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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 checked 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 

evaluation for the Integrated Risk Information System (IRIS). As 
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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 effects 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. 
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SIDS INITIAL ASSESSMENT PROFILE

CAS No. 67-56-1

Chemical Name Methanol

Structural Formula CH3OH

SUMMARY CONCLUSIONS OF THE SIAR

Human Health

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

dermal contact and distributes rapidly throughout 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, the 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 methanol 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 dehydrogenase. 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 relationship 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 μMrange); this also held true for folate-deficient conditions. Also after repeated inhalation of 2.6 mg/L (2000 ml/m3

) 

for 6 hrs/day, 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, these 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 

appear 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 
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ingestion, and fatalities have occurred in untreated 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, LC50 values have been calculated to be 83.2 and 128.8 mg/L after 4 hours. In cats, the LC50 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 LD50s in rabbits range from 15800 to 20000 mg/kg bw. In rats, the 

dermal LD50 is greater than 45,000 mg/kg bw. In monkeys, four daily dermal doses of 400 mg/kg bw eventually 

resulted in death. Oral LD50s in rats range from < 790 to 13,000 mg/kg bw; in mice, the values range from 7,300 

to 10,000 mg/kg bw; in rabbits, the LD50 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 maximization 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), increased 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 possible 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 condition.

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, slight 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 weights 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 test, 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 Aspergillus, 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 Drosophila 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 methanol does 

not have the potential to be mutagenic, not all studies lead to that conclusion. 
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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, respectively). 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 well 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 neurobehavioral 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, decreased 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, malformations, 

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, fully 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 established 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 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 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, 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 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 0C, density 0.79 g/m3, 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 target 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 estimated elimination half-life is calculated to be 

about 17-18 days with a rate constant of 0.93 x 10-2 cm3

/molecule-sec. Methanol is readily biodegradable. 

Bioaccumulation in fish is expected to be low. 

In several 96-hour studies in fish in which methanol 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 
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(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 methanol 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 the 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 body 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 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 metabolism and excretion is noted depending 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 priority for further work, due to its low hazard profile.
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SIDS Initial Assessment Report

1 IDENTITY

1.1 Identification of the Substance

CAS Number: 67-56-1

IUPAC Name: Methanol

Molecular Formula: CH4O

Structural Formula: H3C-OH

Molecular Weight: 32.04

Synonyms: 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/cm3 (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 noctanol/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 • m3

/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 
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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 hydrogen. This 

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

follows:

CO + 2H2 <-> CH3OHCO2 + 3 H2 <-> CH3OH + H2O

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 880°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 unreacted 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 recovery 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 achieve 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:
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Table 2: Production Volumes of Methanol (2000)

Region

Production Volume

(in million tonnes)

North America (based on NAFTA1)Production approx. 7

South America (includes Trinidad) approx. 7

Western Europe approx. 3.3

Eastern Europe approx. 0.5

Former Soviet 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). 

Approximately 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 utilized 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: Main 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 several 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.
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Table 4: Additional Compounds 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, 

insecticides

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 intermediates, 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 addition, 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-made 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. 

The 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 consumer 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 constant 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 
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2.2.3 Stability in Water

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

2.2.4 Transport 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 soil (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 that 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 and 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 Bioaccumulation

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 Kow = - 0.82 to – 0.64) (Hansch and Leo, 1979; Rekker, 1977). 
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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 use 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, processing, quality control, pilot plant, research 

laboratories, filling stations, and others. The 95th 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/m3 (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 mean 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 plants. 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 percent 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 attempts 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 hydrogen 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 being 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-powered 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 HEALTH 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 content 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 CO2 in 

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

recovered as CO2 (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 follows: 

Step 1 Step 2 Step 3

Methanol  Formaldehyde  Formate  CO2 + H2O

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 through a catalaseperoxidase 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 lethal or near-lethal.

Step 2. This step occurs as a two-reaction process. Formaldehyde is catalysed by the NADdependent formaldehyde dehydrogenase in the presence of glutathione to form the intermediate, Sformylglutathione. The intermediate is then hydrolysed 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 CO2 by formylTHF 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 CO2

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/m3) 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 saturation 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 under inhalation exposure is unlikely in 

monkeys (Horton et al. 1992).

After a single exposure of 0.26 mg/L (200 ml/m3

), formate levels remained unchanged in humans, 

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

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

The conversion of formate to CO2 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 other hand, in monkeys exposed 

repeatedly to 900 ml/m3 (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 folatedeficient 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 methanol 

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). Elimination 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  CO2 (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

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Species Comparisons among Humans, Other Primates, and Rodents 

The species-specific kinetics and differences in the metabolic transformation of methanol and 

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

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

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

For doses up to 1.5 mg/L (1200 ml/m3) 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/m3

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

rats to 0.13 mg/L (100 ml/m3

) (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., 1992).

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

Rat1) Mouse Monkey1 Human1

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 (blood) 

[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 3-82

0.65-0.780 approx. 10 approx. 10 approx. 10

1.6 1.0 ±0.1 26.6 ±2.0 approx. 1503 3.2 ±1.2 37.6 ± 8.5 approx. 25

2.6 2.1 ±0.1 79.7 ±6.1 approx. 5004 2.9 ±0.6 64.4 ± 10.7 approx. 50

6.5 no data approx. 400 approx. 20004 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 sources (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 disproportionate 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 involved 
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 accumulation 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 concentrations 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 

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

3.1.2 Acute Toxicity

Studies in Animals

Inhalation. LC50-values in SD rats of 67,000 ml/m3 (6 hours) and 98,000 ml/m3 (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: LC50(2.25 h) = approx. 79 mg/L (Von 

Burg, 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, LC50 values after 6-hour exposures ranged from 26 to 48 mg/L (Von Burg, 1994; Witte, 

1931). A shorter duration (4.5 hours) led to an LC50 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. Eventual 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 LD50 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 (total 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 LD50 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 ophthalmologic changes typically seen in 

humans at high lethal and sub-lethal doses (Gilger and Potts, 1955). 
OECD SIDS METHANOL

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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 putamen 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 LD50 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 LD50. Ten 

minutes 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 response 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 are 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 of 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 untreated 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 acidosis 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 commonly 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 in 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/m3

) or more. 
OECD SIDS METHANOL

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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 visual 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 (Dethlefs 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 of 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 lethal 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 condition, 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 the 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, slight effects on P300 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 subjects 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 applies 

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. The 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 with 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 

Draize 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 cited 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 were 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 conjunctivae 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 

reversibility 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 and 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

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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 (BASF, 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 on 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/m3

) 

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/m3

): 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 timerelated 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. 

Slight 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 exposure 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 peripheral 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).

These effects were not interpreted as significant.

Liver and kidney: Increase in fat granules with doseresponse 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, some 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/trachea: 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 was 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 the 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/m3

) 

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 inhalation [IUCLID 5.4, No. 3]

B6C3F1 mice (m; f)

0.013, 0.13, and 1.3 mg/L 

(10, 100, 1000 ml/m3

) 

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/m3

)

for –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/m3

) 

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

recovery animals 

4 animals 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), necrosis 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

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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 methanol 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., 1991; 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 0.48 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 machines (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 nature. 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 subacute 20-d study in monkeys, 3.9 mg/L (3000 ml/m3

) 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 because 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, 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.

3.1.6 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 observed (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 mutation 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 results (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 

typhimurium

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 

cytotoxicity 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)

Cell 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 L5178Y (+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)(+/-S9)

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: Am. 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 Doses 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/m3)

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 pregnancy 

(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, folatesufficient and folate-deficient conditions300, 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/m3)

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/m3)

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/m3)

negative Campbell 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 (Sex-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 assays 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 methanol 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/m3

) for 24 and 18 months, 20 and 19 

hours per day, respectively. Clinical, ophthalmological, clinical-chemical, hematological and 

pathological/histopathological examination were done (NEDO, 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 through 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 

both 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 testing 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 control 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 increased 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-statistically 

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 performance (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 reproductive toxicity study in rodents. 

Table 11: Animal Studies 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/m3

) 

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 performance: 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 observed: 

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 between 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 

amount 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, and 2.34 

mg/L

(200, 600, 1800 ml/m3

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- to 16-fold), respectively

Blood formate levels: at baseline level at all exposures

Maternal toxicity: none

Reproductive performance (menstrual cycles, conception rate, and livebirth 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 to 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). 

