OECD SIDSETHANOLFOREWORDINTRODUCTIONETHANOLCAS N°: 64-17-5

UNEP PUBLICATIONS

1OECD SIDSETHANOL

SIDS Initial Assessment Report
For
SIAM 19
Berlin, Germany, 19 – 22 October 2004

1. Chemical Name:ETHANOL

2. CAS Number:

64-17-5

3. Sponsor Country:

Czech RepublicContact Point:
Ministry of Environment
Contact Person:
Karel Bláha, Ph.D.
Director
Department of Environmental Risks
PragueSlovak RepublicContact Point:
Centre for Chemical Substances and Preparations
Contact Person:
Peter Rusnak, Ph.D.
Director
Centre for Chemical Substances and Preparations
Bratislava
CEFIC Ethyl Alcohol Group

Co-sponsor Country:

4. Shared Partnership with:
5. Roles/Responsibilities of
the Partners:•

Name of industry sponsor/consortium•

Process used

CEFIC Ethyl Alcohol Group (BP Chemicals Ltd, Sasol Gmbh,
Sodes SA.)
Contact Point:
CEFIC Ethyl Alcohol GroupAve E van Nieuwenhuyse 4
B-1160Brussels
Belgium
Graeme Wallace
Documents drafted by industry consortia then peer reviewed
by sponsor country experts.

6. Sponsorship History•

2

How was the chemical or
category brought into theOECD HPV Chemicals

This substance is an ICCA HPV chemical. Industry approached
the sponsor country. The substance has not been part of any otherassessment although it has been through the US HPV programme
UNEP PUBLICATIONSOECD SIDS
Programme ?
7. Review Process Prior to
the SIAM:

8. Quality check process:9. Date of Submission:

ETHANOL
sponsored by the US Renewable Fuels Association.
The industry sponsor prepared the documents and the Czech andSlovak Competent Authorities reviewed the documents andprovided edits and changes where necessary in an iterative
process with the industry sponsor. All documents were
subsequently updated, based on the comments from Competent
Authorities on the submitted documents received through the
CDG
Data was reviewed against the OECD criteria as describedin the SIDS manual. These criteria were used to select data
for extraction into the SIDS dossier. Original data was sought
wherever possible. Originally reported work was deemed reliable
if sufficient information was reported (according to the manual)to judge it robust. Reviews were only judged reliable if reported
by reputable organisations/authorities or if partners had been directly
involved in their production.
23 July 2004

10. Date of last Update:
11. Comments:

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OECD SIDSETHANOL
SIDS INITIAL ASSESSMENT PROFILECAS No.

64-17-5

Chemical Name

Ethanol

Structural FormulaCH3-CH2-OH

SUMMARY CONCLUSIONS OF THE SIAR

Human Health
The assessment of the substance is focused on its use as industrial chemical. Ethanol consumption in alcoholic
beverages is out of the scope of this report.
Ethanol is readily absorbed by the oral and inhalation routes and subsequently, metabolized and excreted in humans. At
exposures relevant to occupational and consumer exposure during manufacture and use of ethanol containing products,
the alcohol dehydrogenase metabolic route in the liver dominates and does not become saturated. This mechanism
follows first order kinetics. The first step of the metabolic path is the rate-determining step; concentrations of theintermediate metabolite acetaldehyde are very low. Ethanol is not accumulated in the body. Dermal uptake of ethanol isvery low.
Ethanol has a low order of acute toxicity by all routes of exposure. Lowest robust reported values are an inhalation
LC50 of >60,000ppm (114,000 mg/m3), 1 hour, mouse), and an oral LD50 of 8300mg/kg.bw (mouse). Ethanol is amoderate eye irritant but is neither a skin irritant nor a sensitizer.
For repeat dose effects, the lowest reported NOAEL is approximately 2400 mg/kg bw/day from a dietary study with
rats. At higher doses, male rats showed minor changes to organ weights and haematology/biochemistry; female rats
showed minor biochemistry changes and increased length of oestrus cycle along with liver nodules; adverse liver
effects were observed at concentrations of 3600mg/kg.bw/day and above
The balance of evidence is that ethanol is not genotoxic. Negative results from a number of bacterial mutation assays
appear to be reliable. Of the mammalian cell mutation assays a weak mutagenic effect in mouse lymphoma cells
occurred only at very high ethanol concentrations. In vivo tests for chromosome aberrations in both rats and Chinese
hamsters have given negative results. There is very little evidence to suggest that ethanol is genotoxic in somatic cells
and it may have a very limited capacity to induce genetic changes in vivo but under very specific circumstances and atvery high doses achievable in humans only by deliberate oral ingestion.
Evidence of the carcinogenicity of ethanol is confined to epidemiological studies assessing the impact of alcoholic
beverage consumption. These do not indicate any such hazard exists from potential exposure to ethanol in the work
place or from the use of ethanol in consumer products.
No fertility or developmental effects were seen at inhalation exposures up to 16000 ppm (30,400 mg/m3). The lowest
reported NOAEL for fertility by the oral route was 2000 mg/kg bw in rats, equivalent to a blood alcohol concentrationof 1320 mg/l, although this was based on a significant increase in the number of small pups rather than a direct effecton fertility; such direct effects are not seen until much higher doses. Many studies exist examining the developmental
end point for ethanol. However, most use very high doses and few are individually robust enough to allow a NOAEL tobe established. However, the collective weight of evidence is that the NOAEL for developmental effects in animals ishigh, typically >=6400mg/kg bw, compared to maternally toxic effects at 3600 mg/kg bw. The potential for
reproductive and developmental toxicity exists in humans from deliberate over-consumption of ethanol. Blood ethanol
concentrations resulting from ethanol exposure by any other route are unlikely to produce reproductive ordevelopmental effects.

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Environment
The available physicochemical data are adequate to describe the properties of ethanol. The melting point for ethanol is 114oC, the boiling point is 78.3 oC and the log Kow is –0.31. Ethanol has a measured vapour pressure of 57.3 hPa at
20°C. Ethanol has a specific gravity (density) of 0.7864 and a flashpoint of 14 oC. It is fully water miscible at ambient
temperatures. Henry's Law constant is 0.000252.
Ethanol is stable to hydrolysis but is readily biodegradable (74% after 5 days) and is not likely to bioaccumulate(calculated logBCF=0.5). Ethanol is not persistent in the environment. Fugacity-based modelling shows that ethanol
released into the environment will become distributed mainly into air and water. Relative distributions between
compartments based on an emission pattern of 1000:100:10 were 57 % in air, 34 % in water, and 9 % in soil. These
predictions are supported by the limited data available on prevailing concentrations, which shows that ethanol has been
detected in outdoor air and in river water. The total tropospheric half life of ethanol is estimated to be 10-36 hours, withdegradation due to hydroxyl, NOx and SOx radical-mediated photooxidation. As a volatile organic compound in
the atmosphere, ethanol is a potential contributor to tropospheric ozone formation under certain conditions, however its
photochemical ozone creation potential is considered to be moderate to low (40-45 relative to ethylene as 100).
The aquatic toxicity data in fish, invertebrates, and algae indicate a low order of acute toxicity with LC50/EC50 values
greater than 1000 mg/l. The most sensitive species were algae Chlorella vulgaris with a 96hr EC50 of 1000 mg/l and theinvertebrate Artemia Salina with a 24hr LC50 of 1833 mg/l. Valid chronic toxicity data are available for two trophic
levels. The lowest reported NOEC for invertebrates is 9.6 mg/l (10 day reproduction) whilst for plants it is 280mg/l (7day study).

Exposure
Worldwide ethanol production was 25,000 kt in 2001. European production is 1,700 kt. Ethanol is manufactured either
by fermentation of biomass or by the hydration of ethylene in a continuous, closed process; release from production
facilities is low.
Ethanol use falls into four main categories: use as a solvent, in manufacture of chemicals, as a fuel additive and for
potable drink manufacture. Solvent use is mainly in paint and ink manufacture and in pharmaceutical production.
Ethanol is widely used in consumer products, mainly cosmetics, but also detergents, winter deicing and cleaningproducts, including detergents.
There is limited published data on exposure of workers to ethanol but what is available indicates that the vast majorityof exposures are well below current occupational exposure limits (OELs). However, some scenarios were identified in
one of the sponsor countries with the potential for high inhalation exposure (eg pharmaceutical manufacture). Personal
protective equipment should be used in these situations to reduce actual exposure There is no data on consumer
exposure from the use of products containing ethanol, but it is likely that the dominant source of consumer exposure toethanol is through natural sources in foodstuffs and the consumption of alcoholic beverages.

RECOMMENDATION AND RATIONALE FOR THE RECOMMENDATION AND NATURE OF
FURTHER WORK RECOMMENDED

Human Health: The chemical is currently of low priority for further work. The assessment of the substance is focusedon its use as industrial chemical. Ethanol possesses properties that indicate a hazard for human health but these aremanifest only at doses associated with consumption of alcoholic beverages. In the context of an industrial chemical,
these hazards do not warrant further work as they are not likely to result from the manufacture and use of ethanol andethanol containing products.
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:
IUPAC Name:
Molecular Formula:
Structural Formula:
Molecular Weight:
Synonyms:

64-17-5ethanol
C2H5OH
CH3CH2OH
46.07
AlcoholAnhydrolEthyl alcohol
Ethyl hydrate
Ethyl hydroxide
Grain alcohol
JaysolMethyl carbinol
Potato alcohol
SpiritSynasol
TecsolSubstance typePhysical description
Degree of purity
Major impurities
Optional additives

Alcohol
Colourless, clear
95%-99.9%
Up to 5% water
Up to 5% denaturant or organoleptic modifier

1.2

Purity/Impurities/Additives

For Customs and Excise purposes, ethanol is often denatured with up to 1 to 5% of one or moredenaturants to render it unsuitable for human consumption. Further small quantities of organoleptic
modifiers or marker solvents may also be required. Typical denaturants are shown in the Table 1:
Table 1 Additives used for Customs and Excise purposes

6

Examples of commonly used denaturants

Methyl ethyl ketone
Isopropanol
Methanol
Wood naptha
Diethyl phthalate
Ethyl acetate
TolueneCyclohexane

1-5% typical

Typical marker

Tertiary butyl alcohol

0.1%

Typical organoleptic modifier

Bitrex®

10 - 30ppm

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1.3

ETHANOL

Physico-Chemical properties
Table 2

Summary of physico-chemical properties

Property

Value

Physical state

LiquidMelting point-114oC

Boiling point

Comments/Reference
Corcoran (1953)o78.3 C

Ambrose (1970)oVapour pressure

57.3hPa at 19.6 C

Water solubility

Fully miscible

Partition coefficient noctanol/water (log value)
Henry’s law constantFlashpoint
Specific gravity

Ambrose (1970)
Howard (1990), Merck Index (1996)-0.31

Howard (1990)

0.000252 at 15oC

Kavanaugh (1999)

o

14 C

BP Chemicals (1997)o0.7864 at 20 C

Sakurai (1984)

There is no originally reported value available for the partition co-efficient. However, the text bookfrom which the above value is obtained is a frequently cited source for solvent data. This source
was deemed acceptable for the dossier submitted under the US HPV programme.

2

GENERAL INFORMATION ON EXPOSURE

2.1

Production Volumes

Ethanol is one of the most ancient chemical substances, arising from the fermentation processby the action of yeast on sugars. The process is most commonly known as that by which beeror wine is produced but it is also why leavened bread has a small ethanol content.
Total world production was around 25 000 kt in 2001 (CEFIC, 2003). All potable alcohol, anda large proportion of industrial and fuel ethanol, is made by the fermentation process in which
zymase, a yeast enzyme, changes simple sugars (e.g. as found in molasses) into ethanol and carbon
dioxide. Biomass (sugarbeet/cane, molasses, cereals, rice, grain, cellulosics) are used as
the feedstock for fermentation ethanol. Synthetic routes to ethanol also exist, and, althougha process based on acetaldehyde arising from acetylene is quoted, the virtually universal syntheticroute is via the hydration of ethylene to ethanol. The origin of the product, whether via the synthetic
or fermentation route, does not in any way affect its physical, chemical or toxicological properties,
although trace impurities can vary from route to route.
There are literally thousands of alcohol producers in Europe. These include those producing‘neutral’ ethanol, which is used in both industrial applications and in beverages such as gin
and vodka, and those producing spirit drinks containing ethanol (e.g. whisky and wine.) Figures
for production in the Europe Union in 2001 are shown in Table 3. Synthetic ethanol represents
about 30% of total production.

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OECD SIDSETHANOL
Table 3 Production of ethanol in European Union
Ethanol type

EU Production (ktpa)

Agricultural alcohol

790

Synthetic ethanol

500

Wine alcohol

230

Fuel alcohol*

180

TOTAL

1700

* subsidised agricultural ethanol designated for fuel use only
Source: CEFIC Ethyl Alcohol Group, 2003

Figures for the Czech Republic during the years 2000-2 are given in Table 4.
Table 4 Production of ethanol in the Czech Republic
2000

2001

2002

46.0 kt

41.1 kt

49.6 kt

Source: Sponsor country, 2004

US production is around 7 000 kt per year.
2.2

Use Pattern

Applications for ethanol industrially are split into solvent applications (both in industry andfor consumer applications), use as a fuel and further processing. In Europe, the non-potable market
is approximately 830 kt per year and has an approximate value of Euro 250 million (CEFIC, 2003).
The split of end uses is shown in table 5 and then described in more detail in the following subsections:
Table 5 Non potable end uses of ethanol in Europe
Non potable end uses(EU) ktpa

Solvent use

400

Intermediate manuf.

250

Fuel

180

TOTAL

830

Source: CEFIC, 2003. Data for 2001

Note that synthetic ethanol is used in non-potable applications only (non permitted by regulation).
Fermentation alcohols are used in both industrial and potable applications
2.2.1

Use as a solvent

The use of ethanol as a solvent in Europe, includes both domestic and industrial usage (table 6).
Industrially, ethanol is mainly used as a carrier solvent in inks (mainly flexographic) and coatings.
Consumer applications include the use of ethanol in professional cosmetic formulations such as hairsetting sprays and colorants, as well as in consumer cleaning and detergent preparations,
8

UNEP PUBLICATIONSOECD SIDSETHANOLfor example spray cleaners used in kitchens and bathrooms. Most perfumes consist of blendsof predominantly natural essences in an ethanol base. The essences themselves are often extracted
from flowers and barks using ethanol as the process solvent. Ethanol is also found in automotive
deicing products. Ethanol is also used in pharmaceutical and personal care products. Preparations
such as mouthwashes, and cough and cold medicines, are formulated with up to 30% ethanol.
Ethanol is also used as an active biocidal product.
Table 6 European solvent uses of ethanol (CEFIC, 2003)
Industrial

ktpa

Domestic

ktpa

PAINT (industrial)

25

COSMETICS

145

INKS

70

DETERGENTS

25

PHARMACEUTICALS*35

DEICING

40

FOOD*

5

CLEANING

15

FLAVOUR/FRAG5OTHER**

10

EXPLOSIVES

5OTHER**

20

TOTALS

165

235

GRAND TOTAL

400* use in manufacture (processing)** includes use as a biocide

The Scandinavian product register (SPIN, 2003) records the following use of ethanol in registeredpreparations (table 7):
Table 7. SPIN database information
CountryYear

Number of
preparations

Tonnage ofpreparations

FINLAND

2001

1075

86391

NORWAY

2001

808

24754

DENMARK

2001

2470

16587

NORWAY

2000

816

22782

SWEDEN

2000

2260

60551

DENMARK

2000

2149

228769FINLAND

2000

1054

No data

SWEDEN

1999

2265

51874

In consumer
preparations

X

X

X

Solvent use of ethanol in the US, including consumer solvent applications, is approximately
500 ktpa.

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2.2.2

ETHANOL

Use as an intermediate for the production of synthetic chemicals

As a reactive chemical, ethanol in common with all alcohols reacts with acids to produce esters.
Examples include ethyl acrylate, which is used as a reactive diluent in specialised coatings,
and ethyl acetate, which is a widely used solvent in paint and coating formulations. Ethanol is usedin the production of ethylamines, which in turn are reactive industrial chemicals usedin downstream speciality applications including agrochemicals and pharmaceuticals. It can also be
used to make ethoxypropanol, an increasingly used glycol ether solvent in coating formulations(CEFIC, 2003). This list should not be regarded as exhaustive.
350 ktpa of ethanol are used to produce downstream chemicals in the US. Ethyl acrylate production
represents the largest application at just under 30% of the volume (CEFIC, 2003).
2.2.3

Use of ethanol in fuelThe use of ethanol as a component of automotive fuel varies widely throughout the world.
In Europe, use is relatively limited. In the United States ethanol has a different profile. Becauseof the current statutory requirement to move to oxygenated gasoline fuels, ethanol is increasingly
used as a gasoline additive. Total use is around 6 000 ktpa (85% of total US demand). There are
more than 60 US producers, largely using fermentation processes. Total world consumptionof ethanol in fuel applications is around 15 000 ktpa (CEFIC, 2003).
2.2.4

Ethanol use in potable products

The dominant use of ethanol in consumer ‘products’ is in potable products (alcoholic beverages
and vinegar production – around 500 ktpa in Europe and around 900 ktpa in North America)(CEFIC, 2003). Consideration of such uses is outside the scope of this SIAR.
2.2.5

Marketing and distribution of product

Ethanol is distributed neat in closed systems such as marine vessels, tank cars (rail cars), and tanktrucks. Smaller volumes, particularly from distributors, are supplied in closed containers varying
from intermediate bulk containers (IBCs) to containers of 5 litres size. Products containing ethanol,
such as paints, are supplied in closed containers varying from intermediate bulk containers (IBCs)
to small cans and containers of 5 litres (1 gallon) or less (CEFIC, 2003).
2.3

Environmental Exposure and Fate

2.3.1

Sources of Environmental Exposure

Ethanol will enter the environment as emissions from its manufacture, use as a solvent and chemicalintermediate, and release in fermentation and alcoholic beverage preparation. Natural emissions
also occur as a plant volatile, from microbial degradation product of plant and animal wastes and innatural fermentation of carbohydrates (Howard, 1990). Concentrations of ethanol in some edible
products are quite high. (EU, 1996; Howard, 1990; Lerici, 1996; Anonymous, 1986). Releases tothe environment during production and industrial processing at larger production sites areminimized by the use of engineering controls and end-of-pipe abatement systems. Aqueous waste
streams are routinely treated in biodegradation facilities. Any organic wastes from manufacture are
typically incinerated on site or disposed of via specialist waste contractors (CEFIC, 2003). It is
possible that small, farm scale fermentation manufactures may not have such extensive emission
controls but by their nature, volumetric emissions will be low and dispersed.
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UNEP PUBLICATIONSOECD SIDSETHANOL

The largest source of ethanol release to the environment is expected to be from use of ethanol
containing products, including consumer products, where applications are open and engineering
controls to recover and recycle solvent are not always used. The most likely environmental medium
for ethanol release is the atmosphere although fugacity level 3 calculations (see section Error!
Reference source not found.)show that distribution between compartments means that both the air
and water compartments are significant regarding the fate of the substance.
There are no known regional specific factors that would make these potential sources of release
unique. Therefore they should be regarded as common worldwide. However, the relative quantitiesof ethanol release from the use as a fuel additive may vary between regions due to the great
variation in use in this application.
2.3.2

