 lethal cases it

ranged from 9 to 68 mg per cent. In three patients who also

died it ranged from 5.7 to 19 mg per cent. Furthermore,

the methanol concentration in the blood in 23 lethal cases

varied be tween 51 and 274 mg per cent. It becomes obvious

that the mere concentrations of these substances are not

the

only decisive factors in the clinical course of the

poisoning.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

28

- DEC

- 2004 (280)

**Type of experience:** other: review, general (Tephly)

**Remark:** Methanol toxicty in humans and monkeys is characterized by

a

latent period of many hours followed by a metabolic

acidosis

and ocular toxicity. This is not obser ved in most lower

animals. The metabolic acidosis and blindness is apparently

due to formic acid accumulation in humans and monkeys, a

feature not seen in lower animals. The accumulation of

formate is due to a deficiency in formate metabolism which

is, in turn, related, in part, to low hepatic

tetrahydrofolate (H4folate). An excellent correlation

between hepatic H4folate and formate oxidation rates has

been shown within and across species. Thus, humans and

monkeys possess low hepatic H4folate levels, low rates of

formate oxidation and accumulation of f ormate after

methanol. Formate, itself, produces blindness in monkeys in

the absence of metabolic acidosis. In addition to low

hepatic H4folate concentrations, monkeys and humans also

have low hepatic 10

- formyl H4folate dehydrogenase levels,

the enzyme which is the ultimate catalyst for conversion of

formate to carbon dioxide.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

28

- DEC

- 2004 (281)

**Type of experience:** other: review, general (Kavet; Nauss)

**Remark:** Methanol could become a major automotive fuel in the U.S.,

and its use may result in increased exposure of the public

to methanol vapor. Nearly all of the available information

on methanol toxicity in humans relates to the consequence s

of acute, rather than chronic, exposures. Acute methanol

toxicity evaloves in a well

- understood pattern and consists

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

217 of an uncompensated metabolic acidosis with superimposed

toxicity to the visual system. The toxic properties of

methanol are rooted in the factors that govern both the

conversion of methanol to formic acid and the subsequent

metabolism of formate to carbon dioxide in the folate

pathway. In short, the toxic syndrome sets in if formte

generation continues at a rate that exceeds its rate of

metabolism. Current evidence indicates that form ate

accumulation will not challenge the metabolic capacity of

the folate pathway at the anticipated levels of exposure to

automotive methanol vapor.

**Reliability:** (2) valid with restric tions

see remark on selection of literature

28

- DEC

- 2004 (192)

**Type of experience:** other: kinetics, oral (Ferry)

**Remark:** Methanol blood and urine concentrat ions were investigated

in

four volunteers who had ingested small amounts of methanol

(0.2 ml hourly for six hours). The methanol urine

concentration did not exceed 8.0 μg/m l. It was estimated,

that at a MAC value of 200 ppm with a total 8 h ventilatory

volume of 10 m3 and assuming complete absorption and no

exhalation 2.6 g methanol would be absorbed. The highest

urinary concentration attained by oral ingestion of this

amount of methanol at a rate of 0.5 ml hourly in one of the

subjects was 17.6 μg/ml.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (282)

**Type of experience:** other: kinetics, inhalation (Sedivec)

**Remark:** Five volunteers were exposed to constant and suitable

graded

concentration between 103 to 284 mg/m3 of methanol vapours

for a period of 8 h. Pulmonary retention ranged from 53.4

to

61.3 %. Elimination half

- life was calculated to range

between 1.5 to 2.0 h.

**Reliability:** (2) valid with restrictions

aceptable study, meets basic scientific principles

28

- DEC

- 2004 (283)

**Type of experience:** other: review, acute toxicity (Roe 1982)

**Remark:** The minimal lethal dose of methanol in humans has not been

determined. It has been suggested th at about 1 g/kg can

cause death if the patient is untreated and has not

consumed

ethanol. In some clinical cases, the blood methanol content

is low in the last phase of the poisoning. In three such

cases blood methanol concentration was 0.275, 0.277, and

0.194 g/l, respectively. On the assumption that the body

water in diffusion equilibrium with the blood respres ents

about 70 % of the body weight, it has been calculated that

0.19, 0.19, and 0.14 g/kg, respectively, was present in the

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

218 body. Data on the rate of methanol oxidation in humans

probably do not exist. Provided that the rate of methanol

oxidation is the same in man as in rhesus monkeys, the

amount of methanol oxidized during 18 hours (the average

time needed for development of severe acidosis in clinical

cases) would be 0.666 g/kg. It seems reasonable still to

regard 1 g/kg of methanol as the approximate minimal lethal

dose in man.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

28

- DEC

- 2004 (284)

**Type of experience:** other: kinetics, inhalation (Heinrich; Angerer)

**Remark:** Twenty persons occupationally exposed to methanol were

examined according to their methanol levels in blood and

urine and their formic acid excretion. An 8

- h exposure to a

methanol conce ntration of 93 ml/m3 in the air at the

working

area caused average methanol levels in blood and urine of

8.9 +

- 14.7 mg/l and 21.8 +

- 20.0 mg/l, respectively, and a

mean fo rmic

- acid level in urine of 29.9 +

- 28.6 mg/l. Based

on these results a rough estimate shows the corresponding

methanol content in urine to be about 40 mg/l for an 8

- h

exposure at 200 ml/m3.

No correlation between exposure and urinary concentration

of formate could be clearly demonstated, although 15 % of

measured values exceeded the norm

**Reliability:** (2) valid with restric tions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (285)

**Type of experience:** other: review, acute toxicity (Litovitz)

**Remark:** Annual repo rt of the American Association of Poison Control

Centers National Data Collection System. 3 cases of fatal

methanol ingestion were reported. Blood levels were 175,

402, and 39 mg/dl, respective ly.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

28

- DEC

- 2004 (286)

**Type of experience:** other: kinetics, inhalation (Baumann; A ngerer)

**Remark:** A study was carried out among 20 workers employed in a

printing office at three different work places (methanol

concentration: 85, 101, and 134 ppm) to determine whether

concentration of formic acid in blood or urine and the

methanol content of aveolar air permit the estimation of

methanol exposure. The concentration of formic acid in

blood

increased significantly from 3.2 +

- 2.4 mg/ml before to 7.9

+

- 3.2 mg/l after the shift in the exposed workers. In 36

non

- exposed persons, the blood formate levels ranged from 0

- 20 mg /l.

The corresponding concentration in urine was increased

significantly from 13.1 +

- 3.9 mg/l to 20.2 +

- 7 mg/l,

respectively. On the contrary, in the control groups there

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

219 was a small but significant decrease of formic acid

concentration in blood from 5.6 +

- 4.5 mg/l in the morning

to 4.9 +

- 4.2 mg/l in the afternoon.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (287)

**Type of experience:** other: acute toxicity, CNS (Muttray)

**Remark:** Twelve healthy subjects were expose d for 4 h to 200 ppm and

to 20 ppm (control) in an exposure chamber in a cross

- over

design. The EEG was recorded before (reference) and at the

end of each exposure with, the subject's eyes clos ed and

opened and during a choice reaction test (color word stress

test). Spectral power was calculated by fast Fourier

transformation. Subjective symptoms and effects of blinding

with 20 ppm methanol were assessed by questionnaires. The

study was a single

- blind one. During subjects' exposure to

200 ppm, their scores for prenarcotic and irritating

symptoms were n ot different from controls. In the

closed

- eye

condition of subjects, the spectral power of the theta

- band

and of some electrodes of the delta

- band was significantly

less at the end of exposure to 200 ppm, than that of

controls. In the open

- eye condition and during the color

word stress test no significant changes were found. The

changes in the theta

- band suggest a slight excitatory

effect

of 200 ppm methanol. The effect was weak, as scores of

acute

symptoms did not change.

**Reliability:** (2) valid with restrictions

acceptable study meets basic scientific principles

28

- DEC

- 2004 (288)

**Type of experience:** other: acute toxicity (Jacobsen)

**Remark:** Eleven patients concomitantly pois oned with methanol are

described. Their whole blood methanol conc. ranged from

137.2 mmol/l (4.39 g/l) to 7.4 mmol/l (0.24 g/l). The

clinical course in most patients was mild, which was

attributed to the concomitant and subsequent ethanol

ingestion and rapid transport to dialysing units. One

patient suffered permanent visual impairment of one eye,

while the other s recovered completely. Symptoms of

poisoning

were most clearly correlated to the degree of metabolic

acidosis. All patients were dialysed. Ethanol in conc. even

lower than usually recommended may be useful as the only

treatment of patients with blood methanol conc. up to 15

mmol/l (0.5 g/l), provided there is no acidosis or visual

impairment.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (289)

**Type of experience:** other: acute toxicity (Erlanson)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

220 **Remark:** Four cases of methanol poisoning were treated with alkali,

ethyl alcohol, and the haemodialysis. In 3 cases respirator

treatment was also required. Severe acidosis, failing

eye

- sight, and cerebral damage were present at the start of

treatment within 29 to 44 hours of the methanol ingestion.

Three patients, one of whom had consumed 90 g, died despite

correction of the acidosis a nd elimination of methanol.

Autopsy in these cases showed massive necrosis of the brain

and haemorrhages into the putamina. The fourth patient, who

had consumed 80 g methanol, survived but died 1 1/2 year

later from pneumonia; autopsy showed slit

- shaped cysts in

the lateral parts of the putamina. All the patients had

considerable hypopotassaemia.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (290)

**Type of experience:** other: acute toxicity (Naraqi)

**Remark:** An outbreak of acute methyl alcohol intoxication occurred.

Twenty

- eight young men attended a drinking party and drank

methyl alcohol. The amount consumed by each individual

ranged from an equivalent of 60

- 600 ml of pure methanol.

Three had prior ethanol ingestion. All 28 became ill 8

- 36

hours after drinking and were hospitalized. The most

commonly observed clinical syndromes were: acute metaboli c

acidosis, severe visual impairment and acute pancreatitis.

Four died within 72 hours after admission to the hospital.

All had severe metabolic acidosis and visual impairment and

three pancreatitis. Of 24 who recovered, 16 showed no

residual complications, six had bi

- lateral visual

impairment

and had difficulty in speech as well as visual impairment.

A

three month follow

- up examination showed no change in the

findings. Coma, seizures and prolonged acidosis were poor

prognostic signs. The estimated amount of consumed methanol

and the rapidity of the appearance of signs of toxicity

following methanol ingestion did not seem to influence the

outcome of poisoning.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (291)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

221 **Type of experience:** other: worker exposure (Kawai / Yasugi)

**Remark:** In methanol

- exposed male an d female workers there was a

linear relationship between methanol exposure and formate

excretion in the urine after working shift up to an 8

- h

average of 2000 and 4000 ml/m3, respectively. In u nexposed

persons, the baseline urinary excretion of methanol and

formate were 1.9 ±0.8 and 26.2 ±12.2 mg/l urine.

Based upon a regression equation presented by the authors,

the mean added formate concentration in the urine amounted

to about 180 mg/l (males) and 100 mg/l (females) after an

8

- h average of 2000 ml/m3, and to 17 mg/l (males) and about

10 mg/l (femal es) after 200 ml/m3 (Yasugi et al. 1992).

------------------

The exposure

- excretion relationship and possible health

effects of exposure to methanol vapor were studied in 33

exposed workers during the second half of 2 working weeks.

Urinary methanol concentrations were also determined in 91

nonexposed subjects (Kawai et al., 1991). The geometric

mean

value for methanol in urine samples from the latter was <2

mg/l. Among the exposed workers, the methanol level in

urine

samples collected prior to work shift exceeded the 95 %

upper limit of normal. Methanol concentrations in urine at

the end of shift showed a significant correlation between

exposure to

methanol vapour at concentrations of up to 5,500 ppm and

the

level of methanol measured:

Following exposures to mean 1690 ml/m3, more than 100 mg/l

methanol was found in the urine of 8 men, at an average

exposure level of 550 ml/m3, 30

- 100 mg/l methanol were

detected in 6 men. The highest 8

- h exposures from 4000

ml/m3

and higher corresponded to 300 to 500 mg/l (Kawai et al.,

1991).

The calculation indicated that a mean level of 42 mg

methanol/l urine was excreted in the shift

- end urine

samples

following 8 h exposure to methanol at 20 0 ppm.

Highly exposed workers (3500

- 5500 ml/m3) more often

complained of blurred vision and headache during or after

work. Nobody stated to suffer from photophobia.

The examination of the eye fundus failed to reveal retinal

changes. Among three workers employed in that company

between 0.3 and 7.8 years and having been exposed to about

1000

- 1600 and 120 to 36 00 mg/m3 (one case), two showed

retarded pupil reflex and one mild mydriasis (Kawai et al.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

222 1991). Dimmed vision and nasal irritation were among the

most frequent symptoms complained during wo rk.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (292) (293)

**Type of experience:** other: human exposure (Fr anzblau)

**Remark:** Five human subjects were exposed to an atmospheric

concentration of 200 ppm of methanol in a test chamber for

7

h per day for 5 consecutive days. Ambient air in t he

chamber

was monitored continously for methanol, while urine was

monitored for methanol and formic acid. Mean urinary

methanol concentration were increased from baseline at the

end of each exposure session, but returned to baseline in

samples collected 16 h following cessation of exposure. The

concentration of formic acid in morning urine specimens did

not change significantly over the 7 days of the exposure.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (294)

**Type of experience:** other: human exposure (Leaf; Zatman)

**Remark:** The elimination of methanol after doses of 2.5 to 7 .0 ml

has been studied in five human subjects. At any time the

rate of elimination was found to be proportional to the

concentration of methanol in the body.

Blood levels of 47 to 76 mg/l were measured 2 to 3 h after

oral uptake of 71

- 84 mg methanol/kg bw (6.6

- 7.4 ml per

person); methanol disappearance obeyed first

- order kinetics

with a half

- time of about 3 h.

Only a very small fraction of ingested methanol (about 2 %)

was eliminated via the respiratory and urinary routes. The

rates of absorption of methanol by two human subjects

during

exposure to vapour concentrations of 400

- 1000 ppm have

been investigated.

Over short periods the amount of methanol absorbed appears

to be approximately proportional to the duration of

exposure

and to the concentration of vapour in the atmosphere. It is

concluded that accumulation in the body would occur at 3000

ppm an the maximum safe concentration for occupational

exposure is 300 ppm.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

**Type of experience:** other: acute toxicity (Frederick)

**Remark:** A health hazard evaluation was conducted by the National

I nstitute for Occupational Safety and Health (NIOSH) to

determine if vapors from duplicating fluid (99% methyl

alcohol) used in direct

- process spirit duplicating machines

were causing adverse he alth effects among teacher aides, or

had been responsible for the deaths of three former teacher

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

223 aides. Death certificates and autopsy data were obtained

and

evaluated. A s elf

- administered symptom questionnaire was

distributed to current teacher aides (exposed group) and to

a comparison group of teachers. Fifteen

- minute breathing

zone air samples for methyl alcoh ol vapor were collected at

operator stations using an infrared gas analyzer. No

information supported the claim that the three deaths were

related to methyl alcohol exposure. Teacher aides repo rted

significantly more blurred vision, headache, dizziness, and

nausea than the comparison group. Concentrations of

airborne

methyl alcohol ranged from 365

- 3080 ppm; 15 of 21

measurements exceeded the NIOSH

- recommended 15

- minute

exposure limit of 800 ppm. A mean 96% reduction in vapor

concentration was accomplished using inexpensive enclosures

and existing room exhaust systems.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

28

- DEC

- 2004 (296)

**Type of experience:** o ther: human exposure (Batterman)

**Remark:** Dermal exposures to methanol were administered in a

clinical

study designed to compare several biological indicators.

Four subjects were e xposed in five exposure sessions of

varying length. In each session, a sequence of measurements

of methanol conc. in blood, breath, and headspace samples

of

air at exposed and unexposed skin analysed. At exposed skin

samples were highly elevated for at least 8 h following

exposure, indicating the presence of a methanol reservior

in

skin. Afte r exposure, methanol conc. at exposed skin showed

a rapid initial decline, then a slower first order

decrease.

When transfer was restricted, surface conc. at unexposed

skin were similar to levels in breath and were strongly

correlated to methanol conc. in blood.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

28

- DEC

- 2004 (297)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

224 **Type of experience:** other: human exposure (skin) (Dutkiewicz)

**Remark:** Methanol absorption rate through human skin (foremarm,

15

- 60

min.) has been e xamined and a value of 0.192 mg(cm2/min)

was determined. The excretion of unchanged methanol with

urine and exhaled air, after absorption through skin and

administration "per os" amounted to 16 % and 22 % of the

absorbed dose (1.67 g), respectively. It was estimated,

that exposure of one hand to liquid methanol for only 2

min. would lead to the absorption of as much methanol (170

mg), as would be taken up by the lungs fro m an 8 h exposure

to MAC of 50 mg/m3.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (298)

**Type of exp erience:**  other: kinetics, inhalation, human (Batterman)

**Remark:** Relationship between methanol concentration in the blood,

urine, and breath of volunteers exposed to methanol vapor

at 800 ppm for peri ods of 0.5, 1, 2, and 8 h. For methanol,

concentrations are not proportional to the exposure

duration due to metabolic and other elimination processes

that occur concurrently with the exposure. The 0.5 to 2

- h

per iods of exposure were used to estimate the half

- life of

methanol.

Blood data gave a half

- life of 1.44 +

- 0.33 h. Comparable

but slightly more variable results were obtained using

urine

data corrected for the voiding time (1.55 +

- 0.67 h) and

breath data corrected for mucous membrane desorption (1.40

+

- 0.38 h). Methanol concentrations in blood lagged some 15

- 30

min. behind the termination of exposure, and concentrations

in urine were further delayed.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

28

- DEC

- 200 4 (299)

**Type of experience:** other: kinetics, inhalation, human (Batterman)

**Remark:** Experimental study of dermal exposure to methanol in human

volunteers est imating percutaneous absorption. 12

volunteers were exposed to methanol via one hand for

durations of 0 to 16 min in a total of 65 sessions. The

pre

- exposure methanol conc. in blood was 1.7 mg/l, and

subjects had statistically diffe rent mean conc. The maximum

methanol conc. in blood was reached 1.9 h after exposure;

this is comparable to that reached following inhalational

exposure at a methanol conc.

of 200 ppm. Delivery rate from skin into blood lagged

exposure by 0.5 h, and methanol continued to enter the

systemic circulation for 4 h following exposure. The mean

derived absorption rate was 8.1 mg/cm2/h.

**Reliability:** (2) valid with restri ctions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (300)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

225 **Type of experience:** other: kinetics, inhalation, human (Lee)

**Remark:** Accumulati on of formate, the putative toxic metabolic of

methanol, in the blood methanol and the relationship

between

pulmonary intake and blood methanol conc. were investigated

in s ix human volunteers following a 6

- hr exposure to 200

ppm

methanol. At the end of a 6

- hr exposure to 200 ppm at rest,

the blood methanol conc. was increased from a mean of 1.8

μg/ml to 7.0 μg/ml. Under light excercise, the total amount

of methanol inhaled during the 6

- hr exposure period was 1.8

times that inhaled at rest. However, no statistically

significant incr ease in blood methanol conc. was observed

under exercise: the conc. averaged 8.1 μg/ml. Formate did

not accumulate in the blood above its background level

following the 6

- hr exposure to 200 ppm methanol whether

subjects were exposed at rest or during excercise.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

28

- DEC

- 2004 (196)

**Type of experience:** other: kinetics, inhalation (Kawai)

**Remark:** Exposure of 34 male workers to combined toluene, styrene

and

methanol was monitored by personal diffusive sampling of

solvent vapours in breathing zone air, analysis of

shift

- end

blood for the 3 solvents and analysis of shift

- end urine

for

hippuric, mandeli c and phenylglyoxylic acids and methanol.

The exposure of most of the workers was below current

occupational exposure limits. Regression analysis showed

that a linear correlation exists for eac h of the 3 solvents

between any pairs of the concentrations in air, blood and

urine. Namely, toluene, styrene and methanol concentrations

in blood obtained at the end of a shift are linearly

related

to the time

- weighted average intensity of exposure to

corresponding solvents, and also hippuric, mandelic and

phenylglyoxylic acids as well as methanol in shift

- end

urine. The concentrations of hippuric, mandelic and

phenylglyoxylic acids as well as methanol in urine

correlated with the respiratory exposure intensity.

Comparison of the present results with the

exposure

- excretion relationship after occupational exposure

to the individual solvent showed that no modification in

metabolism is induced by the combined exposure when

exposure

is low, as in the present case.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

28

- DEC

- 2004 (301)

**Type of experience:** other: acute toxicity, cns (Dethlefs / Naraqi)

**Remark:** The ocular manifestation of acute methyl alocohol

intoxication were studied in 24 men during an outbreak. The

visual acuity, pupillary reactions to light, fundal

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

226 appearence and visual fields were recorded in all patients

within 72 hours of ingestion of methanol and again three

months later. Nine patients had no ocular abnormality, 7

had only transient ocular abnormalities, and eight had

permanent ocular abnormalites. Complete blindness occurred

in two patients, while sever e visual deficit resulted in

four others. The incidence of permanent ocular

abnormalities

was found to correlate with the incidence of metabolic

acidosis, and with the stat ed volume of methanol consumed.

An inverse correlation was found between stated volume of

methanol consumed and onset of blurred vision.

**Reliability:** (2) valid with restrictions

accept able study, meets basic scientific principles

28

- DEC

- 2004 (302)

**Type of experience:** other: review, reproductive and developmental effects

(Lorente)

**Remark:** Study reviewed a variety of occupations and associated

exposures to complex mixtures in women who gave birth to

infants with and without cleft lip or palate.

No association was found between m ethanol exposure and oral

clefts, but several limitations of the study were raised:

small number of subjects exposed to methanol, lack of

individual exposure data, and confounding by other chem ical

exposures.

Because of these limitations the Panel judged the study

results to be uncertain and concluded there are

insufficient

data upon which t o evaluate the developmental toxicity of

methanol in humans.

**Reliability:** (2) valid with restrictions

acceptable study, meets bsci scientific principles

28

- DEC

- 2004 (303) (233)

**Type of experience:** other: kinetics, nutrition (Davoli)

**Remark:** Aspartam was administered to humans at a single dose of 500

mg per individual in 100 ml tap water.

Four adult volunteers fasted for 8 h and avoided alcohol,

fruits, fruit drinks or vegetable for 24 h.

Blood methanol was measured at 0, 30, 45, 60, 90, 120, and

180 min following ingestion.

The dose of aspartame was representative of the daily

average sugar consumption and corresonded to about 50 mg

methanol = 0.7

- 0.8 mg/kg.

**Result:** Baseline blood methanol: 1.4

- 2.6 mg/ L

Mean incremental increase (maximum after 45 min): <= 1 mg/L

Aspartame consumption by adults at a dose equivalent to the

daily intake of sugar results in methanol blood levels

similar to endogenous levels, in particular when divided in

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

227 smaller fractions over the day.

-------------------

28

- DEC

- 2004 (304) (233)

**Type of ex perience:**  other: kinetics, nutrition (Leon)

**Remark:** This longer

- term study demonstrated that ingesting

aspartame equivalent to a methanol dose of 7.5 mg/(kg bw*d)

per day resulted in blood methanol l evels around 10 mg/LL

in adults.

**Reliability:** (4) not assignable

Secondary literature

28

- DEC

- 2004 (305) (233)

**Type of experience:** other: remark o n selection of literature

**Remark:** Concerning effects in humans several quality reviews have

already been conducted. With a focus on health and safety

issues comprehensive reviews including those of th e World

Health Organization (WHO), 1997, Kavet and Nauss, 1990 and

the Center for the Evaluation of Risks to Human

Reproduction (CERHR), 1988 and single studies cited in

t hese

reviews were selected for this robust summary of methanol.

Relevant recent publications have also been taken into

account.

**Type of experience:** other: review, general

**Remark:** Review of toxicity of methyl aclohol following skin

absorption and inhalation in animal studies.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

(156)

**Type of experience:** other: review, general

**Remark:** Methyl alcohol is an aliphatic alcohol with use in a few

cosmetic formulations as a solvent and denaturant.

Concentrations up to 5% are typically used to denature

ethyl

alcohol in cosmetic products. Methyl alcohol is readily

absorbed through the skin and from the gastrointestinal and

respiratory tracts, is distributed throughout all organs

and

tissues (in direct relation to the body's water

distribution), and is eliminated primarily via the lungs.

Undiluted Methyl alcohol is an ocular and skin irritant.

Inhalation studies showed a no

- effect level for maternal

damage of 10,000 ppm and for teratogenic effects of 5,000

ppm. O verall, methyl alcohol is not considered mutagenic.

Carcinogenicity data were unavailable. The toxicity of

methyl alcohol in humans results from the metabolism of the

alcohol to formate and for mic acid through a formaldehyde

intermediate. Formate accumulation causes metabolic

acidosis

and inhibits cellular respiration. Methyl alcohol toxicity

is time and concentr ation dependent, and its toxic effect

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

228 is

competitively inhibited with ethyl alcohol. Because of the

moderating effect of ethyl alcohol, it was concluded that

methyl alcohol is safe as used to denature ethyl alcohol

used in cosmetic products. No conclusion was reached

regarding any other use of methyl alcohol.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

(306)

**Type of experience:** other: odour

**Remark:** Experimental study in two and seven further subjects for

detect ion of odour thresholds in solvents. For methanol the

odour threshold was 5,900 ppm and for detection of a

distinct odour was 8,800 ppm.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(307)

**Type of experience:** other: odour

**Remark:** In a group of 25 subjects odour threshold was 3,4 ppm.

**Reliability:** (2) va lid with restrictions

basic data given, acceptable restrictions

(308)

**Type of experience:** other: kinetics, inhalation

**Remark:** Urinary formic aci d was studied in fourteen workers exposed

to methanol with 10 +

- 5 years in their current occupation.

Exposure to methanol vapor at 200 ppm methanol produced 80

mg formic acid/g creatinine.

**Rel iability:**  (2) valid with restrictions

acceptable study, meets basic scientific principle

(309)

**Type of experience:** other: kinetics, inhalation

**Remar k:**  Four human subjects were exposed to approximately 200 ppm

of

methanol in a test chamber for six h. Ambient air in the

chamber was monitored for methanol, while urine was

monitored for formic acid. Mean urinary formic acid was

increased from baseline at the end of the exposure session,

but had returned to baseline in samples collected 16 h

followin g cessation of exposure.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(310)

**Remark:** Methanol is sl owly eliminated from the body; hence,

repeated

exposures result in an increasing concentration in blood

and tissue.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(311)

**Remark:** The aim of this study was to determine the reference values

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

229 for methanol in urine of the genera l population

non

- occupationally exposed to the xenobiotic under study.

Thus, urinary methanol was measured in 84 occupationally

non

- exposed subjects from the city of Sao Paulo, Brazil, by

means of headspace gas chromatography with a flame

ionization detector. The results revealed a mean of 2.26

+/

- 1.26 mg methanol/l urine and a geometric mean of 2.10 mg/l

for the studied population. The reference values varied

within the range of 0.50

- 4.78 mg/l (mean +/

- 2 S.D.).

Methanol levels in urine did not differ statistically

between male a nd female subjects. Urinary methanol in the

total population was less than 4.80 mg/l in 95% of cases.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(312)

**Remark:** One hundred and forty

- three workers exposed to one or more

of toluene, xylene, ethylbenzene, styrene, n

- hexane, and

methanol at sub

- occupational exposure limits were examined

for the time

- weighted average intensity of exposure by

diffusive sampling, and for biological exposure indicators

by means of analysis of shift

- end b lood for the solvent and

analysis of shift

- end urine for the corresponding

metabolite(s). Urinalysis was also performed in 20

nonexposed control men to establish the "background level."

Both solvent concentrations in blood and metabolite

concentrations in urine correlated significantly with

solvent concentrations in air. Comparison of blood analysis

and urinalysi s as regards sensitivity in identifying low

solvent exposure showed that blood analysis is generally

superior to urinalysis. It was also noted that estimation

of

exposure i ntensity on an individual basis is scarcely

possible even with blood analysis. Solvent concentration in

whole blood was the same as that in serum in the case of

the

aromati cs, except for styrene. It was higher in blood than

in serum in the case of n

- hexane, and lower in the cases of

styrene and methanol.