Wasting 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 

increase 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 developmental 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/m3) 

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:

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 

At 13 mg/L (not statistically significant): 

Visceral malformations were 2/15 litters with 2/107 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/m3

) for 22.7 hr/d 

(avg), gd 7-17

36 animals/group

Dams: 6.5 mg/L: reduced bodyweight gain; reduced food and water 

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), 

variations (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, thymus, 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/m3

)

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 toxicity: 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 7th 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. Doserelated 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 

Developmental 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-related, 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/m3

);

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 (significant 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 only 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 postimplantation losses (p<0.05) 

LOAEL (develop) = 6.5 mg/L

Bolon et al., 

1993 (2)

Developmental 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)

Methanol 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/m3

) 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 lactation);

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 histomorphology, expression of neural 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 between 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 these 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 folate 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 associated 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, not 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 insignificant (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 experimental 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 developmental 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 unequivocal 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 order 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 doses 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 infusion (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 potassium-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 the 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, 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.
OECD SIDS METHANOL

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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 become 

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 lead 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 

with 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 above 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 methanol 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 residual lesions of the 

putamen and visual and lingual impairment (see 3.1.2: Toxicity Syndrome). 

In rats, LC50 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 LD50 of 17000 mg/kg bw was calculated for rabbits; four daily dermal doses of 

400 mg/kg bw eventually resulted in death. Oral LD50s 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 irritant (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 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 subacute 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 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/m3

) 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. 
OECD SIDS METHANOL

33

In the NEDO (1987) studies of chronic 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, 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, 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 

clastogenicity 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 

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.

3.2.4 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 neurobehavioral 

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 number 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 weights 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 decreased 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 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 associated with 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 definitive 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 ENVIRONMENT

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 in 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 volatilisation 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

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Table 13: Short-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

LC50 (96 h) = 15,400 mg/L 

(measur.)

LC50 (96 h) = 20,100 mg/L 

(measur.)

LC50 (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

EC50 (48 h) > 10,000 mg/L 

(nom.)

2 Kuehn et al., 1989

Algae static

static

Scenedesmus 

quadricauda

Chlorella 

pyrenoidosa

Toxic Threshold (192 h) = 

8,000 mg/L (nom.)

EC50 (10-14 d) = 28,400 

mg/L (nom.)

 2

 2

Bringmann and 

Kuehn, 1978

Stratton and 

Smith, 1988

Bacteria static Pseudomonas 

putida

Toxic 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 static 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 threshold 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 limit 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 relevant 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 as 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, the 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 metabolism 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 currently of low priority for further work due to its low hazard 

profile.
OECD SIDS METHANOL

37

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APPENDIX A: TOXICOKINETIC STUDIES IN ANIMALS 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, methanol air concentrations of 0.26, 0.78, and 2.24 mg/L (200, 600, 1800 

ml/m3) 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/m3 6 hrs/day; 5 

days/week; 1 or 2 weeks) monkeys showed no methanol or formate accumulation (Horton et al., 

1992). 

In rats, three consecutive 6-hour exposures of about 5.85 mg/L (4500 ml/m3

) 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/m3). 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 (Pollack 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/m3

) 

(FS) and 1.2 mg/L (900 ml/m3) (FD) for two hours per session. All doses except the highest might 

be encountered 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 lethality. 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).
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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 urinary 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 corresponded 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 doserelated manner. However, blood 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 which 

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 the 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 in 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 absorption 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/m3) methanol for 30 and 120 minutes, the half-life of methanol in blood 

was 1.44 hours (Batterman et al., 1998). 

After a single exposure of 0.26 mg/L (200 ml/m3) 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 

Dorman (1995) also reported similar results. Higher urinary methanol concentrations of 

approximately 40 mg/L have been predicted for workers exposed to 200 ml/m3 (8-hour shift) 

(Heinrich and Angerer, 1982; Kawai et al., 1991). 

Twenty-six volunteers exposed to 0.26 mg/L (200 ml/m3) 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 watervapor 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 concentrations 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/(cm2

*min) (Dutkiewicz et al., 1980) and 8.1 ±3.7 

mg/(cm2

*h) = 0.135 ±0.062 mg/(cm2*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 an approximate air concentration 

of 0.050 mg/L (38 ml/m3) 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/m3 for one 8-hour working shift (11 

mg/L; corrected for background 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/m3), 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

Revision 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: without 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-docs, registration BPD

 A-docs
OECD SIDS METHANOL

1. GENERAL INFORMATION 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: other: 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
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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 Raffineriegesellschaft 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
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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 ClMol. Formula: ClMethod: 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
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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

Quantity: 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 volumes in the year 2000:

 North America (NAFTA): Production : approx. 7 Mtonnes

 Import : approx. 4 Mtonnes

 -> Total supply: approx. 11 Mtonnes
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 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 : approx. 14%

Exports : approx. 7%

 South America approx. 7 Mtonnes

incl. Trinidad

 Western Europe approx. 3.3 Mtonnes

Eastern Europe 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: 50000 - 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 contact 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 container tightly closed

 (16) Keep away from sources of ignition - No smoking
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 (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-Phrases: (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 inhalation, 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: as 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)
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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

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)

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 potential.

 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. Increasing 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 % are expected.

 Acetic Acid
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 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 search 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 contributed

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 implemented process is methanol to gasoline

(MTG) synthesis.

 Methanol is used to synthesize a large number of other

organic compounds:

 Formic acid preservatives, pickling agents

 Methyl esters of solvents, monomers

organic acids

 Methyl esters of methylation reagents, explosives,

inorganic acids insecticides

 Methylamines 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

Category: 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 process,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
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 technology 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 of 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 location.

 (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 industry

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: other: 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 shaving).

 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 costly flue gas treatment plants but is

not yet economically viable.

12-SEP-2005 (7)

Type: use

Category: other: Other Energy Uses of Methanol
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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 estimate 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 transport.

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: used 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

glycols.

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 advantage 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 removal 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 interfere with further
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 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 to 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 the 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 is 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)
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1.8 Regulatory Measures

-

1.8.1 Occupational Exposure Limit Values

Type of limit: MAK (DE)

 Limit value: 270 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 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: 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 Water Pollution
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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 operating 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)

Labelled 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
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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-SEP-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-SEP-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:
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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: Internal and External

Chapters covered: 5.10

Date of Search: 06-NOV-2002

1.13 Reviews
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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

 Handbook

 (15)

Type: density

Value: = .791 - .792 g/cm3 at 20 degree C

Reliability: (4) not assignable

 Manufacturer / producer data without proof

 (16)
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2.3.1 Granulometry

2.4 Vapour Pressure

Value: = 129 hPa at 20 degree C

Reliability: (4) not assignable

 Manufacturer / producer 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

Method: 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 Partition Coefficient

Partition Coeff.: octanol-water

log Pow: = -.82

Method: other (measured)

Reliability: (2) valid with restrictions

 scientifically verfied data
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 (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.

Reliability: (2) valid with restrictions

 scientifically verfied data

 (23)

2.6.1 Solubility in different media

Solubility in: Water

Value: at 20 degree C

Descr.: miscible

Remark: pH value: neutral

Reliability: (4) not assignable

 Manufacturer / producer data without proof

 (16)

Solubility 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)
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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 temperature

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: because of chemical structure

Reliability: (2) valid with restrictions

 expert judgement
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2. PHYSICAL CHEMICAL DATA ID: 67-56-1

DATE: 26-OCT-2006

69

 (28)

2.11 Oxidizing Properties

Result: no oxidizing properties

Remark: because of chemical structure

Reliability: (2) valid with restrictions

 Expert judgement

 (28)

2.12 Dissociation Constant

-

2.13 Viscosity

Value: = .59 mPa s (dynamic) at 20 degree C

Reliability: (4) not assignable

 Manufacturer / producer data without proof

 (16)

2.14 Additional 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 trioxide, 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

 Manufacturer / 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)
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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 scientifically 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)

Type: 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 test

Reliability: (2) valid with restrictions

 (34)

Type: air

INDIRECT PHOTOLYSIS

 Sensitizer: OH

 Conc. of sens.: 3000000 molecule/cm3
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71

Remark: Atmospheric 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 evaluated 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

concentration 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 PHOTOLYSIS

 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 (Clean Air Act Amendments) with an atmospheric

lifetime of 16-18 days.