Photodegradation

Although ethanol can absorb radiation and is subject to direct photolysis, the principle mechanism
for degradation is likely to be photochemical oxidation in the presence of atmospheric pollutants(photochemical sensitizers) of which the main ones in industrial regions are nitrogen oxides (NOx)and sulfur oxides (SOx). Pseudo first-order half-lives of 15.4 hrs and 13.8 hrs were calculated
for nitrous oxide-mediated indirect photolysis (Yanagihara, 1997) and sulphur dioxide-mediatedindirect photolysis (Hustert, 1978). Ethanol is therefore expected to degrade rapidly in NOx and SOx
polluted atmospheres.
Using the EPA developed model AOPWIN (US EPA, 2002), secondary rate constants for hydroxyl
radical mediated atmospheric photo-oxidation were calculated to be 3.58x10-12 cm3/molecule-sec
for ethanol. Using the standard assumptions of 1.5x106 hydroxyl radicals per cubic centimetreand 12hr/day sunlight, a pseudo first order half-life of around 3.0 days was calculated based onan estimated rate constant. In the presence of hydroxyl radicals, ethanol photodegradation half-life
was 10 hours based on a measured rate constant (Campbell, 1976). These values are different
because of significant differences in the underlying rate constants. In the absence of atmosphericcontaminants and at light wavelengths typical of the troposphere (>290nm) ethanol was notdegraded in the presence of oxygen and water. Photochemical degradation is the rate-limiting step
governing the overall residence time of ethanol in air.
The photochemical tropospheric ozone creation potential (POCP) is 40-45% relative to ethylene
at 100% (Anderson-Skold, 2000; Derwent 1996). The Maximum Incremental Reactivity (MIR) had
an absolute value of 1.34 -1.69 g ozone/g volatile organic carbon (Carter, 2000). This indicates thatethanol would have a low to moderate contribute to tropospheric ozone creation.
2.3.3

Stability in Water

The octanol-water partition coefficient and Henry's Law value suggest that ethanol is unlikely
to bioaccumulate and will volatize from surface waters, off-gas from groundwater and have a highvapour phase retardation. Volatilization from model rivers and lakes were calculated using EPIWIN(US EPA, 2002). Calculated half-lives of volatilization of ethanol from a model river or lake were
3.3 and 38.9 days respectively using a measured Henry's Law constant of 0.000252. Ethanol isconsidered moderately volatile and is stable to hydrolysis (Anbar, 1967; Lyman, 1990.)
2.3.4

Transport between Environmental Compartments

Based on a value of partition coefficient Kow of –0.31, bioaccumulation of ethanol in aquaticorganisms is not expected. Level III distribution modelling showed that most of the ethanol released
to the environment would go primarily to air and water (see table 8), with the rest to the soil,
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OECD SIDSETHANOL

assuming emission ratios of substance to air, water, and soil are 1000:100:10 respectively (Mackay,1996). The ration of the factors were chosen by a weighted summation of emission factors for eachscenario, either using data from the EU Technical Guidance for Risk Assessment (EU, 2003) or byexpert judgement, and then rounding to the nearest order of magnitude with the air emissions set to
1000kg.
Table 8 Environmental distribution of ethanol
Fugacity level III calculations
Relative distributions between compartments based on anemission pattern of 1000:100:10
Air

57%

Water

34%

Soil

9%

Wet deposition (rain-out) is expected to play an appreciable role in the atmospheric removalof ethanol (Howard, 1990)
2.3.5

Biodegradation

In aerobic conditions using adapted wastewater from domestic sewage, degradation was 74% after
5 days rising to 95% by day 15 and in similar conditions in synthetic seawater, ethanol was 45%
degraded after 5 days rising to 75% by day 20 (Price, 1974). Biodegradation in a study to a MITI
protocol showed degradation of 89% after 14 days (>70% after 10 days, CERI, 2004) and >90%
within 10 days (Birch, 1991). Activated domestic sludges were capable of aerobically oxidizing
ethanol, as measured by BOD, which was 37.3% of maximum after 1 day (Gerhold, 1966).
The rate of biodegradation in anaerobic conditions was calculated to be 17.9 ppm ethanol per daywith a total methane recovery of 91% of the theoretical limit (Suflita, 1993).
Biodegradation is the main method of removal of ethanol from water. Ethanol is stable
to hydrolysis. Reaction with hydroxyl radicals in aquatic media will not likely be a significant
process (Anbar, 1969).
It can be concluded that ethanol meets the readily biodegradable criteria.
2.3.6

Bioaccumulation

The bioconcentration factor can be estimated using BCFWIN v2.15 (US EPA ,2002). The resultant
LogBCF=0.5 indicates that ethanol is not likely to bioaccumulate.
2.3.7

Other Information on Environmental Fate

2.3.7.1 Stability in soil
The soil adsorption co-efficient can be predicted using PCKOCWIN v1.66 (US EPA, 2002).
The resultant Koc of 1 indicates that ethanol released to soil would move quickly through the soil.

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2.3.7.2 Measured Environmental Concentrations
There is no recent environmental data on ethanol concentrations.
Ethanol and methanol were detected at Point Barrow, Alaska in 17 of 25 air samples at an average
combined concentration of 0.52 ppb over 24 hours (Cavanagh, 1969). Ethanol was found in groundwater suspected of leachate contamination at 190 ppb at one of 13 sites, and was detected at 58 ppb
in landfill ground water where inorganic levels indicated good or unknown water quality (Sabel,1983). Ethanol was also detected in surface water at a concentration of 4020 ppb in the Hayashida
River in Japan near the site of a leather factory (Yasuhara, 1981).
2.4

Human Exposure

The assessment of the substance is focused on its use as industrial chemical. Ethanol consumption
in alcoholic beverages is out of the scope of this report.
2.4.1

Occupational Exposure

Due to the volatile nature of ethanol, the most significant route of exposure is likely to be
by inhalation. Ethanol manufacturing plants are continuous, enclosed processes with controlled
occupational exposures. Potential exposures can occur during such operations as sample collection,
maintenance of equipment, and loading of trucks and/or rail cars.
Occupational exposure limits in the USA and the main European countries of are in the range 5001000 ppm (1900 mg/m³) over 8 hours (see table 9).
Table 9 Occupational exposure limits
COUNTRY

8hr TWA Hygiene LimitSTEL

US (OSHA)

1900 mg/m³ (1000ppm)

None

US (ACGIH)

1900 mg/m³ (1000ppm)

None

3

Germany (MAK)*

960 mg/m (500ppm)

Peak limit cat. II,1

UK (OES)

1920 mg/m³ (1000ppm)

none

3

Slovak Republic

960 mg/m (500ppm)

1920 mg/m3 (1000ppm)(30 min, 4x per shift)

Czech Republic

1000 mg/m³

3000 mg/m³

Products containing ethanol, such as inks, lacquers, are manufactured in semi-enclosed batchprocesses, usually with extraction systems to control worker exposure. Similarly, processes using
products containing ethanol are also usually semi-enclosed with extraction systems to control
worker exposure. These processes can be continuous or batch in operation.
There is limited data available on measured exposure values, partly due to the perception that actual
exposures are substantially lower than the relatively high OELs and also due to prioritisationof other more hazardous substances. The general data that is available (see appendix) confirms thatthe perception is probably correct in that for the industrial and occupational sectors for which data
have been reported, daily time weighted average ethanol exposures are typically well below
the prevailing regulatory exposure limits. Some detailed measurements from Slovakia confirm
that most industrial exposures are low. Some scenarios were identified with the potential for high

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inhalation exposure (eg pharmaceutical manufacture). Workers used PPE in these situations to limit
actual exposure (Regional Authority of Public Health, Slovakia, 2004).
There is also the possibility of dermal contact with ethanol. It is common practice to recommendthat workers when handling the material wear suitable protective gloves. The half life forthe evaporation of ethanol from skin is 11.7 seconds (Pendlington, 2001) which implies
that continuous immersion would be required for there to be any potential for dermal absorption.
2.4.2

Consumer Exposure

Ethanol is a natural component of many foods including beer, wines, distilled spirits, and a varietyof fruits. Ethanol is also used in small quantities as a diluent in some medicinal products. Ethanol isalso naturally present in a wide range of foods including fruits, yoghurt, ice cream, fruit juices
and leavened bread (Anonymous, 1986; Greubet, 1997; Lerici, 1996;).
Consumers are also widely exposed to ethanol as a component of alcoholic beverages
but discussion of this end use is beyond the scope of this review.
Non-food products containing ethanol include personal hygiene products, fragrances, cosmetics,
adhesives, surface coatings, and inks. For Customs and Excise purposes ethanol spirit intended
for non-beverage applications frequently contains an organoleptic modifiers, denaturants or marker
solvents to render it unpalatable and/or traceable. These are used at levels from ppm up to a few
percent. All routes of exposure (oral, dermal and inhalation) are feasible for these products as
a whole but not all routes apply to all products.
Ethanol is unusual in that it also occurs naturally within the body. This natural burden is thought
to be due to the metabolism of the intestinal microflora and produces blood alcohol concentration(BAC) levels of typically 0.062 to 0.73 mg/l (Sprung, 1981).
2.4.3

Indirect Exposure via the Environment

The measured environmental concentrations of ethanol are low, although the quantity of existingdata is limited. Available information suggests that the greatest source of exposure to ethanol may
be through their presence in alcoholic drinks, foodstuffs and medicines, which are regulated throughdifferent fora and are not considered further in this report.

3

HUMAN HEALTH HAZARDS

3.1

Effects on Human Health

3.1.1

Toxicokinetics, Metabolism and Distribution

Following any route of intake resulting in an elevated blood ethanol level (BEL), metabolism
proceeds in three basic steps. First, ethanol is oxidized within the cytosol of hepatocytes
to acetaldehyde; second, acetaldehyde is rapidly converted to acetate, mainly in the mitochondria;
and third, acetate produced in the liver is released into the blood and is oxidized by peripheral
tissues to acetic acid and ultimately carbon dioxide, and water. The rapid conversion of theintermediate aldehyde means that concentrations are usually very low. However, polymorphism isseen in the acetaldehyde dehydrogenase enzyme meaning that some ethnic groupings are less well
adapted to metabolise this intermediate with resultant higher concentrations, although they still
remain low. At a concentration of 10mM (460 mg/l) ethanol, acetaldehyde levels are <2 µM(88 µg/l) indicating that the human aldehyde dehydrogenase enzyme has a very low Michaelis
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Menton constant Km (Crabb, 1987), meaning that the reaction will proceed significantly at very low
substrate concentrations. The main pathway for ethanol metabolism proceeds via alcohol
dehydrogenase. However, other pathways for ethanol oxidation have been described including a
microsomal ethanol-oxidizing system located in the endoplasmic reticulum and a catalase system
located in the peroxisomes. The rate of hepatic metabolism of ethanol is concentration independent
except at very low or very high concentrations. Blood ethanol in humans decreases more rapidly atconcentrations over 300 mg/dl than at concentrations below this level, possibly due to oxidation by
the microsomal ethanol oxidizing system. The maximum rate of metabolism is 100 - 125 mg/kg
body weight/hour, although tolerant individuals may have higher metabolic rates (up to 175
mg/kg/hour) due to enzyme induction. Adults metabolize 7 - 10 g ethanol/hour reducing blood
ethanol concentrations at a rate of 15 - 20 mg/100 ml/hour. Ethanol is metabolized more rapidly inchronic alcohol abusers (up to 40mg/100 ml/hour) and in children (up to 28 mg/100 ml/hour)(Ellenhorn, 1988). The kidneys and lungs excrete only 5 – 10 % of an absorbed dose of ethanol
unchanged (Ellenhorn, 1988). The major route of excretion of ethanol is in the urine (Conibear,1988). Ethanol is excreted in the un-metabolized form in urine, exhaled air and sweat. Its metabolic
products are also excreted by exhalation and in the urine.
Studies in Animals
In vitro Studies
Ethanol penetration through pig's skin in vitro was greater in occluded cells than in non-occluded
cells (2.19 mg/cm2 and 0.10 mg/cm2 in 24 hours respectively). At the maximum flux underocclusion, the amount of ethanol penetrating from a 1m2 area of skin would give a blood alcohol
level of about 40 mg/l in a 70 kg man. In a comparative human use study, none of the blood
samples taken from sixteen human volunteers exhibited a detectable level of alcohol (Pendlington,2001). Ethanol has a very low octanol:water partition coefficient and this is seen as contributing
to the poor dermal uptake of ethanol in intact human skin. This study suggests that a systemic doseof ethanol is likely to be very low after the use of formulations delivering ethanol to the skin.
Studies in Humans
In a physiologically based pharmacokinetic (PBPK) model the parameters describing MichaelisMenten metabolism of ethanol in the liver were varied using simulation and optimization software.
For each exposure scenario the simulation was run for male workers in 'sitting awake' and 'light
exercise' activity levels. Despite some limitations a reasonably good overall description of the data
was obtained. The model predicted time courses of the mixed venous blood concentrations
for the 12 exposure scenarios. The model predictions are that for men exposed to ethanol, at 0.942
and 1.88 mg/L for 8 hr and for the lower breathing rate in men exposed to 9.42 mg/L, the liver isable to metabolize ethanol at the rate it enters the body. However, for the higher breathing rate
in men exposed to 9.42 mg/L and for men exposed to 37.6 or 63.6 mg/L the rate of ethanol delivery
via breathing exceeds metabolic capacity and ethanol blood levels consequently rise for the durationof the exposure. Men exposed to 20 mg/L ethanol for 4 hr also showed a continued accumulation
during exposure at the higher breathing rate but little or no accumulation at the lower breathing rate
(Conolly, 1999).
Other work has shown a similarly good correlation between inhalation exposure and blood alcohol
concentrations. A group of human volunteers (24 in one experiment, 16 in the second) were
exposed to ethanol vapour concentrations up to 3610 mg/m3 and resultant blood ethanol
concentrations (BEC) measured of between 0.00066 and 0.0056 mg/cm3. Regression analysisof the data shows that BEC = exposure (ppm) × 0.0029 (with a 7% error for 95% confidence).
(Seeber, 1994). Around 60% of inhaled ethanol vapour is absorbed (Lester, 1951; Kruhoffer, 1983).
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Ethanol is eliminated from the body mainly by metabolism in the liver and only minimallyby urinary excretion and pulmonary exhalation. Other tissues such as kidney, stomach
and intestines oxidize ethanol to a small extent. Once absorbed, alcohol is contained in the watercompartment of the body. It is not stored or accumulated to any degree, so that the body burden
at any point in time is a result of recent absorption, usually within the previous 12 hours (Conibear,1988). Such a rate of elimination is supported by the data from Jones and the PBPK modeling work
described above (Conolly, 1999; Jones 1993). In a study involving 130 patients that had abstained
from alcohol intake for 24 hours, physiological ethanol concentrations were below 0.75 mg/ml
with most values in the concentration range 0.1 and 0.2 mg/l (Sprung, 1981).
Conclusion
At exposures relevant to occupational and consumer exposure during manufacture and useof ethanol containing products (ie. Exposure via the inhalation route), the alcohol dehydrogenase
metabolic route in the liver dominates and does not become saturated. This mechanism follows firstorder kinetics. The first step of the metabolic path is the rate-determining step; concentrationsof the intermediate metabolite acetaldehyde are very low. Ethanol is not accumulated in the body.
Dermal uptake of ethanol is very low.
3.1.2

Acute Toxicity

Studies in Animals
Inhalation
In acute inhalation studies, ethanol has shown a low order of acute toxicity. An LC50 value was notachieved at exposures of up to 60,000 ppm for 60 minutes in study in CD-1 mice (Moser, 1985).
Mice in this study experienced moderate ataxia, which reversed after more than 4 hours recovery
period at all exposure levels.
DermalNo acute dermal toxicity was reported in a study in rabbits, LDL0=20,000 mg/kg (Monick, 1968)
and although this study is not experimentally robust, the result is consistent with the findingthat ethanol uptake through intact skin is poor. No other dermal study or reported result has been
identified.
Oral
Ethanol has a low order of toxicity in animals following single oral exposure. Robust figures are:
LD50=8300 mg/kg (oral, mouse) (Bartsch, 1976) and LD50=15010 mg/kg (oral, rat) (Youssef,
1992). An age-related difference is reported (Wiberg, 1970) in which young rats (100 days old)
were less sensitive than old rats (10-12 months old) with an LD50=11,000 mg/kg versus 7,000
mg/kg. The main symptoms of acute exposure are those typical of substances which cause central
nervous system depression e.g. inebriation, gait disturbance and dose-related decrease in response
to painful stimuli, respiratory depression and coma. Deaths were due to cardio-respiratory failure.
Other Routes of Exposure
By other routes, ethanol has, again, shown low acute toxicity with LD50 = 9,710 mg/kg in maleand LD50 = 9,450 mg/kg in female mice receiving i.p. ethanol (Schechter, 1995); LD50 = 9,200
mg/kg in male mice receiving i.p. ethanol (Ho, 1979) and LD50 = 6,710 mg/kg in young (100 daysold) rats and LD50 = 5,100 mg/kg in older (10-12 months) rats receiving i.p. ethanol (Wiberg, 1970).

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Studies in Humans
Oral
Oral consumption of ethanol containing beverages is known to produce symptoms of intoxication(e.g. drowsiness, loss of concentration). However, there is no evidence that such effects can beproduced by inhalation or dermal routes of exposure.
Conclusion
Ethanol has a low order of acute toxicity by all routes of exposure.
3.1.3

Irritation

Skin Irritation
Studies in Animals
There is little evidence of skin irritancy in animal studies. A study conducted to OECD 404
standards in rabbits showed ethanol to be not irritating (Jacobs, 1992) which agrees with an earlier
study (Phillips, 1972).
Studies in Humans
In the form of biocidally active surgical spirit (70-80% ethanol in water), there is a considerable
history of dermal application of ethanol as an antiseptic with no concern for skin irritancy.
Similarly, large amounts of ethanol are used in a variety of cosmetics, personal care and householdcleaning products.
Eye Irritation
Studies in Animals
Available data from animal studies indicates that ethanol is moderately irritating to the eye.
The most recent data indicates that, when assessed in an OECD 405 study, only mild redness
and chemosis remained in by day 7 with all symptoms having disappeared by day 14 (ECETOC,
1998). An older study similarly concluded that ethanol is moderately irritating (Jacobs, 1987).
Studies in Humans
In humans, direct contact of liquid ethanol on the human eye causes an immediate sensationof burning and stinging, accompanied by reflex closure of the eye. The acute discomfort subsides
rapidly, although foreign body type discomfort may be felt for a day or so. Recovery is complete.
Respiratory Tract Irritation
In humans, a concentration of 5000 ppm vapour is quoted as irritating and uncomfortable to breathe
but tolerable (Lester, 1951). Much higher concentrations than this would induce lachrymationand coughing.
Conclusion
Ethanol is moderately irritating to the eyes but not irritating to skin. At high vapour concentrations,in air, ethanol is irritating to breathe.