**Reliability:** (2) valid with restrictions

basic dat a given, acceptable restrictions

(313)

**Type of experience:** other: kinetics, model

**Remark:** Two patients with extremely high blood methanol

concen trations (260 and 282 mg/dl) were successfully

treated

using pharmacokinetic dosing of ethanol, hemodialysis and

supportive measures. Both patients recovered completely

wit hout residual ophthalmologic deficits. Early

hemodialysis

and inhibition of methanol metabolism with effective

ethanol

concentrations were attributed to the patients' full

recovery. Methanol elimination was enhanced by hemodialysis

as evidenced by a decrease in half

- life from eight to two

and a half hours. Methanol dialysance was 98 ml/min. A

dosage regimen for ethanol was devised, utilizing

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

230 dose

- dependent pharmacokinetic parameters and the ethanol

dialysance (100 to 120 ml/min) from these two patients. An

ethanol loading dose of 0 .6 g/kg should be administered to

an adult with an acute methanol ingestion. This dose will

produce a blood ethanol concentration of approximately 100

mg/dl which can be maintained by an ethano l infusion of 66

mg/kg/hour for nondrinkers to 154 mg/kg/hour for chronic

ethanol drinkers. Hemodialysis should be initiated if the

blood methanol concentration is greater than 50 mg/dl. If

hemodialysis is initiated, the ethanol infusion should be

increased by 7.2 g/hour.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(314)

**Type of experience:** other: kinetics, nutrition

**Remark:** Blood methanol concentrations were measured in 24

1

- year

- old

infants administered a spartame, a dipeptide methyl ester

sweetener. The doses studied included a dose projected to

be

the 99th percentile of daily ingestion for adults (34 mg/kg

body weight), a very high use dose (50 mg/kg body weight)

and a dose considered to be in the abuse range (100 mg/kg

body weight). Blood methanol values in infants were

compared

to values o bserved previously in adults administered

equivalent doses of aspartame. Methanol concentrations were

below the level of detection (0.35 mg/dl) in the blood of

10

infants a dministered aspartame at 34 mg/kg body weight, but

were significantly elevated (P less than or equal to 0.05)

after ingestion of aspartame at 50 and 100 mg/kg body

weight. At the latter doses, mean peak blood methanol

concentrations and the area under the blood methanol

concentration

- time curve increased in proportion to dose.

Mean (+/

- SEM) peak blood methanol concentration was 0.30

+/

- 0.10 mg/100 ml at a 50 mg/kg body weight aspartame dose

(n = 6) and 1.02 +/

- 0.28 mg/ml at the 100 mg/kg body

weight

dose (n = 8). Blood methanol values in infants wer e similar

to those observed in normal adults.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(315)

**Type of experience:** other: kinetics, nutrition

**Remark:** Blood methanol concentrations were measured in 30 normal

adult subjects administered aspartame, a dipeptide methyl

ester. The doses studi ed included the 99th percentile of

projected daily ingestion (34 mg/kg body weight) and three

doses considered to be in the abuse range (100, 150, and

200

mg/kg body weight ). Methanol concentrations were below the

level of detection (0.4 mg/dl) in the blood of the 12

normal

subjects who ingested aspartame at 34 mg/kg. They were

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

231 significantly elevated (p less than or equal to 0 .001)

after ingestion of each abuse dose, with the mean peak

blood

methanol concentrations and the areas under the blood

methanol concen tration

- time curve increasing in proportion

to dose. Mean (+/

- SD) peak blood methanol concentrations

were 1.27 +/

- 0.48 mg/dl at the 100 mg/kg dose, 2.14 +/

-

0.35 mg/dl at the 150 mg/kg dose, and 2.58 +/

- 0.78 mg/dl

at

the 200 mg/kg dose. Blood methanol concentrations returned

to predosing levels by 8 h after administration of the 100

mg/kg dose. Methanol was st ill detected in the blood 8 h

after the subjects had ingested aspartame at 150 or 200

mg/kg. Blood formate analyses were carried out in the 6

subjects who ingested aspartame at 200 mg/kg, since recent

studies indicate that the toxic effects of methanol are due

to formate accumulation. No significant increase in blood

formate concentrations over predosing concentrations was

noted. No changes were noted in any of the blood chemistry

profile parameters measured 24 h after aspartame ingestion,

compared to values noted before administration. Similarly,

no d ifferences were noted in ophthalmologic examinations

carried out before and after aspartame loading.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(316)

**Type of experience:** other: review, acute toxicity

**Remark:** The fatal dose in humans is between 2 and 8 ounces (1 ounce

= 28,35g).

**Reliability:** (2) val id with restrictions

see remark on selection of literature

28

- DEC

- 2004 (317)

**Type of experience:** other: review, acute toxicity

**Remark:** Most serious cases of methanol poisoning that have been

reported (many fatal; others involved permanent or

temporary

loss of vision) resulted from the ingestion of methanol in

the belief that i t was ethanol. Ocular disturbance and

blindness in humans resulting from repeated rubbing of the

skin with methanol was also reported.

**Reliability:** (2) valid with restrictions

see rema rk on selection of literature

(135)

**Type of experience:** other: review, acute toxicity

**Remark:** The signs and symptoms most characteristic of methanol

poisoning are visual disturbance and metabolic acidosis.

Permanent damage (residual scotomata) can result even if

complete blindness is avoided.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

(318)

**Type of experience:** other: review, acute toxicity

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

232 **Remark:** Methanol poisoning with typical disturbed vision may appe ar

in the occupational setting, by ingestion after drinking of

methanol instead of ethanol and continous inhalation when

handling the hot product.

**Reliability:** (2) valid with restricti ons

see remark on selection of literature

(319)

**Type of experience:** other: review, acute toxicity

**Remark:** Methanol intoxication causes severe me tabolic acidosis and

can lead to permanent visual damage or death. Methanol,

readily available in common products like antifreeze, is

ingested accidentally or deliberately as a substitute for

ethanol and in suicide attempts. Because it may become

major

fuel source in the 21st century and because industrial uses

are expanding, deliberate and accidental intoxicatio n is

likely to increase. Rapid diagnosis is essential so that

appropriate treatment can be instituted quickly. The

authors

review the pharmacology, clinical and laboratory findings,

and pathology and pathophysiology of methanol intoxication.

In addition, they discuss the differential diagnosis and

treatment of acute intoxication, including the use of

4

- methylpyrazole in preventing the conversion of methanol

to

formate.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(320)

**Type of experience:** other: review, acute toxicity

**Remark:** A 41

- year

- old man ingested orally a large quantity of

methanol and was found dead at home. The presenc e of

methanol in body fluids and tissues was determined by

head

- space gas chromatography. The blood ethanol and

acetone

were negative. Tissue distribution of methanol showe d that

the kidney presented the highest content of methanol (5.13

g/kg) followed by liver (4.18 g/kg), vitreous humor (3.96

g/l), heart (3.45 g/kg), urine (3.43 g/l), pericardial

fluid

(3.29 g/l), blood (2.84 g/l) and finally stomach content

(2.21 g/l).

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(321)

**Type of experience:** other: review, acute toxicity

**Remark:** Methanol ingestion is an uncommon form of poisoning that

can

cause severe meta bolic disturbances, blindness, permanent

neurologic dysfunction and death. While methanol itself may

be harmless, it is converted in vivo to the highly toxic

formic acid. The diagnosis is somet imes elusive and

requires

a high index of suspicion. Because antidotal treatment is

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

233 available it is important to recognize methanol poisoning

promptly. The presence of meta bolic acidosis associated

with

an increased anion gap and increased osmol gap are

important

laboratory findings. Specific therapeutic measures include

correction of the metabolic acidosis with sodium

bicarbonate

and administration of enteral or parenteral ethanol to

competitively inhibit the metabolic breakdown of methanol

to

formic acid. Hemodialysis accelerates the elimination of

both methanol and formic acid and also assists in

correction

of the metabolic acidosis. Experimental data sugg ests that

administration of folic acid may be of benefit by hastening

the metabolism of formic acid to carbon dioxide. Prompt

institution of specific therapy can probably decrease the

morbidity and mortality associated with this form of

poisoning.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(322)

**Type of experience:** other: review, acute toxicity

**Remark:** Review of methanol toxicology incl. composition,

use/sources, acute toxicity, chronic toxicity, target organ

toxicty, absorption

- metablosim

- excretion, genotoxicty,

neurotoxicity, reproductive toxicity, carcinogenicity,

epidemiology, environmetnal fate, environmental toxicity,

and regulatory status

. **Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(152)

**Type of experience:** other: review, acute toxicity

**Remark:** There have been about one hundred cases of amblyopia and

death from inhalation of wood alcohol recorded in the

literature to 1912.

**Reliability:** (2) valid with restrictions

basic data given, acceptable rstrictions

(323)

**Type of experience:** other: acute toxicity

**Remark:** Case report: a 26

- year

- old woman ingested 250 to 500 ml

methanol during the 38 th week of pregnancy. The initial

serum methanol conc. was 230 mg/dl and formate was 33.6

mg/dl. A mild metabolic acidosis was present. Therapy

included ethano l infusion, bicarbonate administration and

three courses of hemodialysis. Delievery occurred six days

after methanol exposure, when methanol was no longer

detected in maternal blood. No further complications were

noted in the mother and her newborn.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(324)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

234 **Type of experience:** other: acute toxicity

**Remark:** Case report of a 38

- year

- old male developing Parkinson

disease after methanol intoxication.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(325)

**Type of experience:** other: acute toxicity

**Remark:** 64 cases of inhalation toxicity were reported. 6 of them

died, 16 were permanently blind, 25 had permanent

impairment of vision, and 8 were temporarily blind.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(326)

**Type of experience:** other: acute toxicity

**Remark:** Clinical and pathologic observations in a group of 320

patients with acute methanol poi soning after ingestion of

bootleg whiskey containing 35 % methanoal and 15 % ethanol

were reported. Amount ingested ranged from 30 ml to 260 ml

methanol. There were 37 deaths. Visual symptoms, occuring

eighteen to forty

- eight hours after ingestion, included

blurred vision, loss of central vision and complete

blindness. Acidosis was present in every severely poisoned

patient. Partial or complete recovery of the initially

reduced visual acuity was observed in most patients.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(327)

**Type of experience:** other: acute toxicity

**Remark:** In four of six patients with methanol intoxication necrotic

changes in the area of putamina were s een. Haemorhages were

also seen in some cases.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(328)

**Type of experience:** other: acute toxicity

**Remark:** 14 patients whose vision was affected by methanol

intoxication were evaluated by a standard

electroretinographic technique. Some increase in m ean

latencies and peak times appeared and the a

- and b

- waves

amplitude to all parameters od stimuli were significantly

below normal range. The changes in the ERG indicate damage

to the outer retinal layers including visual cells. The

usual

ERG observed in methanol poisoning is unlike that found

with

degeneration of the potic nerve fibre or ga nglion cell

layer, and is also unlike that found when damage extends to

the bipolar layer.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(329)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

235 **Type of experience:** other: acute toxicity

**Remark:** A 54

- year

- old woman developed acute necrotizing

pancreatitis

following acute methanol poisoning. She died from multiple

organ failure 54 hours after the ingestion. In a series of

22 consecutive patients admitted with a diganosis of acute

methanol poisoning, evidence of pancreatic damage was found

in 11 patients.

**Reliability:** (2) valid with restrictions

basic data, acceptable restrictions

(330)

**Ty pe of experience:**  other: acute toxicity

**Remark:** Occupational methanol poisoning has frequently caused death

or blindness. Several cases resulted from work in confined

spaces, e.g., varnishing beer va ts. Headache and blurred

vision were reportedly frequent symptoms. It is believed,

that absorption of 8 grams would seriously affect the eyes

and that such a dose could result from inhalation o f 800 to

1000 ppm for 8 hours. Work room concentration of 500 to 600

ppm were found. It is recommended to keep the levels below

1

ppm.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(331)

**Type of experience:** other: acute toxicity

**Remark:** In a suicidal attempt by a 13

- yea r old white girl, methanol

(90

- 240 ml of an antifreeze containing 60 % methanol,

39.25 % potassium phosphate) produced classical immediate

symptoms and permanent damage to the central nervous system

characterized by severe bilateral optic atrophy, rigidity,

spasticity, and hypokinesis.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(332)

**Type of experience:** other: acute toxicity

**Remark:** In two case reports, the characteristic clinical,

computertomograph

- scan and neuropathologica l findings in

acute methanol intoxication are described. The significance

of acidosis for the survival is stressed. After a latency

period of several days, one patient developed the rare

condition of haemorhagic leucocephalopathy with striking

computertomograph

- scan and morphologic findings.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(333)

**Type of experience:** other: acute toxicity

**Remark:** A case is presented of a consultant supervising tank

cleaning prior to methanol loading. He wore positive

pressure breathing apparatus but no protective clothing.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

236 After 2

- 3 hours working in the confined space of the tank,

he worked on deck and continued to wear his methanol

- soaked

clothing which eventually dried out. Visual symptoms of

acute methanol toxicity presented some 8 hours after

exposure. The approriate treatment (with ethanol provided

by

the ship bond) was carried out in the hospital and the

individual recovered completely.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(334)

**Type of experience:** other: acute toxicity

**Remark:** Case report: loss of vision after chronic inhalation and

skin resorption of methanol.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(335)

**Type of experience:** other: acute toxicity

**Remark:** The main toxi c effects of methanol is exerted upon the

nervous system, particularly the optic nerves and possibly

the retinae. The condition can progress to permanent

blindness. Once absorbed methanol is on ly very slowly

eliminated. Coma resulting from massive exposures may last

as long as 2

- 4 days. In the body, the products formed by

its

oxidation are formaldehyde and formic acid, both of which

are toxic. Because of the slow elimination, methanol should

be regarded as cumulative poison. Though single exposures

to

fumes may cause no harmful eff ect, daily exposure may

result

in the accumulation of sufficient methanol in the body to

cause illness. Death from ingestion of less than 30 ml has

been reported. Human sys temic effects: changes in

circulation, cough, dyspnea, headache, lacrimation, nausea

or vomiting, optic nerve neuropathy, respiratory effects,

visual field changes.

**Reliability:** (2) va lid with restrictions

basic data given, acceptable restrictions

(336)

**Type of experience:** other: acute toxicity

**Remark:** Cardinal features in teh mechanism, pathology,

sympthomatology and therapy of methyl alcohol poioning are

reviewed in the light of a series of 23 cases.

The first five of these patients died of respiratory arrest

after several hours of coma. They were sporadic cases who

had ingested relatively hugh amounts of both ethyl and

methyl alcohol. The sixth case was brought to the hospital

in p rofound shock and died within four hours. The remaining

17 cases were all victims of the same batch of liquor that

had killed the six patient, and were treated with alkalies.

None died or susta ined permanent injury.

The two basic mechanisms of poisoning are direct chemical

irritation of tissues and systemic acidosis. The former is

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

237 influenced by the unlimited miscibility of methanol w ith

water and the distribution of water in the tissues of the

body. Acidosis is the most important therapeutic

consideration. Alkalinization should be prompt and

vigorous.

It is suggested that ketosis may play a role in acidosis

and

depression, and that intravenous glucose may be an

important

adjunct to treatment.

**Reliabi lity:**  (2) valid with restrictions

basic data given, acceptable restrictions

(337)

**Type of experience:** other: acute toxicity

**Remark:** A s eries of 49 cases of acute methyl alcohol poisoing with

recovery is reported (no demonstrable eye lesion after five

months). Gastic lavage was not used in any case, because it

is of questionabl e value when poisoning has occurred.

Plasma

extenders were found valuable in combating peripheral

vascular failure. Frequent drainage of the cerebrospinal

fluid does not ap pear essential. The occurrence of

hypopotassaemia during management was definetely

established.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(338)

**Type of experience:** other: acute toxicity

**Remark:** The clincial course of 4 men who drank commercial

duplicating fluid containing 40 % to 60 % m ethyl alcohol is

presented. One of the patients died (ingestion of about 12

ounces), and the necropsy findings were severe, acute

passive congestion of the lungs, kidneys, spleen, liver and

brain, as well as congestion with petechial hemorrhages of

the stomach and segments of the jejumum. Histologic

examination was compatible with the cross findings. The

other me n made a complete recovery.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(339)

**Type of experience:** other: acute toxicity

**Remark:** Report of a common

- source epidemic of methanol poisoning

which involved 18 people, of whom eight died. Of the 18

known cases, 11 were treated with only one death despite

several patients with methanol concentrations of 200 mg or

more per 100 ml blood. Among different types of treatment

used peritoneal dialysis was shown to be effective, with a

clearanc e rate five to ten times that obtained with forced

fluid diuresis.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(340)

**Type of experience:** other: acute toxicity

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

238 **Remark:** Case report of a intoxication in a 55

- year

- old woman after

administration of methanol

- soaked patches. Initial symtome

were blurred vision, decrease of vision, and neuritis of

the

optic nerve. Decrease of vision was permanent.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(341)

**Type of experience:** other: acute toxicity

**Remark:** Report of a child who died after methanol intoxication,

presumably due to p ercutaneous resorption. Methanol soaked

pads had been applied to the child ́s chest for appr. twice

12 hours.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restr ictions

(342)

**Type of experience:** other: acute toxicity

**Remark:** Case report of fuzzy and heavy feeling in the head and

tendency to sway in a wor ker engaged in antimold spraying

to

mildewproof a living room. The antimold agent was diluted

with methanol or ethanol. Since the work generally

continues

for several days, signs and symptoms had persisted.

However,

within 10 days of completing the work, these signs any

symptoms completely disappeared. Examination of the urine

disclosed that methanol in urine was in the normal range

ten

days after discontinuation of the work.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restric tions

(343)

**Type of experience:** other: acute toxicity

**Remark:** Case report on a 50

- year

- old man with methanol conc. of 5

mg/dl after ingestion of alcoholic beverages. The ethanol

level was 275 mg/dl.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(344)

**Type of experience:** other: acute toxicity

**Remark:** Methanol poisoning accounting for several deaths were

investigated in an Ontario registry. Between 1986 to 1991

43 fatalities were fou nd. Suicide attempts account for 21

cases while 22 were classified as accidental. 14 of these

consumed products labelled as methanol or wood alcohol as a

substitue for ethanol. 3 cases ingested methanol being

improperly stored in containers normally associated with

ethanol, and 5 patients were poisoned through consumption

of

liquor from illicit sources.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

239 **Reliabili ty:**  (2) valid with restrictions

basic data given, acceptable restrictions

(345)

**Type of experience:** other: acute toxicity

**Remark:** A pat ient poisoned with methanol was successfully

hemodialyzed with an ethanol

- enriched, bicarbonate

- based

dialysate. Along with a concomitant intravenous infusion of

ethanol, the ethanol

- enriched d ialytic procedure was able

to

maintain an intradialytic plasma ethanol level of 80 to 102

mg/dl. The patient recovered without any sequelae of

methanol intoxication.

**Reliab ility:**  (2) valid with restrictions

basic data given, acceptable restrictions

(346)

**Type of experience:** other: acute toxicity

**Remark:** Ob servations in 323 patients who had ingested bootleg

whiskey containing 35

- 40 % methanol and 4 % ethanol were

reported. 41 deaths occurred. 115 patients showed acidosis

and visual disturbances w hen first seen in the hospital.

Lowest letal dose was 6 ml methanol and highest dose

survival

was 200 ml methanol.

**Reliability:** (2) valid with restrictions

accepta ble study, meets basic scientific principles

(347)

**Type of experience:** other: acute toxicity

**Remark:** 21 cases of percutaneous methanol intoxication in children

were reported. 12 of these children died. The children ́s

cloths soaked in methanol were applied overnight and during

the day with a rate of one to three applications to each

chi ld. Th elapsed time between application and onset of

symptoms varied from 1 to 13 h and the time till diagnosis

varied from 4 to 48 h. The early manifestations were

centralnervous system depres sion and in some cases

hyperpnea

and in some cases severe respiratory depression.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(348)

**Type of experience:** other: acute toxicity

**Remark:** Report of 54 cases of death and blindness from cilombian

spirits and other methylated preparation s. As to the

general

effect: headache was mentioned in 19 cases as a conspicous

symptom; gastric pain in 11; nausea and vomiting in 26;

dilated pupils in 20. the results as to visual disturbance

were 16 total blindness, 3 total blindness of one eye, 15

partial recoveries, 7 recoveries; 10 remaing cases teminted

fatally; sight became dim in three hours in one case , six

hours in 1 case; twelve hours in 2 cases; twenty hours in 1

case; twenty

- four hours in 19 cases; forty

- eight hours in 5

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

240 cases; three days in 2 cases; six days in 1 case; seven

days

in 1 case; sight became lost in twelve hours in 2 cases, in

twenty

- four hours in 10 cases; in thrity hours in 2 cases;

in forty

- eight hours in 3 cases; in three days in 3 cases;

in four days in 3 cases; in five days in 2 cases; in six

days in 1 case; in seventeen days in 1 case. Of the

remaining 21 facts in this connection are not definite.

**Reliability:** (2) va lid with restrictions

acceptable study, meets basic scientific principles

(349) (350)

**Type of experience:** other: acute toxicity

**Remark:** Significant to xicity can result from intentional methanol

inhalation. We report seven cases, involving four patients,

of intentional inhalation of CARB

- MEDIC carburetor cleaner

containing toluene (43.8%), me thanol (23.2%), methylene

chloride (20.5%), and propane (12.5%). Patients arrived at

the emergency department with central nervous system

depression, nausea, vomiting, shortness of breath,

photophobia, and/or decreased visual acuity. Treatment

included correction of acidosis, leucovorin and/or folic

acid, ethanol infusions, and supportive care. Hemodialysis

was n ecessary in three cases. Measured blood methanol

levels

ranged from 50.4 to 128.6 mg/dL. Blood formic acid levels

were 120, 193, and 480 micrograms/mL, respectively, in

thr ee

patients. Ophthalmic examinations revealed hyperemic discs

and decreased visual acuity in one patient. One individual

was found pulseless with several CARB

- MEDIC cans nearby.

Attempts at revival were unsuccessful.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(351)

**Type of experien ce:**  other: acute toxicity

**Remark:** In a large epidemic it was demonstrated that patients

acutely intoxicated with methanol can be triaged

effectively

on the basis of bloodmethanol level, serum bicarbonate

concentration, and assessment of specific clinical

complications. Additional findings such as elevated MCV and

hematocrit may prove useful as markers for nonspecific

toxic

swelling of cells. Ehtanol should be included in the

treatment of all suspected cases, using doses that will

maintain a blood ethanol level of 100 mg/dl until methanol

levels fall to the range of 20 mg/dl.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(352)

**Type of experience:** other: acute toxicity

**Remark:** A 46

- year

- old male presented to the emergency room seven

hours after consuming a large container of sterno. He could

not see and complained of abdominal and back pain. He was

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

241 tachypneic, tachycardic, hypertensive and hypothermic.

Laboratory results were significant for a severe metabolic

acidosis, a serum osmolality of 465 and seru m methanol

level

of 493 mg/dl. Aggressive treatment included ethanol drip,

bicarbonate and hemodialysis. He survived and regained his

eyesight in spite of this degree of el evation of the serum

methanol level.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(353)

**Type of experience:** other: acute toxicity

**Remark:** A 31

- year

- old previously healthy man presented with a 7

- day

history of nausea, intractable vomiting, diffuse abodminal

pain, and progressive b lurred and tunnel vision. The

patient

had visual disturbances and an osmolal gap that suggested

methanol intoxication.

**Reliability:** (2) valid with restrictions

bas ic data given, accpetable restrictions

(354)

**Type of experience:** other: acute toxicity

**Remark:** A 27

- year

- old man admitted to the medical intensive care

unit. A coworker found him standing in the bathroom,

unresponsive and rigid. The patient received standard

emergency department treatment for the unconscious patient

with suspected meth anol poisoning with negative results.

The

, patient had seizure activities with dilated pupils,

C lenched aw, and tonic muscle actitivity. Deep tendon

re flexes wereabsent symetrically. Neurologic al assessment

showed a severely depressed level of consciousness. Initial

blood levels showed a methanol concentration of 234 mg/dl

and an ethanol level of 74 mg/dl.

**Reliability:** (2) v alid with restrictions

basic data given, acceptable restrictions

(355)

**Type of experience:** other: acute toxicity

**Remark:** A 44

- year

- old man was f ound unconscious beneath an elevated

rapid transit right

- of

- way. On admission to the emergency

room, the patient was comatose in metabolic acidosis with

high anion and osmolal gaps. The serum m ethanol was 583

mg/dL. The serum ethanol and ethylene glycol were negative.

The patient was treated with ethanol, bicarbonate, and

hemodialysis. He expired 40 h after admission. The

postmortem methanol concentrations in body fluids were as

follows: bile 175 mg/dL, vitreous humor 173 mg/dL, and

blood

142 mg/dL. Urine was not available for analysis. Postmortem

methanol concentrations in body tissues are given in

decreasing order: brain 159 mg/100 g, kidney 130 mg/100 g,

lung 127 mg/100 g, spleen 125 mg/100 g, skeletal muscle 112

mg/1 00 g, pancreas 109 mg/100 g, liver 107 mg/100 g, and

heart 93 mg/100 g. The total amount of methanol in the

gastric contents was 73 mg.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

242 **Reliability:** (2) valid with restrictions

accepta ble study, meets basic scientific principles

(356)

**Type of experience:** other: acute toxicity

**Remark:** Methyl ethyl ketone is a common solvent but data on

overdose

in humans are scarce. We report a case of co

- ingestion of

methyl ethyl ketone together with methanol associated with

a

hyperosmolar coma without anion gap metabolic acidosis.

Blood levels of methyl ethyl ketone and its metabolite,

2

- butanol, indicated that this solvent did contribute

approximately 20 mosm/L to an observed osmolar gap of 99

mosm/L. At the levels detected, methyl ethyl ketone may

have

inhibited methanol metabolism, contributing to the low

serum

formate (1.3 mmol/L) and normal an ion gap despite a blood

methanol of 67 mmol/L.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(357)

**Remark:** In 10 adult patients acutely intoxicated with methanol, the

base deficit and the total blood CO2 were highly correlated

with blood formate level. No correlation was found between

the same parameters and blood methanol level. Formate

production is the main cause of acidosis in the early

stages

of methanol poisoning. The association of a latency period

before treatment exceeding 10 hours and a blood formate

level above 0.5 g/l (11.1 mmol/l) is predictive of severe

methanol poisoning possibly leading to permanent sequelae.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

28

- DEC

- 2004 (358)

**Type of experience:** other: acute toxicity, cns

**Remark:** Subarachnoid hem orrhage developed in a patient intoxicated

with methanol. Computed tomography performed at the time of

admission suggested this complication. The hemorrhage was

definite and extensive by the 5t h day after admission and

was accompanied by left caudate and pontine hemorrhage, as

well as severe cerebral edema.

**Reliability:** (2) valid with restrictions

basic data given, acceptabl e restrictions

(359)

**Type of experience:** other: acute toxicity, cns

**Remark:** Bilateral putaminal necrosis is characteristic of methanol

poisoning . A 31

- year

- old male alcoholic had headache,

impaired consciousness, neck stiffness, roving eyes with

dilated unreactive pupils, papilloedema, abdominal pain,

vomiting, and severe metabolic aci dosis after a binge.

Abnormalities of the cerebrospinal fluid included an

initial

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

243 pressure of 240 mmH2 O, RBC 286/mm3, WBC 8/mm3, and protein

179 mg/dl. Peritoneal dialysis was performed on the 2nd day

after drinking. A blood test for methanol was not performed

until the 5th day, and its results was negative. However,

computed tomography (CT) on the 3rd day showe d necrosis and

hemorrhage of bilateral putamina and the cerebral cortex,

and post

- contrast enhancement of meninges. On the 22nd day,

a CT revealed further changes: necrosis of bilateral

subcortical white matter, and post

- contrast gyral

enhancement at the otherwise normal

- looking areas of the

cerebral cortex.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(360)

**Type of experience:** other: acute toxicity, cns

**Remark:** A 37

- year

- old man with a history of aclohol abuse was

brought to the emergency department 1 day after having

consumed approximately one half gallon of windshield washer

fluid. The patient was found to be unresponsive, with

shallow respirat ions. Toxicological screening revealed a

blood methanol level of 331 mg/dl (103 mmol/) and an

ethanol

level of 0 mg/dl. MR imaging showed necrosis of the

putamen.