Reliability: (4) not assignable

 secondary literature

 (39)

Type: water

INDIRECT PHOTOLYSIS

 Sensitizer: OH
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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 presence to natural sources.

 (44)

Type of measurement: other

Remark: Owens et al. (1969) identified methanol as one of the

 volatile compounds emitted by alfalfa, 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)
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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 quantity 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 at 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 sediment

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

pharmaceuticals, 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

 associated sediments (no detection limits were reported).

 (48)

Type of measurement: 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 analysed all the samples by GC/MS and the mass spectra from
OECD SIDS METHANOL

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74

 eight 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 atm * 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 adsorption occurred with the two sandy soils tested (percentage 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 adsorption coefficients 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
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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

Henry' 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)
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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 ppm.

 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 added (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 commercial DO meter

fitted with an agitated probe.

The bottles were opened for sampling and DO measurements about

five times during the course of the 20-day test.

Reliability: (2) valid with restrictions
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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 prescribed 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-2006 (65)

Type: aerobic

Inoculum: other bacteria: synthetic sewage

Degradation: = 71.2 % after 5 day(s)

Method: other: Respirometric test (BOD of THOD)

 GLP: 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 substance: 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 concentration 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) valid with restrictions
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78

 acceptable study meets basic scientific principles

 (66)

Type: aerobic

Degradation: = 99 %

Method: OECD Guide-line 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 restrictions

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
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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 impacted 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 design, sufficiently documented, meets

generally accepted scientific principles, acceptable for

assessment

 (70)

Type: aerobic

Inoculum: 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 glass

 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

zones. 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

generally accepted scientific principles, acceptable for

assessment

 (70)

Type: aerobic

Inoculum: activated sludge, adapted

Degradation: = 50 - 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 days 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
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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 degradation

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 sites 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 quotation

 (73)

Type: aerobic

Remark: Biodegradation of methanol in not heavily polluted aerobic

 aquifer material: half-live 58-260 days (location: 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 conditions 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)
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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

Degradation: = 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 Francisco 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 added 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 restrictions

 2.1; accpetable study, meets basic scientific principles

 (77)

Inoculum: other: methylotrophic organisms

Remark: In a survey of methylotrophic 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 formaldehyde 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., Rhodopseudomonas acidophila, Paracoccus denitrificans, Microcyclus

aquaticus, Thiobacillus novellus, 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
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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 radioactivity 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 different

tissue types was the same after 24 hours, and levels remained constant over 72 hours in the liver, kidneys, gills

and intestine but dropped slightly in the blood and muscle.

Bioaccumulation factors of about 1 were proposed for the

fish tissues and organs.

Reliability: (2) valid with restrictions

 (83)

Species: Leuciscus idus (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 methanol 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
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 in a Packard Tri Carb Sample Oxidizer 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

 hours.

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 in

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 place also

 under anaerobic conditions (all electron acceptors besides

oxygen).

Microbial growth on methanol with nitrate as the electron

acceptor is the second most energetically favored 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)
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AQUATIC ORGANISMS

4.1 Acute/Prolonged Toxicity to Fish

Type: flow through

Species: 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

Remark: 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. Diluters

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

recovery 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.

Average 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 initial 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 mean 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
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 methods, sufficiently documented, acceptable for assessment

 (90)

Type: flow through

Species: Pimephales promelas (Fish, fresh water)

Exposure period: 96 hour(s)

Unit: 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 groups 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 delivered 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.

Result: 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 in 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
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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.

Result: 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 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 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 photoperiod 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 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 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 substance: 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)
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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 substance: 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 period: 96 hour(s)

Unit: mg/l Analytical monitoring:
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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 monitoring: 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 serum 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

Method: 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 protein, serum cholesterol.

Result: The treatment led to increased serum cortisol and serum
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 glucose and to decreased liver glycogen, muscle glycogen,

serum protein and serum cholesterol.

Test substance: Methanol, no 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 assignable

 (102)

Type: static

Species: Oncorhynchus mykiss (Fish, fresh water)

Exposure period: 96 hour(s)

Unit: mg/l Analytical monitoring: 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 quotation

 (103)

Type: static

Species: Semolitus atromaculatus (Fish, fresh water)

Exposure period: 24 hour(s)

Unit: mg/l Analytical monitoring: no data

NOEC: < 8000
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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 further 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

 test 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
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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)

Unit: 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: Methanol, 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: no data
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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: 10630

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 Wirkung 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
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 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 capacity: 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 kept 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 accepted

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: other: acute toxicity

 Year: 1979

Test substance: as prescribed by 1.1 - 1.4
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Reliability: (4)not assisnable

 (98)

Type: static

Species: Daphnia 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: Concentrations 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
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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 concentration

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
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 (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: nominal concentration

Reliability: (4) not assignable

 (111)

Species: other: Anodonta imbecilis

Exposure period: 48 hour(s)

Unit: mg/l Analytical 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 monitoring: 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

 Scenedesmus quadricauda was determined in the cell
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 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 nutrient 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

standardized 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 turbidimetrically 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

termination 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% standard 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: graphic evaluation (probability distribution)

 nominal concentration

Reliability: (2) valid with restrictions
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 (115)

Species: Chlorella pyrenoidosa (Algae)

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

containing 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 containing 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 initial 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

concentration 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
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 ECOSAR Class: Neutral Organics

Reliability: (2) valid with restrictions

 (61)

4.4 Toxicity to Microorganisms e.g. Bacteria

Type: aquatic

Species: Pseudomonas 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 beginnenden 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 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 (pH: 7.0).

From the pollutant solution four parallel dilution series

were prepared in 300 ml Erlenmeyer-flasks 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 subsequent 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 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 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-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
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 series of the pollutant having an extinction value at the

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 current 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-flasks 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 subsequent 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 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 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-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 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
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 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 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 limit

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 : = 18756.34

Method: other: Population Growth Inhibition Test

 Year: 1990
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 GLP: no data

Test substance: other TS

Test substance: purchased from Aldrich Chemical Co., Milwaukee, Wisconsin,

 USA with a purity of 95%

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: = 44400 - 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 standard: 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 dose 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
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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: other: 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 from a municipal sewage

 treatment plant

Unit: g/l Analytical monitoring:

EC50: ca. 71.21

Method: other: Activated Sludge Respiration Inhibition Test

Reliability: (4) not assignable

 secondary quotation

 (124)

Type: aquatic

Species: other bacteria: cyanobacteria

Unit: g/l Analytical monitoring:

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,

 ranging from 0.6% to 10% methanol by volume (4.74-79.00

 g/l). The EC50 values were calculated from percentage inhibition values measured daily relative to growth in the

controls.

Anabaena 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)
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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: Rajini 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

Result: = 916 mM

Reliability: (4) not assignable

 (112)

Species: other bacteria: Steptocephalus proboscideus

Exposure period: 24 hour(s)

Unit: g/l Analytical monitoring:

EC50: = 1020
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Result: = 1020 mM

Reliability: (4) not assignable

 (112)

Species: other bacteria: aerobic 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

Reliability: (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

TERRESTRIAL 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, measured 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 dioxide 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
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 produce a decrease in photosynthesis after 5 minutes exposure.

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 Biological Effects Monitoring

-

4.8 Biotransformation and Kinetics

-

4.9 Additional Remarks

Memo: more information see IPCS report No. 196

 (88)
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5.0 Toxicokinetics, 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 substance: 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: 1975

 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.

key 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: The 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
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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 determined 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
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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 assignable

 Secondary literature

25-OCT-2006 (88) (137)

Type: LD50

Species: rabbit

Value: = 14400 mg/kg bw

Method: other: no details inidicated

 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)
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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 showed 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

 Secondary 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 central

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

Strain: 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
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 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 normal 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) and 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 animals (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
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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 retina 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 there 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.
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5. TOXICITY ID: 67-56-1

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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 bodyweight 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
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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

Reliability: (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)

Type: 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)
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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 substance: 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 substance: 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
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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

Species: 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

Remark: 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 mortality

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)
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5. TOXICITY ID: 67-56-1

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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.