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3.1.4

ETHANOL

Sensitisation

Studies in Animals
Skin
Ethanol (75 % v/v) was used as solvent in the induction phase of a Magnusson and Kligman
sensitization test of a polyalkalene glycol. No skin reactions were evoked at challengewith the polyalkalene glycol in 75 % ethanol in either test or control group animals (BP Chemicals,
1984). No increase in ear thickness was recorded following challenge application of ethanol
in a mouse ear swelling test (Descotes, 1988).
Studies in Humans
Skin
A literature review demonstrated that ethanol can be an allergen in immediate and delayed
hypersensitivity by external or internal exposure and can produce subjective irritation, irritantcontact dermatitis and non-immunologic contact urticaria (Ophaswongse, 1994). However, thewidespread use of ethanol in cosmetics and in skin antiseptic formulations suggests that skinsensitization is not an end point of concern.
Conclusion
Ethanol is not considered to have sensitizing properties.
3.1.5

Repeated Dose Toxicity

There are many repeat dose studies in many species reported in the literature using ethanol.
However, these are almost universally carried out to improve the understanding of the risks
associated with the consumption of alcoholic beverages. Characteristically, these are carried out
by the oral route and at high doses, well in excess of 1 g/kg, which limits their value
in characterising the repeat dose toxicity of ethanol at doses relevant to occupational exposureand use of consumer products containing ethanol.
Studies in Animals
Oral
Studies (90 days duration) were carried out in rats and mice to assess whether 5% ethanol
in drinking water would be an appropriate vehicle for a long-term toxicity and carcinogenicity studyof urethane. Based on the water consumption data in the study and averaged body weights overthe exposure period, this equated to doses of at least 4000 mg/kg in rats and 7500 mg/kg in mice.
Data from this study yielded NOAEL values of >5%, for male rats (dose equivalent >4000 mg/kg)
and female mice and <5% for male mice and female rats (dose equivalent <5000 mg/kg). Male rats
showed minor changes to organ weights and haematology/biochemistry; female rats showed minor
biochemistry changes and increased length of oestrus cycle along with liver nodules; male mice
showed increased organ weights and some fatty changes to the liver and a decrease in sperm
concentration at (NTP, 1996). In the ensuing long-term study (2 years duration) in mice, ethanolcaused a marginal exposure-related increase in survival in males but had no effect on the survival
of females. There was evidence of an ethanol-induced reduction in water consumption that wasmore marked in males than in females (NTP, 2002). A good quality supporting study in rats gave
a NOAEL value of <5% , later refined to 2% (approximately 2400 mg/kg), for ethanol in oral feed
ad libitum in rats (Holmberg, 1986). The LOAEL in this study was 3 % (approximately
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3600 mg/kg) due to dose related hepatic yellowing and centrilobular steatosis. This appears to be
the lowest robust oral NOAEL available.
Conclusion
Most data available on ethanol is via the oral route of exposure. Much is at high doses which limits
its value to risk assessment of ethanol as a chemical substance. From the data available, it ispossible to surmise that ethanol is of repeat dose low toxicity by the oral route, with a lowest
reported NOAEL in animals of 2400 mg/kg for rats.
3.1.6

Mutagenicity

All discussion in this section is from in vitro data and in vivo data in animals.
In vitro Studies
Bacterial mutation assays
The results of bacterial mutagenicity assays (McCann, 1975; Blevins, 1982; Blevins, 1983, Hellmer
1992) have generally been negative for ethanol. A very weak positive effect of ethanol in a DNA
repair test in Escherichia coli, but no effect in the Ames test with strains Salmonella typhimurium
TA1535, 1537, 1538, 97, 98 and 100, was found by De Flora. (1984a). There was a weak
but reproducible positive effect in S. typhimurium TA102 but only at ethanol concentrations of 160
and 240 mg/plate, which are concentrations considerably greater than the highest concentration
specified for guideline testing (De Flora, 1984b). At least one positive result has been also reported
but only at concentrations massively above the maximum normally recommended in guideline
study protocols (Hayes 1985). Zeiger. (1992) found negative results at concentrations up to 10
mg/plate in S. typhimurium strains TA97, 98, 100, 104 and 1535 with or without Aroclor-induced
S9, from both rats and Syrian hamsters, at two concentrations. Ethanol is not therefore considered
to be mutagenic to E. coli and S. typhimurium bacteria.
Chromosome aberration tests
No chromosome aberrations were found in human lymphocyte cultures (Banduhn, 1985), orin human lymphoid cell lines at ethanol concentrations of 8 and 16 mg/ml (174 and 348 mM),
(Brown, 1992). Negative results have also been reported with Chinese hamster (CHO) cells (Lin,1989). Darroudi (1987) found chromosome aberrations induced by ethanol only in the presence
an extract of the leaves of maize with an NADPH-generating cofactor mix, metabolising system,a system that induced chromosome aberrations in CHO cells as well as increasing the numberof aberrations induced by 160 mM ethanol.
Many studies for this end point, including most of those above, are incomplete in design,
as currently recommended for screening purposes (OECD, 1997) and individually cannot be
regarded robust. In particular, they generally did not employ metabolic activation and, in somecases did not use a sufficiently wide dose range. However collectively and using a weightof evidence approach, there is little evidence that ethanol is clastogenic in vitro. In those cases
where positive responses were recorded, concentrations were extremely high and it is possiblethat chromosome damage resulted from non-specific effects such as high osmotic pressure
Cell mutation assays
In a study designed to test the conditions under which false positive results may be generated
by the mouse lymphoma assay, ethanol was one of 50 compounds tested (Wangenheim, 1988).
This study showed a small but statistically significant increase in mutants at 4.2 and 34 mg/ml
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without S9, and at 24 mg/ml with S9. However, in no case was the mutant frequency doubled and it
was concluded that a maximum concentration of 20 mM is adequate to detect genotoxins and thathigher concentrations may give false positive results. Ethanol is regarded negative in this assay. No
increase in mutants was found following a 4 hr exposure of S49 mouse lymphoma cells to ethanol
in the presence and absence of S9 (Friedrich, 1983). In the L5178Y mouse lymphoma assay,
ethanol was negative at 35.9 mg/ml, a concentration well above the maximum recommended
for this type of test (Amacher, 1980).
In vivo Studies
Micronucleus assays
When administered in drinking water to rats at 5% (about 4,000 mg/kg) for 10 to 30 days
(Balansky,. 1993) or at 10% and 20% (about 7,850 and 15,700 mg/kg) for 3 or 7 weeks (Tates,
1980), or to mice at up to 40% (about 31,400 mg/kg) for 27 days (Chaubey., 1977), ethanol had noeffect on micronucleus incidence in the bone marrow when administered in the drinking water.
In the study delivering 40% ethanol there was some ethanol-related mortality, which was clearly
above the maximum tolerated dose, recommended as the highest dose for micronucleus studies
in the OECD Guideline No. 474 (OECD, 1997).
Baraona (1981) fed rats for 6 weeks with diet containing ethanol calculated to contribute 36%of the calorific intake, a dosage of about 12 to 16 g/kg/day. The incidence of micronucleated bonemarrow erythrocytes was increased from 0.95 % to 1.30 % in polychromatic cells (PCEs) andfrom 0.67 % to 0.84 % in orthochromatic cells (OCEs), relative to pair-fed controls. The differencein PCEs was statistically significant, but only marginally (p<0.05), and was associated witha decrease in the number of nucleated cells and an increase in the proportion of the nucleated cells
that were in mitosis in the bone marrow. The difference in OCEs was not statistically significant.
An effect on the mitotic spindle may be responsible but the presence or absence of centromeresin the micronuclei induced by treatment was not investigated. Overall, there is no convincing
evidence that ethanol induces micronuclei in the bone marrow of rodents.
Chromosome aberration tests
No chromosome aberrations were found in the bone marrow or peripheral blood lymphocytesof male Wistar rats given 10 or 20% (about 7,850 and 15,700 mg/kg) ethanol in the drinking waterfor 3 or 6 weeks (Tates, 1980). Similarly, in Chinese hamsters, no aberrations were foundin the bone marrow after exposure to ethanol in the drinking water at 10% for 9 weeks (Korte,1979) or at 20% for 12 weeks (Korte, 1981), and no aberrations were observed in the lymphocytesof Chinese hamsters after exposure to 10% ethanol in the drinking water for 46 weeks (Korte,
1981a). Acute in vivo studies of chromosome aberration have not been identified. These studies
of high-dosage ethanol in chronic administration failed to demonstrate an effect attributable toethanol treatment. Therefore it is deemed unlikely that an acute study would show any effect either.
Dominant lethal assay
A large collaborative and robust study involving three laboratories, in which mice were exposedby intubation to the maximum tolerated dose of ethanol (0.64 g/kg) and to 0.16 g/kg and matedfor 8 weeks (James, 1982), failed to yield positive evidence of dominant lethalility for ethanol.
The protocol design used was comprehensive and compliant with OECD test methods.
A high level of dominant lethality has been reported in one series of studies in mice (Badr, 1975;
Badr, 1977). In one experiment, eight male mice, treated with ethanol (1.24 g/kg bodyweight)
by intubation on three consecutive days were mated sequentially to one female mouse per week.
An observed marked reduction in the mean litter size of the ethanol group, on the third week
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of mating only, was taken as evidence of post-implantation loss due to dominant lethal mutations
even though uterine contents were not examined. In another experiment in which mice were treatedwith either 1.24 or 1.86 g ethanol/kg, and the uterine contents examined on gestation day 13 to 15,
there were significant increases in the frequency of dead implants and of the dominant lethal index
at the second and third mating time-points on days 4 to 8 and 9 to 13. This result fails to correlatewith the previous experiment in which the effect was seen only at the 14 to 17-day time-point.
Because of the numerous errors and inconsistencies in the studies, they were assessed as invalid
and not reliable.
In a study deliberately designed to reproduce the effects reported by Badr (1975 & 1977), but usingi.p. injection rather than intubation, Rao (1994) provided evidence that ethanol did not havea significant dominant lethal effect. Some pre-implantation loss observed in this study was thought
to be due to an effect on the fertilization capacity of sperm. The effects of ethanol (approximately
1.26 g/kg/day for 3 days) on the outcome of mating on days 1 to 4, 5 to 8 and 9 to 12 were
investigated further in high numbers Swiss mice. There were markedly fewer pregnant females (by
34% and 30%) at the first two mating times in the treated group and a significant decrease in total
and live implants in the second mating. There was no increase in dead implants from the first two
mating occasions and only a small but statistically significant increase at the third mating. No
dominant lethal effect was found in two other experiments using CBA and C57BL6 micechronically exposed to ethanol in the drinking water.
No dominant lethal effect was found when ethanol was administered to mice included in a liquid
diet as 20% (14 to 17 g/kg) or 30% (24 to 30 g/kg) of dietary calories for 4 weeks (Randall, 1982).
Mean blood levels were 57 mg/dl and 80 mg/dl respectively. In a similar study using ethanol at 28%
of dietary calories (22 to 25.5 g/kg/day), 5-week exposure of mice decreased testicular weight,
reduced fertility and increased pre-implantation losses, foetal mortality and mutation index(Berryman, 1992).
In rats, 15 days of exposure to ethanol, increasing to 58% of dietary calories (estimated ethanol
intake 7.2 to 14.4 g/kg/day), resulted in an increase in early abortions, considered to have bea possible dominant lethal effect (Klassen, 1976). However, only six pregnancies were examinedin the ethanol treatment group and the males had been treated chronically with ethanol such that
quality of the study was impaired. Increases in the frequency of dead implants were also foundwhen male rats were treated with 20% ethanol in drinking water for 60 days prior to mating(Mankes, 1982). Histological examination revealed significant pathological changes in the testesof potential impact on litters. In contrast, Chauhan (1980) found no effect of exposure of rats via
the drinking water (30% ethanol) for 5 weeks. These were studies of small group sizes (10 or less)and of low power for interpretation.
Many studies can be criticised on the grounds of inadequate numbers of animals or on the methods
used to score or evaluate the incidence of early or late foetal deaths or distinguish between earlyand late deaths. Also, the very high ethanol doses used make interpretation of the effects difficult.
The most satisfactory test is the inter-laboratory study performed to OECD guidelines (James,
1982), a study that gave a negative result in mice from which it is concluded that ethanol is negativein the dominant lethal assay in male mice.
Conclusion
Interpretation of the available data on the genotoxicity of ethanol is confounded by experimental
inadequacies. Negative results from a number of bacterial mutation assays appears to be reliable
but studies of chromosome aberration can be criticised for not including an exogenous metabolic
activation system. Of the mammalian cell mutation assays a weak mutagenic effect in L5178Y
mouse lymphoma cells occurred only at very high ethanol concentrations and, when possible
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artefacts are considered, the results of this study, too, can be regarded as negative. In vivo tests
for chromosome aberrations in both rats and Chinese hamsters have given negative results.
The results of the micronucleus and dominant lethal assays are variable and negative only in morerobust studies and the overall weight of evidence favours the conclusion that ethanol does not
induce dominant lethality in assays using standard regulatory methodologies; interpretation of thesestudies is confounded by the effects of ethanol on the fertilizing capacity of sperm. There is
therefore very little evidence to suggest that ethanol is genotoxic in somatic cells and it may havea very limited capacity to induce genetic changes in vivo but under very specific circumstances
and at very high doses achievable in humans only by deliberate oral ingestion (Phillips, 2001).
3.1.7

Carcinogenicity

Many carcinogenicity studies in laboratory animals exist using ethanol as a test substance.
However, these are almost universally carried out to improve the understanding of the risks
associated with the consumption of alcoholic beverages. Characteristically, these are carried out
by the oral route and at high doses, well in excess of 1g/kg, which means they are inadequately
designed and provide too little data to characterise the carcinogenic potential of ethanol at doses
relevant to occupational exposure and use of consumer products containing the substance.
In vivo Studies in Animals
Oral
A recent National Toxicology Program study conducted to GLP standards exposed mice to ethanol
in drinking water at the relatively high doses of 2.5% and 5% for 2 years. It showed only equivocal
evidence of carcinogenic activity of in males based on increased incidences of hepatocellularneoplasms. However, there was no evidence of carcinogenic activity of ethanol in female mice
exposed to either concentration (NTP, 2002). There remains no robust evidence of carcinogenicity
in laboratory animals at doses other than those associated with consumption of alcoholic beverages.
This is consistent with the findings of IARC who concluded that there is inadequate evidence
for the carcinogenicity of ethanol in experimental animals (IARC, 1988)
Studies in Humans
Although no epidemiological studies are available for ethanol per se, there are a large numberof studies (retrospective cohort, prospective cohort and case-control studies) on the effectsof alcoholic beverages, which contain ethanol and water as the two main components. These
epidemiological studies clearly indicate that drinking alcoholic beverages is causally related tocancers of the oral cavity, pharynx (excluding the nasopharynx), larynx and oesophagus. The effectappears to be independent of beverage type. The aetiology is likely to proceed via a mechanism
whereby frequent exposure to high local concentrations of liquid ethanol and its metabolites leads topersistent irritation, and eventually hyperplasia and finally tumor formation. (Greim H, 1999).
For the oral cavity, pharynx, larynx and oesophagus, alcoholic beverage consumption in excess
of 10-40 g ethanol per day is necessary before there is a convincing increase in the relative riskof cancer (Greim H, 1999; UK Dept of Health, 1995). Such a mechanism would not therefore be
relevant to occupational exposure. Drinking alcoholic beverages is also likely to be causally linked
to liver cancer. The liver is the primary site of metabolism and sees high concentrations of ethanol
and its metabolites. Tumor formation is normally associated with cirrhosis, which is in turnnormally seen only following chronic alcohol abuse in excess of 80g ethanol per day (Greim H,
1999; UK Dept of Health, 1995). Such a scenario is not relevant to occupational exposure
to ethanol. Drinking alcoholic beverages is also possibly causally linked to cancer of the breastand large bowel. Should the causal link with breast cancer be proven, the mechanism is believed
to be via disturbance of the hormonal system. There is little or no indication of a causal relation
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with cancer of the stomach, pancreas, lung, urinary bladder, kidney, ovary, prostate, lymphaticor haematopoietic system. There is no convincing evidence that the carcinogenic effectsof alcoholic beverages in humans occurs as a result of the mutagenic effect of ethanol, acetaldehyde
or other beverage constituents (UK dept of Health, 1995). The International Agency for Research
on Cancer has classified alcoholic beverages (but not ethanol) as Group 1 - carcinogenic to humans(IARC, 1988).
The International Life Sciences Institute (ILSI, 1999) has published an extensive reviewof the health issues relating to alcohol consumption. They concluded that ethanol is nota carcinogen by standard laboratory tests. Such tests are the normal measure for the assessmentof industrial chemicals and any chemical that would be expected to present a carcinogenic hazard toworkers or consumers during normal handling and use would be expected to show a positive resultin one or more of such tests.
Conclusion
Taking into account the known information on uptake of ethanol by the inhalation and dermalroutes and the lack of genotoxicity of ethanol, it can be concluded with some confidence thatoccupational exposure to ethanol and the use of ethanol in consumer products does not pose a
carcinogenic hazard.
3.1.8