**Reliabili ty:**  (2) valid with restrictions

basic data given, acceptable restrictions

(361)

**Type of experience:** other: acute toxicity, cns

**Remark:** In two survivors of severe methanol poisoning apart from

blindness Parkinsonian

- like extra

- pyramidal syndrome

characterized by poor voice volume, masked face, mild

tremor, rigidity, bradykinesi a, hyperreflexia, bilateral

Babinski responses, and mild dementia were reported. CT

scan

in both patients demonstrated symmetrical low density

lesion

i n the fronto

- central white matter and putamen bilaterally

EMG showed extensive denervation, mainly involving the

legs,

but normal motor conduction velocities. Autopsy in one

patient who died from suicide 11 months following poisoning

refealed cystic resorption of the putamen and the

peripheral

white matter in addition to widespread neuronal damage

throughout cerebrum, cerebellum, brainstem, and spinal

cord.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(362)

**Type of experience:** other: acute toxicity, cns

**Remark:** Five optic nerves from three cases of methanol intoxication

were examined. The patients died in cardiorespiratory

fai lure thirty hours, two days, and eighteen days,

respectively, after presentation with acute bilateral

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

244 visual loss. The retrolaminary nerve segments showed

circumscribed

dem yelination of loss of myelin staining with relative

preservation of axons. Retinal ganglion cells were spared.

Electromicroscopy in the case with shortest survival

revealed periaxonal intramyel inic spaces behind the lamina

cribosa.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(363)

**T ype of experience:**  other: acute toxicity, cns

**Remark:** The histopathologic effects of methanol on the optic nerve

were studied in four patients. Circumscribed myelin damage

occurred behind the lamina cribrosa in each nerve. Axons

were preserved. Demyelination also occurred in cerebral

hemispheric white matter in one patient. This selective

myelinoclastic effect of methanol metabolism is pro bably

caused by histotoxic anoxia in watershed areas of the

cerebral and distal optic nerve circulations. Juxtabulbar

demyelination may cause optic disk edema in methanol

p oisoning by compressive obstruction of orthograde

axoplasmic flow. Visual loss may be due to disruption of

saltatory conduction. Retrolaminar demyelinating optic

neuropathy is an early morpholo gic correlate of visual loss

in methanol intoxication.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(364)

**Type of experience:** other: acute toxicity, cns

**Remark:** Case report of a 40

- year

- old woman with putaminal necrosis

after methanol intoxication.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(365)

**Type of experience:** other: acute toxicity, cns

**Remark:** Case report of a 62

- year

- old plumber with putamen nec rosis

after severe methanol intoxication.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(366)

**Type of experience:** other: acute toxicity, cns

**Remark:** Case report of a case of acute methanol intoxication with

central nervous system damage producing a Parkinson

syndrome

wi th persitent sequel.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(367)

**Type of experience:** other: acute toxici ty, cns

**Remark:** Report of with areas of decreased attenuation symmetrically

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

245 localised, especially in the putamen in the computerised

tomography in a 62

- year

- old plumber after intake of

methanol

- containing home

- made liquor.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(368)

**Type of experienc e:**  other: review, therapy

**Remark:** Review on the roles of alkaline salts and ethyl alcohol in

the treatment of methanol poisoning.

**Reliability:** (2) valid with restrictions

see remark on selec tion of literature

(369)

**Type of experience:** other: review, therapy

**Remark:** Treatment of methanol poisoning is well established and

consists spe cifically of the administration of alkali and

ethanol, and hemodialysis. Since methanol poisoning often

occur as accidents with a lot of patients involved, it is

not always possible to follow s ophiticated protocols for

treatment. If the patients are awake with no severe

hyperventilation or acidosis, ethanol adminstration is most

important because it is widely available and can be giv en

orally. Under such cicrumstances every person suspected of

having consumed some of the suspect liquid should be given

ethanol orally or intravenously aiming at a blood

c oncentration of about 22 mmol/l. In severely poisoned

patients in coma with pronounced acidosis and

hyperventilation it is vital to correct the acidosis as

soon

as possible . Before any laboratory analyses are available,

at least 200 to 300 mmol of bicarbonate should be infused

intravenously, followed by ethanol infusion. If methods of

determination of methanol or formate are not available, the

magnitude of the osmolal and anion gaps along with the

clinical judgement should provide the basis for treatment.

**Reliability:** (2) valid with restrictions

see remark on selectionof literature

(370)

**Type of experience:** other: review, therapy

**Remark:** Twelve cases of methanol poisoning are reviewed. The

clinical presentation and biochemical features are

described

and the results of treatment with alkali, ethanol and

dialysis reported. The outcome of methanol poisoning

appears

to be related more to the interval between the time of

ingestion and the start of therapy and to the degree of

acidosis than to the initial serum methanol level.

Therefore, early and aggressive treatment with bicarbonate

and ethanol and subsequent institution of hemodialysis are

strongly recommended whenever methanol can be detected in

the blood, especi ally when metabolic acidosis of the

anion

- gap type is present, when mental or visual

disturbances are present, or when more than 30 ml of

absolute methanol has been consumed.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

246 **Reliability:** (2) valid with restrictions

see remark on selection of literature

(371)

**Type of experience:** other: review, therapy

**Remark:** A unique opportun ity presented itself for reviewing and

comparing the results of two different methods of dialytic

therapy for the treatment of methanol poisoning. The group

that was treated with hemodialysis i nitially had a faster

fall in serum methanol level, regained consciousness

faster,

had a shorter hospital stay, and most importantly, had no

residual effects when compared with the group treated with

peritoneal dialysis. In this group, one patient died and

another was permanently blinded.

**Reliability:** (2) valid with restrictions

see remark on selection o f literature

(372)

**Type of experience:** other: therapy

**Remark:** A case report of severe methanol poisoning is reported

which

required 21 hours of hemodialysis to bring their serum

methanol levels down to a nontoxic level.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(373)

**Type of experience:** other: therapy

**Remark:** Nine patients with methyl alcohol poisoning who were

treated

with hemodialysis were described. The time from ingestion

to

dialysis varied from 4 to 100 hours. Predialysis blood

methanol levels ranged from 3 to 570 mg/dl. All patients

were acidotic and had an increased an ion gap. Two patients

died,

seven recovered, but three had permanent visual impairment.

There was little correlation between the blood methanol

level or anion gap and visua l outcome. The interval from

ingestion to treatment appears to be more important than

the

initial biochemical status. Prompt hemodialysis is

recommended, if the blood metha nol level is above 50 mg/dl,

when an amount of methanol exceeding the minimal lethal

dose

(30 ml) is known to have been ingested, when there is

evidence of acidosis or when an abnormality has developed

in

vision, funduscopic examination or mental state. Concurrent

therapy with alkali and ethanol is vital.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(374)

**Type of experience:** other: therapy

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

247 **Remark:** The diagnostic value of determination of the anion and

osmolal gaps was studied in 6 patients poisoned with

methanol and in 5 poisoned with ethylene glycol. Increased

osmolal gap was present on admission in all patients,

wh ereas increased anion gap was present in all except one.

In the methanol

- poisoned patients the mean blood values

were: pH 7.27, anion gap 24 mmol/l, osmolal gap 81

mosmol/kg

H2O, methanol 67 mmol/l, ethanol 11 mmol/l and in the

ethylene glycol

- poisoned patients: pH 6.93, anion gap 38

mmol/l, osmolal gap 35 mosmol/kg H2O and ethylene glycol 24

mmol/l. In the absen ce of alcoholic acidosis or diabetic

coma the finding of a simultaneous increase in both the

anion and osmolal gaps indicates methanol or ethylene

glycol

poisoning. Thus de terminations of the anion and osmolal

gaps

are mandatory whenever facing metabolic acidosis of unknown

etiology.

**Reliability:** (2) valid with restrictions

acceptabl e study meets basic scientific principles

(375)

**Type of experience:** other: therapy

**Remark:** 4

- Methylpyrazole (4

- MP), an inhibitor of alcohol

dehy drogenase, may be useful for the treatment of methanol

and ethylene glycol intoxications. A placebo

- controlled,

double blind, multiple dose, sequential, ascending

- dose

study has been performed to determine the tolerance of 4

- MP

in healthy volunteers. Oral loading doses of 4

- MP were

followed by supplemental doses every 12 h through 5 days,

producing plasma levels in the therapeutic ra nge. A slight,

transient elevation in one or both serum transaminase

values

was observed in 6 of the 15 subjects treated with 4

- MP.

This

effect was not dose related nor apparently mediated through

a hypersensitivity reaction. Serum triglyceride levels were

increased in 30% of 4

- MP treated subjects, but also in 25%

of the placebo subjects. 4

- M P treatment did not produce any

other significant changes in objective clinical parameters

nor in subjective side effects. The results suggest that a

mild, transient increase in liver function tests might be

observed in some subjects treated with multiple doses of

4

- MP. Nevertheless, the slower elimination rate and lesser

degree of toxicity of 4

- MP would make it preferable to

ethanol in therapy of these poisonings.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(376)

**Type of experience:**  other: therapy

**Remark:** 1. Osmolal and anion gaps are helpful in the diagnosis and

evaluation of intoxications with methanol and ethylene

glycol. Reported reference values for osmolal gap and anion

gap are

- 1 (+/

- 6) mosm kg

- 1 H2O and 16 (+/

- 2) mmol l

- 1,

respectively. However, we have repeatedly found unexplained

increased gaps in patients admitted to our department, and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

248 the relevance of the established reference values has been

questioned. 2. Osmolal and anion gaps were determined in an

unselected population of patients consecutively admitted to

an emergency medical department. In the case of unexplained

gaps, the blood samples were analysed with respect to the

presence of alcohols and organic acids. 3. We included all

accessible pa tients admitted during 14 days. Appropriate

blood samples were obtained in 177 patients (88 male, 89

female), with a mean age of 65 years (range 17

- 94). 4. The

mean and (standard deviation) for osmolal and anion gaps in

our material were 5.2 mosm kg

- 1 H2O (7.0) and 12.9 mmol/l

(4.2). Neither methanol nor ethylene

- glycol was detected in

serum from any patients. Small amounts of ethano l were

found

in 5 patients, and high lactate levels explained in part

the

most extensively increased anion gaps. However, the

calculated analytical sta ndard deviation accounted entirely

for the variation in the material.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(377)

**Type of experience:** other: therapy

**Remark:** A patient intentionally ingested an unknown amount of

methanol and was admitted to the hospital 6 h later. On

admission, the m ethanol concentration in blood was

estimated

as approximately of 134 mmol/L, based on the calculation of

the osmolal gap. Intravenous ethanol administration and

hemodialysi s were promptly started. During hemodialysis,

several blood samples were collected for determination of

methanol and ethanol concentrations. Initially, we used gas

chromatography with split

- mod e injection of pretreated

serum

samples; however, methanol concentrations turned out to be

significantly lower than expected, based on calculated

osmolal gap values. Becaus e no explanation for the excess

serum osmolal gap was apparent, we reanalyzed samples,

using

head

- space gas chromatography. The methanol concentrations

measured were signif icantly higher and osmolal gap values

were no longer excessive.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(378)

**Type of experience:** other: therapy

**Remark:** 4

- Methylpyrazole (4

- MP), an inhibitor of alcohol

dehydrogenase, may be useful for the treatment of methanol

and ethylene glycol into xications. A placebo

- controlled,

double blind, multiple dose, sequential, ascending

- dose

study

has been performed to determine the tolerance of 4

- MP in

healthy volunteers. Oral loading doses of 4

- MP were

followed

by supplemental doses every 12 h through 5 days, producing

plasma levels in the therapeutic range. A slight, transient

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

249 elevation in one or both serum transaminase values was

observed in 6 of the 15 subjects treated with 4

- MP. This

effect was not dose related nor apparently mediated through

a hypersensitivity reaction. Seru m triglyceride levels were

increased in 30% of 4

- MP treated subjects, but also in 25%

of the placebo subjects. 4

- MP treatment did not produce any

other significant changes in objective clinical parameters

nor in subjective side effects. The results suggest that a

mild, transient increase in liver function tests might be

observed in some subjects treated with multiple doses of

4

- MP. Nevertheless, the slower elimination rate and lesser

degree of toxicity of 4

- MP would make it preferable to

ethanol in therapy of these poisonings.

**Reliability:** (2) valid with restricti ons

basic data given, acceptable restrictions

(376)

**Type of experience:** other: irritation

**Remark:** In an experimental study, 12 volunteers were e xposed in an

inhalation chamber to 20 and 200 ppm methanol. At 200 ppm

(20 ppm) 5 (3) volunteers reported slight irritation of the

nose and 6 (3) of the throat.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(379)

**Type of experience:** other: irritation

**Remark:** From 1989

- 1993 two workers wit h local skin

- and eye

irritation, as well as malaise, after accidental local and

inhalative exposure to methanol were send to the local

clinic for further treatment.

**Reliability:** (2) v alid with restrictions

basic data given, acceptable restrictions

(380)

**Type of experience:** other: irritation

**Remark:** Local eye contact with liqu id methanol resulted in severe

chemosis and lesions of the cornea, which however receded

in

a few days when properly treated.

**Reliability:** (2) valid with restrictions

acceptable study, meets basic scientific principles

(381)

**Type of experience:** other: sensitisation, skin

**Remark:** In four patients with eczema positive patch

- test reactions

to methanol (10 %) were described.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(3 82)

**Type of experience:** other: review, chronic toxicity

**Remark:** Cases of chronic poisoning from repeated exposure to

methanol

vapor were manifested by conjunctivitis, headache,

giddiness, insomnia, gastric disturbance, and bilateral

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

250 blindness. In one fatal case of occupational methanol

intoxication by inhalation a female worker was exposed

about

1 2 hours. A postevent study of the process evealed methanol

vapor concentrations ranging from 4,000 to 13,000 ppm.

**Reliability:** (2) valid with restrictions

see remark on selection of literature

(383)

**Type of experience:** other: chronic toxicity

**Remark:** A case of chronic methanol poisoning, with marked

diminution

of vision, resul ted apparently from the exposure at 1200 to

8000 ppm for 4 years. Also reports of headaches among

workers exposed at 300 ppm during operation of duplicating

machines were recorded.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(384)

**Type of experience:** other: chronic toxicity

**Remark:** A case of chronic poisoning from methanol vapor with marked

diminution of vision and enlargement of the liver was

reported. The employee was engaged in centrifuging material

which contained 35 to 40 % methanol. Although he had been

employed in this occupation for four years, his symptoms

did

not develop until blackout conditions became necessary with

a resultant decrea se in ventilation. Air analysis at the

work place revealed levels from 1.6 to 10.9 mg/l (appr.

1,200 to 8,300 ppm).

**Reliability:** (2) valid with restrictions

basic data given, acceptabl e restrictions

(385)

**Type of experience:** other: chronic toxicity

**Remark:** A study of the wood heel industry in Massachusetts found

average methan ol vapour concentrations ranging from 160 to

780 ppm, with no definitive evidence of injury to the

exposed workers.

**Reliability:** (2) valid with restrictions

basic data given, acceptabl e restrictions

(386)

**Type of experience:** other: chronic toxicity

**Remark:** A report indicated severe recurrent headaches in workers

exposed to met hanol in concentrations between 200 to 375

ppm.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(387)

**Type of experience:** other: chronic toxicity

**Remark:** From 1978

- 1982 only one new case of methanol

- induced

occupational disease was compensated in Germany.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

251 **Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(388)

**Type of experience:** other: chronic toxicity

**Remark:** Nineteen workers exposed to the fumes of acetone a nd

methanol in a collar fusing plant were studied. Physical

examinations revealed no abnormalities. Blood studies

revealed no disturbances in hematopoesis. Absorption of

ac etone was demonstrated by finding this substance in the

urine of every member of the group. The concentration of

solvents in the workroom atmosphere (methanol: 22 and 25

ppm, acetone: 45 and 40 ppm) was apparently not high enough

to cause pathological changes.

**Reliability:** (2) valid with restrictions

basic data given, acceptable restrictions

(389)

**Type of experience:** other: carcinogenicity

**Remark:** No studies have been reported in the literature on the

carcinogenicity of methanol in humans.

**Reliability:** (2) valid with restrictio ns

see remark on selection of literature

(88)

**Type of experience:** other: review, reproductive and developmental effects

**Remark:** The Panel revie wed four epidemiological studies that

examined folate supplementation and cleft palate (Peer et

al, 1964, Tolarova and Harris, 1995, Shaw et al, 1995, and

Cziezel and Dudas, 1992). The Panel co ncluded that in the

aggregate the studies suggest a reduced risk of oral clefts

following supplementation with vitamins containing folic

acid. The epidemiological findings weakly support the

hypothetical mechanism of action that methanol reduction of

folate activity in the rodent causes oral cleft defects.

However, that extrapolation is difficult due to the various

differences, including different folate levels between

rodents and humans.

No human data on the reproductive toxicity of methanol have

been located.

**Reliability:** (2) valid with res trictions

see remark on selection of literature

(233)

**Type of experience:** other: review, reproductive and developmental effects

**Remark:** No studi es have been reported in the peer

- reviewed

literature on the reproductive and developmental effects of

methanol in humans.

**Reliability:** (2) valid with restrictions

see remark on select ion of literature

(88)

**5.11 Additional Remarks**

**Type:** other: review

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

252 **Remark:** A. The toxic effects of methanol in humans and their

impli cations are reviewed.

The highest estimated exposure to methanol when used as fuel

(EPA 1988) is up to 650 mg/m3.

Acute toxicity in humans affects, as previously reported,

the central nervous system and primarily the optic nerve.

The background level of methanol in blood was measured as

0.73 mg/l (0.32

- 2.61 mg/l) [Sedivec et al. 1981: 31

persons) and the background level of forma te, the major

metabolite, was measured between 3 and 19 mg/l [0.07

- 0.4

mM].

Based on these values a background body burden of 0.5 mg/kg

body weight was estimated and at worst case e xposure

scenarios additional body burdens between 0.03 and 1.1 mg/kg

body weight were calculated (Tab. 6, p. 34).

The clearance of methanol from the body was calculated as

Vmax 48 mg/ kg/h in primates and 30 mg/kg/h in rats.

Metabolism in humans contributed to 96.9 % of the clearance,

whereas renal and pulmonary excretion contributed only to

0.6 and 2.5 % respectively. Over 90 % of the dose was

exhaled as CO2. The half

- time was approx. 2.5

- 3 hours.

The contribution of formate to methanol toxicity is not

clear according to the author.

According to the data rev iewed no toxic effects of methanol

are expected when the substance is used as fuel even at

worst case exposure scenarios.

-----------------

B. The minimal lethal dose to humans i s considered to

range from 0.3 to 1 g/(kg*d). A blood level of 500 mg/l in

acutely poisoned patients generally is regarded as requiring

hemodialysis. This concentration can be achieved by

ingestion of 0.4 ml methanol/kg, which is only 28 ml for an

adult person (70 kg), but just 4 to 6 ml for small children

up to one year old.

Acute methanol intoxication (after single dose) evolves in a

well

- defined pattern: mild depression of the CNS, an

asymptomatic latent period for several hours to >=2 days

(most common 12

- 14 hours).

The toxicity syndrome that emerges consi sts of uncompensated

metabolic acidosis with superimposed toxicity to the visual

system. Clinical symptoms include headache, dizziness,

nausea, and vomiting, abdominal pain, and difficult,

periodic breathing (Kussmaul breathing) which may progress

to coma and death from respiratory failure.

In parallel, visual disturbance to total blindness may

occur.

Ophthalmolog ic examination would reveal hyperemia of the

optic disc, followed by the appearance of oedema projecting

into the surrounding of the retina from the optic disc,

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

253 while the papilla itself is not oedematou s.

The severity of retinal oedema is predictive of restoration

of vision. Pallor of the optic disc is an end

- stage sign of

irreversible effects of the visual system and may appear one

to two months after an acute methanol dosage (or possibly

chronic exposure).

Post

- mortem signs of damage were observed in the basal

ganglia in the brain, specifically the putamen (which area

participates in the control of motor activities). Survivors

of severe methanol intoxication may continue suffering from

residual lesions of the putamen.

-----------------------

**Test substan ce:**  Methanol, no further data.

**Reliability:** (2) valid with restrictions

(192)

**Type:** other: reviews

(390) (88) (192) (233)

**Type:** other: Ocular toxicity / folate / rat

**Remark:** 1. Changes in the electroretinogram (ERG) and the

potassium

- induced Müller cell depolarization (KIMD) of

folate

- reduc ed rats have been observed after oral methanol

administration of 1.5

- 2.5 g/kg whereby formate in blood

increased, concentrations ranging from approx. 6 to 15 mM at

48 h (Lee et al., 1994). Moreover, m ethanol induces a

decrease in retinal ATP, which suggests interference with

oxidative phosphorylation. Formate is known as inhibitor of

mitochondrial cytochrome oxidase in Müller cells (supporting

glial cells).

No effects were noted at 1 g methanol/kg per os despite

folate deprivation. No adverse effects are produced by doses

of up to 3.5 g/kg in rats maintained on normal diet (Garn er

et al. 1995b).

2. However, creating a blood formate profile by infusion

of formate similar to that observed after administration of

a retinotoxic methanol dose failed to alter the ERG. Retinal

levels of formate after infusion were only 16 % of those

measured after methanol administration (Garner et al., 1995.

p. 105).

3. Intraretinal metabolism of methan ol to formate is

necessary for the initiation of methanol

- induced retinal

toxicity. Retinal and vitreous humor formate levels are

approx. 15.3 umol/g and 19.2 mM, resp., after a single oral

methanol dose of 3.0 g/kg bw to folate

- reduced rats. The

enzymes responsible for methanol metabolism, catalase and

aldhyde dehydrogenase, are both present in Müller cells.

4. The acute effect of methanol on the ERG involves a

severe decrease in the b

- wave amplitude at approx. 48 h

following a single dose and has been observed in humans, in

primates and in folate

- reduced rats (Garner et al., 1995, p.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

254 101). The b

- wave is generated by the Müller cell, the

primary glial cell, in response to an increase in

extracellular potassium released by the bipolar cells.

Exactly paralle l, a dose

- related evocation loss of the

potassium

- induced Müller

- cell depolarisation is noted.

5. The electrophysiological as well as histopathological

effects seen after methanol treatment correla te closely to

those induced by alpha

- amino adipic acid, a specific

Müller

- cell toxin.

--------------------------

**Conclusion:** The inhibition of cytochrome oxidase in Müller

- cell

mitochondria by intracellularly generated formate may

explain the mechanism of methanol

- induced retinal toxicity.

------------------ **Reliability:** (2) valid with restrictions

Test procedures i n accordance with accepted standard

methods, sufficiently documented

(391) (392) (393)

**Type:** other: Absorption / excretion / inhalation / human

**Method:** Methanol absorption after inhalative exposure and excretion

in urine was measured in groups of 4

- 5 human volunteers to

be able to extrapolate exposure in the working environment

from excreted urine levels.

Exposure concentrations were approx. 100 to 300 mg/m3.

Retention of methanol via lung was determined from the

concentration difference between inspired and expired air.

Metha nol in urine (8

- h shift) served as a quantitative

measure for retained methanol.

Blood levels were not measured.

------------

**Result:** The mean normal urine level was 0.73 mg/l (range from 0 .32

-

2.61 mg/l), data selected from a control group of 31

individuals.

A mean of 57.7 % of the inhaled dose was retained, range

53.4

- 61.3 %, irrespective of the exposure level of

selected magnitude.

Urine excretion represented nearly 1 % of the retained dose

at a normal diuresis.

Average urine concentrations reached a peak after 8 h (Fig.

3) and were fairly proportional to the exposure levels

(approx. 3.3, 7.0, and 9.5 mg/l). After 18

- 24 h from the

start of exposure (about >= 12 h after termination), urine

methanol has approached baseline level again.

The excretion half

- life was about 1.5 to 2 h.

In cross

- over drinking experiments it could be shown that

the urinary methanol excretion correlated strictly with

diuresis , i.e. irrespective of the urine volume produced at

the same exposure level, the urine concentrations were

identical and were dependent only on the exposure level.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

255 This suggests that methanol distribu tes only passively into

the urine in relation to the blood level.

This also implies that the total quantity excreted into the

urine cannot be the criterion for the exposure level, but

only its concentration.

The mean equation of regression (Fig. 5) between retained

methanol quantitity (body burden) [X in mg] and the

whole

- shift urine concentration [Y in mg/l] could be

formulated as

Y = 0.7470 + 0.00763X

--------------

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accept ed

scientific principles, acceptable for assessment.

(283)

**Type:** other: Absorption / distribution / rat, mouse

**Remark:** In mice exposed to methanol in air at concentrations of

2500 to 10000 ppm (3.3

- 13 mg/l) absorbed the substance

rapidly. Blood methanol concentrations were approximately

1000 mg/l at 1 hour and 3000

- 4000 mg/l at 7 hour s

exposure.

The fractional absorption was higher in mice (approx. 85 %)

than in rats (56

- 87 %) (p. 251).

Comparison of rat and mouse:

The same environmental metha nol vapour concentration leads

to very different blood methanol level and is in mouse

higher although the maximum elimination rate is about twice

as high in mice as in rat (Pollack and Brouwer 1996):

At the teratogenic exposure concentration in mice of 5000

ml/m3, the following blood levels would be achieved

mouse: 2313 +

- 338 mg/l(exposed as group);

rat: 1047 +

- 298 mg/l.

In humans: 224 mg/l.

-----------------

**Test substance:** Methanol, no further data.

**Conclusion:** The same environmental methanol vapour concentration leads

to very different blood methanol level.

Therefore, risk assessment for methanol should be based on

blood concentrations following inhalation exposure, not on

exposure concentrations itself.

The differences between the teratog enic potential of

methanol in rats versus mice, with mice being approximately

twofold more sensitive than rats may be due to the

differences in blood concentrations.

--------------- --- **Reliability:** (2) valid with restrictions

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

256 (394) (395)

**Type:** other: Toxicokinetics / formate / HEI study monkeys

**Method:** Comparative study designed to cl arify methanol and formate

kinetics in blood at low and high exposure concentrations to

methanol in rats and monkeys with extrapolation to humans

(Horton et al. 1992):

Animals: Male F344 rats (190

- 230 g) and young adult male

Rhesus monkeys (Macaca mulatta), 4 rats and 3 monkeys per

test group.

Inhalation exposure: single for 6 h, but in 3 monkeys also

repeated i nhalation (2000 ppm, 6 h/d, 5x/wk, 14 d) (not

documented: see Horton et al., Discussion, p.33)

Test concentrations of methanol: nominal 50, 200, 1200, and

2000 ppm (analytical values were close to nomi nal).

Analytics chamber: continuously by IR gas analyzer for

methanol.

Analytics blood: for methanol GC, for formate enzymatically

coupled to a fluorescence measurement of a reactant (NADH2).

Sampling up to 12 h post

- exposure.

Blood gas analysis for pH, bicarbonate, and pCO2 was

performed.

----------------------

**Result:** Kinetic key data:

Generally, the elimination rate from blood decreased with

increasing concentrations, in rats only at the highest dose

significant. At the lowest concentration, 200 ppm, the

elimination was o nly slightly lower in monkeys than in rats

(approx.

- 30 %), but was distinctly slower at the higher

burdens. No significant increases in blood values above

background were evident at 50 ppm methanol.

T1/2[h]

Rat Monkey

---------------------------------------------

200 ppm 0.8 +

- 0.2 1.1 +

- 0.2

1200 p pm 1.0 +

- 0.1 3.2 +

- 1.2

2000 ppm 2.1 +

- 0.1 2.9 +

- 0.6

---------------------------------------------

Blood levels after 6

- h exposure [ug/ml]:

Rat Monkey

---------------------------------------------

Methanol:

200 ppm 3.1 +

- 0.4 3.9 +

- 1.0

1200 ppm 26.6 +

- 2.0 37.6 +

- 8.5

2000 ppm 79.7 +

- 6.1 64.4 +

- 10.7

---------------------------------------------

Formate: The peak blood levels in methanol

- exposed rats and

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

257 monkeys ranged between 5.4 and 13.2 ug/ml (approx. 0.25

- 0.6 mM) without showing statistically significant

differences to control animals [note: Data are not

documented in Horton et al., 1992: However, Medinsky and

Dorman (1995) pres ent data in Fig. 1 attributed to the

studies by Horton et al. (1992).]