 Mainly 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 substance: 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 scientific principles

25-OCT-2006 (152)

Type: other: narcotic dose

Species: mouse

Exposure time: 3.2 hour(s)

Value: = 40 mg/l

 Year: 1936
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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-OCT-2006 (153)

Type: LC50

Species: cat

Exposure time: 6 hour(s)

Value: = 43.68 mg/l

Method: other: no details reported

 Year: 1994

 GLP: 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.

Reliability: (2) valid with restrictions
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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: Methanol, 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

atrophy 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

Exposure time: 4 hour(s)

Value: = 83.2 mg/l

Method: other: no details reported

 GLP: no data
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120

Test substance: no data

Remark: Substance concentration is given as 64000 ppm.

 non-BASF study

Test 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 the 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 comparing 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-acclimated 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 procedures based on scientific principles, sufficiently

documented, acceptable for assessment

25-OCT-2006 (157)

Type: other: IRT
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5. TOXICITY ID: 67-56-1

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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 when 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 substance: 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 saturation 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 observed 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

atmosphere 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
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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 indicating

effects on the central nervous system occurred in treated

animals.

non-BASF study

Test substance: Methanol, no further data.

Reliability: (4) not assignable

 Secondary source

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 substance: 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 occurred.

non-BASF study

Test substance: Methanol, no further data.
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5. TOXICITY ID: 67-56-1

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123

25-OCT-2006 (163)

5.1.3 Acute Dermal Toxicity

Type: other: lethal dose

Species: rat

Value: >= 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, while 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 eventually 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
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5. TOXICITY ID: 67-56-1

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124

 documentation, acceptable for assessment

25-OCT-2006 (156)

Type: LD50

Species: rabbit

Value: = 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 Acute 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
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5. TOXICITY ID: 67-56-1

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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 substance: other TS

Test substance: Methanol, no further data.

 (148)

Type: LD0

Species: rat

Sex: male
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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: = 120 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.

Value: >= 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 nervous 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
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5. TOXICITY ID: 67-56-1

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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 mortality 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: monkey

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: other

 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 liver (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 pH 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 diluted 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
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No. of Animals: 10

Vehicle: physiol. saline

Doses: 2000, 3000, and 4000 mg/kg bw (as 20- or 25 % solution)

Route of admin.: 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 methanol

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, urine

 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, reagent 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.
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 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 blood 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 and 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-

 methylpyrazole, 50 mg/kg i.v.) prevented the occurrence of

acidosis in the pigtail monkey at a dose which otherwise

routinely produced acidosis.

-----------------------------

Blood analysis identified 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 in 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: Biochemical 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 monkey.

 Results support the view that formic acid is the major agent

to produce acidosis and methanol intoxication in monkeys and
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 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: Methanol, no further data.

 (170)

Type: LD50

Species: mouse

Route of admin.: s.c.

Value: = 4100 mg/kg bw

Test substance: other TS

Test substance: Methanol, 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 substance: Methanol, no further data.

 (107)
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Type: LD50

Species: mouse

Route of admin.: i.v.

Value: = 5660 mg/kg bw

Test substance: other TS

Test substance: Methanol, no further data.

 (171)

Type: LD50

Species: mouse

Route of admin.: i.v.

Value: = 4710 mg/kg bw

Test substance: other TS

Test 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 and severe inflammation of the eyes with blindness

for some days.

Test substance: Methanol, no further data.

 (173)

Type: LDLo

Species: cat

Route of admin.: i.v.

Value: = 118 mg/kg bw
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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

Species: 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)

No. 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.
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Test substance: Methanol, not further specified

Reliability: (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

Test substance: Methanol, no further data.

Reliability: (4) not assignable

 Secondary source

25-OCT-2006 (174)

5.2.2 Eye Irritation

Species: rabbit

Concentration: 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 symptoms.

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

methanol, but which is reversible and, therefore, gives no

rise to classification as eye irritating.

 -----------------

Reliability: (2) valid with restrictions

 Comparable to guideline study and current standards, limited

documentation

, key study

25-OCT-2006 (133)

Species: rabbit

Concentration: undiluted

Dose: .1 ml

Comment: not rinsed

No. of Animals: 6

Result: slightly irritating

Method: OECD Guide-line 405 "Acute Eye Irritation/Corrosion"
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 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: other: 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 prescribed 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 atmosphere 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
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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: Guinea 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 % active 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 parallel

 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 sensitisation 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.
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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, reproductive 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, clinical-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 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.

 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
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 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 eye 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 cells 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

cholesterol, thymol and glucose). This was not considered

treatment-related.

 BRAIN:

In both upper exposure groups, the number of responsive

stellate cells containing increased endoplasmic 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

astroglia 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 dosing, 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

substance 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 (ophthalmencephalon). 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
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 which showed no clear correlation with exposure levels and

exposure 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-positive 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 1000 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 lungs, 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. 28).

 ---------------------------

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 consumer 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 concentration 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, but indicates that long-term exposure to

 1000 ppm methanol is on the borderline to toxicologically

relevant, histological manifestations (authors statement, p.

27).

 There were histological 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.
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 However, under test conditions (continuous exposure), there

was no evidence of irreversible effects arising from

long-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 allow 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

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 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: Fischer 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 period: 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 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 inhalation chamber.

 20 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.

 Analytical control of chamber concentration: Analytical
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 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 dose 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 body-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 development. 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-response 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 observed

were considered spontaneous ones due to aging.

 -------------------------------

Test condition: Exposure time 20 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

 -----------------

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
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 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 current

standards, documentation limited.

, key study

25-OCT-2006 (88) (167)

Type: Chronic

Species: mouse Sex: male/female

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 period: 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 Guideline 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 inhalation 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 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: 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
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 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-weight 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 surviving 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 represent

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 NOAEL, 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-chronic

Species: rat Sex: male/female

Strain: Sprague-Dawley

Route of administration: gavage

Exposure period: 90 days

Frequency of treatment: daily
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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 significantly 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 the 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 substance: 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
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Result: All animals survived. In rats nasal and eye discharge

 (mucoid, nasal, red nasal, lacrimation) was noted in the

treatment 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

ophthalmoscopic 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 effects 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

 weeks) showed no treatment related histopathological

effects.

Inhalation exposure had resulted in some slight

treatment-related signs.

Reliability: (2) valid with restrictions

 Test procedures 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: 10000; 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

=============================================
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 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 exhibited 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 higher 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 changes 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

is 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 noticeable

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 exposures, 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 documented

, key study

25-OCT-2006 (167)

Species: monkey Sex: no data

Strain: Macaca Fascicularis

Route of administration: inhalation

Exposure 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; 2000; 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
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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 focus 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 study

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 recovery 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 nerve

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,

although -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 infiltrations 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
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 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, therefore, 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 significantly 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 where methanol-induced neurotoxic effects may

 become biological relevant (p. 35).

 --------------

Reliability: (2) valid with restrictions

 Comparable to guideline study, also largely meeting current

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, thereafter 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 prescribed 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
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 retrolaminar and intraorbital optic nerve, swelling 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 (Baumbach

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

blood 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.

[note: 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 the 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

mitochondrial 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., 1978).

 ---------------------------

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
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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)

Control 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 histopathological changes in relation to the
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 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 chronaxia 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

Exposure 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 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 exposure 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: Significantly decreased levels of circulating free

 testosterone were observed among rats exposed at 200 ppm for

2 and 6 weeks (32 % of 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 of administration: inhalation
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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: other

 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 exposure 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 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 (195)

Type: Sub-acute

Species: rat Sex: male

Strain: Sprague-Dawley

Route of administration: inhalation

Exposure 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 production 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 animals (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 reduce 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
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 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 treatment

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 the 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 hormone 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)
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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

Remark: 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 used. Chamber methanol

 concentration was measured regularly by IR.

Histopathological examinations included ophthalmoscopy at

the end of exposure, but apparently no microscopical

inspection 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, purity 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).
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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) valid 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 the 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

Method: 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 trend was indicated [(+-)
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 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 systems.