Toxicity for Reproduction

For a novel chemical produced in the quantity in which ethanol is manufactured, there would be
requirements for testing by a relevant route of exposure according to OECD guidelines.
Comparatively few published studies are useful for quantitative risk assessment and few precisely
meet all the requirements of the specified regulatory guideline. Importantly there is only a small
number that have used multiple dose levels in order to clearly demonstrate a lowest observed
adverse effect level (LOAEL) and a no observed adverse effect level (NOAEL). Many do not meetthe criteria for robustness required for inclusion but are nevertheless important for interpretative
purposes and a weight of evidence approach.
Studies in Animals
Effects on Fertility
In laboratory animals there is a paucity of information on potential fertility effects in females.
In one robust 2-generation study in mice, ethanol in drinking water at concentrations up to 15%(equivalent to 20.7 g/kg/day) had no demonstrable effect on fertility (George, 1985). Similarly,
fertility over 7 weeks of treatment with ethanol at 10% or 25% of dietary calorie intake was notaffected (Abel, 1989).
Few studies in males gave information on systemic ethanol exposure, which makes it difficult
to estimate threshold levels for effects and few gave a range of exposures from which to estimate
dose responses and ‘no effect’ levels. However, in one comprehensive study there was no effecton fertility in a group of 20 male rats given 3 g or 2 g/kg ethanol by oral intubation daily for nine
weeks, achieving blood ethanol concentrations of 3380±150 and 1320±50 mg/l, respectively, (Abel,
1993). Although fertility was unaffected, this study did reveal higher incidences of runted pups
in the resulting offspring at the highest exposure level (3g/kg). In another study (Abel, 1995), evenwith daily oral dosing of ethanol at 5 g/kg, there was no effect on male fertility.
In male rats exposed to ethanol by inhalation for 7 hours /day for 6 weeks in a combined fertility
and developmental toxicity study there was no effect on fertility at 16,000 ppm (Nelson, 1985a,
Nelson 1985b, Nelson 1988). All 19 males at this dosage successfully initiated a pregnancy.
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The route of exposure would have resulted in the measured BEL (approximately 500 mg/l) being
steadily maintained throughout the exposures.
An adverse effect on fertility was noted in male rats with administration of ethanol in the diet (10%ethanol derived calories) for 15 days prior to and throughout the mating period (Klassen, 1976).
Only six pregnancies were initiated when the six exposed males were each paired with two
untreated females. However, this study was confounded by general toxicity manifest as ataxia,
lethargy and weight loss during the study period.
Reductions in ovary weight and reductions in oestradiol and progesterone in female rats receiving
liquid diets containing 5% ethanol (36% EDC) for 49 days during the peri-pubertal period havebeen demonstrated (van Thiel, 1976). The reported BEL was relatively low (1100±90 mg/l) but thetiming of the sample (taken 09.00 – 11.00 hours) was probably inappropriate to detect the peaklikely at the usual time of feeding during the previous evening. Irregular cycles and longer oestrous
cycles were noted in rats fed liquid diets containing 5% ethanol (36 % EDC) for 16 weeks but not
after 8 weeks with 8 weeks recovery period (Krueger, 1982). Again, ovarian function was
suppressed in rats that achieved blood alcohol levels of 2500 mg/l (Bo, 1982).
Few studies have given information on systemic exposure to ethanol, so it is difficult to determine
the threshold for any adverse effects. One study demonstrated reduction in testis and epididymis
weights related to effects on spermatogenesis in pubertal male mice given 5% ethanol containing
liquid diets that achieved a BEL of 1600 mg/l (Anderson, 1985). Lower dose levels were notinvestigated, however, virtually all changes observed were found to be reversible.
Male rats exposed to ethanol vapour concentrations of 22, 23, 25 and 27mg/l (11,500 to
14,000ppm) continuously for 3-4 weeks achieved BELs of 94-187 mg/100ml (Rivier, 1983). BELs
of ≥163 mg/mL were associated with inhibition of androgen secretion, but only in those animals
that failed to grow. These results suggest that there is a threshold for adverse effects of around 130
mg/100ml (equivalent to NOAEC inhalation exposure of 23mg/l) by this relevant route of exposure.
Developmental Toxicity
Many of the published studies in laboratory animals have investigated the effects of high dose oral
ethanol intake. High-dose studies are possible because of the low acute toxicity of ethanol,
however, the use of high doses can cause difficulties when interpreting ethanol reprotoxicity data
within the regulatory hazard assessment framework, because doses are often in excess of the
maximum (1 g.kg-1.day-1) recommended in current chemical testing guidelines (e.g OECD 414,
416). Many rodent experimental studies are conducted using a 5% ethanol liquid diet, which
provides 35-36% ethanol-derived calories. The achieved ethanol intake for a pregnant rat with thisdiet is approximately 10-12 g.kg-1.day-1 (i.e. >10 times the limit dose of a standard OECD 414
developmental toxicity study). A reduction in nutrient intake during a critical period of gestation
would reduce foetal/pup weights and cause other postnatal effects.
Assessment of ethanol by inhalation exposure
Most relevant for occupational exposure hazard assessment are a series of inhalation developmental
toxicity studies of ethanol conducted by Nelson et al., (1985a, 1985b, 1988). In the first of thesestudies, the potential teratogenesis of inhalation exposure to ethanol was assessed conventionally(Nelson et al., 1985a) as part of a more extensive study looking at a number of different alcohols,
conducted by the US National Institute of Occupational Safety and Hygiene (NIOSH). Groups of 15
or 16 mated female Sprague-Dawley rats were exposed 7 hours/day throughout gestation (GD1-19,
based on the presence of sperm on GD0) to ethanol concentrations of 0, 10000, 16000
or 20000 ppm. These resulted in average BELs of 0, 27, 420 and 1480 mg/l. Ethanol elicited severematernal toxicity at 20 000 ppm but at the lower exposure levels, dams appeared hyperactive
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after exposures. The authors reported male (but not female) foetal weights to be depressed at
the 16 000 and 20 000ppm exposures, but the differences were small and not significant. There
were also no significant differences in the incidences of external, visceral or skeletal malformations
or variations. These results did not indicate teratogenicity at dose limiting maternal exposures.
Following on from the conventional teratogenesis study, groups of 15 mated female SpragueDawley rats were exposed similarly throughout gestation, but allowed to litter to assess potential
behavioural effects in the offspring (Nelson, 1985b). Ethanol concentrations of 0, 10000
or 16000 ppm were used. There were no differences from controls in maternal weight gain or feed
and water intake. Litter size and birth weights were unaffected, even at 16000 ppm, which would
have been expected had the marginal differences in foetal weight at caesarean evaluationin the other study been a true treatment effect. Offspring survival and growth were unaffected
and there were no postnatal ‘behavioural’ effects at the specified maternal exposures, establishinga developmental toxicity NOAEL at the highest dose tested (16000 ppm).
The results of inhalation studies showed that there was no indication of teratogenicity at dose
limiting, maternally toxic concentrations. Apart from small non-significant differences in foetal
weight of one sex, which were not apparent on pup birth weight, and small inconsistent differencesin some aspects of neurochemical analysis, there was no evidence of developmental toxicity
at 16 000 ppm, with an average steady state BEL of 420 mg/l.
Assessment of ethanol by oral exposure
In a robust study in mice, litter weight was not affected by ethanol-containing dietsbut malformations were significantly increased by maternal diets containing 25% or moreof ethanol-derived calories. Grossly visible abnormalities, external, soft tissue and skeletal
abnormalities affected the limb, eye, brain, heart, urinogenital tract and abdomen of foetuses(Randall, 1979). In rats treated with ethanol by gavage (12.5% v/v in distilled water daily
throughout gestation and gestation plus lactation, learning was impaired in rats of both genderat 9 weeks relative to controls (Vaglenova, 1998). This remained evident in males, but not females,
at 5 months. In offspring treated both pre- and post-natally with ethanol, 60% were poor learners
compared with 33% in sucrose controls. Foetal weights were depressed and skeletal abnormalities
occurred at 100% incidence in two strains of mice treated with ethanol at rates ranging 15 to 30%
of calorie intake with a LOAEL of 15 % (Chernoff, 1977). These effects were primarilyof the occipital bone but also affected the sternum and ribs. Visceral abnormalities affected 36 %,100 % and 100 % of foetuses in the 3 ethanol treated groups. Dilated brain ventricles were the mostfrequent anomaly but open eyelids, exencephaly, gastroschisis and heart defects also occurredin the higher dose groups. in mice (Wier, 1987) is discussed further below.
‘Effect’ and ‘no effect’ levels for teratogenicity of orally administered ethanol were establishedin mice by Wier (1987) using an abbreviated evaluation of uterine contents at term (teratology
probe) and a ‘limited’ postnatal study in which offspring were examined and weighed throughto weaning on PND22. Ethanol dosages of 2.2, 3.6, 5.0, 6.4 and 7.8g.kg-1 were administered daily
by gavage on GD8-14. Post-implantation losses were elicited at 5g.kg-1 and higher. In the postnatal
component of the study, there were no significant effects on pup growth or survival at maternally
toxic dose levels. A NOEL was established at 3.6g.kg-1 along with an embryotoxic LOAEL
of 6400 mg/kg. However, the limited endpoints employed and the lack of BEL information reduced
the value of this work for hazard assessment.
An important consideration in the interpretation of all oral developmental toxicity studies of ethanol
is that pregnant animals exposed to alcohol consume less food than ad libitum non-alcohol subjects,
whether the ethanol is in their drinking water, in a liquid diet or intubated. This meansthat malnutrition may be a confounding variable in such studies.
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A series of older teratogenicity studies of ethanol are included since they followed relatively‘conventional’ designs (Schwetz, 1978). Pregnant CF-1 mice, Sprague-Dawley rats and New
Zealand white rabbits were given 15% ethanol in their drinking water during the period of majororganogenesis, (GD6-15 for mice and rats: GD6-18 for rabbits). BELs (measured in satellite groups
of non-pregnant animals) were about 2000mg/l in mice and 250-500 mg/l in rats and rabbits.
The values for rats and rabbits are considerably lower than expected from other studies
administering such high ethanol concentrations and presumably are related to the observed and noncontrolled decreased liquid intake and decreased maternal body weight in all species. Some skeletal
variants in these studies were probably due to retarded fetal growth consequent to indirect maternal
toxicity.
Conclusion
In extremis, ethanol can elicit adverse effects on the reproductive system and on fertility andfecundability in males and females and can trigger developmental toxicity in females. However, it isclear that this occurs at BELs that can only be achieved by the deliberate oral consumptionof alcoholic beverages. In addition, the role of possible confounding factors is unclear. It can beconcluded that blood ethanol concentrations resulting from ethanol exposure at doses relevant tooccupational exposure and the use of consumer products containing ethanol are unlikely to produce
reproductive or developmental toxic effects (Irvine, 2003).
3.2

Initial Assessment for Human Health

Human health hazards from exposure to ethanol are well known. However these are only evident
at the very high doses associated with the consumption of alcoholic beverages. In the context
of potential exposures resulting from occupational or consumer use of ethanol containing products(i.e. excluding use in beverages), ethanol appears to present a low human health hazard.

4

HAZARDS TO THE ENVIRONMENT

4.1

Aquatic Effects

Valid acute aquatic toxicity data are available for fish, invertebrates, algae, and microorganisms.
Acute Toxicity Test Results
Toxicity to fish
Robust static (Johnson, 1980) and flow-through (Majewski, 1978) studies on freshwater fish gave
LC50 values greater than 1100 mg/l. Values are shown in table 10. A robust static limit test showeda value >100mg/l (Ewell, 1986). A high reliable flow-through study with fathead minnows,
conducted using EPA methodology, gave LC50 values at 1, 24, 48, 72 and 96 hours of 13,480 mg/l
or greater than 18,000 mg/l (Mattson, 1976). There is good agreement between values for staticand flow-through studies despite the fact that static tests are not optimal for studying a relatively
volatile and readily biodegradable compound such as ethanol.

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Table 10 Toxicity to fish
Species

Test period (hr)

LC50 (mg/l)

Salmo gairdneri

96

13,000

Salmo gairdneri

96

11,200Pimephales promelas

96

>100

Pimephales promelas

96

14,200Pimephales promelas

96

13,480

Toxicity to invertebrates
48-hour studies with the pelagic invertebrates Daphnia magna and Cerodaphnia sp. were conductedusing method ASTM Standard E729-80 and gave reliable LC50 values of 12,340 mg/l and5012 mg/l respectively (Takahashi, 1987). In similar studies a range of 24-h EC50 values in excess
of 1000 mg/L have been reported (Bowman,1981; Calleja, 1994). A 24-h LC50 with the marine
invertebrate Artemia salina was 1833 mg/L (Barahona-Gomariz, 1994). See table 11 for all robust
values available.
Table 11 Toxicity to invertebrates
Species

Test and time
periodValue (mg/l)

Ceriodaphnia

LC50 (48hr)

5,012Daphnia magna

LC50 (48hr)

12,340Artemia salina

LC50 (24hr)

1,833

Paramecium caudatum

LC50 (4hr)

5,980

Palaemonetes kadiakensis

EC50 (18hr)

1,000

Daphnia pulex

EC50 (18hr)

2,000

Hyallela azteca

EC50 (18hr)

1,000

Artemia salina

EC50 (24hr)

23,874

Toxicity to aquatic plants
Two robust studies of 96-hr duration each give NOEC values of <500 mg/l for the growth rate end
point in Chlorella vulgaris and Selenastrum capricornutum (El Jay, 1996). Corresponding EC50
values were 1,000 mg/l and 10,000 mg/l. Another study conducted to a high standard
in Chlamydomonas eugametos yielded a 48-hr EC50 of 2000mg/l (Hess, 1980) and a study on
Chlorella pyrenoidosa and a variety of algal species all gave EC50 values greater than 1,000 mg/l
(Cowgill, 1989; Hess, 1980; Stratton, 1987; Stratton, 1988). One study of 5 days durationin Skeletonema costatum gave a NOEC in the range 3,240 to 5,400 mg/l based on cell countand corresponding EC50 values of 10,943-11,619mg/l). The authors (Cowgill, 1989) remarkthat using EPA criteria, ethanol can be judged non-toxic by this test and that ethanol was used asa carbon source stimulating growth of the alga before inhibition began.
All results are shown in table 12.

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Table 12 Toxicity to aquatic plants
Species

Test period (days)

EC50 (mg/l)

Chlorella vulgaris

4

1,000

Lemna gibba

7

4,432

Lemna minor

7

3,690

Selenastrum capricornatum

4

10,000Chlamydomonas eugametos

2

2,000

Skeletonema costatum

4

10,943-11,619

Chlorella pyrenoidosa

10

1,180

Chronic Toxicity Test Results
Several chronic exposure studies have been conducted for periods ranging 4 to 21 days in a varietyof freshwater and marine invertebrates. They consistently yield LC50 values in excess of 100 mg/l
(Cowgill, 1991a; Rayburn, 1997). The lowest reported NOEC is 9.6mg/l for Cerodaphnia sp(Cowgill, 1991a).
Studies of 7 days exposure to ethanol in the higher (vascular) plants Lemna gibba and L. minor,
conducted to EPA OTS 797.1160, gave EC50 values of 4432 mg/l respectively. Equivalent NOEC
values were 280 and 778 mg/l respectively (Cowgill, 1991).
Toxicity to Microorganisms
Only one study has been found giving quantitative data for the bactericidal activity of ethanol.
A study using Pseudomonas putida showed a 16-hour toxicity threshold of 6500 mg/L (Bringmann,
1980).
4.2

Terrestrial Effects

Studies on root growth in onions (Fiskesjo, 1985), germination of lettuce seeds (Reynolds, 1977)
and coleoptile growth and respiration rate in maize (Nashed, 1958) have demonstrated inhibitory
effects at ethanol concentrations in excess of 3000 mg/l. Studies of respiration in potato tuber tissue(Miller, 1935; Rychter, 1979) produced effects of both stimulation and inhibition of respiration at
low concentrations and the toxicological significance of these findings is doubtful. The same is said
for several studies in which plant growth was stimulated, perhaps with beneficial application, forexample in oats (Mer, 1958), girasole (Reichhart, 1979), sugar cane (Clements, 1940) and potato(Guthrie, 1931). The toxicological relevance of these results is uncertain.
In the only study of the effect of ethanol on soil dwelling organisms found, the oligochaete worm
Eisenia foetida showed a 48 hr LC50 of 0.1-1.0 mg/cm2 loading on a filter paper test (Roberts,
1984). The 'dosage', presented in terms of concentration in a given area of substrate is difficult
to interpret but would have equated to the worms being exposed to a theoretical vapour
concentration in the range 200-2000 mg/l.
4.3

Initial Assessment for the Environment

Ethanol is expected to partition to the air and water compartments. It is readily biodegradable, is notexpected to bioaccumulate. Ethanol is predicted to degrade rapidly in atmospheres where NOX
or SOX are present. Valid acute aquatic toxicity data are available for fish, invertebrates, algae,
28

UNEP PUBLICATIONSOECD SIDSETHANOLand microorganisms. The most sensitive species were algae Chlorella vulgaris with an EC50
of 1000 mg/l and the invertebrate Artemia Salina with an LC50 of 1833 mg/l. Chronic data isavailable for invertebrates and algae. In the long term tests the most sensitive species was clearly
the invertebrate Ceriodaphnia with a NOAEC of 9.6mg/l. The data available on terrestrial species isdifficult to extrapolate to a the context of a hazard assessment but does suggest that ethanol isof low toxicity.

5

RECOMMENDATIONS

It is recommended that ethanol be considered as low priority for further work because of very low
toxicity to humans and the environment at conceivable exposures likely to result from the
manufacture and use of ethanol and ethanol containing products.

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6

ETHANOL

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Lerici, C.R., Nicoli, M.C. (1996). Chemical and physico-chemical properties affecting the quality
and stability of bakery products. Adv. Food Sci 18(5/6):229-233.
Lester D. & Greenberg L.A. (1951). The Inhalation of Ethyl Alcohol by Man. Q. J Stud Alcohol
12:167-178.
Lin, Y-C., Ho, I-C., Lee, T-C. (1989). Ethanol and acetaldehyde potentiate the clastogenicity ofultraviolet light, methane methanesulfonate, mitomycin C and bleomycin in Chinese hamster ovary
cells. Mutat. Res. 216:93-99.
Lyman, W., Reehl, W., Rosenblatt, D. (1990) Handbook of Chemical Property Estimation Methods:
Environmental Behaviour of Organic Compounds. American Chemical Society, Washington, DC.
34

UNEP PUBLICATIONSOECD SIDSETHANOL

Majewski, H., Klaverkamp, J. and Scott, D (1978) Acute mortality and sub-lethal effects of acetone,
ethanol and propylene glycol on the cardiovascular and respiratory systems of rainbow trout (Salmo
gairdneri) Water Res. 13:217-221.
Mankes, R.F., LeFevre, R., Benitz, K.-F., Rosenblum, I. et al. (1982). Paternal effects of ethanol in
the Long Evans rat. J. Toxicol. Envir. Hlth 10:871-878.
Mattson, V., Arthur, J. and Wallbridge, C. (1976). Acute toxicity of selected organic compounds toFathead Minnows. U.S. EPA Environmental Reasearch Lab., Duluth, Minnesota, EPA 600/3-76097.
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in the Salmonella/microsome terst: Assay of 300 chemicals. Proc Nat Acad Sci 1975;72(12):51355139.
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Mer, C.L. (1958) Growth-Promoting Effect of Ethanol on Oat Seedlings Nature; 182:1812-1813.
Merck (1996) The Merck Index 12th edition, Merck & Co., Inc., Rahway, New Jersey.
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the Respiration of Potato tubers. Contr Boy T;7:1-17
Moen, B.E. and Hollund, B.E. (2000). Exposure to organic solvents among car painters in Bergen,
Norway. Ann Occup Hyg 44(3):185-189.
Monick, J.A. (1968) in Alcohols. Their Chemistry Properties and Manufacture. Reinhold 1969,
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halothane and ethanol in mice: effects of exposure duration. Toxicol Appl. Pharmacol 77:285-291.
Muiswinkel, W-J van, Kromhout, H., Onos, T. and Kersemaekers, W. (1997). Monitoring andmodelling of exposure to ethanol in hairdressing salons. Ann Occup Hyg 41(2):235-247.
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UNEP PUBLICATIONS

35

OECD SIDSETHANOL

Nelson B, Brightwell W, MacKenzie-Taylor D, Burg J & Massari V. (1988). Neurochemical, but
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Rivier C & Vale W. (1983) Influence of ethanol on reproductive functions of the adult male rat as afunction of body weight. Alcohol Clin. Exp. Res. 7(2):210-212.
Roberts, B.L., H.W. Dorough (1984). Relative Toxicities of Chemicals to the Earthworm Eisenia
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Sabel GV, Clark TP (1983) Volatile Organic Compounds as indicators of municipal solid wasteleachate contamination. Ann Madison Conf Appl Res Pract Munic Ind Waste, p108.
36

UNEP PUBLICATIONSOECD SIDSETHANOL

Sakurai M, Nakagawa T (1984), Densities of dilute solutions of water in n-alkanols at 278.15,
288.15, 298.15, 308.15 and 318.15K. Partial molar volumes of water in n-alkanols. J Chem
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combination in mice: implications for cocaethylene formation. Pharmacol Biochem Behav
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UNEP PUBLICATIONS

37

OECD SIDSETHANOL

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women in a 1981 national survey. Alcohol. Clin. Exp. Res. 8(5):451-458
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tests: V. Results from the testing of 311 chemicals. Environ Mol Mut 19;2-141

38

UNEP PUBLICATIONSOECD SIDSETHANOL

ANNEX
Summary of general occupational exposure values
Operation andlocation

NumberofSamples

Averagemeasuremen
t

Range

Comments

Reference

Hairdressing
salons,Netherlands

1955.7ppm0.05 – 30 ppm

28 salons in Wageningen andRotterdam. 114 people sampledover two seasonsMuiswinkel, 1997
(Ann Occ Hyg 41(2)Hairdressing
salons,Norway10

10ppm

2.1 – 19 ppm

6 salons in Bergen

Hollund, 1998 (AnnOcc Hyg 42(4)Liquid ink
manufacture,
UK

3

30ppm

18 – 51 ppm

Open top manufacture typical ofhighest exposures likely to be
seen

Coates Inks internaldata, 2000

Flexographic
printing, UK

6

52ppm

10 - 110 ppm

Greatest exposure in
mixing/application areas

BP Amoco Chemicals,
Darton, 1999

Flexographic
printing, UK

15

69ppm

19 – 177 ppm

Greatest exposure in washoff
area.