Blood acidosis: no signs of acidosis which is in line with

the formate blood levels in normal range.

------------ -------------------------

Repeated inhalation in monkeys (data not shown: see Horton

et al., 1992, Discussion, p. 33), carried out as

confirmatory test to examine accumulation toxicology:

In blood samples collected after 1 and 2 weeks of exposure,

no accumulation of methanol or formate.

--------------------------------------

Prediction of in

- vivo time

- course data by the PBPK model:

Below 1500 mg/m3 (1200 ppm), all three species (rat, monkey,

human) would exhibit similar post

- exposure blood levels of

methanol which would be proportional to atmospheric

concentrati ons. Above 1200 ppm, increase in blood would

become non

- linear for rat and monkey, but remain linear for

human.

------------------

**Test substance:** Methanol purity >98%

**Conclusion:** Compari son of monkey blood

- methanol elimination rate

constant with those derived for humans (HEI, 1987) and rats

shows an interspecies similarity in the overall elimination

of methanol after low doses (Horton et al., 1992,

Discussion).

The dose of methanol required to saturate the

folate

- dependent metabolism of formate, calculated from an

equation derived by the Health Effects Institute (H EI,

1987), is about 250 mg/kg. If one assumes a 6

- kg monkey

breathes 118.8 l/h and 100% absorption of methanol occurs, a

dose equivalent to 308 mg/kg is absorbed during a 6

- h

exposure to 2000 ppm methanol(2600 mg/m3).

While this amount of methanol could overwhelm the formate

metabolism pathway if given as a bolus, the saturation of

metabolism with this amount divided over 6 h is unl ikely

(Horton et al., 1992, Discussion).

------------------------------------ **Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scie ntific principles, acceptable for assessment.

(396) (397) (252)

**Type:** other: Absorption / inhalation / human

**Method:** Six human volunteers (from 29

- 55 years) were sub jected to

a controled diet

- regimen (without obvious

methanol

- delivering nutrition and additives) throughout the

study and exposed to 200 ppm (260 mg/m3) methanol for 6

hours. Five indi viduals were each tested at rest or at light

exercise.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

258 ------------

**Remark:** Blood methanol concentrations increased from 1.8 ug/ml (mean

endogenous level) to 7.0 ug/ml at rest and to 8.1 ug /ml

under light exercise (increase in mean pulmonary ventilation

at a factor of about 2.7 from average 10.5 to 26.6 L/min and

increase in respiratory rate at a factor of about 1.7 from

11.2 to 18.6 breathes/min).

Blood formate levels did not increase.

Blood levels at rest (from Tab. III and V):

Test Methanol[ug/ml] Formate [ug/ml]

person pre

- post

- pre

- post

-

exposure exposure

--------------------------------------------------

1 2.53 5.44 8.11 8.12

2 3 .57 7.55 10.57 13.06

3 1.32 7.93 10.35 7.24

4 0.57 8.08 9.08 8.98

5 1.12 5.83 7.31 6.12

--------------------------------------------------

Mean 1.82 6.97 9.08 8.70

+

- SD +

- 1.21 +

- 1.24 +

- 1.26 +

- 2.38

------------------------------------------------- -

Blood levels at exercise (from Tab. IV and VI):

Test Methanol[ug/ml] Formate [ug/ml]

person pre

- post

- pre

- post

-

exposure exposure

--------------------------------------------------

1 1.99 9.82 5.36 8.90

2 2.00 7.40 9.28 11.14

3 3.15 7.4 7 9.35 9.54

4 0.54 6.40 9.07 9.91

5 1.95 9.57 10.83 8.10

--------------------------------------------------

Mean 1.93 8.13 8.78 9.52

+

- SD +

- 0.93 +

- 1.49 +

- 1.82 +

- 1.02

--------------------------------------------------

The results show that no formate mediated toxicity

is expected when workers are exposed to 200 ppm methanol,

the current Occupational Safety and Health Administration

8

- hr time

- weighted average permissible exposure limit.

**Test substance:** Methanol, no further data.

**Conclusion:** At 200 ppm methanol or below, methanol blood levels may

increase 3.5 to 4

- fold above the endogenous methanol

concentration in human blood, while the formate level

remains unchan ged, irrespective of physical activity whether

at rest or light exercise.

This was confirmed e.g. by D ́Alessandro et al. (1994) in

similar experiments on human volunteers (comp. also Medinsky

and Dorman, 1995).

--------------------- **Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

259 scientific principles, acceptable for assessment.

(398) (196) (252)

**Type:** other: Risk Assessment Model / inhalation

**Method:** A "multicompartment biologically based dynamic" inhalation

model based on kine tic methanol inhalation data from rats

(Horton et al., 1992), monkeys (Dorman et al., 1994) and

humans (Sedivec et al., 1981; Osterloh et al., 1996;

Batterman et al., 1998) was developed to describe the time

evolution of methanol and its metabolites in the whole body

and in accessible biological matrices.

---------------

**Result:** Predictions from simulations (PBPK modelling) of continuous

inhalation of 200 ppm methanol in humans for 5 days

(Bouchard et al. 2001, p. 177/178):

Based on the following assumptions:

- a negligible background burden of methanol,

- an absorption fraction of 0.577 (Sedivec et al. 1981),

- a pulmonary ventilation rate of 10.8 L/min (Sedivec et

al. 1981; Batterman et al. 1998),

- an apparent distribution volume for m ethanol of 0.7 l/kg

(corresponding to human body fluid),

- an apparent distribution volume for formate of 4.6 l/kg

(estimations by Bouchard et al. 2001, Tab. 2, p. 173

an d p. 180),

- a daily urine volume of 1.5 l,

near steady state will be reached in 20 h. After 5 d,

methanol in blood and urine is estimated at 5.5 mg/l (171

umol/l) and 8.1 mg/l (252 umol/l); formate in blood and

urine is 1.16 mg/l (3.5 mol/l) and 1.5 mg/l (31.7 umol/l =

0.97 mg/g creatinine or 2390 umol/mol creatinine).

This shows that exposure concentrations of <500 ppm are not

sufficient to raise formate levels significantly, while

methanol increases.

The model, adapted to kinetic data in humans exposed acutely

to methanol, predicts that 8

- h inhalation expo sures ranging

from 500 to 2000 ppm, without physical activities, are

needed to increase concentrations of blood formate and

urinary formic acid above reported background values

(4.9

- 10 .3 and 6.3

- 13 mg/l, resp.).

Therefore, acc. to authors, blood and urinary methanol

levels are the most sensitive biomarkers of absorbed

methanol.

---------------------

Pulmonary retention: Using the experimental human data of

Osterloh et al. (1996), Sedivec et al. (1981) and Batterman

et al. (1998), the best fit in the model for the average

absorption fraction was higher than that given by Sedivec et

al. (1981), namely about 80 % and corresponded to the

retention of 79 % given by Batterman et al. 1998 (Bouchard

et al., 2001, p. 177).

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

260 ----- ------------ **Reliability:** (2) valid with restrictions

(299) (399) (251) (397) (277) (283)

**Type:** other: Absorption / distribution / inhalation / rat

**Method:** Whole

- body exposure sys tem:

1. Adult female Long

- Evans rats (n = 12) were exposed to

4500 ppm methanol for 3 consecutive days once for 6 h. Blood

methanol levels were measured 1 and 6 h during and 1

- h post

exposure as wel as background levels before each exposure.

2. Adult female Long

- Evans rats (n = 12) were exposed to

4500 ppm methanol during pregnancy and lactation along with

the progeny (gd 6 thr ough post

- natal day[PND] 21).

(see also other entry under 5.8.2)

--------------------------

**Result:** exposure time 1 h 6 h 1 h post exp.

[in approx. averages mg methanol/ml]

-----------------------------------------------------

day 1 0.25 0.90 0.95

day 2 0.40 0.60 0.60

day 3 0.30 0.50 0.60

-----------------------------------------------------

After the first day, the levels were higher which can be

explained with adaptation to the new environment (higher

activity) . The pre

- exposure levels (1 h before first

exposure and 17 h after each previous exposure interval)

were non

- detectable.

During pregnancy and lactation, blood levels in dams were in

the range of 0.50 +

- 0.07 to 0.55 +

- 0.09 mg/ml. But in

offsprings, the level was about 2x that of dams, at 1.26

+

- 0.23 mg/ml, and gradually decreased to that of the dams

after 52 PND.

----------------------

**Test substance:** Methanol, HPLC grade

**Reliability:** (1) valid without restriction

1c: Test procedures in accordance with accepted standard

methods, well documented

(241)

**Type:** other: Absorption / distribution / oral / rat

**Result:** After oral administration of 100 and 2500 mg methanol/kg to

female SD rats, gastrointestinal abs orption was 100 % within

minutes (abs. half

- life 1.5 min and 7.6 min, respectively)

(p. 16).

The maximum elimination rate is about twice as high in mice

as in rat: 117. +

- 3 and 60.7 +

- 1.4 mg/(kg*h) (Pollack and

Brouwer 1996, Tab. 7, p. 21; Ward et al. 1995).

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

261 After inhalation, the mean fractional respiratory absorption

of methanol in rats and mice was found to about 85 %,

respectively, at exposure concentrations from 1000 to 5000

ml/m3, and tended to be lower (60

- 70 %) at 10000

- 20000

ml/m3 in rats, but not in mice (Pollack and Brouwer 1996,

Tab. 12, p. 27; Ta b. 15, p. 33).

Blood levels in rats were about 800 (calc.)

- 1200 mg/l

(measured) at 5000 ml/m3 (8 h) (Fig. 37, p. 34).

Blood levels in mice were about 3000 mg/l at 5000 ml/m3(8 h)

( Fig. 37, p. 34).

------------------- **Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

(395) (400)

**Type:** other: Toxicokinetics / formate / folate deficiency / HEI

study monkeys

**Method:** Pharmacokinetic study to evaluate the contribution of

methanol

- derived formate to the endogenous formate pool in

nonprimates under folate

- sufficient and folate

- deficient

condition (Medinsky et al., 1995, 1997):

Four 12 year old female cynomolg us monkeys (Macaca

fascicularis) were used: lung

- only exposure under

isoflurane

- anaesthesia. Exposure time = 2 h.

14C

- radiolabelled methanol was used as atmospheric tracer

material.

Exposure concentrations/situation: nominal 10, 45, 200, and

900 ppm (normal folate) and 900 ppm (folate

- deficient) in

the every same monkeys two months later after

folate

- deficient diet

. Measured concentrations corresponded

well to nominal ones.

(Achieved folate deficiency corresponded to stage

- III and

greater.)

Air analytics of methanol by GC.

Blo od analysis of methanol and formate by HPLC and liquid

scintillation spectroscopy: The methanol

- dependent fraction

of formate was estimated by using 14C

- radiolablelled

methanol.

--- -------------------------

**Remark:** It is unclear why the endogenous animal

- specific total

formate blood levels were not determined in the same test

animals. This restriction is not considered to invalidate

the overall result and conclusion.

------------------------

**Result:** Methanol levels in blood:

Average 2

- h concentrations were 0.65 +

- 0.3, 3.0 +

- 0.8, 21

+

- 16, 106 +

- 84, and 211 +

- 71 uM following 10, 45, 200, 900

and 900 ppm(deficient).

AUCs were linearly related to the inhaled methanol dose, the

T1/2 were largely unaffected by the methanol concentrations

and ranged from mean t1/2 of 0.56, 0.88, 0.62, and 0.95 h,

respectively, with 1.31 h under folate deficiency showing a

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

262 positive increasing trend, although not significant (Dorman

et al., 1994; Medinsky et a l., 1997).

Formate levels in blood:

Average 2

- h concentrations were 0.07 +

- 0.02, 0.25 +

- 0.09,

2.3 +

- 2.9, 2.8 +

- 1.7, and 9.5 +

- 4.7 uM following 10, 45,

200, 900 and 900 ppm(deficient).

The endogenous formate blood level was not measured, but

assumed to be 0.1 to 0.2 mM (taken from other sources). The

range of the author ́s group is given as 0.29 to 0.56 mM (see

Disc ussion).

----------------------

**Test substance:** Radiolabelled methanol from Sigma Chemical Corporation, >98%

pure, methanol HPLC pure.

**Conclusion:** The results suggest that low

- dose short

- term exposure to

methanol would not result in toxic elevated methanol or

formate levels in blood even under folate deficiency,

because, even under folate deficiency, the methanol

- related

formate levels remai ned at a factor of 10 to 100 below

endogenous blood concentrations and are several orders of

magnitude lower than levels of formate known to be toxic (>7

mM) [see also: Medinsky and Dorman, 1995, p. 709 ].

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

(25 1) (401) (252)

**Type:** other: Metabolism / formate / monkey ip

**Method:** 8 weeks old male Fischer 344 rats and male monkeys (Macaca

fascicularis) received 14C

- labeled methanol per i.p.

injection in do ses of 25; 125; 600 and 3000 mg/kg. 5 animals

per dose group were tested (monkeys were repeatedly used).

Blood was collected after 6; 12; 24; 30 and 48 hours.

The following parameters were analysed:

- formic acid

- organic acid

- folic acid

- pH

- biochemistry (glucose, blood urea nitrogen, Na, K, Cl).

-----------------

**Result:** Rats:

In 25 to 600 mg/kg dosage groups no abnormality was seen and

there were hardly any individual difference in blood

radioactivity.

In the 3000 mg/kg group, however, animals grew weak

immediately after administriation, with one death during 48

hours. Moreover, somewhat serious toxic symptoms, such as

difficulty in blood collection and diarrhea, were seen. The

disappear ance of radioactivity in blood delayed markedly

when compared with the other groups. The recovery was noted

in only on animals which had shown a decrease of the blood

level within 48 hours.

In animals of 25 to 600 mg/kg dosage groups, the major

excretion route was exhalation: 70

- 80 % of administered dose

was excreted as CO2 and methanol. The excretion of methanol

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

263 increased with incre ased dosage.

The excretion into urine and feces was very low: 6.1

- 8.3 %

and 2.1

- 2.7 % respectively. The excretion was almost

completed within 48 hours.

At 3000 mg/kg dosage group 2 ani mals died within 24 hours,

and one additional animal within 48 hours after

administration. The excretion was very slow: about 22 % was

excreted into exhaled breath as CO2 or methanol and about 8

% in urine. There were hardly any feces, and therefore the

fecal excretion of radioactivity remained very few.

Monkeys:

The major route of excretion was exhalation at 25 to 600

mg/kg, 50 % of the administered dose being excreted

as CO2. Even after 48 hours, total excretion remained low:

about 60 % (probably due to the test procedure: loss of CO2

during anesthetization and blood collection). Methanol

excretion was 0.82

- 3.8 %, urinary excretion 5.9

- 7.5 % and

excretion via feces 0.35

- 0.53 %.

At 3000 mg/kg the excretion were 25 % as CO2, 9 % as

methanol, 2 0 % via urine, the excretion was delayed in this

group.

------------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

metho ds, sufficiently documented

(167)

**Type:** other: Absorption skin / human

**Method:** a) Exposure of 0.19

- 0.21 ml methanol to the skin of 6

hu man volunteers by means of a closed application glass

chamber (11.2 cm2), absorption was measured from 15 to 60

min p.a., 22 determinations in total. Residual available

methanol was washed off with wat er and determined

sprectrophotometrically (Dutkiewicz et al. 1980).

b) Exposure of one hand to methanol (Dutkiewicz et al.

1980).

---------------

**Result:** a) Following occl uded exposure in 6 volunteers, average skin

absorption rate of methanol is reported to be at 0.192

mg/cm2/min (Dutkiewicz et al. 1980).

b) Exposure of one hand to liquid methanol for 2 min would

result in a body burden of 170 mg, similar to that resulting

from exposure to an approximate air concentration of 50

mg/m3 (38 ml/m3) for 8 h (Dutkiewicz et al. 1980) (see also:

IPSC/WHO, 19 97).

c) In 12 humans one hand of whom was exposed to liquid

methanol for 2 to 16 min, the average uptake through the

skin was 8.1 +

- 3.7 mg/(cm2*h) = 0.135 +

- 0.062 mg/(cm2*min)

(Batter man and Franzblau 1997).

Full exposure of one hand for 16 min resulted in a blood

level equivalent to that reached after inhalation of 400

ml/m3 for one 8

- h working shift with a maximal blood level

of some 11 mg/l (corrected for background value) (Franzblau

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

264 and Batterman 1995; Batterman and Franzblau 1997).

-------------

**Test substance:** Methanol, no further data.

**Conclusion:** The percutaneous absorption may be considerable on direct

contact of skin to methanol and constitute a substantial

part of the body burden in relation to inhalation.

Both independent sources arrived at similar quantita tive

kinetic results for skin permeation in humans and appear to

be reliable.

--------------- **Reliability:** (2) valid with restrictions

Special study design, sufficiently docum ented, meets

generally accepted scientific principles, acceptable for

assessment

24

- DEC

- 2004 (300) (390) (298) (402)

**Type:** other: metabolism / mouse / FDH defic iency

**Method:** In normal (CB6

- F1) and FDH

- deficient mice (NEUT2), acute

methanol toxicity and formate elimination after formate i.p.

application were compared. The homozygous NEUT2 mice lack

c ytosolic 10

- formytetrahydrofolate dehydrogenase (FDH).

14CO2 from exhaled air was collected after treatment of mice

with a low (5 mg/kg) or a high dose (100 mg/kg) of

14C

- formate. For discrimination bet ween catalase

- dependent

and other biotransformation routes, 3

- aminotriazole (3

- AT: 1

g/kg i.p.) was employed as selective inhibitor of catalase.

---------------------

**Result:** Acute LD50 wer e virtually similar in either strain at about

6000 mg/kg methanol.

At 5 mg/kg formate, the difference in CO2 formation in the

NEUT2 strain was about one half of the normal mice as

exp ected. However, the inhibitor 3

- AT failed to reduce CO2

generation in either case.

At 100 mg/kg formate, CO2 exhalation was similar in both

strains (which is in line with the congruent acute toxicity

in either strain), but also inhibited by 3

- AT to the same

extent.

There was no significant difference in formate

- related CO2

release in either strain after 3

- AT inhibition, although th e

FDH

- deficient strain should

- in theory

- be unable to oxidise

formate under inhibitory condition.

---------------------

**Conclusion:** At low formate burden (here 5 mg/kg), the function of the

catalase is poor or not operating, but is active at high

doses (here 100 mg/kg). The folate

- dependent FDH system

appears to function under normal metabolic conditions.

The residual meta bolic capacity of the FDH

- deficient mouse

strain to oxidise formate is apparently a folate

- independent

alternate oxidation system not yet characterised.

Therefore, there appear to be at least three met abolic

pathways in mice for the oxidation of formate arising from

methanol or other sources:

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

265 1. the common cytosolic folate

- dependent

10

- formyltetrahydrofolate dehydrogenase (FDH) pathway,

2. the catalase

- dependent route, and

3. a yet unknown route, likely folate

- independent.

-------------------- **Reliability:** (2) valid with restrictions

Test proce dures in accordance with accepted standard

methods, sufficiently documented

27

- DEC

- 2004 (403)

**Type:** other: Toxicokinetics / formate / folate deficiency / monkey s

**Method:** Sodium formate metabolism was studied in monkeys (Macaca

fascicularis) after i.v. infusion usine 14C

- radiolabelled

formate /doses 1, 2.5, 5 and 10 mmol/kg). Tests were

performed un der folate

- sufficient and folate

- deficient

conditions.

----------------

**Result:** At the highe rdose levels of formate, the rate of formate

metabolism to CO2 was less than that observed in rat (Fig.

3). At lower doses the rate of oxidation to CO2 was about 25

% of that in rat.

The catalase

- peroxidase pathway was shown to play no role in

the oxidation of formate in monkeys (using catalase

inhibitor) (Fig 4).

The half

- life of formate elimination was about 38 min at 2.5

mmol/kg formate, in the absence or presence of a catalase

inhibitor.

Folate depletion reduced formate metabolism significantly

while folate supplementation accelerated it (Fig. 6

- 9).

There was a striking increase in blood formate following

methanol application under dietary folate deficiency

(approx. 5 vs. 2 mEq/l blood after 0.5 g methanol/kg).

-------------

Elimination half

- lives for formate from blood ranged from

some 30

- 50 min in the mon key and about 12

- 23 min in

rats, depending on the doses (Clay et al. 1975, p. 57).

**Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific princip les, acceptable for assessment.

24

- DEC

- 2004 (168) (404)

**Type:** other: Metabolism / in vitro / formate / various species

**Method:** The folate

- dependent formate oxidation and the a ctivities

of several folate

- dependent enzymes were measured in vitro

with liver homogenate and liver cytosolic fraction of rat

(male SD), mouse (Swiss

- Webster), monkey (female Cynomolgus,

Macaca fascicularis) and humans (from victims of traumatic

head injuries, without evident liver diseases).

----------------

**Result:** Human and monkey liver tissue showed much lower

10

- form yltetrahydofolate dehydrogenase activity, the enzyme

catalyzing the final step of formate oxidation to CO2, about

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

266 25 % and 37 %, respectively, than in rat liver.

Furthermore, total folate and H4folate i n human liver were

only 40 and 50 %, respectively, of that observed in rat

liver.

In monkey liver, H4folate was comparable to that in humans,

but total folate was comparable to that i n rat.

Mouse liver contained much higher H4

- and total folate

levels than rat and monkey.

The

The in

- vitro rates of formate oxidations were approx. 30 to

40 mg/(kg bw*h) in monkey, about 75

- 80 mg/(kg*h) in rats,

and approx. 300 mg/(kg*h) in mice [humans not shown].

--------------

**Test substance:** Methanol, no further data.

**Conclusion:** The authors explain the higher toxi city of methanol in

humans and monkeys compared to rodents by the low H4folate

levels as well as the reduced enzymic capacity of formate

oxidation.

--------------- **Reliability:** (2) valid with restrictions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

24

- DEC

- 2004 (405)

**Type:** other: Ocular toxicity / folate / human / rat

**Method:** Comparative biochemical study to quantify folate and

10

- formyl tetrahydrofolate dehydrogenase in human and rat

retina and thus explain the hig her ocular toxicity of

methanol towards human being.

-------------

**Result:** The cell

- specific localisation of the enzyme, 10

- FDH, was

found to be similar in rat and human retina, preferenti ally

located in the Müller

- cell type, the principal glia of the

retina (by immunohistochemistry).

The amount of 10

- FDH found in cytosolic as well as in the

mitochondrial fraction, was about 3x higher in humans than

in the SD rats (Western blot analysis).

However, the retinal folate levels were lower in humans

(about 14 % of that in rats), compared with the high folate

liver pools, the retina contains very much less folate.

---------------------

**Conclusion:** Formate oxidation was found to be about 50 % lower in human

than in rat retina (Eells et al, 1995). This is in line wi th

this finding that lower folate levels in human retina may

limit conversion of formate into CO2 and result in higher

ocular toxicity in humans.

------------------ **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

(406) (407)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

267 **Type:** other: Risk Assessment / Reproductive parameters

**Remark:** A range of toxicological investigations, especially

teratogenicity studies, was evaluated to perform a risk

assessment for consumer and occupational ex posure

scenarios. The author came to the conclusion that exposure

to

the current TWA (time weighted permissible exposure limit)

does not pose any risk.

-------- --------

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

(249)

**Remark:** The authors evaluated published data of formate produc tion

in humans and animals after methanol exposure. They came to

the conclusion that even high methanol exposure up to 2.6

mg/l did not produce toxic blood formate levels and

therefore exposure to methanol likely to be encountered

during normal use of methanol

- based fuels (up to 0.06 mg/l)

will no pose an unacceptable risk.

--------------

**Test substance:** Methanol, no further da ta.

(408)

**Type:** other: absorption skin human

**Remark:** Penetration test in male human voluteers: Methanol was applied

to the skin for 4 to 6 h on a soaked cotton patch under

occlusive

conditions for 3 days. It was absorbed through the human skin

at

significant amounts, and excretion into the urine increased

after 2h with the maximum after 6 h.

-----------

**Test substance:** Methanol, no further data.

29

- DEC

- 2004 (409)

**Remark:** Review on important studies for essen tial endpoint, intended

to perform a risk assessment for the general human

population: Reference concentrations are derived.

The study by NEDO (1987) on monkeys is employed as central

basis for this estimation.

--------------------

**Result:** A 1

- h reference concentration (RfC) was derived to be at

about 100 mg/m3, taking into account sensitive people. A

very conservative c hronic RfC was obtained and proposed at

0.38 mg/m3, based on the assumption that 10 ppm has to be

used as NOAEL and 100 ppm as the LOAEL for "neurotoxic

effects". This appears to be not in compliance wi th the

authors ́ observations and is not suggested in the report.

--------------------------------- **Reliability:** (2) valid with restrictions

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

268 ( 167) (179)

**Remark:** Conclusions on the role of formate:

"From the studies of formate in rodents in vivo and in

vitro... it appears that formate is toxic to the developing

embryo at concentrati ons far in excess of those achieved

after teratogenic methanol exposure in the mouse, but within

the range of levels achieved in humans after acute high

- dose

methanol poisoning. Thus, although methanol appears to be

the proximate murine teratogen, formate is still of concern

in terms of potential developmental toxicity in humans." (p.

386)

Conclusions on the role of folic acid:

"These results (note: including studies on folate

- deficient

diet) suggest that dietary folate intake may be an important

consideration in assessing the risk of methanol

- induced

development al toxicity." (p. 386)

(264)

**Type:** other: Absorption / inhalation / rat

**Method:** A flow

- through inhalation chamber system (coupled with the

barometric method of evaluation of the tidal volume) is

introduced which allows the on

- line monitoring of the

essential ventilation parameters, the tidal repiration

volume and respiration frequency of the test animal, under

unrestrained conditions as well as in

- and efflux vapor

concentrations. This enables to obtain more accurate and

thus representative data about extraction of the inhalant

from air flow. Concomitant blood sampling via a fixed

cannula further allows to analyse blood levels at any time

interval.

Female SD rats were used, exposed to 1000, 5000, 10000,

15000, and 20000 ppm methanol for 8 h.

-------------------

**Result:** Based on this technique, the mean ventilation volume for 8 h

was 280

- 230 L/kg b.w. (mean 250 L/kg), a slight downward

trend discernible with increasing exposure concentration.

The respiration frequency was found to be significantly

lower than commonly published (about

- 20 to

- 30%), while the

tidal volumes were sim ilar.

At 5000 ppm, an extraction (retention) factor (absorption)

of about 0.9, at 15000 ppm of about 0.6 resulted from the

plots of cumulative mass vs. exposure time. No data given at

the lower exposures.

The highest blood concentrations were at about 3500 mg/L (No

exposure

- blood correlation is shown.).

---------------------

**Conclusion:** Based on these results, the respira tory uptake of methanol

by rats can be assumed to be approx. 100 % at exposure

concentrations of <5000 ppm.

**Reliability:** (2) valid with restrictions

(410)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

269 **Type:** other: Ocular toxicity / rat model

**Method:** Nonprimate model in rats in which the folate

- dependent

formate oxidation (on the level of methionine synthetase)

has been select ively inhibited by treatment with N2O in

inhaled air.

Animals: Male Long

- Evans rats (250

- 300 g)

Methanol administered i.p. at an initial dose of 4 g/kg

followed by a supplemental d ose of 1 or 2 g/kg 12 h later.

Thus, blood formate levels were from 7

- 15 mM or 4

- 6 mM

for 30

- 40 h.

The "non

- intoxicated" controls received 4 and 2 g/kg

methanol without N20, the non

- treated controls either the

vehicle or N2O in air.

Retinal function was assessed by flicker electroretinography

(ERG). Histopathology of retina and optic nerve was carried

out by light microscopic visualisation.

---------------------

**Result:** Methanol intoxicated rats developed formic acidemia,

metabolic acidosis and visual toxicity within 36 hours.