 -------------

Reliability: (2) valid with restrictions

 Comprehensive test programme using test procedures in

accordance with accepted standard methods, but limited

documentation

 --------------

, key study

25-OCT-2006 (202) (203)

Type: Ames test

System of testing: Salmonella typhimurium TA1538, TA98, TA1537, TA1535

Concentration: <= 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 restrictions

 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

 fraction of KC500(polychlorinated biphenyl)-pretreated rats.

 non-BASF study

Test substance: Methanol, 99.6 % pure, no further data.
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Reliability: (2) valid with restrictions

 Comprehensive test programme using test procedures in

accordance with accepted standard methods, sufficiently

documented

25-OCT-2006 (205)

Type: DNA damage and repair assay

System 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 intermediate 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-, lexA-).

 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 method" on nutrient broth agar).

 --------------------

 The initial concentration was governed by the solubility or by

the toxicity of the TS as inferred from preliminary testing.

Starting 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 well. 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
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 presence of S9 and in a positive result in the absence of S9

at 20 mg/well (Flora et al., 1990, 1984). But 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 than solely "genotoxicity".

(Note: A similar result was obtained with ethanol at somewhat

lower concentrations).

 --------------

Reliability: (2) valid with restrictions

 2c: Meets generally accepted scientific standards, well

documented with methodological restrictions (see Conclusions)

, key study

25-OCT-2006 (203) (206)

Type: other: Micronucleus 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 substance: 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 documented, 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
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Method: Comparative study including a broad spectrum of chemicals.

 Exposure time was 18 hours, no further details of method are

reported.

 Report refers to:

Schmuck, 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 dishes).

 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

given 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 procedures

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

Method: 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, but test substances appeared to be added every day "up

to a final concentration of 1 %".

 -----------
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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 procedure 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, 100 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, HPLC 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 mitomycin, MMN,

 cyclophosphamide and dichromate were highly active.

 ---------------

Test substance: Methanol from Merck, Germany, p.a., no further data.
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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 testing: 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)

Metabolic 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 increases 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

standards, 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

 ------------------
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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 maximum 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 further data.

Reliability: (4) not assignable

 Only abstract, no detailed data.

, key study

25-OCT-2006 (213)

Type: other: Mitotic recombination in ascomycetes

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 activation: 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 requirements and classified as mitotic cross-over,

 non-disjunctional diploids and haploids.

 ----------------------

 Treatment of conidia (10 exp^5/ml) with the TS at early

germination, 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 survival and
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 detection of mitotic segregants.

 ---------------------

Remark: Approx. 3 % chromosomal malsegregation was induced.

 The result was statistically significant at two concentrations

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 aneuploidies 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-related 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

primary 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 mutation 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.

 ---------------------
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Remark: There was no increase 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 for 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 methodological 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: 1977

 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
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 Year: 1983

 GLP: no data

Test substance: no data

Remark: non-BASF study

Test substance: Methanol, no further data.

Reliability: (4) not assignable

 Secondary source

25-OCT-2006 (218) (219)

Type: Bacterial 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 activation: 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 special 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. Bacteria

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" effect.

25-OCT-2006 (220)
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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 polychromatic 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 standards,

sufficient documentation, acceptable for assessment,
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 however, missing positive control

25-OCT-2006 (222)

Type: Micronucleus assay

Species: 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 induction 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.

 ---------------------

Test 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

documentation, 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

Method: 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 mouse 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).

 ----------------
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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

Species: 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

Result: 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 evaluated 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 certain chemicals, and
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 because methanol oxidation via formate to CO2 depends on

tetrahydrofolate.

 ---------------------

 key study

non-BASF study

Result: 4/10 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 current 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.
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 1. Peripheral blood MN: Blood smears 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 on 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 in the frequency of aberrations per cell and
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 percent aberrant cells and no toxic effects were induced by

the treatment:

Aberration rate per cell (including chromatid and chromosome

breakage 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 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/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: Comprehensive 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 electronmicroscopy: 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 synaptonemal

complex.

 -------------

 Spermatocytes of the pachytene substage of the meiotic

prophase after 5 days of last exposure were isolated, and

isolates fixed for electronmicroscopy: 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.
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 --------------------

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 pos. 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: Methanol, no further data

Reliability: (4) not assignable

 Only abstract, no detailed data

25-OCT-2006 (226) (88)

Type: Drosophila SLRL test

Species: Drosophila 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-line 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) close 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
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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 the 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: Fischer 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: 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:

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 concentration: 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
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 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 for 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

been 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 and 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 CHANGES:

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) Adematoids = 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
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 -----------------------------------------

 C 2/50 3/51 2/49 7/51

 =========================================

 -- = not examined

 A) Papillary adenomas (males)

B) Adematoids = hyperplastic changes (pre-neoplastic) (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 (chromaffinomas) 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

compared 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 somewhat 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 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 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 largely meeting current

standards, documentation limited, acceptable for assessment.

, key study

26-OCT-2006 (88) (167) (180)
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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 mg/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-term 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.

 Analytical 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 general 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 control 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 tumors exclusively observed in the 1000-ppm group were
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 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-tumors 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)

Species: 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

Type: 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
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 male: 60 d (approx. 1200 h)

 female: 60 d (approx. 1200 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

Method: 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 evidence

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).

 Pathohistology:

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
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 week after weaning (p. 187/188).

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 obtained 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 " : approx. 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

may 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 occurring 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 migration

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 the 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 respective

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
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 weeks. This was still found in females necropsied

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 control 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 copulation 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 generations 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 obvious 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

progeny 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
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 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 Period: 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 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 them 26 in-utero
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 treated offsprings (see other entry: 5.8.2).

 ------------------------------------

Remark: key 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

 GLP: 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

spermatogenesis 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 evaluated per

animal.

 -----------

Remark: non-BASF study

Result: In the methanol-treated animals, a slight, but statistically

 insignificant increase in sperms abnormalities (mean factor

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.
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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 Sex: 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 were

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, 2001, 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
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 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 reproduction:

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-related 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, respectively).

 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
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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 estimations 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 0.001 at 980 ppm.

 ------------------------

Reliability: (2) valid with restrictions

 Comparable to guideline study and current standards,

sufficiently documented, acceptable for assessment

, key study

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 reproduction 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 used 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 concentration values
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 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 which even

 may have been higher than in studies using the same exposure

concentration, but shorter exposure times.

 --------------------

 key study

non-BASF study

Result: 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 number 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).

 Morphological 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 with 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. about 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

costotransversarium" (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 the 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 5000 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, mortality 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
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 and opening of eyelid for both sexes and descensus testis

for males were significantly 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 functional 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 (missing 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 F1 (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 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 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 induce 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 toxicity

is considered to be 1000 ppm.

 --------------------

Reliability: (2) valid with restrictions

 Comparable to guideline study and current standards, also

largely meeting current standards, documentation limited,

acceptable for assessment

, key study

26-OCT-2006 (167) (180)

Species: mouse Sex: female

Strain: 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
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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

5000 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 levels in human blood upon kinetic
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 saturation.

 ----------------

 key study

non-BASF study

Result: Mean blood levels:

 approx. 100 ug/ml (1000 ppm), 540 (2000 ppm), 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 exencephaly 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 ribs 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).

The 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 data

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
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189

Method: Study was to elucidate time-related 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 principles, acceptable for

assessment

26-OCT-2006 (238) (239)

Species: mouse Sex: female

Strain: CD-1

Route of administration: 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 prescribed 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 (neural

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
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 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 defect 30 (3.6) 65 (14.7)

Cleft palate 50 (14.6) 88 (50.4)

-------------------------------------------------------

At 5000 ppm no difference from control: no neural tube

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 in

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 significant 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 detrimental

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 methanol. 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
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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 dams 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 exposed 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 difference 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 indices

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., 1997).

 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
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 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 responded 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 development/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, previously 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: prebreeding 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 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 (see also entry: 5.8.1)

 - reproductive toxicity

 - developmental neurotoxicity

 Specific aims of the study were to determine whether
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 - 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 them 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 600 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

thereafter.

 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

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.

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.

 The results or behavioral assessments of offspring did not

indicate methanol-exposure effects on most domains of early
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 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 effects 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,

many 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 effect 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 over

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 females 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 used 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 offsprings 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 function 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
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 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 increase

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 levels 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 1800 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, Appendix 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
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Route 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 fetal 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 intestinal 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 period 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.

 Folate 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 erys (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 irrespective 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

weight 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 folate-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, folate deficiency failed to produce significant

malformations (in accordance with previous reports: e.g.
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 Heid et al., 1992):

cleft palate (5/215 vs. 0/282) and execephaly (2/215 vs.