BP Amoco Chemicals,
Darton, 1992

Vehiclerefinishing,
Norway16

1.2ppm

0.8 - 8.1 ppm

Ethanol a small component ofsolvent mixtures. Ranges areaverages of 3-6 individual
measurements

Moen, 2000 (Ann Occ
Hyg 44(3)Woodcoating,
Sweden38

17ppm(3 – 70 ppm)

Study primarily looking atformaldehyde exposures. Range
data only given for total solvent
concentration.

Alexandersson, (Archof Env Hlth, 1988)

Electrotechnicalcompany,
Slovakia

3
3 ppm

1.2- 5.8 ppm

Operator - soldering, mechanicalrepair (personal sampling)

Regional Authority of
Public Health,Slovakia, 2004

Wood
processing
company,
Slovakia

1
104 ppm

86-162 ppm

Painter - spraying with spray gun
under exhaust ventilation(personal sampling)

Regional Authority of
Public Health,Slovakia, 2004

Bus Transportcompany,
Slovakia

2
53 ppm5-302 ppm

Painter - spraying of paint withspray gun (personal sampling)

Regional Authority of
Public Health,Slovakia, 2004

Pharmaceutic
al company,
Slovakia

16
591 ppm

5,9 - 3432
ppm

Chemist – operator (personal
sampling)

Regional Authority of
Public Health,Slovakia, 2004

Pharmaceutic
al company,
Slovakia

21111 ppm

60-2441 ppm

Service engineer (personal
sampling)

Regional Authority of
Public Health,Slovakia, 2004

Pharmaceutic
al company,
Slovakia

2

Foreman (personal sampling)
356 ppm

4,8-1260 ppm

Regional Authority of
Public Health,Slovakia, 2004

Pharmaceutic
al company,
Slovakia

8
127 ppm

1.5-851 ppm

background in the hall (stationary
sampling)

Regional Authority of
Public Health,Slovakia, 2004

*Values expressed as ppm have been converted

UNEP PUBLICATIONS

39

OECD SIDSETHANOL

Note (data from measurements made in the Slovak Republic): Long-term stationary and personalmeasurements were applied. Time of sampling was >70% of the shift time, each measurement
represents the whole shift exposure (Ref.: Regional Authority of Public Health, Slovakia, 2004.)

40

UNEP PUBLICATIONSOECD SIDSETHANOLIUCLID
Data Set
Existing Chemical
CAS No.
EINECS NameEC No.
TSCA Name
Molecular Formula

:
:
:
:
:
:

ID: 64-17-5
64-17-5
Ethanol
200-578-6
Ethanol
C2H6O

Producer related part
Company
Creation date

: BP Chemicals Ltd.
: 15.04.1994

Substance related part
Company
Creation date

: BP Chemicals Ltd.
: 15.04.1994

StatusMemo::

Printing date
Revision date
Date of last update
Number of pages

:
:
:
:

Chapter (profile)
Reliability (profile)
Flags (profile): Chapter: 1, 2, 3, 4, 5, 6, 7, 8, 10: Reliability: without reliability, 1, 2, 3, 4
: Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE),Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SIDS

19.11.2004
15.04.1994
19.11.2004
340

UNEP PUBLICATIONS

41

OECD SIDS
1. GENERAL INFORMATION
1.0.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

APPLICANT AND COMPANY INFORMATION

Type
Name
Contact person
Date
StreetTown
Country
Phone
Telefax
Telex
Cedex
Email
Homepage

:
:
:
:
:
:
:
:
:
:
:
:
:

CEFIC
Mr Graeme Wallace
Av. E. van Nieuwenhuyse 41160 Bruxelles
Belgium+32 (0) 2 676 7410
+32 (0) 2 676 7216

gwa@cefic.be

26.09.2003
Type
Name
Contact person
Date
StreetTown
Country
Phone
Telefax
Telex
Cedex
Email
Homepage

:
:
:
:
:
:
:
:
:
:
:
:
:

cooperating company
BP Chemicals Ltd.
Mr Jeff Kelsey
Chertsey Road
TW16 7LN London
United Kingdom+44 (0)1932 762577+44 (0)1932 764147

kelseyj@bp.com

26.09.2003
1.0.2

LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR

1.0.3

IDENTITY OF RECIPIENTS

Remark:Ethanol is being notified under the biocides directive as a basic substance.
It is in use in all EU Member States.

16.01.2004
1.0.4

DETAILS ON CATEGORY/TEMPLATE

1.1.0

SUBSTANCE IDENTIFICATIONIUPAC Name
Smiles Code
Molecular formula
Molecular weight

42

:
:
:
:

Ethanol
CCO
C2H6O
46.07

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATIONPetrol class
08.01.2002
1.1.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

GENERAL SUBSTANCE INFORMATION

Purity type
Substance typePhysical status
PurityColour
Odour

:
:
:
:
:
:

typical for marketed substance
Organic
Liquidca. 95 - 99.9 % w/w
Colourless
Alcoholic18.01.2002
1.1.2

SPECTRA

1.2

SYNONYMS AND TRADENAMES

Absolute ethanol

AlcoholAnhydrol30.04.2004
Ethyl alcohol

Ethyl hydrate

Ethyl hydroxide

Fermentation Alcohol
30.04.2004
Grain alcohol

Jaysol30.04.2004
Methylcarbinol

Molasses Alcohol

UNEP PUBLICATIONS

43OECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

30.04.2004
Potato Alcohol
30.04.2004
Spirit30.04.2004
Synasol
30.04.2004
Tecsol30.04.2004
1.3

IMPURITIES

PurityCAS-NoEC-No
EINECS-Name
Molecular formula
Value

:
:
:
:
:
:

7732-18-5
231-791-2
Water
H2O
ca. .1 - 5 % w/w

Purity type
CAS-NoEC-No
EINECS-Name
Molecular formula
Value
Function of additive

:
:
:
:
:
:
:

ca. 1 - 5 % v/v
other: Denaturants or organoleptic modifiers

Remark:Reliability:

18.01.2002
1.4

44

ADDITIVES

For customs and excise reasons, ethanol is usually denatured
when supplied into non beverage applications. Permitted
denaturants vary between EU member states and a large numberof permitted formulations exist depending on end use anduser preferences. There is no comprehensive list ofdenaturants authorised under EU legislation. Common
denaturants include wood naptha, methanol, isopropanol,
methyl ethyl ketone, ethyl acetate, cyclohexane and acetone,
etc. Typical concentration ranges are 1 to 5%, however,quantities can be lower than 0.5% and exceed 20% without
altering the essential characteristics of ethanol for nonbeverage applications. Bitrex is also often used at aconcentrations of 10's ppm as an organoleptic modifier.
This information should not be regarded as an exhaustive
list of denaturants or concentrations used.
(2) valid with restrictions

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION
Flag
20.08.2003

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Critical study for SIDS endpoint

Quantity:= 1700000 - tonnes produced in 2001

Remark:There are literally thousands of alcohol producers in Europe. These
include those producing 'neutral' ethanol, which is used in both industrial
applications and in beverages such as gin and vodka, and those producing
spirit drinks containing ethanol (e.g. whisky and wine.) Figures for
production in the Europe Union in 2001 are shown in this and subsequent
records. Synthetic ethanol represents about 30% of total production.

1.5

TOTAL QUANTITY

Ethanol type
EU Production (tpa)
Agricultural alcohol
790 000
Synthetic ethanol
500 000
Wine alcohol
230 000
Fuel alcohol
180 000
TOTAL
1 700 000(1) valid without restriction

Reliability
12.10.2004:

Quantity:= 46000 - tonnes produced in 2000

RemarkReliability
12.10.2004::

Figures for production in Czech republic(1) valid without restriction

Quantity:= 41100 - tonnes produced in 2001

RemarkReliability
12.10.2004::

Figures for production in Czech republic(1) valid without restriction

Quantity:= 49600 - tonnes produced in 2002

RemarkReliability
12.10.2004::

Figures for production in Czech republic(1) valid without restriction

Quantity:= 7000000 - tonnes produced in 2001

RemarkReliability
12.10.2004::

Figures for production in the USA(2) valid with restrictions

Quantity:= 25000000 - tonnes produced in 2001

RemarkReliability
12.10.2004::

Worldwide production(2) valid with restrictions

1.6.1

(1)

(2)

(2)

(2)

(1)

(1)

LABELLING

UNEP PUBLICATIONS

45

OECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

LabellingSpecific limits
Symbols
Nota
R-Phrases
S-Phrases

:
:
:
:
:
:

as in Directive 67/548/EEC
No
F, , ,,,(11) Highly flammable(2) Keep out of reach of children(7) Keep container tightly closed(16) Keep away from sources of ignition - No smoking

Reliability
20.08.2003:(1) valid without restriction

Classified
Class of danger
R-Phrases
Specific limits

:
:
:
:

as in Directive 67/548/EEC
highly flammable(11) Highly flammable

Reliability
20.08.2003:(1) valid without restriction

:
:

Type
Use in closed system

1.6.2

CLASSIFICATION

1.6.3

PACKAGING

1.7

USE PATTERN

Type of use
Category
12.10.2004
Type of use
Category

(1)
:
:

Type
Non dispersive use

12.10.2004
Type of use
Category(1)::

Industrial
Personal and domestic use

12.10.2004
Type of use
Category

(1)
:
:

Type
Wide dispersive use

12.10.2004
Type of use
Category(1)::

Industrial
Basic industry: basic chemicals

12.10.2004
Type of use
Category

46

(1)
:
:

Industrial
Chemical industry: used in synthesis

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

12.10.2004
Type of use
Category(1)::

Industrial
Fuel industry

12.10.2004
Type of use
Category(1)::

Industrial
Paints, lacquers and varnishes industry

12.10.2004
Type of use
Category(1)::

Industrial
Public domain

12.10.2004
Type of use
Category

(1)
:
:

Use
Anti-freezing agents

12.10.2004
Type of use
Category

(1)
:
:

Use
Cosmetics

12.10.2004
Type of use
Category

(1)
:
:

Use
Fuel

12.10.2004
Type of use
Category

(1)
:
:

Use
Intermediates

12.10.2004
Type of use
Category

(1)
:
:

Use
Solvents

12.10.2004
Type of use
Category

(1)
:
:

Use
Biocide12.10.2004(1)

Type of use
Category::

Use

Remark12.10.2004:

Beverages.

1.7.1

(1)

DETAILED USE PATTERNUNEP PUBLICATIONS

47

OECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

5 Personal / domestic use
5 Anti-freezing agents
No extra details necessary
No extra details necessary
Available

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

5 Personal / domestic use
15 Cosmetics
No extra details necessary
No extra details necessary
Available

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

5 Personal / domestic use
48 Solvents
No extra details necessary
No extra details necessary
Available

12.10.2004

48

(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document:

5 Personal / domestic use
39 Biocides, non-agricultural
No extra details necessary
No extra details necessary
Available

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
:
:
:
:
:

01

Human hygiene biocidal products

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup::

Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:

6 Public domain
39 Biocides, non-agricultural
No extra details necessary
No extra details necessary
Available
02 Private area and public health area disinfectants andother biocidal products

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

6 Public domain
39 Biocides, non-agricultural
No extra details necessary
No extra details necessary
Available
03 Veterinary hygiene biocidal products

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application

:
:
:
:
:

6 Public domain
39 Biocides, non-agricultural
No extra details necessary
No extra details necessary
Available
04 Food and feed area disinfectants

UNEP PUBLICATIONS

49

OECD SIDS
1. GENERAL INFORMATION
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

14 Paints, lacquers and varnishes industry
48 Solvents
Solvent based

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

Available

12.10.2004
Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: yes:
Private use:
Recovery
:

:
:
:
:
:
:

Remark12.10.2004

50

(1)

:

3 Chemical industry: chemicals used in synthesis
48 Solvents
No extra details necessary
No extra details necessary
Available

Pharmaceutical processing(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing
: :

:
:
:
:
:
:

15/0 other
18 Explosives
No extra details necessary
No extra details necessary
not available

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION
Private use
Recovery::Remark12.10.2004:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Damping agent(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

Remark12.10.2004:

15/0 other
26 Food/feedstuff additives
No extra details necessary
No extra details necessary
not available

Food flavourings, fragrances, beverages(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

15/0 other
27 Fuels
No extra details necessary
No extra details necessary
not available

12.10.2004(1)

Industry category
Use category
Extra details on use category:::

Emission scenario document
Product type/subgroup
Tonnage for Application
Year
Fraction of tonnage for application
Fraction of chemical in formulation
Production: :Formulation: :Processing: :Private use:
Recovery
:

:
:
:
:
:
:

2 Chemical industry: basic chemicals
55/0 otherNo extra details necessary
No extra details necessary
not available

UNEP PUBLICATIONS

51

OECD SIDS
1. GENERAL INFORMATION
Remark12.10.2004
1.7.2

:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Raw material(1)

METHODS OF MANUFACTURE

Origin of substance
Type

:
:

Synthesis
Production

Remark:All potable alcohol, and a large proportion of industrial and fuel ethanol, ismade by the fermentation process in which zymase, a yeast enzyme,changes simple sugars (e.g. as found in molasses) into ethanol and carbon
dioxide. Biomass (sugarbeet/cane, molasses, cereals, rice, grain,cellulosics) are used as the feedstock for fermentation ethanol. Syntheticroutes to ethanol also exist, and, although a process based on
acetaldehyde arising from acetylene is quoted, the virtually universal
synthetic route is via the hydration of ethylene to ethanol.
(1)12.10.2004

1.8

REGULATORY MEASURES

1.8.1

OCCUPATIONAL EXPOSURE LIMIT VALUES

Type of limit
Limit value::

OES (UK)
1920 mg/m3

Reliability
12.10.2004:(1) valid without restriction

Type of limit
Limit value::

MAK (DE)
960 mg/m3

Remark:Reliability
12.10.2004:

Peak limit category II,1
Carcinogen category 5
Pregnancy group C
Mutagen group 2(1) valid without restriction

Type of limit
Limit value::

other: ACGIH PEL
1900 mg/m3

Reliability
12.10.2004:(2) valid with restrictions

(3)

(4)

(5)

Type of limit: other: Czech republic
Limit value: 1000 mg/m3
Short term exposure limit value
Limit value: 3000 mg/m3
Time schedule:
Frequency: TimesReliability
12.10.2004

52:(1) valid without restriction(2)

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type of limit
Limit value::

other: NIOSH PEL
1900 mg/m3

Reliability
12.10.2004:(2) valid with restrictions(5)

Type of limit: other: OSHA PEL
Limit value: 1900 mg/m3
Short term exposure limit value
Limit value: 0Time schedule:
Frequency: TimesReliability
12.10.2004:(2) valid with restrictions(5)

Type of limit: other: Slovak republic
Limit value: 960 mg/m3
Short term exposure limit value
Limit value: 1920 mg/m3
Time schedule: 30 minute(s)Frequency: 4 times
Reliability
12.10.2004
1.8.2

:

(1) valid without restriction(6)

ACCEPTABLE RESIDUES LEVELS

Proposed residues level
Maximum residue level::

Remark:Reliability
08.08.2003

:

None
mg/kg
Ethanol occurs naturally and is endogenously produced. Humanconsumption of ethanol in alcoholic beverage can be high. A
meaningful and controllable ARL may therefore be impossible
to establish.
(4) not assignable

1.8.3

WATER POLLUTION

1.8.4

MAJOR ACCIDENT HAZARDS

1.8.5

AIR POLLUTIONClassified byLabelled byNumberClass of danger

:
:
:
:

other: US EPA

Remark:Reliability:

No entry for ethanol in Hazardous Air Pollutants list, Air
Toxics Website(1) valid without restriction

other: not listed as toxic

UNEP PUBLICATIONS

53

OECD SIDS
1. GENERAL INFORMATION
Flag
12.10.2004
1.8.6

:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Critical study for SIDS endpoint(7)

LISTINGS E.G. CHEMICAL INVENTORIES

Type
Additional information::

EINECS
Listed as ethanol, 2005786

:
:

TSCA
Listed as ethanol, molecular formula C2H6O::

Annex I, Council Regulation (EEC) No. 793/93
Listed as ethanol, ethyl alcohol, Notes r59::

Council Directive (EEC) No. 76/769
Listed as ethanol, ethyl alcohol, Notes r59

:
:

IARC
Listed as ethanol.

17.01.2002
Type
Additional information
22.09.2003
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
26.09.2003
Type
Additional information

(8)
:
:

other: Canada, Transportation Dangerous Goods, Schedule II
Listed as ethanol or ethanol solutions, No. UN 1987::

other: DOT UN/NA
NA1987, Hazard Class 3.

::

Annex I, Council Regulation (EEC) No. 793/93
Index number 603-002-00-5. Risk Phrases 11, Symbol F::

other: FDA Direct Food Additives
Se 21 CFR 184.1293.

::

other: FDA Everything Added to Food in the US
EAFUS document 421:

other: FEMA Generally Recognised as Safe List

17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type

54

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION
Additional information

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Listed as Ethanol. TRGS 900 Lim value 1000 ml/m3 (ppm) = 1900 mg/m3.
FEMA no. 2419::

other: HPV Chemicals
Fully sponsored::other: Japan - Examined Substances/National Inventory
Listed as ethanol. ENCS No. (2)-202::

other: Korea, National Inventory (KECI)
Listed as ethanol, ethyl alcohol No. KE-13217
ECL 2-858::

other: NIOSH Health effects
Listed as ethyl alcohol (ethanol). Health effects nihe267::

other: NIOSH OELs
Listed as ethyl alcohol, ni11.

::

other: NTP
Listed as ethanol for toxicological testing.

::other: OSHA
OELs see 29 CFR 1910.1000 (Subpart Z)::

other: UK HSE EH40
Listed as ethanol. OELs

:
:

AICS
Registered under CAS number::

other: Norway
YL number = 1163

:
:

CHINA
Listed26.09.2003
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information

26.09.2003
Type
Additional information
22.09.2003
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
17.01.2002
Type
Additional information
26.09.2003
Type
Additional information
26.09.2003
Type
Additional information

UNEP PUBLICATIONS

55

OECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

26.09.2003
Type
Additional information

:
:

PICCS
Listed as CAS number::

other: Switzerland
BAG number = G-1158, list category 1/not listed

26.09.2003
Type
Additional information
26.09.2003
1.9.1

DEGRADATION/TRANSFORMATION PRODUCTS

Type
CAS-NoEC-No
EINECS-NameIUCLID Chapter

:
:
:
:
:

degradation product64-19-7
200-580-7
acetic acid

Remark
17.01.2002:

By oxidation.