Histopathological effect on retinal structure:

In the high

- dose group (7

- 15 mM blood formate vs.

methanol

- treated control with 0.5 to 2 mM formiate),

prominent vacuolation in the photoreceptors near the

junction of inner and outer segments, with accumulation of

densely stained material in the inner segments near the

outer limiting membrane. Mitochondrial swelling and

disruption was noted in the re tinal pigment epithelium,

photoreceptor inner segments and optic nerve (Eells et al.,

2000; Seme et al., 2001). Ultrastructural studies by

electronmicroscopy revealed that the retinal morphology (as

represented by the mitochdrial

- rich, inner segment of the

photoreceptor) was similar to the control after recovery of

72 h, but subtle photoreceptor changes were still present as

a spacin g between the cell nuclei of the outer nuclear layer

which suggests residual histological alterations from

formate

- induced, previous edema (Seme et al., 2001).

In the low

- dose group (4

- 6 mM formate i n blood), no

histopathological changes were apparent at the

light

- microscopic level (Wallace et al., 1997).

However, visual dysfunction was already visible in

functional dignostic, oc curring as reductions in the flash

evoked cortical potential (FEP) and in electroretinogram

(ERG) at formate concentrations lower than those associated

with morphological changes and provide functional evidence

of direct retinal toxicity in methanol poisoning (Wallace et

al, 1997). Rod

- and cone

- mediated ERG responses were

attenuated in a formate

- and time

- dependent manner (Seme et

a l., 1999, 2001).

Biochemical effects:

Retinal ATP, ADP, and GSH were significantly depleted

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

270 following methonol

- treatment under inhibition of formate

oxidation after 72 and 144 h with GSH levels about 1/2 of

controls, and after recovery still decreased, while energy

metabolites showed no difference from the control values

(Seme et al., 2001).

---------------

**Test substance:**  Methanol, HPLC grade from Sigma

**Conclusion:** Functional tests provide functional evidence of direct

retinal toxicity in methanol poisoning at stages not yet

pronounced in histopathological changes.

Hypothetical mechanism: Formic acid binds to cytochrome aa3

and inhibits cytochrome oxidase activity with inhibition

constant values between 5

- 30 mM, which is in the range of

concentra tions found in the retina and vitreous humor of

methanol

- intoxicated rats. This may explain the effect on

mitochondria and resulting visual dysfunction (Eells et al.,

2000).

The rat m odel appears to be a useful tool to also elucidate

the sequelae of methanol intoxication in humans who are more

sensitive than rodents.

---------------------- **Reliability:** (2) valid with restrict ions

2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

(411) (412) (413) (414)

**Type:** Biochemical or cellular interactions

**Remark:** The cytotoxicity of methanol on C6 rat glioma cells was

measured by various methods: trypan blue exclusion, protein

synthesis, neutral red assay and induction of heat shock

protein synthesis. Methanol showed a very low toxicity with

first toxic effects in all test systems at concentrations

between 1 and 10 Mol (ca. 32

- 320 g/l).

**Test substance:** Methanol from Sigma Chemical s, Germany, no further data.

(415)

**Type:** Biochemical or cellular interactions

**Remark:** The cytotoxicity of methanol was tested in chicken embryo

forebrain neuron cultures using the MTT and the neutral red

assay to predict acute toxicity.

LC50 in the MTT assay was 482.2 uMol and in the neutral red

assay 340.6 uMol. The toxicity of methanol was low in

comparison with the other 49 test substances.

**Test substance:** Methanol from BDH, analar analytical grade, no further data.

(416)

**Type:** B iochemical or cellular interactions

**Remark:** In neuronal cell cultures (pheochromocytoma PC12) alcohols

in combination with neuronal growth factor leads to greatly

enhanced neurite extension and cell differenti ation. The

effect increases with increasing chain length, methanol

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

271 being the less potent substance. The role of this effect in

alcohol toxicity on brain is not understood.

**Test substance:** Methanol, no further data.

(417)

**Type:** Biochemical or cellular interactions

**Remark:** Rat hepatocytes were exposed to methanol for 20 hours at

concentra tions of up to 39.55 ug/ml. The treatment had no

effect on LDH

- leakage, MTT

- reduction, cellular protein

content and neutral red uptake.

**Test substance:** Methanol, no further data.

(418)

**Type:** Distribution

**Remark:** After oral administration of a single dose of methanol to

dogs (1.97 g/kg) about 10 % was excreted unchanged in the

urine. The methanol concentration in the organs was nearly

half as high as that found in the urine. About 20 % of the

administered dose was excreted in urine as formic acid.

Formic acid concentrations in tissues was about half the

serum concentration.

After oral administration of 2.38 g/kg methanol to rabbits

10 % were excreted unchanged in urine. No further details

are reported.

--------------

**Test substance:** Methanol, no further data.

(88) (419)

**Type:** Distribution

**Remark:** The examination of the toxicokinetics of intravenously

administered methanol to female rats as a single dose of 50

or 100 or 2500 mg/kg resulted in a markedly non linear

elimination of methanol from the systemic circulation

suggestive of a significant capacity limited rate of

elimination. The data from the 2500 mg/kg group was

described by a kinetic model incorporating parallel

first

- order and saturable elimination processes, a portion

of this apparent linear elimination pathway was due to

renalexcretion of the unchanged alcohol.

**Test substance:** Methanol, no further data.

(88) (420 )

**Type:** Metabolism

**Remark:** The metabolism of methanol (and ethanol) by catalase and

alcohol

- dehydrogenase was studied in alcohol

- dehydrogenase

positive and negative deer mice. Metabolism by catalas e was

the predominant pathway for both alcohols but both enzymes

contributed to the total metabolism (Bradford et al., 1993).

Methanol metabolism in alcohol

- dehydrogenase deficient deer

mice was inhibited by 4

- methylpyrazole, which inhibited

fatty acyl CoA

- synthetase, thus reducing the availabilty of

H2O2 for catalase dependent alcohol metabolism (Bradford et

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

272 al., 1992).

-----------------------

**Test substance:** Methanol, no further data.

(421) (422)

**Type:** Toxicokinetics

**Remark:** In monkeys, four daily skin application o f 0.5 ml (daily

dose 1.56 g/kg) could be lethal. Rabbits proved to be

distinctly more insensitive to methanol either dermally

applied or also after inhalation exposure. Individual

vari ation was nonetheless large in either species.

**Test substance:** Methanol, no further data.

(156)

**Type:** Toxicokinetics

**Method:** Comparative toxicity and meta bolic study

1. in normal mice a) under folate

- sufficient, b) under

folate

- deprived conditions, and

2. in a C57BL mouse strain suffering catalase deficiency

(catalase considered to be the prime enzyme oxidising

formate) as well as a strain with normal catalase activity.

Methanol and formate levels in blood as well as urinary

formate excretion were compared with acute toxicity (L D50)

under respective conditions after oral administration of

methanol (2

- 8 g/kg bw).

----------------------

**Remark:** Those mice with catalase deficiency showed a somewhat lower

LD50(oral and i.p.) than those with normal catalase level,

irrespective of folate status. Folate

- and catalase

deficient mice excreted considerably higher amounts of

formate into the urine (30. 8

- 40.8 %), with significantly

higher blood formate levels than in those normal, but

formate

- deficient mice (15 to 17 umol/ml vs. 0.8

- 1.4

umol/ml (24 h after 6 and 5 g/kg methanol oral).

In no case, folate deficiency alone could not explain the

poor decrease in LD50 which was unexpected in relation to

the pronounced increase in the formate level with 24 h.

A low supplementation wi th methionine (0.2%) had no

influence on the LD50 under catalase

- and folate

- deficient

conditions, while at 1.8 % methionine, there was an apparent

LD50

- decrease from 7.1 g/kg to 6.4 g/kg.

-----------------

**Test substance:** Methanol, no further data.

**Conclusion:** The authors concluded that the formate concentrations in

plasma do not correlate well with methanol

- induced

lethalities in catalas e

- deficient, but folate

- sufficient

mice or folate

- deficient mice. In addition, urinary

excretion, not oxidation, is the primary means to get rid of

high levels of formate.

The result s in mice with catalase or folate deficiency

- acc.

to authors

- appear to correspond to conditions in

methanol

- poisoned monkeys: high formate level at low pH in

blood.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

273 ------------- ------

(137)

**Type:** Toxicokinetics

**Remark:** The pharmacokinetics of methanol was compared in Fisher

rats and Rhesus monkeys. After i.v. appl ication to rats 96.6

% were metabolized, 2.6 % exhaled and only 0.8 % excreted in

urine. The kinetic parameters measured or cited were: AUC at

2000 ppm 623.6 (rats) and 479.8 (monkeys) ug x h/ml,

t1/2(half

- life) 2.1 hours in rats and 2.9 hours in monkeys,

Vmax15.41 mg/h x kg, Km 33.92 mg/l.

**Test substance:** Radiolabelled Methanol from Sigma Chemicals Corporation, no

further data.

(397)

**Type:** other: Absorption / distribution / inhalation / monkey

**Method:** Comprehensive study design in analogy to Segment

- I study

limited to exposure of fem ale monkeys (Macaca fascicularis)

and qualitative/clinical signs of toxicity. It comprised

- methanol pharmacokinetics

- maternal toxicity (see other entry: 5.8.2)

- reproductiv e toxicity (see other entry: 5.8.2)

- developmental neurotoxicity (see other entry: 5.8.2).

Groups of 11

- 12 adult female monkeys were exposed to

methanol vapor in the concentrations of 0; 200; 600 or 1800

ppm, daily for 2.5 hour before breeding, during breeding

and during pregnancy.

Blood kinetics for methanol, blood concentration of formate

as well as folate were determined 2x/w eek 30 min after the

2.5

- h exposure in female monkeys during the prebreeding

period, after 87 d of exposure and 2x during pregnancy

(Kinetic studies 1 to 4). The number or animals was 9 to 10

per group.

---------------------

**Result:** Methanol: No significant differences in the blood

concentrations between the 4 phases (measured 0.5 h

post

- exposure, n = 11 or 12 (non

- pregn ant); n = 9 or 10

(pregnancy) (Fig 7, Tab. 11, Fig. 14, Part I):

Baseline range approx. 2 ug/ml [approx. 0.06 mM]

200 ppm: range approx. 5 ug/ml [approx. 0.15 mM]

600 ppm: range approx. 10 ug/ml [approx. 0.30 mM]

1800 ppm: range approx. 35

- 40 ug/ml [1mM

- range]

At 1800 ppm, after 5 h elimination, the residual methanol

level was near baseline (max. 2

- fold higher).

The mean estimated elimination half

- lives (for 600 and 1800

ppm) ranged between about 60 to 90 min.

Formate: Irrespective of concentration levels or exposure

intervals, there was no evidence of a significant increase

above the background range of about 0.15

- 0.30 mM (approx.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

274 7 to 14 ug/ml) (Fig. 8, Fig. 15, Tab. 12, Part I).

Folate in serum: There was no significant shift in the

folate levels during pregnancy, independent of exposure

(Tab. 13, Part I).

**Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, well document ed

(423)

**Type:** other: Ocular toxicity / animal model

**Remark:** 1. In a preliminary study, F

- 344 rats fed control and

folate

- deficient diet s and exposed to 1050 mg/m3 methanol (20

h/d, 7 d/wk, 13 weeks) showed degeneration of the retina and

optic nerve under both conditions, but not yet after 4 eeks of

exposure.

However, Long

- Evans rats fed either diet failed to develop any

ocular lesion.

The authors suggest that spontaneous degeneration was specific

for F

- 344 rats and are therefore not suitable for ocular

toxicity studies. (Lee et al. 1991).

2. To create an animal model for human methanol toxicity

folate

- deficient rats (male Long

- Evans) were exposed to

methanol (single oral administration of 2

- 3 .5 g/kg) and

formate accumulation in blood was measured.

Methanol concentrations in blood did not greatly differ

between normal and folate deficient rats, while formate

accumulated in folate

- deficient but not in normal rats.

Inhalation of 1200 or 2000 ppm for 6 hours led also to

increased blood formate levels only in folate deficient

rats.

If in fact formate accumu lation is responsible for methanol

toxicity the animal model may be suitable to test these

effects. (Lee et al. 1994)

-------------

**Test substance:** Methanol, no further data.

(424) (425)

**Type:** other: Ocular toxicity / formate /monkey

**Method:** Test design to elucidate the mechanism of methanol

- induced

toxicity syndrome and ocular toxicity: here influence of

formate (McMartin et al., 1978).

Four male rhesus monkeys (Macaca mulatta) received buffered

Naformate i.v. to a loading of 1.25 mmol/kg bw., with a mean

infusion rat e of 3.1 mEq/kg/h. This procedure produced no

acidosis.

(see also other investigations of this group under 5.4)

----------------

**Result:** After 10 h, all animals accumulated formate in blo od between

10 to 30 mEq/L. Blood pHs were maintained between 7.4 and

7.6. Pupillary reflexes were rapidly altered, and in most

animals no response to light was observed between 24 and 48

h. Ophthalmology revealed marked optic disc edema (mainly in

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

275 the prelaminar region, central portion of the proximal part

of the optic nerve without significantly reaching to the

distal part.

The retina including the ganglion

- cell layer were completely

normal. (McMartin et al., 1978)

-----------

**Conclusion:** These results with formate were completely consistent with

those findings after methanol

- intoxication (see other

entries of this group under 5.4).

Formic acid and not formaldehyde (McMartin et al., 1979) has

to be considered the causative agent for ocular damage

within the methanol

- intoxication syndrome, irrespective of

acidosis.

The mechanism of formate toxicity may be seen in the

inhibition of oxidative phosphorylation by formate based on

the on findings that this substance is an efficient

inhibitor of cytochrome oxidase.

------------- **Reliability:** (2) valid with restrictions

(187) (426)

**Type:** other: Review

**Remark:** The reviewer describes the effects of methanol intoxication

in humans. Methanol affects primarily the central nervous

system involving the vis ual system up to optic atrophy. The

minimal lethal dose in humans is reported to be 80 g.

Methanol causes a variety of histopathological changes in

the central nervous system and especially in the optic

nerve.

**Test substance:** Methanol, no further data.

(427)

**Type:** other: Review

**Remark:** The toxicity, kinetics and metabolism of methanol i s

reviewed. According to the reviewer methanol is readily

resorbed by all exposure routes and metabolized in the liver

firstly to formaldehyde and subsequently to CO2. It is of

moderat e acute toxicity with the central nervous system and

especially the optic nerve as primary target organs.

Methanol was not carcinogenic in rats and mice in

inhalation carcinogenicity studies but in repr oductive

toxicity studies it was teratogenic and embryotoxic in rats

and mice.

----------------

**Test substance:** Methanol, no further data.

(428)

**Type:** other: Review

**Remark:** Potential Health Effects of Gasoline and Its Constituents: A

Review of Current Literature (1990

- 1997) on Toxicological

Data

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

276 (429)

**Type:** other: Risk Assessment Model / inhalation

**Remark:** Quantitative Models of Risk Assessment for Developmental

Neurotoxicants, based on inhalation d ata in SD rats from

Nelson et al., 1995:

The use of continuous foetal weight data was as sensitive as

foetal brain malformation data in estimating excess risk to

methanol inhalation ex posure in developing rat. Excess risk

(assumed to be the additional risk that foetal weight is

below the first percentile of control animals) was estimated

by using a logistic model: 0.1 at 16000 ppm, 0.01 at 5400

ppm, and 0.001 at 980 ppm.

------------

(430) (234) (235)

**Type:** other: Toxicokinetics / ethanol influence

**Method:** Com parative study with ADH inhibitors: Male CD

- 1 mouse i.p

injected 1 g/kg methanol followed by the inhibitor (i.p.).

Elimination of methanol from blood was determined (t/2, AUC,

mean elemination rate).

**Re sult:**  Ethanol (4 g/kg) doubled the elimination half life as

compared with control (approx. 9 vs. approx. 5 h); chloral

hydrate (0.4 g/kg) had no significant impact on methanol

elimination. But in combination (0.2 g/kg) with ethanol (4

g/kg), methanol elimination was retarded at a factor of >3

as compared with the control, which corresponded to about a

twofold decrease of the elimination rate as compared with

ethanol alone.

(431)

**Type:** other: absorption

**Remark:** In shortly described experiments, monkeys (normal or folate

deficient) exposed to low levels of methanol (10

- 900 ppm =

0.013

- 1.17 mg/l) showed a methanol

- derived increment of

formate in blood which were 10

- 1000 fold lower than those

levels norma lly present in the body.

It is concluded that exposure to methanol at the threshold

limit value of 200 ppm (0.26 mg/l) is unlikely to pose an

unacceptable risk in healthy or even moderately

folate

- deficient individuals.

------------------

**Test substance:** Methanol, no further data.

**Reliability:** (2) valid with restrictions

24

- DEC

- 2004 (432)

**R emark:**  Acute methanol poisoning in men and monkeys: latency,

acidosis, ocular toxicity, not observed in most other

species tested. Formic acid as likely causative agent is

discussed: formate a ccumulation mainly due to limitation/low

level in hepatic tetrahydrofolate along with a low

10formyl

- THF dehydrogenase.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

277 ---------

**Test substance:** Methanol, no further data.

(281)

**Method:** In anesthetised dogs, the acute cardiovascular effect of

methanol was investigated after i.v. infusion (20

- %

solution).

---- ---------

**Remark:** Methanol distributed rapidly in the body fluid.

Methanol concentrations of 1.3

- 2.0 mg/ ml blood reduced

cardiac stroke volume, cardiac output per minute, systemic

blood pres sure and blood flow through the Arteria femoralis

and Ateria carotis. On the other hand, peripheral

resistan ce increased. Death caused by cardiac arrest came

about at a methanol concentration in blood of 4 mg/ml.

--------------

**Test substance:** Methanol, no further data.

(433)

**Remark:** In anesthetised dogs, methanol retention was about 85 % of

inhaled methanol at exposure concentrations from 360 to 960

mg/m3. Absorption was dependent on ventilation rate.

**Test substance:** Methanol, "pure", no further data.

(434)

**Remark:** In vitro study on enzyme preparations and in vivo:

The influence of glutathione and other SH

- compounds on the

metabolic inactivation of the alcohol and aldehyde

dehydrogenase by aldhyde and methanol was examined by

poisoning male adult mice by an i.p. dose of 8.6 g/kg

methanol. After 10 min, 1, 2 or 3 hours, the mice were

treated either with 4 mg/kg 2,4

- dimercaptopropanol, 120 mg

cysteine or 27 mg mercaptoethanol.

Mean survival time of methanol

- treated control animals was

no longer than about 10 hours. But following preventive

treatme nt, mice survived considerably longer: the earlier

the protective agent was applied, the longer was the

survival up to 12

- fold compared with the control with a few

treated animals alive for 2 months (en d of experiment).

**Test substance:** Methanol, from Fisher Scientific Comp., no further data.

**Conclusion:** These findings suggest that part of methanol intoxication in

mice may be caused by formaldehyde

- exerted inactivation of

alcohol and aldehyde dehydrogenase, as SH

- reagents are also

effective in the mitigation of poisoning.

--------------

**Method:** In this study, enzyme activities responsible for the

oxidation of methanol to formate were investigated in liver

homogenate and the eye of Long

- Evans rats.

----------------------

**Remark:** There was no evidence of an alcohol dehydrogenase

- dependent

metabolism of methanol in the liver and in the eye.

In liver, conversion of methanol was catalised by an

acid

- labile catalase and acid

- stable alcohol

- oxidising

enzyme. In the eye, there was onl y acid

- labile catalase

activity.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

278 Km value(catalase, eye) = 366 mM,

Km value(catalase, liver) = 1.06 mM,

Vmax value Werte(catalase, eye) = 19.8 umol/h/g,

Vmax value Werte(catalase, liver) = 18.3 umol/h/g

Km value(alc

- oxid. enzyme, liver) = 147 mM,

Vmax value(alc

- oxid. enzyme, liver) = 30.5 umol/h/g tissue.

Metab. capacity(tissue weight xvmax)[eye] = 2.7 umol/h

Metab. capacity(tissue weight xvmax)[liver catal.] = 255

umol/h

Metab. capacity (tissue weight x vmax)[liver alc.oxid.] =

425 umol/h.

------------------------------

**Test substance:** Methanol, no further data.

**Conclusion:** The results demonstrate that acid

- labile catalase is the

sole active system in rat eye to metabolise methanol, while

in liver both, catalase and an acid

- sta ble alcohol

- oxidising

enzyme system significantly contribute to the biooxidation

of methanol to formaldhyde.

------------------

(436)

**Remark:** The toxicity of methanol in relation to formate metabolismus

was investigated in isolated hepatocytes (Sprague

- Dawley

rats):

In isolated hepatocytes, formate arising from methanol is

further oxidised by folate

- dependent pathways (10

- formyl

- THF

synthetase and 10

- formyl

- THF dehydrogenase) rather than by

the catalase

- peroxidase reactions.

In these experi ments, oxidation rate of methanol to CO2 was

significantly lower than found in vivo. After addition of

methionine, formation of CO2 from methanol increased, which

was ascribed to a restoration and stim ulation of the formate

oxidation and is based on the fact that in in

- vitro liver

preparations, the methionine content is at a factor of about

10 lower than in vivo.

In hepatocytes from folate

- deficient rats, oxidation to

formate was poor or absent even after addition of

5

- formyltetrahydrofolate or folate, but could be restored by

addition of methionine.

The functio nal role of methionine remains to be elucidated.

--------------

**Test substance:** Methanol, radiolabelled from New England Nuclear, no

furtherdata.

(437)

**Method:** Methanol was administered by gavage to normal and

folate

- deficient rhesus monkeys (Macaca fascicularis) at a

dose of 3000 mg/kg and 2000 mg/kg bw, respectively.

Formalde hyde and formate were analyesd in blood, urine, and

in the cerebral fluid, the orbit fluid of the eye, liver,

kidney, brain, and optic nerve.

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

279 -----------------

**Result:** There was no evidence of any formaldehyde increase at a time

period when signs of acidosis and methanol intoxication

occurred, while significant increases in formate were found

in blood and other body fluids.

For verification, one untreated monkey was infused

14C

- radiolabelled formaldehyde (1mmol/kg bw, 0.2 M solution)

over 3

- 4 min. The half

- life time was 1.5 min.

-------------------

**Test substance:** Methanol, radiolabelled from New England Nuclear, no

further data.

**Conclusion:** Formaldehyde cannot be considered as the main factor in the

development of methanol

- induced toxicity.

**Reliability:** (2) valid w ith restrictions

(426)

**Remark:** Human alcohol dehydrogenase exhibited a 10

- fold higher Km

value for methanol associated with a 10

- fold lower activity

than for ethanol.

This striking difference suggests that there are different

kinetic mechanisms of dehydrogenation of either alcohol.

**Test substance:** Methanol, purified, from Merck, Germany, no further data.

(438)

**Remark:** Formic acid arising from methanol is eliminated at different

rates by different species: most rapidly in rats with T/2 of

approx. 12 min, then in rabbits with T/2 of about 32 min.,

dog with T/2 of 77 min., and human with T/2 of about 55 min.

Higher folate levels in blood appear to correlate to lower

formate leve ls. Pretreatment of dogs with amethopterin

(folic

- acid antagonist) induced a pronounced increase in

formate of plasma and urine. On contrast, prophylactic

administration of folic acid completely preven ted

formic

- acid accumulation in blood.

-------------------

**Test substance:** Methanol, no further data.

(439)

**Test substance:** Methanol, n o further data.

(279) (230) (429) (440) (441) (442) (443) (444) (384) (88) (445)

(446) (447) (233) (326) (284) (448) (449) (450) (451)

**Remark:** In alcohol

- free persons, the endogenous methanol blood level

ranges from 0.5

- 1 mg/l (1.5 mg/l not exceeded), the urine

level from 0.32

- 2.61 mg/l, and the endogenous breath level

between 0.21

- 0.79 ug/l. Long

- term ingestion of significant

amounts of ethanol (drinks, wine etc.), the background

methanol levels may shift to higher values due to cumulation

of endogenously formed methanol, for example through

competitively inhibition of methanol by ethanol.

----------------

**Test substance:** Methanol, no further data.

(452)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

280 **Remark:** Laquer thinner containing methanol, which is used for

in halation abuse, affected male fertility with a

significant decrease in testicular and prostatic weights and

plasma testosterone levels. Methanol exposure alone did not

produce such effects.

**Test substan ce:**  Methanol, no further data.

(453)

**Remark:** The metabolism of formate was compared in monkeys and rats.

Formate oxidation was 50 % lower in monkeys than in rats.

The activity of a range of folate

- dependent enzymes in

liver tissue as well as the hepatic H4folate levels were

compared. H4folate levels were 40 % lower in monkeys than in

rats. Enzyme activities were partly higher and partly lower

in monkeys than in rats, and their specific contribution to

methanol toxicity could not be clarified.

**Test substance:** Methanol, no further data.

(454)

**Remark:** A range of publications with respect to the role of formate

toxicity in methanol toxicity in humans are reviewed. The

difference of methanol toxicity in primates and rodents is

suspected to derive from formate accumulation due to lower

formate metabolism. In fact humans and monkeys possess low

hepatic H4folate levels and low rat es of formate oxidation.

Formate itself produces blindness in monkeys, the major

toxic effect of methanol.

**Test substance:** Methanol, no further data.

(281)

**Remark:** The role of liver catalase activity and H2O2 formation on

methanol oxidation in the liver is discussed. According to

the author the methanol oxidation is mainly dependent on

the rate of H2O2 generation, despite the fact that catalase

plays a role in methanol metabolism in liver tissue.

---------------

**Test substance:** Methanol, no further data.

(455)

**Remark:** The pigtail monkey was tested for effects of toxic methanol

exposure to establish an animal model for human methanol

toxicity. Blood acidosis was measured a s major toxic effect

of methanol and the responsible acid was identified.

In pigtail monkeys but not in rhesus monkeys significant

blood acidosis was produced by a single i.p. dose of 4 g/kg

bodyweight methanol. Blood concentration of formate was

correlated with acidosis, no other organic acid was found

at increased levels in acidotic monkeys.

**Test substance:** Methanol, reagent grade, no further data

. **Reliability:** (2) valid with restrictions

(168)

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

281 **Remark:** Male rats were exposed to methanol at 200 ppm (0.26 mg/l)

for up to 6 weeks (8 hours/day , 5 days/week) did not show

an effect on serum testosterone levels. Exposure to 800 ppm

(1.04 mg/l) for up to 13 weeks had no effect on testes to

body weight ratio and had no effect on testicular morpho logy

in normal and folate deficient rats. A greater incidence of

testicular degeneration was seen only in 18 month old

folate

- deficient rats exposed to 800 ppm methanol for 13

weeks (2 0 hours/day, 7 days/week, 12

- 13 rats/group). The

results are equivocal.

**Test substance:** Methanol, no further data.

(456)

**Remark:** Male rats were ex posed to methanol at 200 ppm (0.26 mg/l)

for 6 hour/day for up to 1 week. Compared to controls

methanol treatment for 1 week led to decreased plasma

levels of testosterone, luteinizing hormone and

corticosterone. The changes were not statistically

signficant.

**Test substance:** Methanol, no further data.

(195)

**Remark:** 81

- 88 % of inh aled methanol was retained in the

respiratory tract in dogs. The inhaled fraction was related

to the concentration in air with a 73 % retention at 390

mg/m3 and a 90 % retention at 720 mg/m3.

**Test subst ance:**  Methanol, no further data.

**Reliability:** (2) valid with restrictions

(434)

**Remark:** Methanol is metabolised in rodents and primates in

diff erent ways and different speed. In rodents methanol is

primarily metabolised to formaldehyde by the

catalase

- peroxidase system. The Vmax value was determined at

30 mg/kg/hour. In humans and monkeys the alcohol

dehydrogenase oxidizes methanol to formaldehyde more

rapidly. The Vmax value in primates is 48 mg/kg/hour.

Formaldehyde is finally oxidized to CO2 by folate dependent

enzymes. Primates have less folate than rodents and

therefore the detoxication of methanol may be limited by the

last steps of metabolism in primates.