1/282 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-def.) 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).

 -----------

Test 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 procedures in accordance with accepted standard

methods, sufficiently documented

, key study

26-OCT-2006 (244) (224) (245) (246)

Species: rat Sex: female

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 (day18), 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.
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 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 bodyweight were modestly

 but statistically lower after treatment. No increase in

postnatal mortality.

No other test revealed significant effects of methanol

treatment.

Test substance: Methanol, 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 per

 group. The following parameters were examined: maternal body

weightand clinical signs, gross, skeletal and visceral

malformations in fetuses, histopathology and scanning

electon microscopy in fetuses.

 --------------

Remark: non-BASF study

Result: Maternal toxicity was seen as clinical signs and reduced

 bodyweight, in fetuses the treatment induced reduced

bodyweight 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 Baker

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
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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

Doses: 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 evaluated.

 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 following 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, because 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

Result: 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-methylpyrazole 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 affected 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 folate 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
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 proximate teratogen in pregnant CD-1 mice exposed to high

concentration of methanol vapor, because

1. no formate 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 doses 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 evidence 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). But 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;

Dorman 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: until 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 methanol

 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 methanol 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
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 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

that 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

 Test 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 mg/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)
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 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 birth

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

 neurotoxic 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/kg 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/Screening: 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 0.67

 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 changes

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.

Reliability: (4) not assignable

 Only short communication, abstract

26-OCT-2006 (256)

Species: rat Sex: female

Strain: Long-Evans

Route of administration: drinking water
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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 bodyweight (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: other 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

performed 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 significantly 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

publication.

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

Duration 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 alizarin 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 % decrease).

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 19.5 %), 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 and

anophthalmia.

 ------------

Test substance: Methanol, HPLC grade
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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 fetuses, 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

 ribs 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) invalid

 Unsuitable test design (see Conclusions)

26-OCT-2006 (261)

Species: rat Sex: female

Strain: other: Holtzman

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: yes, concurrent vehicle
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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

bodyweight, 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 determined 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 parameters 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 inhibition of the DCR. However, this

 apparent potential impact on decidua development was not
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 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 parturition

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, concurrent 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 description 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 weights 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 cranio
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 facial defects (Rogers et al., 1996).

 -----------------------

Remark: non-BASF study

Result: 1.7 g/kg methanol induced 30 % resorptions and midfacial

 deficiencies and micro/anophthalmia 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, cebocephaly, 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 arches. 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 Sex: 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 day 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.
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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

proliferation, 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, somite 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 embryos 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 induced 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
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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: Rat 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 significantly 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 in 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 of

 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.
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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 stages 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 concentration-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 exposure 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
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 GLP: no data

Test substance: as prescribed by 1.1 - 1.4

Method: Screening test including developmental stages of

 embryogenesis:

The following parameters were examined: percentage of live

embryos, cephalic dysraphism, prosencenphalic lesions,

branchial arch hypoplasia.

Remark: non-BASF study

Result: Methanol 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

Frequency 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 interfere 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., 1998b).

 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 individual

toxicities, suggesting antagonistic interaction (Andrews et

al., 1998a).

 A standard methanol dose-response line has been constructed,

based on the developmental scoring system. Combinations 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
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 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 dose

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 test

 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 FR20

 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 running 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.

 STATISTICS:

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 of experience: other: review, general

Remark: Humans (and non-human primates) are uniquely sensitive to
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 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

beverages and from methanol-containing products. Although

 ingestion dominates as the most frequent route of

poisoning,

inhalation of high concentrations of methanol vapor and

percutaneous 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 methanol 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 subsequent 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.

 The 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 permanent 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 interindividual 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 ingestion 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 symptoms 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 photophobia 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
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 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 in

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 effects 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 200 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
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 methanol

 and formate levels; visual qualities (color discrimination

 and contrast sensitivity); and neurophysiological (auditory

evoked potentials) and neurobehavioral qualities.

Exposure to methanol increased serum concentrations and

urinary excretions of methanol, but did not affect formate

levels. Overall visual, neurophysiological, and

neurobehavioral test outcomes were not significantly

affected, unless certain between-subject variables are

considered. Slight effects on P-300 amplitude and Symbol

Digit testing were noted.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

24-DEC-2004 (278)

Type of experience: other: review, general (Becker)

Remark: Methanol poisoning is an uncommon 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 swift laboratory confirmation

is

essential in managing this poisoning. Methanol poisoning

may

occur in sporadic or epidemic circumstances. Chronic

exposure 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 administration 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 evidence

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 alkalinization to counteract metabolic

acidosis. Folate should be administered to enhance

metabolic

breakdown of formate. Alcoholic patients may especially
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 susceptible to methanol 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 between 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 observed 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 formate 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 consequences

of acute, rather than chronic, exposures. Acute methanol

toxicity evaloves in a well-understood pattern and consists
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 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 formate

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 restrictions

 see remark on selection of literature

28-DEC-2004 (192)

Type of experience: other: kinetics, oral (Ferry)

Remark: Methanol blood and urine concentrations 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/ml. 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 that 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 respresents

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
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 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 concentration 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 formic-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 restrictions

 acceptable study, meets basic scientific principles

28-DEC-2004 (285)

Type of experience: other: review, acute toxicity (Litovitz)

Remark: Annual report 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, respectively.

Reliability: (2) valid with restrictions

 see remark on selection of literature

28-DEC-2004 (286)

Type of experience: other: kinetics, inhalation (Baumann; Angerer)

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
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 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 exposed 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 closed 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 not 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 poisoned 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 others 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)
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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 and 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 metabolic

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)
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Type of experience: other: worker exposure (Kawai / Yasugi)

Remark: In methanol-exposed male and 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 unexposed

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 (females) 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 200 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 3600 mg/m3 (one case), two showed

 retarded pupil reflex and one mild mydriasis (Kawai et al.
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 1991). Dimmed vision and nasal irritation were among the

most frequent symptoms complained during work.

Reliability: (2) valid with restrictions

 acceptable study, meets basic scientific principles

28-DEC-2004 (292) (293)

Type of experience: other: human exposure (Franzblau)

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 the

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

 Institute 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 health effects among teacher aides, or

had been responsible for the deaths of three former teacher
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 aides. Death certificates and autopsy data were obtained

and

evaluated. A self-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 alcohol 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 reported

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: other: human exposure (Batterman)

Remark: Dermal exposures to methanol were administered in a

 clinical

study designed to compare several biological indicators.

Four subjects were exposed 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. After 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)
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Type of experience: other: human exposure (skin) (Dutkiewicz)

Remark: Methanol absorption rate through human skin (foremarm,

 15-60

min.) has been examined 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 from 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 experience: 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 periods 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 

periods 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-2004 (299)

Type of experience: other: kinetics, inhalation, human (Batterman)

Remark: Experimental study of dermal exposure to methanol in human

 volunteers estimating 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 different 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 restrictions

 acceptable study, meets basic scientific principles

28-DEC-2004 (300)
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Type of experience: other: kinetics, inhalation, human (Lee)

Remark: Accumulation 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 six 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 increase 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, mandelic 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 each 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
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 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 severe 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 stated 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

 acceptable 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 methanol 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 chemical

exposures.

 Because of these limitations the Panel judged the study

results to be uncertain and concluded there are

insufficient

data upon which to 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
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 smaller fractions over the day.

 -------------------

28-DEC-2004 (304) (233)

Type of experience: 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 levels around 10 mg/LL

in adults.

Reliability: (4) not assignable

 Secondary literature

28-DEC-2004 (305) (233)

Type of experience: other: remark on 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 the 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

these

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. Overall, 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 formic acid through a formaldehyde

intermediate. Formate accumulation causes metabolic

acidosis

and inhibits cellular respiration. Methyl alcohol toxicity

is time and concentration dependent, and its toxic effect
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 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

 detection 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) valid with restrictions

 basic data given, acceptable restrictions

 (308)

Type of experience: other: kinetics, inhalation

Remark: Urinary formic acid 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.