Type
CAS-NoEC-No
EINECS-NameIUCLID Chapter

:
:
:
:
:

combustion products
Oxides of carbon

17.01.2002
1.9.2

COMPONENTS

1.10

SOURCE OF EXPOSURE

Source of exposure
Exposure to the::other: endogenous ethanol
Substance

Method:Blood samples were collected from 130 subjects that had not consumed
alcohol over the previous 24 hours, from 10 test subjects on 4 differentdays to ensure abstinence from ethanol consumption and from a total of 30
patients receiving treatment for metabolic illnesses, in some cases
receiving treatment for withdrawal from alcohol.

Remark:Result:Samples taken with a 'Koller venule' with the addition of NaF. Blood
alcohol levels were determined with a 2-column chromatograph andheadspace technique - details given in a separate reference (Bonte, 1981).
All of the individual measurement values are presented in the reference.
From these it is possible to estimate that the endogenous ethanol range
covering 95% of the population is 0.062 to 0.73mg/l.
In the first group, physiological ethanol concentrations were all below 0.75
mg/ml with most values in the concentration range 0.1 and 0.2 mg/l.
In the second and third group, values were essentially the same as in

56

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

group 1.
(2) valid with restrictions

Reliability
12.10.2004:

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

Remark:Result:Reliability
12.10.2004:

This paper is primarily about the role of ethanol produced
during dough fermentation in the production of Maillard
reaction products and their role as antioxidants. Ethanol
production during bread making is discussed.
About 1.6 to 2.8 g ethanol is produced per 100 g flour.
Some is utilized in the Maillard reaction therefore baked
bread has a lower ethanol content than this.
(2) valid with restrictions

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

Remark:Reliability
12.10.2004:

Human exposure to alcohol from different foods and drinks istabulated:
Level (g/100g)
Alcohol-free beer
0.2-0.33
Malt beer
1.16-1.28
Apple juice
0.14
Grape juice
0.17
Kefir
0.5
Black Forest gateaux0.5-1.0
Praline with alcohol filling
6.95
Ice-cream (alcohol/fruit)
0.74-2.44
Tiramisu
1.0
Wine saurkraut
0.1-0.8
Worcestersauce
0.79(4) not assignable

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

Result:Alcohol content of food products:

(9)

(10)

(11)

Product:
A-1 sauce
Canada Dry Ginger Ale
Corr's Grapefruit Soda
Corr's Lemon Tangerine soda
Dr Pepper
Gerstel Brau (sample 1)
Gerstel Brau (sample 2)
Grey Poupon Mustard
Haagan-Daz Rum-Raisin icecream
Haagan-Daz vanilla icecream
Heinz catsup
Hunt's Snack Pak Vanilla Pudding
Kikkoman Soy Sauce
Kikkoman Teriyaki
Knott's Berry Farm Strawberry Pres.
Knudsen Black Cherry Juice
Knudsen Cherry cooler

UNEP PUBLICATIONS

%vol
0.04
0.08
0.17
0.21
0.03
0.53
0.51
0.06
0.35
0.48
0.04
0.04
2.3
3.0
n/d
0.10
0.19

57

OECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

Knudsen Cider and Spicen/dKnudsen Hibiscus Cooler
0.06
Kroger Vanilla extract
35.7
Lea and Perrins Worcestershire Sauce 0.16
Martinelli Sparkling Cida
0.05
S&W Ripe Olives
n/d
S&W Sauerkraut
0.15
Schweppes Tonic
0.08
Scope Mouthwash
18.9%
Seven-Up
0.07
Smucker's Strawberry Jellyn/d
Spice Islands Red Wine Vinegar
0.24
Spice Islands White Vinegar
0.24
Sprite0.06
Texas Select malt Beverage
0.24
Vick's NyQuil (25% label)
25.1
Vlasic Kosher Dill Pickles
0.04(4) not assignable

Reliability
12.10.2004:

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

RemarkReliability
12.10.2004::

Quality requirement for all orange juices: ethanol less than 3g/l.
(4) not assignable

Source of exposure
Exposure to the::

Remark:m the SPIN product register of the Scandinavian countries (Finland, Sweden,
Denmark) showing the number of registered products for the year 2001 in
Use (NACE). Only products with total registration >10te shown.

(12)

(13)
Human: exposure of the consumer/bystander
Substance

Country
FIN
FIN
N
FIN
DK
DK
N
DK
FIN
FIN
FIN
FIN
N
DK
FIN
FIN
DK
FIN
N
DK
DK
DK
FIN
DK
FIN
N
DK
DK
DK
DK
N

58

Category of use

Number of
products

Pulp, paper and paper products
14
Chemicals and chemical products
13
Chemicals and chemical products
88Radio, television and communication equipment 19
Chemicals and chemical products
115
Publishing, printing and reproduction media
123
Construction
159
Construction
272
Research and development
30
Manufacture of food products and beverages
15
Supporting transport activities
6
Sale, maint. repair of motor vehicles
78
Health and social work
8
Private households with employed persons
186
Publishing, printing and reproduction media
121
Wood and products except furniture
18
Manufacture of furniture
375
Health and social work
31
Land transport; transport via pipelines
10
Other business activities
119
Manufacture of wood prods. except furniture
285
Sewage and refuse disposal
5
Land transport; transport via pipelines
7
Health and social work
79
Construction
70
Manufacture of wood prods. except furniture
52
Sale, maintenance, repair of motor vehicles
310
Manufacture of rubber and plastic products
183
Fabricated metal products, except machinery
339
Manufacture of food products and beverages
426
Sale, maintenance, repair of motor vehicles
90

UNEP PUBLICATIONS

Preparation
quantity(tonnes)42261.016956.1
12272.3
12239.811342.1
6814.73414.43025.32797.52515.02500.02418.12367.72216.71528.91422.11314.11193.11114.01010.7
934.6
607.9
476.0
443.2
406.4
367.0
347.5
339.5
304.1
258.0
248.5

OECD SIDS
1. GENERAL INFORMATION
N
FIN
N
FIN
DK
FIN
DK
FIN
N
N
DK
DK
DK
DK
N
DK
FIN
N
DK
FIN
DK
DK
DK
FIN
DK
DK
N
N

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
Publishing, printing, reproduction media
24
Forestry
4
Manufacture of furniture; manufacturing n.e.c. 82
Fabricated metal products, except machinery
19
Manufacture of other transport equipment n.e.c 239
Other business activities
41
Manufacture of basic metals
25
Manufacture of furniture; manufacturing n.e.c. 37
Retail trade, except of motor vehicles
11
Manufacture of other transport equipment n.e.c 144
Medical, precision and optical instruments etc 19
Retail trade, except of motor vehicles
43
All kinds of activities
48
Public administration and defence
34
Manufacture of food products and beverages
20
Manufacture of machinery and equipment
61
Extra-territorial organisations and bodies
26
Education
4
Agriculture and horticulture14
Non-metallic mineral products
7
Motor vehicles, trailers and semi-trailers
56
Pulp, paper and paper products
18
Wholesale trade and commission trade9
Motor vehicles, trailers and semi-trailers
17Radio, television and communication equipment
21
Manufacture of elec. machinery and apparatus
77
Fabricated metal prods, except machinery and eq. 47
Other service activities
21

189.9
186.1
180.2
174.1
172.9
156.8
155.3
132.3
131.7
110.4
98.3
90.5
87.9
84.1
78.1
65.0
63.0
58.7
40.1
35.5
28.7
20.2
17.2
16.0
15.3
14.1
12.8
10.7

Reliability
12.10.2004:

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

Remark:Data from the SPIN product register of the Scandinavian countries(Finland, Sweden, Norway, Denmark) showing the number of registeredproducts for the year 2001 in Industrial Use (national) category. Only
products with a tonnage >10 are shown.

(4) not assignable(14)Country
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK

Product type

Number of
products
Paints, varnishes and ink
61
Printing of books and offset printing
9
Private households with employed persons
186
Other printing works n.e.c.
20
Manufacture of pharmaceuticals
6
Printing and service activities
35
General construction of buildings
48
Bricklaying contractors
30
Industrial cleaning
42
Construction
10
Chemicals and chemical products
19
Manufacture of furniture76
Perfumes and toilet preparations
14
Varnishing/acid washing of furniture, etc. 191
Manufacture of chairs and seats
74
Publishing
12
Manufacture of wood products, ex. furniture 45
General repair shops
127
Manufacture of plastic packing goods
40
Printing of newspapers
9
Publishing, printing and reproduction media 40
Other printing works
11
Floor and wall covering and floor planing
27
Manufacture of soap and detergents
4
Manufacture of other products of wood n.e.c 28
Manufacture of other chemical products n.e.c 6
Hospital activities
21
Building and repairing of ships and boats
41
Manufacture of plastic products
126

UNEP PUBLICATIONS

Tonnage of
product
5448.03299.12216.71197.41123.71105.01028.4
1019.4
770.5
677.6
524.0
446.9
406.2
302.1
273.7
270.2
238.7
218.2
200.5
193.8
178.9
175.7
175.2
174.5
170.3
151.4
134.1
129.0
120.6

59

OECD SIDS
1. GENERAL INFORMATION
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK
DK

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
Treatment and coating of metals
228
General cleaning activities
70
Painting
133
Manufacture of other kitchen furniture
95
Specialized cleaning activities
38
All kinds of activities
48
Office and shop furniture except of chairs
63Household furniture/varnishing of furniture 60
Dispensing chemists
4
Dentists
7
Motor vehicle painters
130
Production of mineral waters and drinks
118
Manufacture of agricultural machinery
4
Treatment and coating of metals
10Central heating radiators and boilers
9
Manufacture of other fabricated metal prods
9
Manufacture of builders carpentry/joinery
137
Manufacture of other products of wood
9
Casting of iron
6
Manufacture of ice cream96
Administration of the state
9Fire service activities
8
Home nursing activities/general health care
7Home help
7
Manufacture of electronic equipment
28
Residential nursing homes/sheltered homes
7
Day institutions for elderly people
5
Manufacture of food products and beverages
32
Basic iron and steel and of ferro alloys
9
Manufacture of cast iron tubes
5
Casting of steel
5
Manufacture of cocoa; chocolate and sugar
102
Retail sale of automotive fuel19
Other motor vehicle services
24
Human and health activities
22
Fabricated metal prods, except machinery
96
Manufacture of chairs except of upholstery
15
Building and repairing of ships
113
Corrugated paper and paperboard etc.
14
Serigraphic printing
16
Plastic plates/sheets/hoses/film/tubes etc.
4
Building/repairing of boats
80
Joinery installation
46
Motor vehicles, trailers and semi-trailers
34
Manufacture of machinery and equipment
46

Reliability
12.10.2004:

Source of exposure
Exposure to the::

Human: exposure of the consumer/bystander
Substance

Remark:Data from the SPIN product register of the Scandinavian countries(Finland, Sweden, Norway, Denmark) showing the number of registeredproducts for the year 2001 in Use category UC62. Only products with total
registration >10te shown.

(4) not assignable(14)Country
N
DK
DK
N
N
DK
DK
DK
DK
DK
DK
N
N
N
N

60

116.7
108.3
100.2
97.6
93.0
87.9
85.3
79.6
77.6
77.5
67.5
53.2
52.2
51.8
51.8
51.8
51.4
47.7
47.0
33.8
32.3
32.1
32.1
32.1
30.2
29.7
29.4
28.0
27.2
27.0
27.0
25.6
21.5
21.3
20.7
20.2
19.9
18.9
17.4
17.2
16.3
15.9
12.6
12.2
11.5

Application

Number ofprods reg.
Solvents
47
Solvents
158
Cleaning/washing agents306
Non-agri. pesticides/preservatives
11
Cleaning/washing agents
127
Reprographic agents130
Non-agri. pesticides/preservatives
58
Paints, laquers and varnishes
700
Process regulators
84
Anti-freezing agents
16
Surface treatment
99
Paints, laquers and varnishes
392
Reprographic agents
29
Surface treatment
20
Process regulators
21

UNEP PUBLICATIONS

Tonnage ofprods reg.
16323
10174
3820
3605
2102
2024
1031
891
867
865
661
364
193
177
146

OECD SIDS
1. GENERAL INFORMATION

Reliability
12.10.2004
1.11

:

DK
DK
N
DK
DK
DK
DK
DK

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
Adhesives, binding agents
136
Surface-active agents
18
Adhesives, binding agents
54
Others15
Food/feedstuff flavourings/nutrients 331
Impregnation materials
18
Anti-set-off and anti-stick agents
11
Cosmetics
31

139
134
90
81
50
38
23
19

(4) not assignable(14)

ADDITIONAL REMARKS

Memo

:

Conversion factors

Remark

:1 mg/m^3 = 0.52 ppm
1 ppm = 1.92 mg/m^3

26.09.2003

(15)

Memo

:

Threshold Odour Concentration (Air)RemarkReliability
22.06.2004::

50% of threshold odour concentrations are below 100 mg/m^3(4) not assignable

Memo

:

Threshold Odour Concentration (Water)

Remark:Reliability
22.06.2004:

50% of Threshold Odour Concentrations are at approximately
800 mg/l.
(4) not assignable

1.12

(15)

(15)

LAST LITERATURE SEARCH

Type of search
Chapters covered
Date of search:::

External
422.11.2002

08.01.2002
Type of search
Chapters covered
Date of search

(16)
:
:
:

External
409.01.2002

11.01.2002
Type of search
Chapters covered
Date of search

(17)
:
:
:

External
518.01.2002:

Fertility

18.01.2002
1.13

REVIEWS

Memo

UNEP PUBLICATIONS

61

OECD SIDS
1. GENERAL INFORMATION

Remark:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Studies in mice and rats generally have not shown an effect on
reproductive performance.
When female C57Bl/Crgl mice were given 10% ethanol (v/v) in water as thedrinking fluid before mating, throughout gestation and lactation, no
significant effect on reproductive capacity was seen (Thiessen D.D. et al.
1966. Q. J. Stud. Alcohol 27, 591, cited in IARC, 1988).
When female Wistar rats were given 20-25% of the calories consumed as
12% ethanol in a sucrose solution as the drinking fluid before mating andthroughout gestation and lactation, there was no effect on reproductive
performance (Oisund J.F. et al. 1978. Acta pharmacol. toxicol. 43, 145,cited in IARC, 1988).
Exposure of male rats [strain unspecified] to ethanol in utero or as
neonates by administration of a liquid diet containing ethanol (36% of total
calories) resulted in adverse effects on gonadal growth and development
and disturbances in their sexual behaviour and performance when adult(Parker S. et al. 1984. Neurobehav. Toxicol. Teratol.6, 289, cited in IARC,
1988).
Mating of female Holtzman rats fed a liquid diet containing 5% ethanol for
16 weeks with untreated males resulted in no adverse effect on fertility,
litter size or neonatal body weight (Krueger W.A. et al. 1982. Pharmacol.
Biochem. Behav. 17, 629, cited in IARC, 1988).
Studies in mice and rats have shown effects on the testis and on otherreproductive tissues.
In a study in which male C57BL/6J mice were given 5 or 6% (v/v) ethanol
in a liquid diet for 70 days or 35 days, respectively, there was a significant
decrease in testicularweight and in seminal vesicle/prostate weight, an
increase in the frequency of germ-cell desquamation, inactive
seminiferous tubules, inhibition of in-vitro fertilisation of mouse oocytes by
epididymal spermatozoa, as well as a significant decrease in the total
number of motile sperm. During a ten-week recovery period, improvement
was greater in the group given 5% than in those given 6% ethanol(Anderson R.A. et al. 1985. Alcohol 2,479, cited in IARC, 1988). Preparation of sperm from the cauda epididymis
five weeks after oral administration of ethanol (1, 2 or 4 ml/kg bw) to male
(CBA x Balb/c)F1 mice
five times daily did not show sperm anomalies (Topham J.C. 1980. M.
Res. 69, 149, cited in IARC, 1988). Addition of ethanol to ram
spermatozoa (0.62M; 15 µl in 0.4 ml semen
samples containing2-dioxy-D-glucose) inhibited sperm motility (Mayevsky
A. et al. 1983. Archs Toxicol. Suppl.6, 295, cited in IARC, 1988).
There is evidence in vitro and in vivo that ethanol is toxic to animal andhuman Leydig cells and seminiferous tubules (Gavaler J.S. & Van Thiel
D.H. 1987. M. Res. 186,
269; Van Thiel D.H. et al. 1983. Pharmacol. Biochem. Behav. 18, 317,bothcited in IARC, 1988). Male Sprague- Dawley rats maintained on a liquid
diet containing 6% ethanol (95%v/v) for one week followed by four weeks on a 10% ethanol liquid dietshowed adverse effects on sex organs (testes, seminal vesicles, ductules)
as well as a significant
decrease in serum testosterone levels (Klassen R.W. & Persaud T.V.N.
1978. Int. J. Fertility 23, 176, cited in IARC, 1988). Male Sprague-Dawley
rats that received an intraperitoneal injection of 2.5 g/kg bw ethanol

62

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATIONETHANOL
ID: 64-17-5
DATE: 19.11.2004

showed a significant decrease in the levels of luteinizing hormone andtestosterone and marked attenuation of testicular steroidogenesis (Cicero
T.J. et al. 1979. J. Pharmacol. exp. Ther. 208, 210, cited in IARC, 1988.)
Exposure of male rats [strain unspecified] to ethanol in utero or as
neonates by administration of a liquid diet containing ethanol (36% of total
calories) resulted in adverse effects on gonadal growth and development
and disturbances in their sexual behaviour and performance when adult(Parker S. et al. 1984. Neurobehav. Toxicol.Teratol.6, 289, cited in IARC,
1988). Subcutaneous administration of 7.9 g/kg bw ethanol to female CD
rats inhibited ovulation, primarily by blocking ovulatory surges of luteinizing
hormone (Kieffer J.D. & Ketchel M.M. 1970. Acta endocrinol. 65, 117, citedin IARC, 1988). Blood levels of luteinizing hormone varied with dose andtiming of treatment, but ethanol administered by intraperitoneal injection
increased the secretion of prolactin by female Wistar rats (Alfonso M. et al.
1985. Gen. Pharmacol. 16, 43, cited in IARC, 1988). Exposure ofSprague-Dawley rats to ethanol (average, 11.6 g/kg bw) in utero altered
the adult patterns of luteinizing hormone secretion in male and female
offspring, indicating an effect on the central mechanisms that control
secretion of pituitary luteinizing hormone (Handa R.J. et al. 1985. Life Sci.
37, 1683, cited in IARC, 1988). Administration of 5% ethanol (36% of total
calories) in a liquid diet to female Wistar rats for 49 days decreased ovarian
weight by 60% and significantly
decreased plasma oestradiol-17ß levels and the development of oestrogentarget organs (Van Thiel D.H. et al. 1978. J. clin. Invest. 61, 624, cited in
IARC, 1988).
Ovarian function in female Holtzman rats, 20 days of age, was suppressedby feeding of liquid diets containing 5% ethanol (36% of total caloric intake)for up to 55 days, in which blood ethanol concentrations averaged 2.5 g/l,
but not by 2.5% ethanol (Bo W.J. et al. 1982. Anat. Rec. 202, 255, cited in
IARC, 1988).
Vaginal opening was delayed in female Holtzman rats fed a liquid dietcontaining 5% ethanol for eight or 16 weeks. Among rats treated for 16
weeks, irregular oestrous cycles and cycles longer than those in control
were observed. Mating of these females with untreated males resulted in
no adverse effect on fertility, litter size or neonatal body weight (Krueger
W.A. et al. 1982. Pharmacol. Biochem. Behav. 17, 629, cited in IARC,
1988).
In female macaque monkeys that administered ethanol to themselves
intravenously on a schedule of reinforcement used for food acquisition,
providing 2.9-4.4 g ethanol/kg
bw/day for 3-6.5 months, amenorrhoea, atrophy of the uterus, decreased
ovarian mass and significant decreases in luteinizing hormone levels wereobserved (Mello N.K. et al. 1983. Science 221, 677, cited in IARC, 1988).
In female rhesus monkeys infused intravenously with 2-4 g/kg bw ethanol
after spontaneous onset of labour or following the induction of labour byinfusion of oxytocin, partial suppression of labour was observed only inpreterm animals with irregular uterine contractions (Horiguchi T. et al.
1971. Am. J. Obstet. Gynecol. 109, 910, cited in IARC, 1988).
(2) valid with restrictions(18)

Reliability
22.06.2004

:

Memo

:

Developmental effects.