**Test substance:** Methanol, no further data.

(457)

**Remark:** 1. A high dose of methanol (single 3 g/kg, oral) to male

Wistar rats gave some evidence of increased LPO with a

transient time

- dependent increase in MDA in liver and serum

(about +20 % and 40 %, respectively, reversed after 7 d) and

wth transient decrease in components of the antioxidant

system: GSH in liver after 12 h and 2 d, recovered at day 5

(in between no data), various enzymes also showed a

transient decrease (Cu,Zn

- Superoxide dismutase

,GSH

- Peroxidase, GSSG

- Reductase).

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

282 Blood methanol peak was ca. 2300 ug/ml after 6 h and rapidly

declined (Skrzydlewska et al. 1997, 1998).

-------------

2. After oral doses (1.5, 3, 6 g/kg bw) to male Wistar

rats, the activity of proteases (elöastase and cathepsin G)

were incresed from 12 h to 5 d in relation to the methanol

dose. Concomitantly, the activity of their inhibitors was

reduced. Similar disorders in the

proteolytic

- antiproteolytic balan ce in favour of protease

action can also be observed in human acute intoxications

with methanol. The further implications are addressed: blood

platrelet action, protein degradation, release of

anaphylotoxins and other peptides (e.g. kinins) incuding

vasoactive mediators, activation of other proteases etc.

(Skrzydlewska et al. 1999).

----------------

3. The effe ct of methanol intoxication (6 g/kg bw) on

protein

modification in the liver of rats was investigated.

An increase in free radicals were determined 6 and 12 hours

after intoxication.

Malondialdehyde and carbonyl groups in protein were

increased; the levels of amino groups and sulphydryl groups

and the amount of tryptophan in proteins were decreased,

whereas the amou nt of bi

- tyrosine was increased.

According to the authors changes in the protein structure

resulted both from free radical action and formaldehyde

generation during methanol intoxication (Skrzydlewska e t

al., 2000).

(458) (459) (460) (461) (462) (463)

**Remark:** The PBPK approach was used to develop a model of methanol

disposition during gestation in rats and mice. To validate

this model, concentrations of methanol in the dams and the

conceptus were determined after methanol exposure of rats on

GD 14 and 20 and of mice on gd 18.

In pregnant anim als, conceptal/maternal AUC and Cmax

- ratios

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

283 decreased with increasing dose at both gd 14 and 20 and at

gd 18 in the mouse. Additionally, the conceptal/maternal

diffusion

- constant ratio consistently dec reased with

increasing dose in pregnant rats and mice.

These results are consistent with earlier observations that

methanol limits its own delivery to the conceptus.

It was further s hown that the maximal rate of methanol

elimination (vmax) in vivo decreased at term in both species

with 65 to 80 % of that in non

- pregnant rodents. This was

also found in vitro in rodent liver homogena tes. This

appears to suggest that not only altered disposition but

also a change in metabolic capacities makes the difference

in gestation. Fetal rodent liver was capable of metabolizing

methanol in vitro, but only at a rate of <5 % of respective

adult livers. (Ward and Pollack, 1996; Pollack and Brouwer,

1996)

**Reliability:** (2) valid with restrictions

(395) (464) (465)

**Method:** In

- vitro study on human hepatic enzyme fraction to examine

various inhibitors of ADH with methanol as substrate

**Result:** In methanol intoxication, cimetidine may potential ly be a

better antidote than 4

- methylpyrazole because of its

slightly higher inhibitory actitivity in particular at

higher methanol concentrations.

(466)

**Remark:** According to Caprino and Togna, 1998:

"Intoxication [of methanol] has delayed onset characterized

by acidosis, mental confusion, ocular toxicity with visual

disturbance [recently suggested as due to intraretinal

metabolism of methanol, rather than to elevated blood

formate levels (Garner et al., 1995a,b)], reversible or

permanent blindness, and, in seve re cases, death. These

effects represent a classic example of lethal synthesis in

which toxic metabolites can cause fatality after

characteristic latency period."

............

"...no interactive effects between methanol and gasoline

were found in a short

- term inhalation study in rats (Poon et

al., 1995), and a study on perinatal exposure to low

concentrations of me thanol resulted... in no significant

abnormalities in the brain of treated rats (Stern et al.,

1996)."

"There is little evidence from available information of

human health effects fro m low

- level exposure which

demonstrates that methanol vapors from motor vehicle fuel

can cause acute adverse effects to health. Lee et al. (1992)

demonstrated that after 6

- hr exposure to current methano l

threshold limit value (TLV) of 200 ppm, the formate does not

accumulate in blood in human subjects at rest and during

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

284 exercise. This was confirmed by other authors (D ́Alessandro

et a l., 1994; Osterloh et al., 1996)."

(429) (391) (392) (196) (277) (467) (241)

**Remark:** Compilation of human case reports on acute methanol

intoxications with a collection of the blood leve ls found in

sublethal and lethal events at various time intervals after

ingestion.

A provisional nomogram has been derived, beginning at a 3

- h

concentration of 20 mM and ending at a 1 30

- h concentration

of 0.5 mM. Concentrations at any time

- point above this

straight line result either in life

- threatening poisoning

or a lethal outcome.

----------------

(468)

**Result:** After oral application of methanol (3 g/kg bw), the

thiobarbituric acid

- reactive substances (TAS) indicating

lipid peroxidati on were significantly increased in brain

tissue from 6 h to 7 d after treatment, also activities of

liver GSH peroxidase and GSH reductase were reduced after 6

h already.

N

- Acetylcyst eine exerted an protective antioxidant effect in

the brain and liver of methanol

- treated rats: TAS remained

on the background

- level after prior i.p. treatment with

N

- acetylcysteine (150 mg/kg). It resul ted in a lower degree

of parenchymal damage (Kasacka and Skrzydlewska, 2001).

Furthermore, concomitantly, methanol

- induced GSH

- depletion,

which was significant after 24 h and steadily increased

until 7 d, could be mitigated, and enzyme activities

involved in the removal of reactive oxygen species were also

similar to the untreated control while significantly

increased without protec tive counteraction.

A Trolox

- Derivative, an analog of alpha

- tocopherol, is

introduced as a beneficial antioxidant which prevents or

reduces methanol

- induced LPO in Wistar rats (Skrzydlewska

and Farbiszewski, 1997).

-------------------

(469) (470) (471) (472)

(250) (251) (397) (196) (252) (264)

**Method:** Co mprehensive study programme on three species (rat, mouse,

monkey) including metabolic, pharmacokinetic and short

- , long

- term and reproduction studies and carcinogenicity

studies:

Here two

- generation reproduction study in rats (see other

entry, 5.8.2).

In 9

- week old F1 pups, blood methanol was measured, but not

formate (p. 191). Exposure: continuous for 19 to 20 h/d.

-------------------------

**Remark:** Blood levels of methanol measured in the F1 offsprings (age

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

285 9 weeks) (NEDO, 1987, p. 191):

controls (baseline): approx. 2

- 3 ug/ml

10 ppm methanol: approx. 3

- 3.5 ug/ml

100 ppm " : approx. 1

- 4.2 ug/ml

1000 ppm " : approx. 53 (males)

- 100 (females) ug/ml.

There no data on formate.

------------- **Reliability:** (2) valid with restrictions

Test procedures in accordance with accepted standard

methods, sufficiently documented

(167)

**Remark:** Under in

- vitro conditions, in the presence of liver

microsomes or liver nuclei fraction, hydroxymethyl radicals

were formed from methanol. This bioactivation was observed

only under anaerobic condi tions which also resulted in the

production of formaldhyde. Formaldehyde was the primary

intermediate during aerobic incubation. But both

formaldehyde

- generating processes appear to be different,

because the anaerobic one was strongly inhibited by

diphenylene iodonium, while the aerobic one was not.

(473)

**Result:** Male rats were expose d to methanol vapour at concentrations

of 200, 5000 or 10000 ppm (0.26, 6.5 or 13 mg/l) for 6 hours

and killed immediately after exposure or at 24 hours. Rats

were divided in acclimated (2 weeks prior t o handling) and

non

- acclimated groups. At 6 h, sham

- exposed rats already

showed an increase in the level of serum LH. Effects on

serum hormone levels (luteinizing hormone, follicle

sti mulating hormone, testosterone, prolactine) were

inconsistent with partly no effects and partlycontradictory

results. According to the author, methanol treatment led

only to increased prolactin levels i n serum under both

acclimated and non

- acclimated conditions.

**Test substance:** Methanol, no further data.

(157)

**Result:** The oral administration of 14C

- methanol to rats is reported

to have resulted in covalent binding to hemoglobin. A linear

relationship from 10 to 100 umol/kg was found.

------------------- **Reliability:** (4) not assignable

Only abstract, no detailed data.

(474)

**Remark:** In clinical cases, the methanol level is low in the last

phase of poisoning.

In Rhesus monkeys given a single methanol dose of 1 g/kg, 37

mg(kg*h) was oxidized (Makar et al, 1968).

### OECD SIDS METHANOL

5. TOXICITY ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

286 Provided that the rate is the same in man, the amount of

methanol oxidized during 18 h (t he average time needed for

development of severe acidosis in clinical cases) would be

0.7 g/kg.

Acc. to Roe (1982), it seems reasonable to regard 1 g/kg of

methanol as the approximate minimal lethal dose in man.

**Reliability:** (4) not assignable

Secondary source

(475) (284)

**Remark:** In Rhesus monkeys given a single methanol dose of 1 g /kg, 37

mg(kg*h) was oxidized (Makar et al, 1968).

**Reliability:** (4) not assignable

(475) (284)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

287 (1) BASF Specification METHANOL; according to "Standard

Specification Methanol" ASTM D 1152

- 84.

(2) BASF AG, Safety data sheet METHANOL PURE, 19.05.2005

(3) GESTIS

- Substance Database (Information system on hazardous

substances of the Berufsgenossenschaften), 29.08.2005

(4) BASF AG, Product Details METHANOL TECHN.II, 25.08.2005

(5) IMPCA Methanol Reference Specifications, April 1999

(6) Commission Directive 2004/73/EC, 29 April 2004 (29th

adaption to the technical progress of 67/548/EEC)

(7) Ullmann's Encyclopedia of Indus trial Chemistry, Sixth

Edition, 2000 Electronic Release

(8) World Methanol Analysis 2001, CMAI

(9) "Beyond the Internal Combustion Engine: The Promise of

Methanol Fuel Cell Vehicles," Prepared for the Methanol

Institute by Brea kthrough Technologies Institute, January

2001.

(10) The Methanol Institute, 2003

(11) List of MAK

- and BAT

- values 2004 (Notification 40),

23.07.2004, WILEY

- VCH Verlag GmbH, Weinheim, Germany

(12) Catalogue of Substances Hazardous to Water

- Umweltbundesamt

Berlin, 31.08.2005

(13) National Chemical Inventories, 2005 Issue 1

(14) Ariel WebInsight® 3.2 Chemical Database, 18.08.2005

(15) The Merck Index, 11th edition, p. 939, 1989

(16) BASF AG, Safety data sheet Methano l pure, 03.11.1999

(17) Yaws C.L. et al., Chemical Engineering, p. 115

- 118, July

1990

(18) Kirk

- Othmer, Encyclopedia of Chemical Technology, 4th.

Edition, Vol. 1, p. 828, 1991

(19) Deutsche Forschungsgemeinschaft, Senatskommission zur

Prüfung gesundheitsschädlicher Arbeitsstoffe, Mitteilung 29,

MAK

- und BAT

- Werte

- Liste 1993

(20) Daubert T. E. and Danner R. P., Physical and Thermodynamic

Properties of Pure Chemicals Data Compilations, Washington,

D. C., Taylor and Francis, 1989

(21) Hansch C., Leo A., Substituent Constants for Correlation

Analysis in Chemistry and Biology, John Wiley & Sons, New

York, 1979

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

288 (22) Sangster J., J. Phys. Chem. Ref. Data, Vol. 18, No. 3, p.

1150, 1989

(23) Rekker R.F., The Hydrophobic Fragmental Constant, Elsevier

Scientific Publishing Company, Amsterdam, 1977

(24) Commercial Solvents Corporation (CSC), Industrial Chemicals

Department, Methanol

(25) BASF AG, Sicherheitstechn ische Kennzahlen, unveröffentlichte

Untersuchung, Probe Nr. 72/0239, 17.05.1972

(26) BASF AG, Sicherheitstechnische Kenndaten, unveröffentlichte

Untersuchung, TKM/SIK 78/1255, 15.01.1979

(27) ARCO Chemical Company, Technical Literature Methanol, 1983

(28) BASF AG, Sicherheitstechnik, interne Mitteilung, 02.11.1999

(29) Redeker T., Schön G., Sicherheitstechnische Kennzahlen

brennbarer Gase und Dämpfe, 6. Nachtrag, 1990

(30) Mathis E., Chemie für Labor und Betrieb, Volume 2 9, p. 490

-

495, 1978

(31) Atkinson,R., J. Phys. Chem. Ref. Data Monograph 1, p.160,

(1989)

(32) Atkinson,R. et al., Atmospheric Environment 26A(7),

1187

- 1230, (1992)

(33) Lemaire,L. et al., An assessment of test methods for p hoto

- degradation of chemicals in the environment, Chemosphere

11(2), 119

- 164, (1982)

(34) Kloepffer,W. et al., Quantitative Erfassung der

Photochemischen Transformationsprozesse in der

Troposphaere, Chemiker

- Zeitung 110(2), 57

- 61, (1986)

(35) Kelly,T.J. et al., Environ. Sci. Technol.28(8), 378A

- 387A,

(1994)

(36) Atkinson, R. (1994): J. Phys. Chem. Ref. Data Monograph 2

(37) Environmental Hazard Assessment: Methanol (Draft), EPTS/21D,

February 1993

(3 8) Barnes,I. et al., Reactivity studies of organic substances

towards hydroxyl radicals under atmospheric conditions,

Atmos. Env. 16(3), 545

- 550, (1982), zitiert nach:

Environmental Hazard Assessment: Methanol (Entwurf),

EPTS/21 D, February 1993

(39) Kao,A.S., J. Air Waste Manage. Assoc.44, 683

- 696, (1994)

(40) Guesten,H. et al., Atmos. Environ.15, 1763

- 1765, (1981)

(41) Kloepffer,W., Kohl,E.

- G., Ecotoxicology and Environmental

Safety 22, 67

- 78, (1991)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

289 (42) Lyma n,W.J. et al., Handbook of Chemical Property Estimation

Methods, McGraw

- Hill, New York, 7

- 4, (1982)

(43) Graedel,T.E. et al., Atmospheric chemical compounds,

Sources, occurrence and bioassay, Academic Press Inc.,

zitiert nach Enviro nmental Hazard Assessment: Methanol

(Entwurf), EPTS/21D, February 1993

(44) Holzer,G. et al., Collection and analysis of trace organic

emissions from natural sources, J. Chr. 142, 755

- 764,

(1977), zitiert nach Environmental Hazard A ssessment:

Methanol (Entwurf), EPTS/21D, February 1993

(45) Isidorov, V.A. et al.: Volatile organic compounds in the

atmosphere of forests, Atmos. Env. 19(1), 1

- 8, 1985 (cited

in: Environmental Hazard Assessment: Methanol

(dr aft), EPTS/21D, February 1993)

(46) Owens, L.D. et al.: Identification of plant volatiles that

simulate microbial respiration and growth in soil.

Phytopathol., 59, 1468

- 1472, 1969 (cited in Environmental

Hazard Assessment: Methanol (Entwurf), EPTS/21D, February

1993)

1993

(47) Jonsson, A. et al.: Measurements of some low

molecular

- weight oxygenated, aromatic, and chlorinated

hydrocarbons in ambient air and in vehicle emissions.

Environment

International, 11, 383

- 392, 1985 (cited in:

Environmental Hazard Assessment: Methanol (draft), EPTS/21D,

February 1993)

(48) Jungclaus,G.A. et al., Organic compounds in an industrial

wastewater: A case study of their environmental impact,

Environ. Sci. Technol. 12(1), 88

- 96, (1978), zitiert nach:

Environmental Hazard Assessment: Methanol (Entwurf),

EPTS/21D, February 1993

(49) Mohr, D.H., King, C.J.: Identification of polar organic

compounds in coal

- ga sification condensate water by gas

chromatography

- mass spectrometry analysis of

high

- performance liquid chromatography fractions. Environ.

Sci. Technol., 19(10), 929

- 935, 1985 (cited in:

Environmental Hazard Assessment: Methanol (draft), EPTS/21D,

February 1993)

(50) Pellizzari,E.D. et al., Bull. Environ. Contam. Toxicol.

28(3), 322

- 328, (1982)

(51) Coleman, E.C. et al.: Isolation and identification of

volatile compounds from baked potatoes. J. Agric. F ood

Chem., 29, 42

- 48, 1981 (cited in: Environmental

Hazard Assessment: Methanol (draft), EPTS/21D, February

1993)

(52) Kinlin,T.E. et al., Volatile components of roasted filberts,

J. Agric. Food Chem. 20(5), 1020

- 1028, (1972) , zitiert nach:

Environmental Hazard Assessment: Methanol (Entwurf), EPTS/

21D, February 1993

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

290 (53) Lovegren,N.V. et al., Volatile constituents of dried

legumes, J. Agric. Food Chem. 27(4), 851

- 853, (1979),

zitiert nach: Enviro nmental Hazard Assessment: Methanol

(Entwurf), EPTS/21D, February 1993

(54) Howard,P.H. (ed.), Handbook of Environmental Fate and Expo

- sure Data for Organic Chemicals, Volume II, Solvents,

Lewis Publishers Inc., Michigan, 310

- 317 (1990)

(55) BASF AG, Department of Product Safety, unpublished

calculation, 10 Oct. 2002

(56) Lokke, H.: Leaching of ethylene glycol and ethanol in

subsoils. Wat. Air Soil Poll., 22, 373

- 387, 1984 (cited in:

Environmental Hazard Assessment: Methanol (Draft), EPTS/21D,

February 1993

(57) BASF AG, department of ecology, unpublished calculation,

09.11.2004

(58) Lyman,W.J. et al., Handbook of Chemical Property Estimation

Methods, McGraw

- Hill, New York, (198 2)

(59) Gaffney,J.S. et al., Environ. Sci. Technol., Vol. 21, No. g,

519

- 524, 1987

(60) BASF AG, department of ecology, unpublished calculation,

20.12.1998

(61) BASF AG, Department of Product Safety, unpublished

calculation, 17.04.2003

(62) Kempa,E.S., Oesterreichische

- Abwasser

- Rundschau, Folge 2,

20

- 25, (1976)

(63) Lamb,Ch.B., Jenkins,G.F., Proceedings of the 8th Industrial

Waste Conf., Purdue University, 326

- 339, (1952)

(64) Price,K.S. et al., JWPCF 4 6(1), 63

- 77, (1974)

(65) Wagner,R., Vom Wasser 47, 241

- 265, (1976)

(66) Matsui, S. et al.: Experience of 16 years' operation and

maintenance of the Fukashiba industrial wastewater treatment

plant of the Kashima petrochemical complex

- I I.

Biodegradability of 37 organic substances and 28 process

wastewaters. Wat. Sci. Technol., 20(10), 201

- 210, 1988

(67) HUELS AG, DIN

- Sicherheitsdatenblatt 'Methanol', (15.03.93)

(68) Takemoto,Sh. et al., Suishitsu

- Odaku

- Kenkyu 4(2), 80

- 90,

(1981)

(69) Hoechst AG; unveroeffentlichte Untersuchung (RWL 18.06.73);

(1973)

(70) Pugh, L.B.; et al.: Bioremediation of contaminated soils and

groundwater at a former solvent storage site

Biotechnol. Ind. Waste Trea t. Biorem.

page 195

- 211 (1996)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

291 (71) Swain,H.M., Somerville,H.J., Microbial metabolism of metha

- nol in a model activated sludge system, J. Appl. Bacteriol.

45, 147

- 151, (1978), zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

(72) Goldsmith,Jr.,C.D., Biodegradation of methanol and tertiary

butyl alcohol in previously uncontaminated subsurface

systems (groundwater, gasohol, Virginia, Pennsylvania,

New York), Dis s. Abstr. Int. B 46(11), 3767, (1985),

zitiert nach: Environmental Hazard Assessment: Methanol

(Entwurf), EPTS/21D, February 1993

(73) Novak,J.T. et al., Biodegradation of methanol and tertiary

butyl alcohol in subsurface systems, W at. Sci. Technol. 17,

71

- 85, (1985), zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

(74) Van Beelen,P., Stygologia 5(4), 199

- 212, (1990)

(75) Bekes,J. et al., Decomposition of methanol under t he con

- ditions of an anaerobic fermentation, Proceedings of the

Hungarian 15th Annual Meeting on Biochemistry, 27

- 28,

(1975), zitiert nach: Environmental Hazard Assessment:

Methanol (Entwurf), EPTS/21D, February 1993

(76) Ch ou,W.L. et al., Acclimation and degradation of petro

- chemical wastewater components by methane fermentation,

Biotechnol. Bioeng. Symp. No.8, 391

- 414, (1978)

(77) Oremland,R.S. et al., Methane production and simultaneous

sulphate re duction in anoxic, salt marsh sediments, Nature

296, 143

- 145, (1982)

(78) Hanson, R.S.: Ecology and diversity of methylotrophic

organisms. Adv. Appl. Microbiol., 26, 3

- 39, 1980 (cited in:

Environmental Hazard Assessment: Methanol

(Draft), EPTS/21D, February 1993)

(79) Amanchukwu,S.C. et al., Hydrocarbon degradation and utili

- sation by a palm

- wine yeast isolate, FEMS Microbiol. Lett.

57(2), 151

- 154, (1989), zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

(80) Frankenberger,Jr.,W.T., Troeh,F.R., Bacterial utilisation of

simple alcohols and their influence on saturated hydraulic

conductivity, Soil Sci. Soc. Am. J. 46(3), 535

- 538, (1982),

zi tiert nach: Environmental Hazard Assessment: Methanol

(Entwurf), EPTS/21D, February 1993

(81) Okpokwasili,G.C., Amanchukwu,S.C., Petroleum hydrocarbon

degradation by Candida species, Environment International

14(3), 243

- 247, (1988), zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

(82) Urakami,T., Yano,I., Methanol

- utilising Mycobacterium

strains isolated from soil, J. Gen. Appl. Microbiol. 35,

125

- 133, (1989), zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

292 (83) Gluth,G. et al., Accumulation of pollutants in fish, Comp.

Biochem. Physiol. 81C(2), 273

- 277, (1985)

(84) Freitag,D. et al., Chemosphere 14, 1589

- 1 616, (1985)

(85) Hansch,C., Leo,A.J., Medchem. Project Issue No.26, Claremont

CA, Pomona College, (1985)

(86) Lyman,W.J. et al., Handbook of Chemical Property Estimation

Methods, McGraw

- Hill, New York, 5

- 4, (1982)

(87) Geyer, H. et a l., Chemosphere, Vol. 13, No. 2, 269

- 284,

(1984)

(88) IPCS/WHO: Methanol, Environmental Health Criteria 196,

International Programme on Chemical Safety, World Health

Organization, Geneva, 1997

(89) Malcolm Pirnie, Inc., Evaluat ion of the fate and transport

of methanol in the environment, prepared for the American

Methanol Institute, January 1999

(90) Poirer S.H. et al.: Bull. Environ. Contam. Toxicol., 37,

615

- 621, (1986)

(91) Call, D.J. et al.: Toxici ty and metabolism studies with EPA

priority pollutants and related chemicals in freshwater

organisms, EPA

- 600/3

- 83

- 095, PB83

- 263665 (1983)

(92) Veith, G.D. et al.: cited in: Bishop, W.E. et al.: Aquatic

Toxicology and Hazard Assessm ent, 6th Symp. ASTM, STP 802,

90

- 97, Philadelphia (1983)

(93) Brooke L.T. et al.: Acute toxicities of organic chemicals to

Fathead Minnows (Pimephales promelas), University of

Wisconscin

- Superior, supported by U.S.EPA through

cooperative agreements 806864 and 809234, (1984)

(94) Hermens, J. et al.: Ecotox.Environ.Safety, 6(3) 302

- 310,

1982 (cited in UK Dept. of Environment Draft: Environ.Hazard

Assessment Methanol, Feb. 1993)

(95) Koenemann, H.: Toxicology, 19 (1981) 209

- 221; zitiert

im UK Dept. of Environment Draft: Environ.Hazard Assess

-

ment Methanol, Feb. 1993 (zur Verfuegung gestellt von

ICI)

(96) Portmann, J.E. et al.: Shellfish Information Leaflet No.22,

2nd edition (197 1); zitiert im UK Dept. of Environment

Draft: Environ.Hazard Assessment Methanol, Feb. 1993

(zur Verfuegung gestellt von ICI)

(97) Bengtsson, B.E. et al., Chemosphere 8 13(5/6), 613

- 622

(1984)

(98) Linden et al., Chemosphere 8 (1 1/12), 843

- 851 (1979)

(99) Gluth, G. and Hanke, W.: Exotoxicology and Environmental

Safety 9, 179

- 188 (1985).

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

293 (100) Gluth, G. and Hanke, W.: Comp. Biochem. Physiol. 79C, 39

- 45

(1984)

(101) Hoechst AG: unveroeffentlichte Untersuchung, R WL 18.06.1973,

zitiert im Euclid Datensatz der Hoechst AG, 26.05.1994

(102) Juhnke, I. et al., Z. Wasser Abwasser Forsch., 11(5),

161

- 164 (1978);

(103) Johnson, W.W. et al.: Handbook of acute toxicity of

chemicals to fish and aquat ic invertebrates, 1980 (cited in

UK Dept. of Environment Draft: Environ. Hazard Assessment

Methanol, Feb. 1993)

(104) Gillette, L.A. et al.: Water Environ. Research 24,

1397

- 1401(1952)

(105) Nishiuchi, Y. and Hashimoto, Y.: Botyu

- Kagaku 32, 5

- 11

(1967)

(106) Huels, unveroeffentlichte Untersuchung (29.11.87)

Mitteilung vom 23.03.93

(107) NIOSH: Dangerous Properties of Industrial Materials Report,

(1985)

(108) Kuehn,R. et al., Water Research 23(4), 495

- 499, (1989)

(109) Bringmann,G., Kuehn,R., Zeitschrift fuer Wasser

- und

Abwasser

- Forschung, 10(5), 161

- 166, (1977)

(110) Bringmann,G., Kuehn,R., Zeitschrift fuer Wasser

- und

Abwasser

- Forschung, 15(1), 1

- 6, (1982)

(111) Ewell,W.S. et al., Env iron. Toxicol. and Chemistry 5,

831

- 840, (1986)

(112) Calleja, M.C.; et al.: Arch. Environ. Contam. Toxicol. 26,

69

- 78 (1994)

(113) Keller,A.E., Bull. Environ. Contam. Toxicol. 51(5), 696

- 702,

(1993)

(114) Bringmann,G., Kuehn,R ., Vom Wasser 50, 45

- 60, (1978)

(115) Krebs F., DGM 35, H.5/6, 161

- 170, 1991

(116) Stratton,G.W., Smith,T.M., Interaction of organic solvents

with the green alga Chlorella pyrenoidosa, Bull. Environ.

Contam. Toxicol. 40(5), 736

- 742, (1988 )

(117) Bringmann,G., Gesundheits

- Ingenieur 94(12), 366

- 369, (1973)

(118) Bringmann,G., Kuehn,R., Zeitschrift fuer Wasser

- und

Abwasser

- Forschung, 10(3/4), 87

- 98, (1977)

(119) Bringmann,G., Zeitschrift fuer Wasser

- und Abwasser

-

forschu ng 11(6), 210

- 215, (1978)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

294

(120) Schultz,T.W. et al., Ecotoxicology and Environmental Safety

19, 243

- 253, (1990)

(121) Bringmann,G., Kuehn,R., Zeitschrift fuer Wasser

- und

Abwasserforschung 1, 26

- 31, (1980)

(122) Jennings et al. 2001, A ssessing chemical toxicity with the

bioluminescent photobacterium (Vibrio fischeri): a

comparison of three commercial systems. Wat. Res. 35:

3448

- 3456

(123) Hooper,A., Terry,K., J. Bacteriol. 115 (2), 480

- 485, (1973)

(124) Kilroy, A.C., Gray, N.F.: The toxicity of four organic

solvents commonly used in the pharmaceutical industry to

activated sludge, Water Research 26(7), 887

- 892, 1992 (cited

in: Environmental Hazard Assessment: Methanol (Draft),

EPTS/21D , February 1993)

(125) Stratton,G.W., Toxic effects of organic solvents on the

growth of blue

- green algae, Bull. Environ. Contam. Toxicol.