Reliability: (2) valid with restrictions

 acceptable study, meets basic scientific principle

 (309)

Type of experience: other: kinetics, inhalation

Remark: 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

following cessation of exposure.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (310)

Remark: Methanol is slowly 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
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 for methanol in urine of the general 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 and 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 blood 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 urinalysis 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 intensity 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

aromatics, 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 data given, acceptable restrictions

 (313)

Type of experience: other: kinetics, model

Remark: Two patients with extremely high blood methanol

 concentrations (260 and 282 mg/dl) were successfully

treated

using pharmacokinetic dosing of ethanol, hemodialysis and

supportive measures. Both patients recovered completely

without 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
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 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 ethanol 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 aspartame, 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 observed 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 administered 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 were 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 studied 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
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 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 concentration-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 still 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 differences 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) valid 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 it 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 remark 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
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Remark: Methanol poisoning with typical disturbed vision may appear

 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 restrictions

 see remark on selection of literature

 (319)

Type of experience: other: review, acute toxicity

Remark: Methanol intoxication causes severe metabolic 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 intoxication 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 presence 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 showed 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 metabolic 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 sometimes elusive and

requires

a high index of suspicion. Because antidotal treatment is
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 available it is important to recognize methanol poisoning

promptly. The presence of metabolic 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 suggests 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 ethanol 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)
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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 poisoning 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 seen. 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 mean

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 ganglion 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)
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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)

Type 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 vats. 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 of 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-year 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 neuropathological 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.
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 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 toxic 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 only 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 effect, 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 systemic effects: changes in

circulation, cough, dyspnea, headache, lacrimation, nausea

or vomiting, optic nerve neuropathy, respiratory effects,

visual field changes.

Reliability: (2) valid 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 profound 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 sustained permanent injury.

The two basic mechanisms of poisoning are direct chemical

irritation of tissues and systemic acidosis. The former is
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 influenced by the unlimited miscibility of methanol with

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.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (337)

Type of experience: other: acute toxicity

Remark: A series 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 questionable value when poisoning has occurred.

Plasma

extenders were found valuable in combating peripheral

vascular failure. Frequent drainage of the cerebrospinal

fluid does not appear 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 % methyl 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 men 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

clearance 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
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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 percutaneous 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 restrictions

 (342)

Type of experience: other: acute toxicity

Remark: Case report of fuzzy and heavy feeling in the head and

 tendency to sway in a worker 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 restrictions

 (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 found. 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.
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Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (345)

Type of experience: other: acute toxicity

Remark: A patient poisoned with methanol was successfully

 hemodialyzed with an ethanol-enriched, bicarbonate-based

 dialysate. Along with a concomitant intravenous infusion of

ethanol, the ethanol-enriched dialytic 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.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (346)

Type of experience: other: acute toxicity

Remark: Observations 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 when 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

 acceptable 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

child. 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 depression 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 preparations. 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
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 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) valid with restrictions

 acceptable study, meets basic scientific principles

 (349) (350)

Type of experience: other: acute toxicity

Remark: Significant toxicity 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%), methanol (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 necessary 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

three

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 experience: 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
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 tachypneic, tachycardic, hypertensive and hypothermic.

Laboratory results were significant for a severe metabolic

acidosis, a serum osmolality of 465 and serum 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 elevation 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 blurred and tunnel vision. The

patient

had visual disturbances and an osmolal gap that suggested

methanol intoxication.

Reliability: (2) valid with restrictions

 basic 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 methanol poisoning with negative results.

The, patient had seizure activities with dilated pupils,

 Clenched aw, and tonic muscle actitivity. Deep tendon

 reflexes wereabsent symetrically. Neurological 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) valid with restrictions

 basic data given, acceptable restrictions

 (355)

Type of experience: other: acute toxicity

Remark: A 44-year-old man was found 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 methanol 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/100 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.
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Reliability: (2) valid with restrictions

 acceptable 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 anion 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 hemorrhage 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 5th 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, acceptable 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 acidosis after a binge.

Abnormalities of the cerebrospinal fluid included an

initial
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 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 showed 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 respirations. 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.

Reliability: (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, bradykinesia, hyperreflexia, bilateral

Babinski responses, and mild dementia were reported. CT

scan

in both patients demonstrated symmetrical low density

lesion

in 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

failure thirty hours, two days, and eighteen days,

respectively, after presentation with acute bilateral
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 visual loss. The retrolaminary nerve segments showed

circumscribed

demyelination of loss of myelin staining with relative

preservation of axons. Retinal ganglion cells were spared.

Electromicroscopy in the case with shortest survival

revealed periaxonal intramyelinic spaces behind the lamina

cribosa.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (363)

Type 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 probably

caused by histotoxic anoxia in watershed areas of the

cerebral and distal optic nerve circulations. Juxtabulbar

demyelination may cause optic disk edema in methanol

poisoning by compressive obstruction of orthograde

axoplasmic flow. Visual loss may be due to disruption of

saltatory conduction. Retrolaminar demyelinating optic

neuropathy is an early morphologic 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 necrosis

 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

with persitent sequel.

Reliability: (2) valid with restrictions

 basic data given, acceptable restrictions

 (367)

Type of experience: other: acute toxicity, cns

Remark: Report of with areas of decreased attenuation symmetrically
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 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 experience: 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 selection of literature

 (369)

Type of experience: other: review, therapy

Remark: Treatment of methanol poisoning is well established and

 consists specifically 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 sophiticated 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 given

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

concentration 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, especially 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.
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Reliability: (2) valid with restrictions

 see remark on selection of literature

 (371)

Type of experience: other: review, therapy

Remark: A unique opportunity 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 initially 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 of 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 anion 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 visual outcome. The interval from

ingestion to treatment appears to be more important than

the

initial biochemical status. Prompt hemodialysis is

recommended, if the blood methanol 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
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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,

whereas 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 absence 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 determinations of the anion and osmolal

gaps

are mandatory whenever facing metabolic acidosis of unknown

etiology.

Reliability: (2) valid with restrictions

 acceptable study meets basic scientific principles

 (375)

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 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 range. 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-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 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
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 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 patients 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 ethanol were

found

in 5 patients, and high lactate levels explained in part

the

most extensively increased anion gaps. However, the

calculated analytical standard 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 methanol concentration in blood was

estimated

as approximately of 134 mmol/L, based on the calculation of

the osmolal gap. Intravenous ethanol administration and

hemodialysis were promptly started. During hemodialysis,

several blood samples were collected for determination of

methanol and ethanol concentrations. Initially, we used gas

chromatography with split-mode injection of pretreated

 serum

samples; however, methanol concentrations turned out to be

significantly lower than expected, based on calculated

osmolal gap values. Because no explanation for the excess

serum osmolal gap was apparent, we reanalyzed samples,

using

head-space gas chromatography. The methanol concentrations

measured were significantly 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 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 range. A slight, transient
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 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-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 restrictions

 basic data given, acceptable restrictions

 (376)

Type of experience: other: irritation

Remark: In an experimental study, 12 volunteers were exposed 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 with 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) valid with restrictions

 basic data given, acceptable restrictions

 (380)

Type of experience: other: irritation

Remark: Local eye contact with liquid 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

 (382)

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
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 blindness. In one fatal case of occupational methanol

intoxication by inhalation a female worker was exposed

about

12 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, resulted 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 decrease 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, acceptable restrictions

 (385)

Type of experience: other: chronic toxicity

Remark: A study of the wood heel industry in Massachusetts found

 average methanol 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, acceptable restrictions

 (386)

Type of experience: other: chronic toxicity

Remark: A report indicated severe recurrent headaches in workers

 exposed to methanol 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.
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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 and

 methanol in a collar fusing plant were studied. Physical

examinations revealed no abnormalities. Blood studies

revealed no disturbances in hematopoesis. Absorption of

acetone 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 restrictions

 see remark on selection of literature

 (88)

Type of experience: other: review, reproductive and developmental effects

Remark: The Panel reviewed 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 concluded 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 restrictions

 see remark on selection of literature

 (233)

Type of experience: other: review, reproductive and developmental effects

Remark: No studies 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 selection of literature

 (88)

5.11 Additional Remarks

Type: other: review
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Remark: A. The toxic effects of methanol in humans and their

 implications 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 formate, 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 exposure

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 reviewed 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 is 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 consists 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.

 Ophthalmologic 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,
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 while the papilla itself is not oedematous.