Method:A large number of literature references were reviewed. These
are listed below as 'sources'. The overall summary of thisreview is presented in the Remarks paragraphs.

UNEP PUBLICATIONS

63

OECD SIDS
1. GENERAL INFORMATION
Result64:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

The purpose of this document is to review recent (generally post 1990)
scientific literature on the developmental effects of ethanol exposure andthe research on potential mechanisms of adverse effects. The review
focuses on the confounding influences, such as nutritional defects, andalso examines the evidence for thresholds for adverse effects of ethanol
exposure on development. Consideration of thresholds is of particularimportance, to place the observed effects related to high ethanol exposuresfrom excessive beverage consumption in context with the low exposuresencountered with normal handling and use of ethanol in the occupationalsetting.
Ethanol is a commonly used industrial solvent, reactant and intermediary
product, potential exposure to which in the occupational setting is by theinhalation and dermal routes. It is unique amongst chemicals in that it ispresent in many consumer goods and pharmaceutical preparations and it
has been consumed by humans for millennia as a beverage and as aconstituent of foods.
Excessive consumption of ethanol containing beverages increases the riskof adverse health effects, can be addictive and is associated withwidespread social problems. A range of specific congenital malformations,known as the foetal alcohol syndrome (FAS), has been identified in a low
percentage of children born of 'heavy drinking' / alcoholic mothers.
Numerous animal studies and human epidemiology studies have confirmed
an association with ingestion of high levels of ethanol during pregnancy
and FAS, and also of lesser alcohol related birth defects (ARBDs) andalcohol related neurodevelopment disorders (ARNDs). There are conflicting
results in human epidemiology studies as to threshold levels of ethanol
consumption in relation to adverse pregnancy outcomes, reflecting
confounding influences such as smoking history, nutritional status andsocioeconomic status. Other factors also contribute to the variability ofthese studies, including difficulty inverifying intake of ethanol, different
patterns of consumption and polymorphism in ethanol metabolism. There is
general agreement that full expression of FAS occurs only with the chronicmaternal ingestion of at least 2g/kg per day ethanol (approximately 3-4oz/
90-120g ethanol per day for an average sized woman). his would beachieved by drinking about 10-12 'UK' drinks or 6-8 'US standard' drinks
and is associated with a blood alcohol level (BAL) of around 150200mg/dL. There is, however, less agreement as to the threshold for othereffects (ARBDs and ARNDs). In recent concerted European epidemiology
studies (EUROMAC), no cases of FAS were identified amongst 8448
pregnancies where the mothers consumed 'moderate' amounts of ethanol.
Although 'binge' drinking (i.e. 5 or more 'standard' drinks per occasion) at
critical stages of intrauterine development may be related to adverse
effects, the studies found an ethanol intake of around 120g per week
(averaged at 2 'standard' drinks per day) to be generally without adverse
effect.
Some adverse effects on children's cognitive function at up to 14 years ofage were observed in the 'Seattle' longitudinal epidemiology studies atlower average weekly maternal ethanol consumption. However, these
averages included 'binge' drinkers, who constituted 25% of the participating
mothers. Differentiation of results according to pattern of intake is critical,
since it is the number of drinks per occasion and the attendant higher peak
BAL, rather than a relatively constant lower BAL, which appears to be themajor risk factor for ARBDs. There is also evidence of a threshold foradverse effects in animal studies. In a recently reported study of pregnant
rats, carefully controlled for nutritional deficiencies, the effects of gastricintubation during pregnancy of 'moderate' levels of ethanol were estimated.
The authors found that doses of 0.15 and 0.30g/kg ethanol (achieving
BALs of up to 150mg/dL) were below the threshold for adverse effects on
neonatal birth weight or on subsequent learning of weaned pups in passive
avoidance tests.

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION

Conclusion:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

In an earlier primate study in pregnant Macaques, it was found the BAL
must be greater than 140mg/dL in order to produce neurologicalimpairment or dysmorphology in offspring. This was achieved with dosagesof at least 1.2g/kg/day. The authors also concluded that a dose of
0.6g/kg/day to the Macaque, which achieved a BAL of 51-71mg/dL, waswithout developmental toxicity. Although there have been occasional
studies in animals reporting effects at lower ethanol exposure levels,
usually there are questions about the validity of the BAL. It is generally
found that BALs of around 100mg/dL during pregnancy in laboratory
animals are without significant changes in offspring behaviour.
One of the more important findings from animal research is that nutritional
aspects of prenatal ethanol exposure are inseparable from ethanol's
teratogenic effects. Research inanimals has demonstrated that inadequate
maternal diets can exacerbate the effects of ethanol and has confirmedthat ethanol can directly and indirectly compromise nutritional status.
However, although reducing nutrient deficiencies appears to mitigate someeffects of lower doses of ethanol, foetal alcohol effects do not appear to beeliminated by dietary supplement beyond nominally adequate diets. The
underlying basic mechanism by which ethanol consumption leads to FAS
and ARBDs remains unknown. However, since ethanol apparently affects
CNS development at all stages, itis highly unlikely that a single mechanism
could be responsible for all of the varied effects that have been observed.
Ethanol affects several physiological reactions that potentially can alter
embryogenesis. These include altered metabolism with consequentfunctional changes of various nutrients in the foetus, hypoxia withinterrupted blood supply to foetal organs, free radical production and ahost of other actions. It is also possible that ethanol produces FAS throughan interaction with certain nutrients, including vitamins and minerals.
Research into the elucidation of mechanisms of effect continues and will
ultimately refine the process of risk assessment. However for the present
time, animal studies using the inhalation route of administration are themost relevant for occupational risk assessment. In conventional rodent
developmental toxicity studies, investigating the effects of inhaled ethanol
during pregnancy, there was no significant developmental toxicity, even athigh doses, which induced maternal intoxication (narcosis), and with which
BALs of 148-193mg/dL were achieved. Small quantities of ethanol are
rapidly metabolised. It isonly after the ingestion of large quantities, suchas from beverages, that metabolism is saturated and the high BALs,
associated with toxicity, are achieved. Ethanol absorbed by inhalation ismetabolised similarly and, unless exposure is significant, no noticeable
effects are observed. In a recently reported human pharmacokinetic
modelling study, with normal breathing of ethanol vapours of up to
5,000ppm (10 times the maximum workplace concentration [MAK]), theliver was able to metabolise ethanol at the rate it entered the body.
The BALs achieved through abuse of alcoholic beverages, and associatedwith developmental toxicity, would not be reached in the occupationalsetting. Nor is the abuse situation of relevance in the directed use of any
other consumer product. For example a 30mL dose of a cold medicine
which, for a consumer product contains a high (25%) ethanol
concentration, would only be expected to give a peak level of 15-18mg/dL,
even if all of it was absorbed instantaneously.
Based on abuse of alcoholic beverages,
ethanol appears to be a human developmental toxicant at highintake levels and to elicit adverse health outcomes on themother. However, the role of possible confounding factors
such as nutrition and socioeconomic status is unclear.
Furthermore, the threshold at which adverse effects areachieved is not exceeded by a significant margin in theoccupational setting or in consumers using ethanol
containing non-beverage products.

UNEP PUBLICATIONS

65OECD SIDS
1. GENERAL INFORMATION66

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Reliability
10.08.2003

:

Memo

:

Mutagenicity

Method:Remark:Mutagenicity studies in vitro and in vivo were studied in
the context of both industrial exposure to ethanol andexposure through consumption of alcoholic beverage. The main
findings of the review are presented in the following
remarks.
The available data on ethanol from standard genotoxicity test methods areincomplete. There is clear evidence that ethanol is not a bacterial or
mammalian cell mutagen but in vitro assays for chromosome aberration,although mostly negative, have generally not included exogenous
metabolic activation. Evidence from the use of ethanol as a vehicle control
suggests that it is not mutagenic or clastogenic in vitro.

(2) valid with restrictions(19)

Reported tests for chromosome aberration induction in vivo are all negativeand only a minority of micronucleus tests are positive. Conflicting results
have been reported for the dominant lethal assay, although aninterlaboratory study performed to OECD guidelines was negative. There issome evidence that ethanol induces SCE in vivo and can also act as ananeugen at high doses. Many in vivo studies were designed to model
alcoholism and used very high doses, sometimes for long periods.
Outcomes may have been affected by disturbances of metabolism giving
rise to secondary
effects.
It is concluded that there is no robust evidence that ethanol is a genotoxic
hazard according to the criteria normally applied for the purpose ofclassification and labeling of industrial chemicals. Some degree ofgenotoxicity may result from excessive alcohol drinking, butthis is notconsidered relevant to any conceivable exposureobtainable by either
inhalation or dermal exposure in the workplace.
(2) valid with restrictions(20)

Conclusion:

Reliability
25.08.2003

:

Memo

:

Carcinogenicity

Remark:Drinking of alcoholic beverages:
Although no studies are available for ethanol per se, there are a large
number of epidemiological studies (retrospective cohort, prospective
cohort and case-control studies) on the effects of alcoholic beverages,
which contain ethanol and water as the two main components.
These epidemiological studies clearly indicate that drinking alcoholic
beverages is causally related to cancers of the oral cavity, pharynx(excluding the nasopharynx), larynx and oesophagus. The effect appears
to be independent of beverage type. Drinking alcoholic beverages is also
likely to be causally linked to liver cancer and possibly to cancer of thebreast and large bowel. There is little or no indication of a causal relation
with cancer of the stomach, pancreas, lung, urinary bladder, kidney, ovary,
prostate, lymphatic or haematopoietic system.

Reliability
26.09.2003

:

Memo

:

The International Agency for Research on Cancer has classified alcoholic
beverages as Group 1 - carcinogenic to humans.
(2) valid with restrictions(18)
Allergic contact dermatitis

UNEP PUBLICATIONSOECD SIDS
1. GENERAL INFORMATION
Remark:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Literature review demonstrates that ethanol can be an allergen in
immediate and delayed hypersensitivity by external or internal exposureand can produce subjective irritation, irritant contact dermatitis and nonimmunologic contact urticaria.
Allergic contact dermatitis was confirmed by positive patch testing at low(1%) concentration of challenge. Ethanol can also produce subjective
irritation contact dermatitis but neither phototoxiciuty nor photoallergy has
been documented. Contact dermatitis caused by ethanol may be
misdiagnosed or overlooked.
(4) not assignable(21)

Reliability
26.09.2003

:

Memo

:

Sensible Drinking - Health Effects

Remark:Conclusion:

Reliability
10.08.2003:

This Report of a Department of Health Inter-Departmental Working Group
looks at the beneficial long term effects of alcohol, the harmful effects ofalcohol including its influence on all cause mortality and examines the caseof women and alcohol consumption. It gives general public health advice
and provides recommendations for future action.
This is an excellent source of material examining theimplications of non occupational exposures to alcohol.
(2) valid with restrictions
(22)

Memo

:

Health Issues Related to Alcohol Consumption

Remark:Reliability
16.01.2004:

A comprehensive review of the literature to 1999 covering moderatedrinking as a concept, assessment of alcohol consumption, and effects ofalcohol on genetics, body weight, cardiovascular system, pregnancy,
breast cancer, bone, CNS, cancers of the digestive tract and larynx and theliver.
(2) valid with restrictions
(23)

Memo

:

Moderate Drinking: Concepts, definitions and public health significanceRemark:Reliability
12.10.2004:

The concept of moderate drinking is reviewed with consideration ofmoderate as non-intoxicating, as statistically normal, as non-injurious, as
problem-free, and as optimal. Many different quantitative definitions are
considered, all converted to similar units of g/day. For those sources whichcarry definitions of light, moderate and heavy drinking, the range quoted
from multiple references and multiple national definitions for moderate is
4.5 to 50 g/day at the lower end and 24-80g/day at the upper end. The
recommended level of alcohol intake for 'problem free' drinking is
recommended as 24-60g/day for men and 12-36g/day for women. For the
average man, the optimal level of ethanol consumption is 10-19g/day whilst
a non-injurious level is 30-40g/day. For women, these levels are halved
and do not apply in cases of pregnancy where abstinence is recommended
as the safest choice (whilst accepting that occasional light drinking 1-2
drinks/week may have no adverse effects.)(2) valid with restrictions(24)

UNEP PUBLICATIONS

67OECD SIDS
2. PHYSICAL CHEMICAL DATA
2.1

MELTING POINT

Value
Sublimation
MethodYear
GLP
Test substance

:
:
:
:
:
:

= -114 °C

Method:RemarkReliability
Flag
11.09.2002:::

Freezing and melting points determined in a stirred cell designed to protect
contents from contact with atmosphere. The cell was surrounded with aclear glass dewar flask which provided uniform changes in temperaturewhen the assembly was immersed in cooling or warming baths. The
temperature in the cell was measured with a copper-constantan
thermocouple inserted into a thermoouple well which contained n-propyl
alcohol as a thermal conducting medium. The thermocouple wascalibrated by measuring the freezing points of purified materials. Freezing
points of benzene, water, carbon tctrachloride, mercury, chloroform andtoluene were determined and a correction curve plotted. From this curve a
correction was applied to the freezing points of the mixtures being studied.
Cooling was using liquid nitrogen.
The authors noted that ethanol was prone to supercooling.
(2) valid with restrictions
Critical study for SIDS endpoint
(25)

Value
Decomposition
Sublimation
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

= -114.1 °C
no, at °C
No
other

Remark:Reliability
16.01.2004:

Method not specified.
Value after Corcoran, J., Kruse, H. and Skolnik, S. (1953) Thermal analysisof the systems hydrazine-methanol and hydrazine-ethanol. J. Phys.
Chgem. 57:435-437.
(2) valid with restrictions
(26)

2.2

68

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

1953
no dataother TS:USI absolute

BOILING POINT

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 78.3 °C at 1013.25 hPa
No
other
1970
no dataother TS

Method:Comparative ebulliometry. The apparatus used is described in detail in
one of the references along with a detailed description of its operation. The
two boilers (reference plus test substance) are connected by a commonpressure line. Platinum resistance thermometers are used to measure thetemperature using a Mueller bridge. Water was used as the reference
substance. The method is reported to be repeatable to within a few
thousandths of a degree.

UNEP PUBLICATIONSOECD SIDS
2. PHYSICAL CHEMICAL DATA
Result:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

The results consist of two sets of readings for the thermometers, one forthe sample and one for the standard. The data was processed bycomputer to establish the best fit Antoine and Kirchoff equations. The
source of the reference data for water is quoted and any necessary
corrections are described.
The result is quoted as 351.443K and is corrected for the freezing point ofwater of 273.15K
Riddick JA, Bunger WB, Sakano TK (1986), Techniques of Chemistry, Vol
II, Organic Solvents, Physical Properties and Methods of Purification, Wiley
Samples purified before use, including drying in vapour phase. Fraction
molarity purity 0.9995(2) valid with restrictions
Whilst old and not to an OECD protocol, the method and technique is wellreported.
Critical study for SIDS endpoint(27) (28)

Source:Test substance:

Reliability:

Flag
09.11.2004

:

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 78 °C at 760 hPa
No
other
1951
no dataas prescribed by 1.1 – 1.4

Source:Reliability
09.11.2004:

Budavari, S., (ed.) (1996). The Merck Index. 12th Ed.
Merck&Co: Whitehouse Station, NJ.
(2) valid with restrictions

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 78.5 °C at

RemarkReliability
09.11.2004::

Data generated by SG Redwood (UK) Ltd, ISO9002 No Q4856(2) valid with restrictions

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 78.2 °C at
No
other

RemarkReliability
29.09.2003::

Method not specified.
(4) not assignable

2.3

(29)

other: IP71 section 1/97
1997
other TS: double rectified absolute alcohol(30)

no datano data(31)

DENSITY

Type
Value
Method
Year

:
:
:
:

Density= .7864 g/cm? at 25 °C
other
1984

UNEP PUBLICATIONS

69

OECD SIDS
2. PHYSICAL CHEMICAL DATAETHANOL
ID: 64-17-5
DATE: 19.11.2004

GLP
Test substance::no dataother TS

Method:Density determined using an oscillating tube densitometer DMA 60/61(from Paar). The density is determined by measuring the natural vibration
frequency of a U shaped glass tube filled with the test fluid. The density isrelated to the period of oscillation of the instrument but can be calculated
relative to a known substance, in this case water, by the equation:
Ds - Dw = A x (Ts2 - Tw2)
where Ds and Dw are densities of substance and water, and Ts2 and Tw2
are the oscillation periods to the power of 2 of the test substance and water
respectively. The constant A can be determined by making measurementswith water and air. (Density values for water from reference Kell (1975)

70

The temperature was maintained within 0.002K and measured using acalibrated quartz thermometer. Water content was checked using a Karl
Fisher titration apparatus.
Results available at 5, 15, 25, 35 and 45C to an accuracy of 0.01C and 2E6 for density.
Riddick JA, Bunger WB, Sakano TK (1986), Techniques of Chemistry, Vol
II, Organic Solvents, Physical Properties and Methods of Purification, Wiley
Ethanol dried over a molecular sieve 3A then fractionally distilled. Water
reference deionised then distilled using a quartz still.
(2) valid with restrictions
Not to a standard OECD protocol but reported in detail and considered
reliable
Critical study for SIDS endpoint(32) (33)Result:Source:Test substance:

Reliability:

Flag
11.11.2004

:

Type
Value
MethodYear
GLP
Test substance

:
:
:
:
:
:

relative density= .7896 at 20 °C
other: IP365/97
1997

RemarkReliability
11.11.2004::

Test performed by SG Redwood (UK) Ltd. ISO9002, no Q4856(2) valid with restrictions

Type
Value
MethodYear
GLP
Test substance

:
:
:
:
:
:

relative density= .789 at 20 °C
other

Reliability
29.09.2003:(4) not assignable

Type
Value
MethodYear
GLP
Test substance

:
:
:
:
:
:

other TS: double rectified absolute alcohol

(30)

(31)
Density= .7892 - .7896 at 20 °C
other: ASTM D4052
2003
other TS

UNEP PUBLICATIONSOECD SIDS
2. PHYSICAL CHEMICAL DATA
RemarkTest substance
Reliability:::

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Sales specification for ethanol.
>99.9% ethanol(2) valid with restrictions
Routine property measured to standard method.