38(6), 1012

- 1019, (1987)

(126) Blum,D.J.W., Speece,R.E., A database of chemical toxicity

to environmental bacteria and its use in interspecies

comparisons and correlations, JWPCF 63(3), 198

- 207, (1991)

(127) Rajini, P.S. et al.: Cytotoxicity of certain organic

solvents and organophosphorus insecticides to the ciliated

proto zoan

Paramecium caudatum. Microbios, 59, 157

- 163, 1989 (cited in:

Environmental Hazard Assessment: Methanol)

(Entwurf), EPTS/21D, February 1993

(128) Kaiser, K.L.E., Palabrica, V.S.: Photobacteria phosphoreum

toxicity data ind ex. Wat. Poll. Res. J. Canada 26(3),

361

- 431, 1991 (cited in: Environmental Hazard Assessment:

Methanol (draft), EPTS/21D, February 1993)

(129) Schiewe,M.H. et al.: Use of a bacterial bioluminescence

assay to assess toxicity of conta minated marine sediments.

Can. J. Fish. Aquat. Sci. 42(7), 1244

- 1248, 1985 (cited in:

Environmental Hazard Assessment: Methanol)

(Entwurf), EPTS/21D, February 1993

(130) Van Haut,V.H., Prinz,B., Beurteilung der relativen Pflanzen

- schaedlichkeit organischer Luftverunreinigungen im LIS

-

Kurzzeittest, Staub

- Reinhalt. Luft 39, 408

- 414, as quoted

in IRPTC Database, zitiert nach: Environmental Hazard

Assessment: Methanol (Entwurf), EPTS/21D, February 1993

(131 ) Nikolayevsky, V.S., Miroshnikova, A.T.: The air pollution

levels permissible for plants. Gig. Sanit., 4, 16

- 18 (in

Russian) (as quoted in US EPA, 1976 (cited in:

Environmental Hazard Assessment: Methanol (Draft) EPTS/21D,

Febr uary 1993)

(132) US EPA, Assessment of methyl alcohol as a potential air

pollution problem, Volume II, Patterson,R.M., Bornstein,

M.I., Garshick,E., prepared for the US EPA by GCA Cor

-

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

295 poration, Massachusetts, NTIS Publication No. PB

- 258 354,

(1976), zitiert nach: Environmental Hazard Assessment:

Methanol (Entwurf), EPTS/21D, February 1993

(133) BASF AG, Dept. of Toxicology: Short

- term tests with

methanol. Unpublished data, (XXIV 436), 03 July 1975

(134) Kimur a E.T. et al.: Acute toxicity and limits of solvent

residue for sixteen organic solvents. Toxicol. Appl.

Pharmacol., 19, 699

- 704, 1971 (cited in: DGMK 1982)

(135) Rowe, V. K., McCollister, S. B., Alcohols in: Patty's

Industrial Hygie ne and Toxicology, 3rd. rev. Ed., Vol. 2C,

Toxicology, G.D.Clayton and F.E.Clayton (Eds.), John Wiley &

Sons, New York, 4528

- 4541, (1982)

(136) Smyth H.F. et al.: The single dose toxicity of some glycols

and derivatives. J.Ind.Hyg.T oxicol., 23, 259, 1941 (cited

in: Rowe V.K. and McCollister S.B.: Patty's

Industrial Hygiene and Toxicology, Vol. II, 1981)

(137) Smith E.N. and Taylor R.T.: Toxicology, 25, 271

- 287, (1982)

(138) Munch J.C. , Industr. Med., 41, 31

- 33, (19 72) cited in: DGMK

1982

(139) Gilger A.P. and Potts A.M.: Studies on the visual toxicity

of methanol, V. The role of acidosis in experimental

methanol poisoning. Amer. J. Ophthalmol., 39, 63

- 86, 1955

(cited in: DGMK 1982)

(1 40) Cooper, J.R. and Felig, P.: The biochemistry of methanol

poisoning, II. Metabolic acidosis in the monkey. Toxicol.

Appl. Pharmacol., 3, 202

- 209, 1961

(141) Gilger, A.P. et al., Am. J. Ophthalmol., 42, 244

- 252, 1956

(cited in Coo per and Felig, 1961)

(142) Gilger, A.P. et al., Am. J. Ophthalmol., 48, 153

- 161, 1959

(cited in Cooper and Felig, 1961)

(143) Potts, A.M.: The visual toxicity of methanol, VI. The

clinical aspects of experimental methanol poisoning treate d

with base. Amer. J. Ophthalmol., 39, 86

- 92, 1955 (cited in

DGMK 1982)

(144) Potts, A.M. et al.: Studies on the visual toxicity of

methanol, VIII. Additional observations on methanol

poisoning in the primate test object. Amer . J. Ophthalmol.,

40, 76

- 83, 1955 (cited in DGMK 1982)

(145) Welch H. and Slocum G.G.: J.Lab.Cin.Med., 28, 1440, (1943)

cited in: Rowe V.K. and McCollister S.B.: Patty's

Industrial Hygiene and Toxicology, Vol. II, (1981)

(146) Deic hmann W.B.: J.Ind.Hyg.Toxicol., 30, 373, 1948,

cited in: Rowe V.K. and McCollister S.B.: Patty's

Industrial Hygiene and Toxicology, Vol. II, (1981)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

296

(147) BASF AG, Department of Toxicology, unpublished study,

X/304

- 307, 17 Jan. 1961

(148) RTECS, Update code 8909, (1990)

(149) Dorman, D.C. et al., Fund. Appl. Toxicol., 20, 341

- 347, 1993

(150) BASF AG, Dept. of Toxicology: Bestimmung der akuten

Inhalationstoxizität LC50 von Methanol bei 4stuendiger

Exposition an Sprag ue

- Dawley

- Ratten. Unpublished report,

(80/158), 20 Nov. 1980

(151) BASF AG, Dept. of Toxicology, Bestimmung der akuten

Inhalationstoxizität LC50 von Methanol bei 6stuendiger

Exposition an Sprague

- Dawley

- Ratten. Unpublished report,

(80/158), 07 Aug. 1980

(152) Von Burg, R.: J. Appl. Toxicol. 14(4), 309

- 313 (1994)

(153) Mashbitz N.L. et al.: J. Industr. Hyg. Toxicol., 18, 117,

1936 (cited in: DGMK 1982)

(154) Lehmann K.B. and Flury F., Toxicology and hygiene of

industrial solvents., Williams und Wilkins, Baltimore, 1943

(cited in: DGMK 1982)

(155) Witte R.: Dissertation Wuerzburg 1931,

(cited in: DGMK 1982)

(156) McCord, C.P.: Toxicity of methyl alcohol (methanol)

following skin absorpt ion and inhalation. Industrial and

Engineering Chemistry, 23, 931

- 936, 1931

(157) Cooper, R.L. et al.: Effects of inhaled methanol on

pituitary and testicular hormones in chamber acclimated and

non

- acclimated rats. Toxicology 71, 69

- 81, 1992

(158) BASF AG, Dept. of Toxicology, unpublished results, 78/646,

10 Jan. 1980

(159) BASF AG, Dept. of Toxicology: Short

- term test with methanol.

Unpublished results, (X/306), 17 Jan. 1961

(160) BASF AG, Dept. of Toxicology, unp ublished data, (XXIV 436),

03 July 1975

(161) Flury F. and Zernik F.: Schaedliche Gase, J. Springer

Verlag, Berlin, 345

- 348, (1931) (cited in: DGMK 1982)

(162) Eulner, H.H. and Gedicke, K.H.: Ueber die Hautresorption von

Methylalko hol. Sammlung Vergiftungsfaelle, 15, 409

- 414, 1955

(cited in: DGMK 1982)

(163) Loewy A. and van der Heide R.: Biochem. Zschr., 65, 230,

1914 (cited in: DGMK 1982)

(164) Eulner, H.H. and Gedicke, K.H.: Ueber die Hautresorption von

M ethylalkohol. Sammlung Vergiftungsfaelle, 15, 409

- 414, 1955

(cited in: DGMK 1982)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

297 (165) Carnegie

- Mellon Institute of Research, unpubl. data,

cited in: Rowe V.K. and McCollister S.B.: Alcohols, p. 4531.

Patty's Industrial Hygiene and Toxicology, Vol. II,

4527

- 4708, 1981

(166) Gilger A.P. et al.: Amer. J. Ophthalmol. 35, (Suppl.),

113

- 126, (1952),

cited in Forschungsbericht der Deutschen Gesellschaft

fuer Mineraloelwissenschaft und Kohlechemie e.V.(1982)

(167) NEDO: Toxicological Research of Methanol as a Fuel for

Power Station, Summary Report on Tests with Monkeys, Rats

and Mice. New Energy Development Organization, Tokyo, 1987

(168) Clay, K.L. et al.: Experimental methanol toxicity in the

primate: Analysis of metabolic acidosis. Toxicol. Appl.

Pharmacol., 34, 49

- 61, 1975

(169) BASF AG, Dept. of Toxicology, unpublished results, (X/306),

17 Jan. 1961

(170) Goldenthal E.I.: Toxicol. Appl. Pharmacol., 18, 185

- 207,

1971, cited in: DGMK 1982

(171) Zagradnik R et al.: Farmakol. Toksikol, 25, 618, 1962,

cited in: Rowe V.K. und McCollister S.B.: Patty's

Industrial Hygiene and Toxicology, Vol. II, (1981)

(172) EHP, Environmental Health Perspectives 61 , 321 (1985),

cited in: RTECS 1996

(173) Marc

- Aurele J. and Schreiner G.E.: J. Clin. Invest., 39,

802

- 807, 1960, (cited in: DGMK 1982)

(174) NIOSH (1982). Registry of Toxic Effects of Chemical

Substances, Edition July 1982 [cited i n: DGMK 1982]

(175) Jacobs, G.A.: OECD eye irritation tests on three alcohols,

in: Acute Toxicity Data. J. Am. Coll. Toxicol., 1, 56

- 57,

1990

(176) Carpenter C.P. and Smyth H.F.: Am.J.Ophthalmol., 29, 1363,

1946, cited in: Rowe V.K . and McCollister S.B.: Patty's

Industrial Hygiene and Toxicology, Vol. II, (1981)

(177) BASF AG, Dept. of Toxicology (1979a): Bericht zur Prüfung

auf sensibilisierende Wirkung am

Meerschweinchen

- Maximierungstest (Magnusson und Kligm an).

Unpublished data, (78/333), 11 April 1979

(178) BASF AG, Dept. of Toxicology (1979b): Bericht zur

vergleichenden Prüfung auf sensibilisierende Wirkung am

Meerschweinchen

- Maximierungstest (Magnusson und Kligman).

Unpublish ed data, (78/333), 11 July 1979

(179) Vyskovil, A. and Viau, C., Environ. Toxicol. Pharmacol., 9,

9

- 18, 2000

(180) Takeda, K. and Katoh, N.: Long

- term effects of methanol

vapor at low concentration. Proc. of the 8th Int. Symp.

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

298 Alco hol Fuels, Tokyo, Japan, (1988)

(181) US EPA, Office of Solid Waste, Washington, DC: Rat oral

subchronic toxicity study with methanol, 1986 (cited in:

US EPA/IRIS Database 1995)

(182) US EPA/IRIS Database: Methanol, Update through October 1995

(183) Andrews J.J.C., Terrill, J.B., and Bolte, H.F. (1987):

Subchronic inhalation toxicity of methanol. J. Toxicol.

Envir. Health, 20,

117

- 124, (1987)

(184) Baumbach, G.L. et al.: Methyl alcohol poisoning

IV. Alterati ons of the morphological findings of the retina

and optic nerve. Arch. Ophthalmol., 95, 1859

- 1865, Oct. 1977

(185) Hayreh, M.S. et. al.: Methyl alcohol poisoning. Arch.

Ophthalmol., 95, 1851

- 1858, Oct. 1977

(186) Martin

- Amat, G. et al., Metab. Syst., 2, 419

- 428, 1977

(cited in DGMK 1982)

(187) Martin

- Amat, G., McMartin, K.E., Hayreh, S.S., Hayreh, M.S.

and Tephly, T.R.: Methanol poisoning: Ocular toxicity

produced by formate. Toxicol. Appl. Pharmacol., 45, 201

- 208,

1978

(188) Martin

- Amat, G., Tephly, T.R., McMartin, K.E. et al.: Methyl

alcohol poisoning II. Development of a model for ocular

toxicity in methyl alcohol poisoning using the Rhesus

monkey. Arch. Ophthalmol., 95, 1847

- 1850, Oc t. 1977

(189) McMartin, K.E., Makar, A.B., Martin G., Palese, A. and

Tephly, T.R.: Methanol poisoning I. The role of formic acid

in the development of metabolic acidosis in the

monkey and the reversal by 4

- methylpyrazol. Biochem. Me d.,

13, 319

- 333, 1975

(190) White L. et al.: Biochemical and cytological studies of rat

lung after inhalation of methanol vapours. Toxicology

Letters, 17, 1

- 5, 1983

(191) Chao C.T.: Gig. Sanit., 24, 7, (1959),

cited in Kavet R. and Nauss K.M., Critical Review in

Toxicology, 21, (1990)

(192) Kavet, R. and Nauss, K.M.: The toxicity of inhaled methanol

vapors. Critical Reviews in Toxicology 21(1), 21

- 50, 1990

(193) Ubaydullayev, R., USSR Lit. Air Pollut. Rel. O ccup. Dis.

- A

Survey, 17, 39, 1963 (cited in Kavet, R. and Nauss, K.M.,

Critical Reviews in Toxicology, 21, 1990)

(194) Cameron A.M. et al.: Circulating concentrations of

testosterone, luteinizing hormone and follicle stimulating

hormone in male rats after inhalation of methanol. Arch.

Toxicol. Suppl., 7, 441

- 443, 1984

(195) Cameron, A.M. et al.: Circulating steroids in male rats

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

299 following inhalation of n

- alcohols. Arch. Toxicol. Suppl. 8,

422

- 424, 1985

(196) Lee, E.W., Terzo, T.S., D ́Arcy, J.B., Gross, K.B. and

Schreck, R.M.: Lack of blood formate accumulation in humans

following exposure to methanol vapor at the current

permissible exposure limit of 200 ppm. Am. Ind. Hyg. Assoc.

J. 53(2), 99

- 104, 1992

(197) Cooper R.L. et al.: The Toxicologist, 10, Nr. 843, (1990)

(198) Rao K.R. et al.: Indian. J. Med. Res., 65, 285

- 292, (1977)

(199) Sayers R.R. et al.: Report of Investigations RI 3617, US

Dept. of Interior, Bur eau of Mines, 1942 (cited in: Kavet R.

and Nauss K.M.: Critical Review in Toxicology, 21, 1990)

(200) Andrews, J.J.C. et al.: Subchronic inhalation toxicity of

methanol. J. Toxicol. Environ. Health, 20, 117

- 124, 1987

(201) DeFlora, S.: St udy of 106 organic and inorganic compounds in

the Salmonella/microsome test. Carcinogenesis, 2, 283

- 298,

1981

(202) DeFlora S., Camoirano, A., Zancchi, P. and Bennicelli, C.:

Mutagenicity testing with TA97 and TA102 of 30 DNA

- damagin g

compounds, negative with other Salmonella strains. Mutat.

Res., 134, 159

- 165, 1984a

(203) DeFlora, S.; Zanacchi, P.; Bennicelli, C.; Camoirano, A.;

Basso, C.; Bagnasco, M.; Izzotti, A. Badolati, G.S.:

Genotoxicity, Biotransf ormation, and Interactions of Marine

Pollutants as Related to Genetic and Carcinogenic Hazards 5,

3

- 32, 1990

(204) Gocke E.; King, M.

- T.; Eckhardt, K. and Wild, D.:

Mutagenicity of cosmetics ingredients licensed by the

Europea n Community. Mutat. Res., 90, 91

- 109, 1981

(205) Shimizu H., Suzuki, Y., Takemura, N., Goto, S., Matsushita,

H: The results of microbial mutation test for forty

- three

industrial chemicals. Jpn. J. Ind. Health, 27, 400

- 419, 1985

(206) DeFl ora, S.; Zanacchi, P.; Camoirano, A.; Bennicelli, C.;

Badolati, C.: Genotoxic activity and potency of 135

compounds in the Ames reversion test and in a bacterial

DNA

- repair test. Mutat. Res., 133, 161

- 198, 1984b

(207) Lasne C., Gu, Z .W., Venegas, W and Chouroulinkov, I.: The in

vitro micronucleus assay for the detection of cytogenetic

effects induced by mutagen

- carcinogens: Comparison with the

in vitro sister

- chromatid exchange assay. Mutat. Res., 130,

273

- 282, 1984

(208) Fritzenschaf, H., Kohlpoth, M., Rusche, B., Schiffmannn, D.:

Testing of known carcinogens in Syrian hamster embryo (SHE)

micronucleus test in vitro; correlations with in vivo

micronucleus formation and cell transforma tion. Mutation

Research 319, 47

- 53 (1993)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

300

(209) Schmuck, G., Lieb, G., Wild, D. and Schiffmann, D. and

Henschler, D.: Characterisation of an in vitro micronucleus

assay with Syrian hamster embryo fibroblasts. Mut. Res.,

203, 3 97

- 404, 1988

(210) Obe, G. and Ristow, H.: Chromosomal damage by alcohol in

vitro and in vivo. Mutat. Res., 56, 211

- 213, 1977

(211) Ragan, D.L. and Boreiko, C.J.: Initiation of C3H/10T1/2 cell

transformation by formaldhyde. Cancer Letter s, 13, 325

- 331,

1981

(212) Strobel, R. and Greb, W.: Reproducibility of and experiences

with the BHK cell transformation assay. Toxicology, 19,

55

- 66, 1981

(213) McGregor, D.B., Martin, R., Riach, C.G. and Caspary, W.J.

(19 85): Optimisation of a metabolic activation system for

use in the mouse lymphoma L5178Y tk+tk

- mutation system,

Environ. Mutagen., 7(Suppl. 3), 10 (abstr.)

(214) Crebelli, R, Conti, G, Conti, L, and Carere, A (1989): A

comparative st udy on ethanol and acetaldehyde as inducers of

chromosome malsegregation in Aspergillus nidulans. Mutation

Research, 215, 187

- 195

(215) Abbondandolo A.; Bonatti, S.; Corsi, C.; Corti, G.; Fiorio,

R.; Leporini, C.; Mazzaccaro, A.; Nie ri, R.: The use of

organic solvents in mutagenicity testing. Mutat. Res. , 79,

141

- 150, (1980)

(216) Florin I., et al., Toxicology, 15, 219

- 232, 1979

(217) Simmon V.F. et al.: Progress in Genetic Toxicology, Scott

D., Bridges B.A. , Sobels F.H. eds., Elesier/North Holland

Biomedical Press, (1977)

(218) Heidelberger, C., Freeman, A.E., Pienta, R.J. et al.: Cell

transformation by chemical agents

- a review and analysis of

the literature. Mutat. Res., 114, 283

- 38 5, 1983

(219) Pienta, R.J. et al., Int. J. Cancer, 19, 642

- 655, 1977

(cited in: Heidelberger, C. et al., Mutat. Res., 114,

283

- 385, 1983)

(220) Hayes, S. et al.: Mutation Research 231, 151

- 163 (1990)

(221) Pereira, M.A.; Chang, L.W.; Mc Millan, L.; Ward, J.B.;

Legator, M.S.: Battery of short

- term tests in

laboratory animals to corroborate the detection of human

population exposures to genotoxic chemicals. Environ.

Mutagen., 4, 317, 1982

(222) Campbell, J.A. e t al.: Evidence that methanol inhalation

does not induce chromosome damage in mice. Mutation Research

260, 257

- 264, 1991

(223) Gocke E., et al.: Mutagenicity of cosmetics ingredients

licensed by the European Community. Mutat. Res., 9 0, 91

- 109,

1981

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

301 (224) Fu, S.S., Sakanshi, T.m., Rogers, J.M., Hong, K.H. and Keen,

C.L.: Influence of dietary folic acid on the developmental

toxicity of methanol and the frequency of chromosomal

breakage in the CD

- 1 mouse. Re prod. Toxicol., 10, 455

- 463,

1996

(225) American Petroleum Institute/TSCATS, OTS0000812, Doc. I. D.

FYI

- OTS

- 0791

- 0812, 19 July 1991, SRI Project 1409

- C01

- 90

(226) Chang, LW, McMillan, L, Wynne, BR, Pereira, MA, Colley, RA,

Ward, JB , and Legator, MS (1983): The evaluation of six

monitors from the exposure to formaldehyde in laboratory

animals. Environ. Mutagen., 5, 381

- 387 [cited in IPCS/WHO,

1997]

(227) Gocke E., King, M.

- T., Eckhardt, K. and Wild, D.:

Mutagenicity of cosmetics ingredients licensed by the

European Community. Mutat. Res., 90, 91

- 109,

1981

(228) O ́Loughlin, K. et al.: Environ. Mol. Mutagen. Suppl. 20, 47

(1992)

(229) Maltoni, C., Minardi, F, Soffritti, M., Lefemine , G.:

Long

- term carcinogenicity bioassays in industrial chemicals

and man

- made mineral fibers, at the Bentivoglio (BT)

Laboratories of the Bologna Institute of Oncology: Premises,

Programs, and Results. Toxicol. Ind. Health, 7, 63

- 94, 1991

(230) Burbacher, T. et al: Reproductive and Offspring

Developmental

Effects Following Maternal Inhalation Exposure to Methanol

in Nonhuman Primates

Part I: Methanol Disposition and Reproductive Toxicity in

A dult Females;

Part II: Developmental Effects in Infants Exposed Prenatally

to Methanol;

Health Effects Institute (HEI), Research Report No. 89,

October 1999

(231) Ward, J.B., Hokanson, J.A., Smith, E.R. et al.: Sperm count,

morphology, and fluorescent

body frequency in autopsy service workers exposed to

formaldehyde. Mutat. Res., 130, 417

- 424, 1984

(232) Nelson, B.K. and Brightwell, W.S. et al.: Teratological

assessment of methanol and ethanol at h igh inhalation levels

in rats. Fund. Appl. Toxicol., 5, 727

- 736, 1985

(233) NTP (2003): NTP

- CERHR Monograph on the Potential Human

Reproductive and Developmental Effects of Methanol. NIH

Publication No. 03

- 4478. September, 2003

(23 4) Slikker W. Jr. and Gaylor D.W.: Handbook of Developmental

Neurotoxicology (Slikker, W. and Chang, L.W., eds.), Chapter

42, 727

- 731, Acad. Press, 1998

(235) Slikker, W. and Gaylor, D.W., Adv. Occup. Med. Rehabil.,

Vol. 3, 191

- 197, 1997,

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

302

(236) Rogers J.M. et al.: Toxicologist, p. 344, (1991)

(237) Rogers, J.M., Mole, M.L., Chernoff, N., Barbee, B.D.,

Turner, C.I., Logsdon, T.R. and Kavlock, R.J.: The

developmental toxicity of inhaled methanol in the CD

- 1

mouse , with quantitative dose

- response modeling for

estimation of benchmark doses. Teratology, 47, 175

- 188, 1993

(238) Connelly, L.E. and Rogers, J.M.: Methanol causes

posteriorization of cervical vertebrae in mice. Teratology

55, 138

- 144 , 1997

(239) Connelly, L.E. and Rogers, J.M.: Teratology 49, 393 (1994)

(240) Bolon, B., Dorman, D.C., Janszen, D., Morgan, K.T., and

Welsch, F.: Phase

- specific developmental toxicity in

mice following maternal methanol inhalation. Fund. Appl.

Toxicol. 21, 508

- 516, 1993

(241) Stern, S., Cox. C., Preston, R. et al.: Perinatal methanol

exposure in the rat

II. Behavioral effects in neonates and adults. Fund. Appl.

Toxicol., 36, 163

- 176, 1997

(242) Stern, S., Re uhl, K., Soderblom, S. et al.: Perinatal

methanol exposure in the rat I. Blood methanol

concentration and neural cell adhesion molecules. Fund.

Appl. Toxicol., 34, 36

- 46, 1996

(243) Weiss, B., Stern, S., Soderblom, S.C. et al.: Deve lopmental

neurotoxicity of methanol

exposure by inhalation in rats. Health Effects Institute

(HEI), Res. Report No. 73, 1996

(244) Fu, S.S. et al.: Teratology 51, 162 (1995)

(245) Heid, M.K. et al.: Folate deficiency alone does not produce

neural tube defects in mice. J. Nutr., 122, 1198

- 1200, 1992

(246) Hong, K.H. et al.: Plasma formate levels in methanol treated

dams are not markedly influenced by dietary folate.

Teratology, 55, 57, 1997 (abstr.)

(247) Sta nton, M.E. et al.: Fundamental and Applied Toxicology

28,100

- 110 (1995).

(248) Bolon, B. et al.: Teratology 49, 497

- 517 (1994)

(249) Dorman, D.C. and Welsch, F.: CIIT Activities (Chemical

Industry Institute of Toxicology) 16(2), 1

- 7 (1996 )

(250) Dorman, D.C. et al.: Teratology 52, 30

- 40 (1995)

(251) Dorman, D.C., Moss, O.R., Farris, G.M., Janszen, D.; Bond,

J.A., Medinsky, M.A.: Pharmocokinetics of inhaled

[14C]methanol and methanol

- derived [14C]formate in normal

a nd folate

- deficient cynomolgus monkeys. Toxicology and

Applied Pharmacology, 128, 229

- 238, 1994

(252) Medinsky, M.A. and Dorman, D.C., Recent development in

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

303 methanol toxicity. Toxicology Lett., 82/83, 707

- 711, 1995

(253) Sakanashi, T.M. e t al., Teratology 49, 368, 1994 (abstr.)

(cited in: Dorman and Welsch, 1996)

(254) Rogers J.M. and Mole M.L: Critical periods of sensitivity to

the developmental toxicity of inhaled methanol in CD

- 1

mouse. Teratology, 55, 364

- 372, (1 997)

(255) Infurna, R. and Weiss, B.: Teratology 33, 259

- 265 (1986)

(256) Ryan, B.M. et al.: Teratology 49, 399 (1994)

(257) TSCATS, OTS0534807, Doc. I. D. 88

- 920000356, 8E, 12

- 09

- 91,

AMOCO Corporation.

(258) Youssef A. F. et al.: Reproducti ve Toxicology, 11 (4),

503

- 510, (1997)

(259) Youssef A.F.: The Toxicologist, 11, 1349, (1991)

(260) Youssef, A.F. et al.: Teratology 43, 467 (1991)

(261) De

- Carvalho, R.R. et al.: Brazilian J. Med. Biol. Res. 27,

2915

- 2923 (1994)

(262 ) Cummings, A.: Toxicology 79, 205

- 214 (1993)

(263) Abel, E.L. and Bilitzke, P.J.: Am. J. Obstet. Gynecol. 166,

433 (1992)

(264) Rogers J.M. and Daston G.P.: Alcohols: Ethanol and Methanol,

in Handbuch Exp. Pharmacol., Vol. 124, Drug Toxi city in

Embryonic Development II, 333

- 405, (1997)

(265) Rogers, J.M. et al., Teratology 53, 100, 1996

(266) Rogers, J.M.: Teratology 51, 195 (1995)

(267) Barbee, B.D. et al.: Teratology 57, 246 (1998)

(268) Abbott, B.D. et al.: Toxicology 97 , 159

- 171 (1995)

(269) Andrews, J.E. et al.: Teratology 43, 461 (1991)

(270) Andrews, J.E. et al.: Toxicology 81, 205

- 215 (1993)

(271) Brown

- Woodman, P.D.C. et al.: Teratology 52, 233

- 243 (1995)

(272) Deglitz, S.J. et al., Teratology, 55, 58, 1997 (abstr.)