 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 substance: 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-reduced 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, methanol 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 (Garner

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 methanol 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.
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 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 parallel, 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 correlate 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 in 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.

Methanol 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.
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 This suggests that methanol distributes 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 accepted

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 hours

 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 methanol 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 teratogenic 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
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 (394) (395)

Type: other: Toxicokinetics / formate / HEI study monkeys

Method: Comparative study designed to clarify 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 inhalation (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 nominal).

 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 only 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 ppm 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
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 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) present 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

concentrations. 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: Comparison 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 (HEI,

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 unlikely

(Horton et al., 1992, Discussion).

 ------------------------------------

Reliability: (2) valid with restrictions

 2e: Study well documented, meets generally accepted

scientific principles, acceptable for assessment.

 (396) (397) (252)

Type: other: Absorption / inhalation / human

Method: Six human volunteers (from 29 - 55 years) were subjected 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 individuals were each tested at rest or at light

exercise.
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 ------------

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 7.47 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 unchanged, 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
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 scientific principles, acceptable for assessment.

 (398) (196) (252)

Type: other: Risk Assessment Model / inhalation

Method: A "multicompartment biologically based dynamic" inhalation

 model based on kinetic 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 methanol 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

 and 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 exposures 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).
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 -----------------

Reliability: (2) valid with restrictions

 (299) (399) (251) (397) (277) (283)

Type: other: Absorption / distribution / inhalation / rat

Method: Whole-body exposure system:

 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 through 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 absorption 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).
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 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; Tab. 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 cynomolgus 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.

 Blood 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
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 positive increasing trend, although not significant (Dorman

et al., 1994; Medinsky et al., 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

Discussion).

 ----------------------

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 remained 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.

 (251) (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 doses 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

disappearance 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
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 increased with increased 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 animals 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, 20 % via urine, the excretion was delayed in this

group.

 -------------------

Reliability: (2) valid with restrictions

 Test procedures in accordance with accepted standard

methods, sufficiently documented

 (167)

Type: other: Absorption skin / human

Method: a) Exposure of 0.19 - 0.21 ml methanol to the skin of 6

 human 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 water and determined

sprectrophotometrically (Dutkiewicz et al. 1980).

 b) Exposure of one hand to methanol (Dutkiewicz et al.

1980).

 ---------------

Result: a) Following occluded 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, 1997).

 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)

 (Batterman 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
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 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 quantitative

kinetic results for skin permeation in humans and appear to

be reliable.

 ---------------

Reliability: (2) valid with restrictions

 Special study design, sufficiently documented, meets

generally accepted scientific principles, acceptable for

assessment

24-DEC-2004 (300) (390) (298) (402)

Type: other: metabolism / mouse / FDH deficiency

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

cytosolic 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 between 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 were 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

expected. 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 the

 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 metabolic 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 metabolic

pathways in mice for the oxidation of formate arising from

methanol or other sources:
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 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 procedures in accordance with accepted standard

methods, sufficiently documented

27-DEC-2004 (403)

Type: other: Toxicokinetics / formate / folate deficiency / monkeys

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 under 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 monkey 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 principles, acceptable for assessment.

24-DEC-2004 (168) (404)

Type: other: Metabolism / in vitro / formate / various species

Method: The folate-dependent formate oxidation and the activities

 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-formyltetrahydofolate dehydrogenase activity, the enzyme

 catalyzing the final step of formate oxidation to CO2, about
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 25 % and 37 %, respectively, than in rat liver.

Furthermore, total folate and H4folate in 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 in 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 toxicity 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 higher 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, preferentially

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 with

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)
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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 exposure

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 production

 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 data.

 (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 essential 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 chronic 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 with the

authors ́ observations and is not suggested in the report.

 ---------------------------------

Reliability: (2) valid with restrictions
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 (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 concentrations 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

developmental 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 similar.

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 respiratory uptake of methanol

 by rats can be assumed to be approx. 100 % at exposure

concentrations of <5000 ppm.

Reliability: (2) valid with restrictions

 (410)
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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 selectively 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 dose 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 retinal 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 spacing 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 in blood), no

 histopathological changes were apparent at the

light-microscopic level (Wallace et al., 1997).

 However, visual dysfunction was already visible in

functional dignostic, occurring 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

 al., 1999, 2001).

 Biochemical effects:

Retinal ATP, ADP, and GSH were significantly depleted
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 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

 concentrations 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 model 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 restrictions

 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 Chemicals, 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: Biochemical 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 differentiation. The

effect increases with increasing chain length, methanol
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 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

 concentrations 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 catalase 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
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 al., 1992).

 -----------------------

Test substance: Methanol, no further data.

 (421) (422)

Type: Toxicokinetics

Remark: In monkeys, four daily skin application of 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

variation was nonetheless large in either species.

Test substance: Methanol, no further data.

 (156)

Type: Toxicokinetics

Method: Comparative toxicity and metabolic 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 (LD50)

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 with 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 catalase-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 results 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.
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 -------------------

 (137)

Type: Toxicokinetics

Remark: The pharmacokinetics of methanol was compared in Fisher

 rats and Rhesus monkeys. After i.v. application 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 female monkeys (Macaca fascicularis)

and qualitative/clinical signs of toxicity. It comprised

 - methanol pharmacokinetics

 - maternal toxicity (see other entry: 5.8.2)

 - reproductive 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/week 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-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 increase

above the background range of about 0.15 - 0.30 mM (approx.
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 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 documented

 (423)

Type: other: Ocular toxicity / animal model

Remark: 1. In a preliminary study, F-344 rats fed control and

 folate-deficient diets 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 accumulation 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 rate 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 blood 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
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 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 visual 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 is

 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

moderate 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 reproductive

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
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 (429)

Type: other: Risk Assessment Model / inhalation

Remark: Quantitative Models of Risk Assessment for Developmental

 Neurotoxicants, based on inhalation data 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 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 0.001 at 980 ppm.

 ------------

 (430) (234) (235)

Type: other: Toxicokinetics / ethanol influence

Method: Comparative 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).

Result: 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 normally 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)

Remark: 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 accumulation mainly due to limitation/low

level in hepatic tetrahydrofolate along with a low

10formyl-THF dehydrogenase.
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 ---------

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 pressure and blood flow through the Arteria femoralis

and Ateria carotis. On the other hand, peripheral

resistance 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

treatment, 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 (end 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 only acid-labile catalase

 activity.
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 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-stable 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 experiments, 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 stimulation 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 functional 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.

Formaldehyde and formate were analyesd in blood, urine, and

in the cerebral fluid, the orbit fluid of the eye, liver,

kidney, brain, and optic nerve.
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 -----------------

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 with 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 levels. 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 prevented

formic-acid accumulation in blood.

 -------------------

Test substance: Methanol, no further data.

 (439)

Test substance: Methanol, no 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)
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Remark: Laquer thinner containing methanol, which is used for

 inhalation 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 substance: 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 rates 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 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

at increased levels in acidotic monkeys.

Test substance: Methanol, reagent grade, no further data.

Reliability: (2) valid with restrictions

 (168)
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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 morphology

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 (20 hours/day, 7 days/week, 12 - 13 rats/group). The

 results are equivocal.

Test substance: Methanol, no further data.

 (456)

Remark: Male rats were exposed 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 inhaled 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 substance: Methanol, no further data.

Reliability: (2) valid with restrictions

 (434)

Remark: Methanol is metabolised in rodents and primates in

 different 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).
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 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 balance 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 effect 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 amount 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 et

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 animals, conceptal/maternal AUC and Cmax-ratios
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 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 decreased 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 shown 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 homogenates. 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 potentially 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 severe 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 methanol 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 from 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 methanol

 threshold limit value (TLV) of 200 ppm, the formate does not

accumulate in blood in human subjects at rest and during
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284

 exercise. This was confirmed by other authors (D ́Alessandro

et al., 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 levels 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 130-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 peroxidation 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-Acetylcysteine 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 resulted 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 protective 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: Comprehensive 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
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 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 conditions 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 exposed 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 to 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

stimulating hormone, testosterone, prolactine) were

inconsistent with partly no effects and partlycontradictory

results. According to the author, methanol treatment led

only to increased prolactin levels in 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).
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 Provided that the rate is the same in man, the amount of

methanol oxidized during 18 h (the 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)
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