(273) Andrews, J.E. et al.: Teratology 57, 231

- 232, 1998

(274) Andrews, J.E. et al.: Teratology 58, 54

- 61 (1998a)

(275) Ebron

- McCoy, M.T. et al., Teratology, 49, 393, 1994 (abstr.)

(276) Youssef, A.F., Weiss, B., Cox, C. (1993): Neurobehav ioral

toxicity of methanol reflected by operant running.

Neurotoxicol Teratol.,15, 223

- 227 (cited in NTP (2003)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

304

(277) Osterloh, J.D. et al.: Serum concentrations of methanol

after inhalation at 200 ppm. J. Occup. Environ. Med., 38,

571

- 576, 1996

(278) Chuwers, P., Osterloh, J., Kelly, T., D ́Alessandro, A., et

al. (1995): Neurobehavioral effects of low

- level methanol

vapor exposure in healthy human volunteers. Environ. Res.,

71, 141

- 150

(279) Becker, C.E .: Methanol poisoning. J. Emerg. Med., 1, 51

- 56,

1983

(280) Roe, O.: The metabolism and toxicity of methanol. Pharmcol.

Rev., 7, 399

- 412, 1955

(281) Tephly, T.R.: Mini review

- The toxicity of methanol. Life

Sci., 48, 1031

- 1041, 1 991

(282) Ferry, D. et al.: Methanol monitoring. Int. Arch. Occup.

Environ. Health 47, 155

- 163, 1980

(283) Sedivec, V., Mraz, M. and Flek, J.: Biogical monitoring of

persons exposed to methanol vapours. Int. Arch. Occup.

Environ. H ealth, 48, 257

- 271, 1981

(284) Roe, O.: Species differences in methanol poisoning. Crit.

Rev. Toxicol., 10, 275

- 286, 1982

(285) Heinrich, R., Angerer, J.: Occupational chronic exposure to

organic solvents. X. Biological monitoring paramet ers for

methanol exposure. Int. Arch. Occup. Environ. Health, 50,

341

- 349, 1982

(286) Litovitz T.L. et al.: Annual report of the American

Association of Poison Control Centers National Data

Collection System. Am. J. Emerg. Me d., 6, 479

- 515, 1988

(287) Baumann, K. and Angerer, J.: Occupational chronic exposure

to organic solvents. VI. Formic acid concentration in blood

and urine as an indicator of methanol exposure. Int. Arch

Occup. Environ. Health, 42, 241

- 249, 1979

(288) Muttray A., Int. Arch. Occup. Environ. Health, 74, 43

- 48,

(2001)

(289) Jacobsen, D. et al.: Studies on methanol poisoning. Acta

Med. Scand., 212, 5

- 10, 1982a

(290) Erlanson, P. et al.: Severe methanol intoxication. A cta Med.

Scand., 177, 393

- 408, 1965

(291) Naraqi, S. et al.: An outbreak of acute methyl alcohol

intoxication. Aust. N. Z. J. Med., 9, 65

- 68, 1979

(292) Kawai, T., et al.: Methanol in urine as a biological

indicator of occupational exposure to methanol vapor. Int.

Arch. Occup. Environ. Health, 63, 311

- 318, 1991c

(293) Yasugi, T. et al.: Formic acid excretion in comparison with

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

305 methanol excretion in urine of workers occupationally

exposed to methanol. Int. Arch . Occup. Environ. Health, 64,

329

- 337, 1992 (cited from DFG 1999, p. 7)

(294) Franzblau, A., et al.: Absence of formic aid accumulation in

urine following five days of methanol exposure. Appl. Occup.

Environ. Hyg. 8, 883

- 888, 1993

(295) Leaf, G. and Zatman, L.J.: A study of the conditions under

which methanol may exert a toxic hazard in industry. Brit.

J. Ind. Med. 9, 19

- 31, 1952

(296) Frederick, L.J. et al.: Investigation and control of

occupational hazards a ssociated with the use of spirit

duplicators. Am. Ind. Hyg. Assoc., 45, 51

- 55, 1984

(297) Batterman, S., A., et al; Int. Arch. Occup. Environ Health

68, 268

- 274, (1996)

(298) Dutkiewicz B., Kónczalik, J., Karwacki, W.: Skin absorption

and per os administration of methanol in men. Int. Arch.

Occup. Environ. Health, 47, 81

- 88, 1980

(299) Batterman, S.A. et al.: Breath, urine, and blood

measurements as biological exposure indices of short

- term

inhalation exposure to methanol. Int. Arch. Occup. Environ.

Health, 71, 325

- 335, 1998

(300) Batterman, S.A. and Franzblau, A.: Time

- resolved cutaneous

absorption and permeation rates of methanol in human

volunteers. Int. Arch. occup. Environ. Health, 7 0, 341

- 351,

1997

(301) Kawai, T. et al.: Int. Arch. Occup. Environ. Health, 63,

429

- 435, 1991b

(302) Dethlefs, R. and Naraqi, S.: Ocular manifestatons and

complications of acute methyl alcohol intoxication. Med. J.

Australia , 2, 483

- 485, 1978

(303) Lorente C. et al.: Maternal occupational risk factors for

oral clefts. Scan. J. Work Environ. Health, 26, 137

- 145,

2000

(304) Davoli, E. et al. (1986): Serum methanol concentrations in

rats and in men after a single dose of aspartame. Food Chem.

Toxicol., 24, 187

- 189 (cited in NTP 2003)

(305) Leon, A.S., Hunninghake, D.B., Bell, C., Rassin, D.K.,

Tephly, T.R. (1989): Safety of long

- term large doses of

aspartame. Arch Intern Med. 149: 2318

- 24 [cited in NTP,

2003]

(306) Lanigan S., Int. J. Toxicol., 20, Suppl. 1, 57

- 85, (2001)

(307) May, J.; Staub

- Reinhalt. Luft 26, 385

- 389, (1966)

(308) Ubaidullaev, R.; Gig. Sanit. 31, 9

- 12, 1966 (cited in:

IPCS/WHO 1997)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

306

(309) Liesivuori, J., Savolainen, H.; Am. Ind. Hyg. Ass. J. 48,

32

- 34, (1987)

(310) Franzblau, A., et al; Appl. Occup. Environ. Hyg. 7, 467

- 471,

(1992)

(311) Henderson, Y., Haggard, H., W.; Noxious Gases, Reinhold

Publishing Corp., New York, 218, (1943)

(312) Passarelli M., M., et al., Sci. Total Environ., 243/244,

349

- 352, (1999)

(313) Kawai T, et al.: Int. Arch. Occup. Environ. Health, 63,

223

- 234, 1991a

(314) McCoy H. C., Am. J. Med., 67, 804

- 807, (1979)

(315) St egink L. D., et al., J. Nutr., 113, 1600

- 1606, (1983)

(316) Stegink L. D., et al., J. Toxicol. Environ. Health, 7,

281

- 290, (1981)

(317) Gosselin, R., et al., Clinical Toxicology of Commercial

Products, 5th Ed., Section III Therapeutics I ndex, Williams

& Wilkins, Baltimore, 275

- 279, (1984)

(318) National Institute for Occupational Safety and Health;

Criteria for a recommended Standard

- Occupational Exposure

to Methyl Alkohol. DEHW(NIOSH) Pub.No. 76

- 148 (1976); NTIS

Pub.No. PB

- 273

- 806, National Technical Information Services,

Springfield, VA, (1976)

(319) Thiess, A., M.; Internist 15, 424

- 431, (1974)

(320) Suit P., Esres M.L., Cleve. Clin. J. Med., 57, 464

- 471,

(1990)

(321) Pla A., et al., F orensic Sci. Int. 49, 193

- 196, (1991)

(322) Kruse J. A., Intensive Care Med., 18, 391

- 397, (1992)

(323) Tyson, H. H., JAMA, 63, 915

- 922, (1914)

(324) Hantson P., et al., Clin. Toxicol. 35, 187

- 191 (1997)

(325) Verslegers W., et al., Acta neurol. be lg. 88, 163

- 171 (1988)

(326) Posner, H. S.; J. Toxicol. Environ. Health 1, 153

- 171,

(1975)

(327) Benton, C., D., Calhoun, F., P.; Trans. Amer. Acad. Ophthal.

Oto

- Lar. 56, 875

- 885, (1952)

(328) Aquilonius, S. M., et al., J. Comp. Ass. To mogr., 4,

425

- 428, (1980)

(329) Ruedemann, A. D.; Am. J. Ophthalmol. 54, 34

- 53, (1962)

(330) Hantson P., Mahieu P., Clin. Toxicol. 38, 297

- 303, 2000

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

307 (331) Mc Nally, W., D.; Toxicology, Industrial Medicine Publisher,

Chicago, 615, (1937)

(332) Guggenheim, M. A., et al., Arch. Neurol., 24, 550

- 554,

(1971)

(333) Henze, T., et al., Nervenarzt, 57, 658

- 661, (1986)

(334) Downie, A., et al., Occup. Med., 42, 47

- 49, (1992)

(335) De Schweinitz, G., E.; Ophthalmic. Rec. 10, 289

- 296, (1901)

(336) Lewis, R., J.; Sax ́s Dangerous Properties of

Industrial Materials, Van Nostrand Reinhold Company, New

York, 9th Ed., 2174, (1996)

(337) Keeney, A., H., Mellinkoff, S., M.; Ann. Int. Med. 34,

331

- 338, (1951)

(338) To nning, D., et al., Canad. Med.Ass. J., 74, 20

- 27, (1956)

(339) Kaplan, K.; Am. J. Med. Sciences 244, 170

- 174, (1962)

(340) Kane, R. L., et al., Arch. Environ. Health, 17, 119

- 129,

(1968)

(341) Eulner, H., H.; Arch. Toxikol. 15, 73

- 76, (1954)

(342) Kahn, A., Blum, D., J. Pediatrics, 94, 841

- 843, (1979)

(343) Kudo, Y., et al; Int. Arch. Occup. Environ. Health 68,

513

- 515, (1996)

(344) Tintinalli J. E., Ann. Emerg. Med. 26, 393 (1995)

(345) Liu J. J., et al., Clin. Toxicol. 37, 69

- 7 3 (1999)

(346) Chow M. T., et al., Am. J. Kidney Dis. 30, 568

- 570 (1997)

(347) Bennett, I. L., et al., Medicine (Baltimore), 32, 431

- 463,

(1953)

(348) Gimenez E. R., et al., Clin. Toxicol., 1, 39

- 48, (1968)

(349) Buller F., Wood C. A., JAMA, 43, 1058

- 1062, (1904)

(350) Wood C. A., Buller F., JAMA, 43, 972

- 977, (1904)

(351) Frenia M. L., Schauben J. L., Ann. Emerg. Med., 22,

1919

- 1923, (1993)

(352) Swartz R. D., et al., Medicine (Baltimore), 60, 373

- 382,

(1981)

(353) Pami es R. J., J. Florida M. A., 80, 465

- 467, (1993)

(354) Vogt M. J., et al., Chest, 103, 262

- 263, (1993)

(355) King M. L., Heart Lung, 21, 260

- 264, (1992)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

308

(356) Wu Chen N. B., et al., J. Forensic Sci., 30, 213

- 216, (1985)

(357) Price E., A., et al., C lin. Toxicol., 32, 79

- 84, (1994)

(358) Mahieu P., et al., Human Toxicol., 8, 135

- 137, (1989)

(359) del Cario

- O ́Donovan R., Glay J., Can. Assoc. Radiol. J., 43,

299

- 301, (1992)

(360) Hsieh F.

- Y., et al., Chin. Med. J. (Taipei), 49, 283

- 288,

(1992))

(361) Glazer M., Dross P., A. J. R., 160, 1105

- 1106, (1993)

(362) McLean, D. R, Ann. Neurol, 8, 161

- 167, (1979)

(363) Sharpe, J. A., et al., Can. J. Neurol. Sciences, 5, 350

- 351,

(1978)

(364) Sharpe J. A., et al., Neurology (Ny), 32, 1093

- 1100, (1982)

(365) Pelletier J., J. Neurol. Neurosurg. Psych., 55, 234

- 235,

(1992)

(366) Dotzauer G., Br. Med. J., II, 929

- 930, (1978)

(367) Ley C. O., Gali F. G., Eur. Neurol. 22, 405

- 409 (1983)

(368) Aquilonius, S. M., et al., Br it. Med. J., 2, 929

- 930, (1978)

(369) Roe O., Q. J. Stud. Alcohol., 11, 107

- 112, (1950)

(370) Jacobsen D., McMartin K. E., Med. Toxicol., 1, 309

- 334,

(1986)

(371) Pappas S., C., Silverman M., Can. Med. Assoc. J., 15,

1391

- 1394, (1982)

(372) Keyvan

- Larijarni H., Tannenberg A. M., Arch. Intern. Med.,

134, 293

- 296, (1974)

(373) Burgess E., Pharmcotherapy, 12, 238

- 239, (1992)

(374) Gonda A., et al., Am. J. Med., 64, 749

- 758, (1978)

(375) Jacobsen D., et al., Acta Med. Scand., 212, 17

- 20, (1982b)

(376) Jacobsen D., et al., J. Emerg. Med., 8, 455

- 461, (1990)

(377) Aabakken L., et al., Human Toxicol., 13, 131

- 134, (1994)

(378) Demedts P., et al., Clin. Chem., 40, 1587

- 1590, (1994)

(379) Muttray A., et al., Allergologie 20, 528

- 529 (1997)

(380) BASF AG, Works Medical Care Service. Unpublished

communication, 1993

(381) Thies, O.: Zbl. Gewerbehyg. Unfallverhuet., 15, 303

- 308,

1928

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

309 (382) Fregert, S., et al., Acta Derm.

- Venerol., 49, 493

- 497,

(1969)

( 383) Browning, E.; Toxicity and Metabolism of Organic Solvents,

Elsevier Publishing Co., Amsterdam, 311

- 323, (1965)

(384) Henson, E., V.; J. Occup. Med. 2, 497

- 502, (1960)

(385) Humperdinck, K.; Arch. Gewerbepathol. Gewerbehyg. 10,

569

- 57 4, (1941)

(386) Massachusetts Division of Occupational Hygiene; Health

Hazards of Wood Heel Covering, Boston, 1937 (cited in:

ACGIH, Documentation of the Threshold Limit Values, 6th Ed.,

Cincinnati, Ohio, 905, 1991)

(387) Kingsley, W., H., Hirsch, F., C.; Compen. Med. 40, (7

- 8),

(1954

- 1955)

(388) Zerlett, G.; Ergo

- Med. 8, 6

- 7, (1984)

(389) Greenburg L., et al., J. Ind. Hyg. Toxicol., 20, 148

- 154,

(1938)

(390) DFG Commission for the Investigation of Health Hazards of

Chemical Compounds in the Work Area: Methanol, in:

Occupational Toxicants: Critical Data Evaluation for MAK

Values and Classification of Carcinogens (Greim, H., ed.),

Wiley

- VCH, 1999

(391) Garner, C.D., Lee, E.W., Louis

- Fe rdinand, R.T.: Müller cell

involvement in

methanol

- induced retinal toxicity. Toxicol. Appl.

Pharmacol., 130, 101

- 107, 1995a

(392) Garner, C.D., Lee, E.W., Terzo, T.S., Louis

- Ferdinand, R.T.:

Role of retinal metabolism in metha nol

- induced retinal

toxicity. Toxicol. Environ. Health, 44, 43

- 56, 1995b

(393) Lee, E.W., Garner, C.D. and Terzo, T.S.: A rat model

manifesting methanol

- induced

visual dysfunction suitable for both acute and long

- term

exposure studies. Toxicol. Appl. Pharmacol., 128, 199

- 206,

1994

(394) Perkins, R.A., Ward, K.W. and Pollack, G.M.: Comparative

toxicokinetics of inhaled

methanol in the female CD

- 1 mouse and Sprague

- Dawley rat.

Fund. Appl. Toxicol. 28 , 245

- 254, 1995

(395) Pollack, G.M. and Brouwer, K.L.R.: Maternal

- fetal

pharmacokinetics of methanol. Health Effects Institute

(HEI), Res. Report No. 74, 1996

(396) HEI (Health Effects Institute): Automotive Methanol Vapors

and Hum an Health: An Evaluation of Existing Scientific

Information and Issues for Future Research. Cambridge,

MA/USA, 1987 (cited in: Horton et al., 1992)

(397) Horton, V.L., Higuchi, M.A. and Rickert, D.E. :

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

310 Physiologically based pharmacok inetic

model for methanol in rats, monkeys, and humans. Toxicol.

Appl. Pharmacol. 117, 26

- 36, 1992

(398) D ́Alessandro, A. et al.: Formate in serum and urine after

controlled methanol exposure at the threshold limit value.

Envi ron. Health Persp., 102, 178

- 181, 1994 (cited in:

Medinsky and Dorman, 1995)

(399) Bouchard, M. et al.: A biologically based dynamic model for

predicting the disposition of methanol and its metabolites

in animals and humans. Toxicol. Sci., 64, 169

- 184, 2001

(400) Ward, K.W. et al.: Comparative toxicokinetics of methanol in

the female mouse and rat. Fundamental and Applied Toxicology

26, 258

- 264, 1995

(401) Medinsky, M. Dorman, D.C., Bond, J.A., Moss, O.R., Janszen,

D.B. and Everitt, J.I: Pharmacokinetics of methanol and

formate in female Cynomolgus monkeys exposed to methanol

vapors. Health Effects Institute, Res. Report No. 77, 1997

(402) Franzblau, A. and Batterman, S.A. et al.: Breath monitor ing

of inhalation and dermal exposure to methanol. Appl. Occup.

Environ. Hyg., 10, 833

- 839, 1995 (cited in DFG Commission,

1999)

(403) Cook, R.J. et al.: Methanol toxcity and formate oxidation in

NEUT2 mice. Arch. Biochem. Bio phys., 393, 192

- 198, 2001

(404) McMartin, K.E., Martin

- Amat, G., Makar, A.B., and Tephly,

T.R: Methanol poisoning V. Role of formate metabolism in

the monkey. J. Pharmacol. Exp. Ther., 201, 564

- 572, 1977

(405) Johlin, F.C. et al.: Studie s on the role of folic acid and

folate

- dependent enzymes in human methanol poisoning. Mol.

Pharmcol., 31, 557

- 561, 1987

(406) Eells, J.T. et al., Int. Toxicol., 7, 61, 1995 (cited in:

Martinasevic et al., 1996)

(407) Martinasevic, M.K., Green, M.D., Baron, J. and Tephly, T.R.:

Folate and 10

- formyltetrahydrofolate dehydrogenase in humans

and rat retina: Relation to methanol toxicity. Toxicol.

Appl. Pharmacol., 141, 373

- 381, 1996

(408) Medinsky, M.A. and Dorman, D.C.: Chemical Industry

Institute of Toxicology (CIIT) 14(7), 1

- 8 (1994)

(409) Tada O. et al.: J. Science of Labour, 51, 143

- 153, (1975)

(410) Perkins, A.P. Ward, K.W. and Pollack, G.M.: Ventilation of

ambulatory rats exposed to methanol vapor in flow

- through

exposure system: measurement and input to toxicokinetic

models. Inhal. Toxicol., 8, 143

- 162, 1996

(411) Eells, J.T. et al., NeuroToxicol., 21, 321

- 330, 2000

(412) Eells, T. et al., Toxicol. Appl. Pharmacol., 140, 58

- 69,

1991

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

311 (413) Seme, M.T. et al., J. Pharmacol. Exp. Therap., 289, 361

- 370,

1999

(414) Wallace, K.B. et al., Fund. Appl. Toxicol., 38, 23

- 37, 1997

(415) Neuhaus

- Steinmetz, U. et al.: Molecular Pharmacology 45,

36

- 41 (1993)

(416 ) Weiss, M.T. and Sawyer T.W.: Toxic. in Vitro 7(5), 653

- 667

(1993)

(417) Wooten, M.W. and Ewald, S.J.: Brain Research 550, 333

- 339

(1991).

(418) Fautrel, A. et al.: Toxic. in Vitro 5(5/6), 543

- 547 (1991)

(419) Lund, A.: Acta Pharmacol. 4, 99

- 212, 1948, (cited in:

IPCS/WHO 1997)

(420) Pollack, G.M. et al.: Fund. Appl. Toxicol. 21, 105

- 110, 1993

(cited in: IPCS/WHO 1997)

(421) Bradford, B.U. et al.: Archives of Biochemistry and

Biophysics 303(1), 172

- 176 (1993)

(422) Bradford, B.U. et al.: Molecular Pharmacology 43, 115

- 119

(1992)

(423) Burbacher, T.M.: Environmental Health Perspectives

Supplements 101(suppl. 6), 133

- 141 (1993)

(424) Lee, E.W. et al.: Evaluation of methanol toxicity on the

visual system: spontaneous degeneration of the retina and

optic nerve in the F

- 344 rat. Toxicologist, 10, 155, 1990

(425) Lee, E.W., Garner, C.D. and Terzo, T.S.: Animal model for

the study of methanol toxicity: Comparison of folate

- reduc ed

rat responses with published monkey data. J. Toxicol.

Environ. Health, 41, 71

- 82 (1994)

(426) McMartin, K., Martin

- Amat, G., Noker, P.E., Tephly, T.R.:

Lack of a role for formaldehyde in methanol poisoning in the

monkey. Bi ochem. Pharmacol., 28, 645

- 649, 1979

(427) Finnegan, K.T.: in Chang, L.W. and Slikker, W. eds.

Neurotoxicology, Approaches and Methods, 641

- 655, Academic

Press (1995)

(428) Costantini, M.G.: Environmental Health Perspectives

Supple ments 101(suppl. 6), 151

- 160 (1993)

(429) Caprino, L. and Togna, G.I.: Potential health effects of

gasoline and its constituents. A review of current

literature (1990

- 1997) on toxiciological data. Environ.

Health Persp., 106(3), 115

- 125, March 1998

(430) Nelson, B.K. et al.: Teratological assessment of methanol

and ethanol at high inhalation levels in rats. Fund. Appl.

Toxicol., 5, 727

- 736, 1985

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

312

(431) Faci, A. et al., Toxicology, 131, 1

- 7, 1998

(432) Dorman, D.: CI IT Activities 13(10), 6

- 7 (1993)

(433) DeFelice A. et al.: Toxicol. Appl. Pharmacol., 38, 631

- 638,

(1976)

(434) Egle J.L. und Gochberg B.J. : Amer. Ind. Hyg. Ass. J.,

369

- 373, May (1975)

(435) Guerri C. et al.: Physiol. Chem.& Physics, 8, 543

- 550,

(1976)

(436) Moore T.B. and Lee E. W.: The Toxicologist, 7, 233, 1987

(437) Billings R.E. and Tephly T.R., Biochem. Pharmacol., 28,

2985

- 2991, 1979

(438) Pietrusko R.: Biochem. Pharmacol., 24, 1603

- 1607, (1975)

(439) Rietb rock N. et al.: Biochem. Pharmacol., 20, 2613

- 2622,

(1971)

(440) Coudoré, F. et al., Toxicologie du méthanol et de

l ́éthylene

- glycol. Lyon Pharmaceutique, 48, 343

- 353, 1997

(441) Delbrueck W.

- R.: in Erdoel und Kohle

- Erdgas

- Petrochemie

, 36(10), October 1983

(442) DGMK (Deutsche Gesellschaft fuer Mineraloelwissenschaft und

Kohlechemie e.V.): Wirkung von Methanol auf Mensch und Tier,

DGMK

- Projekt 260

- 07, Hamburg, Dec. 1982

(443) Documentation of the threshold limi t values and biological

exposure indices, 5th edition, American Conference of

Governmental Industrial Hygienist Inc., pp. 372, (1986)

(444) Esthers E. et al.: in "Patty's Industrial Hygiene and

Toxicology", 3rd revised edition, 2262

- 2269, (1981)

(445) Lundberg P.: Scientific Basis for Swedisch Occupational

Standards VI, Vetenskaplig Skriftserie, 32, 115

- 121, (1985)

(446) Marcus, W.L.: Drinking Water Health Advisory, Office of

Drinking Water, U.S. EPA, J. Environ. Pat hol., Toxicol.,

Oncol., 12, 115

- 138, 1993

(447) Moeschlin S.: "Klinik und Therapie der Vergiftungen", 6.

Auflage, Georg Thieme Verlag Stuttgart, New York, 259

- 262,

(1980)

(448) Teisinger J.: "Expositionstests in der

Industri etoxikologie", VEB Verlag Volk und Gesundheit

Berlin, 167

- 168, (1984)

(449) Treon J.F.: Industrial Hygiene and Toxicology, 2nd revised

edition, 1412

- 1421, (1962)

(450) United Nations Environment Programme, Centre of

International P rojects, GKNT, Moskau (1983)

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

313 (451) Wimer W.W. et al.: "Alcohols Toxicology", Noyes Data

Corporation, Mill Road, Park Ridge, New Jersey 07656, 5

- 23,

(1986)

(452) Gilg T. et al.: Blutalkohol, 24, 321

- 332, (1987)

(453) Witorsch, R.J. et al. : Reproductive Toxic Effects of

Alcohol, Tobacco and Substances of Abuse

(454) Black, K.A. et al.: Proc. Natl. Acad. Sci. 82, 3854

- 3858

(1985)

(455) Oshino, N. et al.: Biochem. J. 131, 555

- 567 (1973)

(456) Lee, E. et al.: Toxicology and Industrial Health 7(4),

261

- 275 (1991)

(457) DFG Commission for the Investigation of Health Hazards of

Chemical Compounds in the Work Area: Methanol, Sammelkapitel

MAK

- Werte und Schwangerschaft, in: Occupational Toxicants:

Cr itical Data Evaluation for MAK Values and Classification

of Carcinogens Wiley

- VCH, Suppl. 1995, Lieferung 21

(458) Skrzydlewska E. et al., J. Appl. Tox., 20, 239

- 243, 2000

(459) Skrzydlewska, E. and Farbiszewski, R., Arch. Toxicol., 71,

741

- 745, 1997a

(460) Skrzydlewska, E. and Farbiszewski, R., Free Rad. Res., 27,

369

- 375, 1997b

(461) Skrzydlewska, E. and Farbiszewski, R., J. Toxicol. Environ.

Health, Part A, 53, 637

- 649, 1998a

(462) Skrzydlewska, E. et al., Comp. Bio chem. Physiol., Part C,

120, 289

- 294, 1998b

(463) Skrzydlewska, E. et al., J. Toxicol. Environ. Health, Part

A, 57, 431

- 442, 1999

(464) Ward K.W. et al.: Tox. Appl. Pharmacol., 145, 311

- 322,

(1997)

(465) Ward, K.W. and Pollack, G .M., Drug Metabol. Dispos., 24,

1062

- 1070, 1996

(466) Dawidek

- Pietryka, K. et al., Arch. Toxicol., 72, 604

- 607,

1998

(467) Poon, R. et al., Toxicol. Ind. Health, 11, 343

- 361, 1995

(cited from Caprino and Togna, 1998)

(468) Ekwall , B. et al.: MEIC Monographs on time

- related, high

survived and lethal blood concentrations of chemicals from

acute, human poisonings, No. 8, provisional draft May 1995

(469) Farbiszewski, R. et al., Toxicology, 156, 47

- 55, 2000

(470) Kas acka, I. and Skrzydlewska, E., Roczniki Akademii

Medycznej w Bialymstoku, 46, 133

- 144, 2001

### OECD SIDS METHANOL

6. REFERENCES ID: 67

- 56

- 1

DATE: 26

- OCT

- 2006

314

(471) Skrzydlewska, E. and Farbiszewski, R., Drug Alcohol Dep.,

57, 61

- 67, 1999

(472) Skrzydlewska, E. and Farbiszewski, R., Fund. Clin.

Pha rmacol., 11, 460

- 465, 1997c

(473) Castro, G.D. et al., Toxicol. Lett., 129, 227

- 236, 2002

(474) Pereira, M.A. et al.: Battery of short

- term tests in

laboratory animals to corroborate the detection of human

population exposures to genotoxi c chemicals. Environ.

Mutagen., 4, 317, 1982

(475) Makar, A.B. et al.: Methanol poisoning in monkey. Mol.

harmacol., 4, 471, 1968 (cited in Roe 1982)