18.10.2004

(34)

2.3.1

GRANULOMETRY

2.4

VAPOUR PRESSURE

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 57.26 hPa at 19.6 °C
Noother (measured)1970
no dataother TS

Method:Result:

Test substance:

Reliability:

Flag
11.11.2004:

Comparative ebulliometry. The apparatus used is described in detail in
one of the references along with a detailed description of its operation. The
two boilers (reference plus test substance) are connected by a commonpressure line. Platinum resistance thermometers are used to measure thetemperature using a Mueller bridge. Water was used as the reference
substance. The method is reported to be repeatable to within a few
thousandths of a degree. The substance was first boiled at a pressure ofaround 15kNm-2 to check for consistency in boiling temperature.
Multiple measurements of vapour pressure at temperatures between
approximately 20C and 93C (i.e. above the boiling point).
Samples purified before use, including drying in vapour phase. Fraction
molarity purity 0.9995(2) valid with restrictions
Whilst old and not to an OECD protocol, the method and technique is wellreported.
Critical study for SIDS endpoint(27) (28)

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 78.7 hPa at 25 °C

Method:Scatchard equilibrium still. Ethanol was fractionated in a 5 foot column
packed with glass helices and then treated with magnesium ethylate. The
final product of d(sup 25)(sub 4) 0.78506 was kept under its own vapour
pressure in a sealed container over magnesium ethylate and samples werewithdrawn by vacuum distillation.

other (measured)1948
no dataother TS: Commercial absolute

Vapour pressure was measured using an inverted U-tube manometer and
12 mm diameter tubing read with a M901 Gaertner cathetometer at a
distance of 250 m.

Result:Static measurements were made by vapour pressure cell connected
directly to the manometer. Agreement between
methods was within 0.2 mmHg.
Value was recorded as 59.03 mmHg and converted.

UNEP PUBLICATIONS

71

OECD SIDS
2. PHYSICAL CHEMICAL DATA72

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Source:Reliability
11.11.2004

:

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

Remark:ResultReliability
11.11.2004

:
:

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 66.66 hPa at 25 °C

RemarkReliability
11.11.2004::

Method not specified.
(4) not assignable

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

RemarkReliability
11.11.2004

:
:

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 49 - 56 at °C

RemarkReliability
11.11.2004::

Value is mmHg.
(4) not assignable

Value
Decomposition
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 179.35 hPa at 40 °C
Noother (measured)
1979
no dataother TS

Method:Method designed to pressure isotherm data for binary mixtures.

U.S. Environment Protection Agency High Production Volume,
Chemical Right to Know Program.
(2) valid with restrictions(35)= 66.3 hPa at 21.2 °C
other (measured): see remark

Test performed at Sheffield Hallam university using andisoteniscope and internal method QP58
Vapour pressure 76.0 hPa @ 24.4 deg C(2) valid with restrictions(30)

other (measured)
no data(36)= 18

at 38 °Cno data

Value is Reid vapour pressure in psi.
(4) not assignable
(37)

1999
no data(37)

UNEP PUBLICATIONSOECD SIDS
2. PHYSICAL CHEMICAL DATA

Reliability
11.11.2004
2.5:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Temperatures measured to 0.01C.
(4) not assignable(38)

PARTITION COEFFICIENT

Partition coefficient
Log pow
pH value
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

octanol-water= -.31 at 25 °C

Method:Test method and date are not known.

other (measured)
1985
no datano data

There is no mention of surface activity, dissociative properties or of watersolubility.
(2) valid with restrictions
This value has been accepted by U.S. Environment Protection Agency
High Production Volume, Chemical Right to Know Program.
Critical study for SIDS endpoint(35)

Reliability:

Flag
05.10.2003:

Partition coefficient
Log pow
pH value:::octanol-waterat °C

RemarkReliability
19.10.2004::

QC reviewed value(2) valid with restrictions

Partition coefficient
Log pow
pH value:::octanol-water= -.16 - -.32 at °C

RemarkReliability::

Value expressed as log Kow and presumed calculated.
(4) not assignable
Secondary source(39)

29.09.2003(40)

Partition coefficient
Log pow
pH value:::octanol-water= -.32 at 25 °C

RemarkReliability
19.10.2004::

No details of method available.
(4) not assignable

2.6.1

(41)

SOLUBILITY IN DIFFERENT MEDIA

Solubility inValue
pH value
concentration
Temperature effects

:
:
:
:
:

Water
> 10000 mg/l at 25 °C
=0
at °C

UNEP PUBLICATIONS

73

OECD SIDS
2. PHYSICAL CHEMICAL DATA74

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Examine different pol.
pKa
Description
StableDeg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Reliability
Flag
19.10.2002::

Solubility inValue
pH value
concentration
Temperature effects
Examine different pol.
pKa
Description
Stable

:
:
:
:
:
:
:
:
:

Water
at °C

Reliability
16.01.2004:(2) valid with restrictions

Solubility inValue
pH value
concentration
Temperature effects
Examine different pol.
pKa
Description
Stable

:
:
:
:
:
:
:
:
:

Water
at °C

Reliability
29.09.2003:(4) not assignable

Solubility inValue
pH value
concentration
Temperature effects
Examine different pol.
pKa
Description
Stable

:
:
:
:
:
:
:
:
:

Water
at °C

RemarkReliability
29.09.2003::

Ethanol is stable in water. pKa is irrelevant.
(4) not assignable

Solubility inValue
pH value
concentration
Temperature effects

:
:
:
:
:

16 at 25 °C

Other
1900
no datano data(2) valid with restrictions
Critical study for SIDS endpoint
(35)

at °C
at 25 °C
Miscible

(26)

at °C
at 25 °C
Miscible

(42)

at °C
0 at 25 °C(1)other: ether, acetone, benzene
at °C
at °C

UNEP PUBLICATIONSOECD SIDS
2. PHYSICAL CHEMICAL DATAETHANOL
ID: 64-17-5
DATE: 19.11.2004

Examine different pol.
pKa
Description
Stable::::at 25 °C
miscible

Reliability
29.09.2003:(4) not assignable

Solubility inValue
pH value
concentration
Temperature effects
Examine different pol.
pKa
Description
Stable

:
:
:
:
:
:
:
:
:

RemarkSource::Reliability
11.11.2004

:

2.6.2

(5)
at 20 °C
at °C
at 25 °C
Miscible

Described as infinite solubility of ethanol in water and water in ethanol.
Riddick JA, Bunger WB, Sakano TK (1986), Techniques of Chemistry, Vol
II, Organic Solvents, Physical Properties and Methods of Purification, Wiley(4) not assignable(43)

SURFACE TENSION

Test type
Value
Concentration
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

Ring method= 24.5 mN/m at 20 °C

Remark:Reliability
29.09.2003:

Test performed by Sheffield Hallam University using atorsion balance and internal method QP55.
(2) valid with restrictions

2.7

other: See remark
1997
other TS: double rectified absolute alcohol(30)

FLASH POINT

Value
Type
MethodYear
GLP
Test substance

:
:
:
:
:
:

= 14 °C
closed cup
other: Abel closed cup. IP170/95
1997

RemarkReliability
16.10.2003::

Test performed by SG Redwood (UK) Ltd. ISO9002. no Q4856(2) valid with restrictions

Value
Type
Method
Year

:
:
:
:

other TS: double rectified absolute alcohol(30)= 13 °C
closed cup
other

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75

OECD SIDS
2. PHYSICAL CHEMICAL DATA
GLP
Test substance::no data

Reliability
16.10.2003:(4) not assignable
(31)

2.8

AUTO FLAMMABILITY

2.9

FLAMMABILITY

Result
MethodYear
GLP
Test substance

:
:
:
:
:

highly flammable
other

Remark:Reliability
29.09.2003:

Based on flashpoint and Dangerous Substances Directive
AnnexI classification.
(2) valid with restrictions

2.10

no data(44)

EXPLOSIVE PROPERTIES

Result:Other

RemarkReliability
29.09.2003::

Explosivity range 3.3 - 19% in air by volume(4) not assignable
(5)

2.11

OXIDIZING PROPERTIES

2.12

DISSOCIATION CONSTANT

Acid-base constant:

pKa = 15.9 @ 20 deg C

Reliability
29.09.2003:(4) not assignable

2.13

(5)

VISCOSITY

Test type
Test procedure
Value
Result
MethodYear
GLP
Test substance76

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

:
:
:
:
:
:
:
:

other: not specified= 1.22 - mPa s (dynamic) at 20 °C
other: IP71 1/97
1997
other TS: double rectified absolute alcohol

UNEP PUBLICATIONSOECD SIDS
2. PHYSICAL CHEMICAL DATA
RemarkReliability
29.09.2003
2.14

:
:

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test performed by SG Redwood (UK) Ltd. ISO9002, no. Q4856.
(2) valid with restrictions(30)

ADDITIONAL REMARKS

Memo

:

Henry's Law ConstantRemark:Henry's constant = 0.000252 at 15C.
Substances with values for Henry's Constant <0.05 are:SourceReliability::

19.10.2004

1. unlikely to volatize from surface waters,
2. unlikely to off-gas from groundwater,
3. have a high vapour phase retardation.
U.S. EPA (1999) URL http://www.epa.gov/oar/caaac/mtbeethan.pdf.
(4) not assignable
No reference to published experimental work is given so the reliability ofthis value cannot be evaluated. However, the validity of this study,
published by the U.S. EPA, is highly dependent upon this value for Henry's
Law Constant.
(40)

Memo

:

Henry's Law ConstantRemark:Calculated value from HENRYWIN (EPIWIN suite)----------+---------------------------------------------+---------+---------CLASS | BOND CONTRIBUTION DESCRIPTION| COMMENT| VALUE----------+---------------------------------------------+---------+---------HYDROGEN | 5 Hydrogen to Carbon (aliphatic) Bonds || -0.5984
HYDROGEN | 1 Hydrogen to Oxygen Bonds|| 3.2318
FRAGMENT | 1 C-C|| 0.1163
FRAGMENT | 1 C-O|| 1.0855
FACTOR | * Non-cyclic alkyl or olefinic alcohol|| -.2000----------+---------------------------------------------+---------+---------RESULT | BOND ESTIMATION METHOD for LWAPC VALUE |TOTAL | 3.635----------+---------------------------------------------+---------+---------HENRYs LAW CONSTANT at 25 deg C = 5.67E-006 atm-m3/mole= 2.32E-004 unitless

Reliability
05.01.2004:--------+-----------------------------------------------+------------+-------|GROUP CONTRIBUTION DESCRIPTION| COMMENT |VALUE--------+-----------------------------------------------+------------+-------|
1 CH3 (X)|
| -0.62|
1 CH2 (C)(O)|| -0.13|
1 O-H (C)|| 4.45--------+-----------------------------------------------+------------+-------RESULT | GROUP ESTIMATION METHOD for LOG GAMMA VALUE |TOTAL | 3.70--------+-----------------------------------------------+------------+-------HENRYs LAW CONSTANT at 25 deg C = 4.88E-006 atm-m3/mole= 2.00E-004 unitless(2) valid with restrictions(45)

UNEP PUBLICATIONS

77

OECD SIDS
2. PHYSICAL CHEMICAL DATA78

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Memo

:

Henry's Law ConstantRemark:Reliability
19.11.2004:

Reference cites 3 values: an experimental log value of -3.59 (with noreference) and two calculated values using group and bond contribution
methods described in the reference. These are 3.70 and 3.72 respectively.
(4) not assignable(46)

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS

3.1.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

PHOTODEGRADATION

Type:Light source:
Light spectrum:
Relative intensity:
Conc. of substance:
INDIRECT PHOTOLYSIS
Sensitizer:
Conc. of sensitizer:
Rate constant:Degradation:
Deg. product:
Method
:
Year
:
GLP
:
Test substance:
Method:Result:Reliability:

Flag
19.10.2004:

Type
Light source
Light spectrum
Relative intensity
Spectrum of substance

:
:
:
:
:

Conc. of substance:
INDIRECT PHOTOLYSIS
Sensitizer:
Conc. of sensitizer:
Rate constant:Degradation:
Deg. product:
Method
:
Year
:
GLP
:
Test substance:
Remark

:

Air
Other
ca. 345 - 355 nm
based on intensity of sunlight
2 mg/l at 30 °C
other: Nox
1 mg/l= .045 cm³/(molecule*sec)= 20 % after 5 hour(s)other (measured): Photodegradation Test
1977
no datano data
Light source: UV fluorescent lamps.
Analytical methods: GC and UV spectroscopy.
Controls: unclear.
Light intensity 700 muW/cm^2.
Ethanol was irradiated for 5 - 6 hours in a 12 cubic metre "smog chamber"at 55% relative humidity. The amount of ethanol present was measured by
GC.
Indirect photolysis; t1/2 15.4 hours. % degradation results other than halflife: A 20% decrease in ethanol concentration was observed after 2 hr.
Rate constant calculated (1st Order assumed) 0.045 hr^-1 and half life 15.4
h. Ethanol ranked low on authors reactivity scale.
NO depletion rate: 2.3ppb/min(2) valid with restrictions
No data on hydroxy radical concentrations given.
Critical study for SIDS endpoint(47)
Airother: High pressure mercury lamp with water cooled pyrex filter> 290 nm
= 125 based on intensity of sunlightlambda (max, >295nm) : 182 nm:
epsilon (max):epsilon (295)
at 25 °C
other: O2, NOx, water
cm³/(molecule*sec)% afterother (measured)
1978
no dataother TS: analytically pure
Ethanol was irradiated in a 4 litre or 20 litre reactor in the presence ofcorresponding amounts of water, nitrogen dioxide and sulphur dioxide insynthetic air. The test concentrations used were 100, 500 and 1000 ppm(mg/l). Irradiation was for up to 10 hours. Samples were taken for analysis

UNEP PUBLICATIONS

79

OECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

every hour and the reduction in ethanol concentration was measured by
GC. Light intensity 125W; 25-30 degree C; samples were taken hourly.
Concentration of substance: 100, 500, 1000ppm.
Relative intensity in Watts relative to sunlight.
Result:Original paper in German.
No degradation of ethanol occurred in the presence of oxygen after
irradiation at >290 nm, but degradation was evident at >230 nm.
After irradiation for 4 hours in the presence of nitrogen dioxide, 30%
degradation occurred at 100 ppm, 55% at 500 ppm and 80% at 1000 ppm.
Half life calculated to be 13.8 hours.

Conclusion:

Reliability:

No degradation was seen in the presence of water at >290 nm.
Ethanol is not degraded in the troposphere by photodegradation in theabsence of sensitisers to promote indirect degradation.
(2) valid with restrictions
No data on hydroxy radical concentrations given. No data available onconcentration of sensitisers used.

19.10.2004

(48)

Type
:
Light source:
Light spectrum:
Relative intensity:
Conc. of substance:
INDIRECT PHOTOLYSIS
Sensitizer:
Conc. of sensitizer:
Rate constant:Degradation:
Deg. product:
Method
:
Year
:
GLP
:
Test substance:
Result:

Test condition:

Reliability:

19.10.2004

80

Air
Nm
based on intensity of sunlightat 19 °COH
= .0000018 cm³/(molecule*sec)% afterother (calculated)
1976
no datano data
A values for the half-life in a typical urban sunlit atmosphere was calculated
from the rate constant for the reaction of the hydroxyl radical with ethanol
vapour, assuming hydroxyl radical concentrations in such atmospheres tothe of the order of 10E-14 mol.dm-3. The calculated halflife was 10 hours.
The rate constant was 1.8 +/- 0.2 x 10^-9 dm^3mol^-1s^-1Hydroxyl radicals were generated in a reaction vessel in a "dark system" bychain reaction in a hydrogen peroxide/nitrogen dioxide/carbon monoxide
substrate mixture. No light source is used in this system.
(2) valid with restrictions
No detailed methodology provided.
(49)

Type
Light source
Light spectrum
Relative intensity

:
:
:
:

Air

Remark:This is a critical review of the available photodegradation data. Values
quoted are:Nm
based on intensity of sunlight

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS

Conclusion:

Reliability:

18.10.2004
Type
Light source
Light spectrum
Relative intensity
Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Method:Remark:ResultReliability
18.10.2004

:
:

Type
Light source
Light spectrum:::

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Rate constant k (E-12 cm3.mol-1.s-1)[temperature(K)]3.2+/-0.4 [292] Campbell et al
3.74+/-0.37 [296+/-2] Overend et al
2.62+/-0.36 [298] Ravishankara et al
3.5+/-0.6 [295+/-2] Cox et al
2.07[300] Meier et al
3.0 +/-0.6 [298] Lorenz et al
3.66 +/-0.42 [303+/-2] Kerr et al
3.4 +/-0.17 [293] Grenhill et al
3.33 +/-0.23 [296] Wallington et al
3.26 +/-0.14 [293] Hess et al
Following a critical review of the available data, the author concluded thatthe study by Hess et al (1988) was the preferred value forrecommendation. This study measured rate constants over thetemperature range 293-750K. An overall rate constant was calculated fromthe Hess data using a unit weighted least squares regression, yielding a
rate constant of 3.27E-12 at 298K with an estimated uncertainty of +/-20%.
(2) valid with restrictions
Whilst this is a secondary source, the data within it are critically evaluated.
The data reviewed presents a consistent picture and a weight of evidence
approach also confirms the conclusions reached.
(50)
Air
Nm
based on intensity of sunlightother (calculated)
2000
no dataas prescribed by 1.1 - 1.4
SAPRC-99 chemical mechanism linked with EKMA box model descriptionsof air pollution episodes in 39 urban locations. The EKMA approach
involves use of single-cell box models to simulate how ozone formation in
one day episodes is affected by changes in ROG and NOx inputs. Suchsingle-cell models cannot represent realistic pollution episodes in great
detail but they can represent dynamic injection of pollutants, time-varying
changes of inversion heights with entrainment of pollutants from aloft as
the inversion height increases throughout the day, and time-varying
photolysis rates, temperatures, and humidities. Thus, they can be used tosimulate a wide range of the chemical conditions which affect ozone
formation from reactive VOCs and NOx. These are the same as thoseaffecting VOC reactivity. The incremental reactivity is the change in ozone
formed caused by adding, in this case, ethanol to the initial and emittedbase reactive organic gas mixture in a scenario, divided by the amount of
VOC added.
Type: VOC reactivity scale.
This is an update of the SPARC-90 mechanism of Carter (1990) andincorporates recent reactivity data on the Maximum Incremental Reactivity(MIR) Scale from a wide variety of VOCs.
Absolute Value:
1.34 - 1.69 gm ozone/g VOC(4) not assignable(51) (52)Air
Nm

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81

OECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS
Relative intensity
Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:

based on intensity of sunlightYesother (calculated)
2000
no dataas prescribed by 1.1 - 1.4

Method:Remark:Calculated using the Harwell Photochemical Trajectory Model using the
updated IVL photochemical scheme, updated using the oxidation
mechanism of the acetates, as described in the Master Chemical
Mechanism oxidation mechanism. The model follows chemicaldevelopment in air parcels as they travel across from continental Europe to
the UK. Parcel dimensions are 10km squares to the top of the boundary
layer, the latter being 300m at 06:00 rising to 1300m by 14:00, falling fromearly evening back to 300m. The chemical inventory of the parcel is statedin the reference (95 hydrocarbons plus methane.) The model contains 771
thermal chemical reactions, with rate constants obtained from Atkinson(1989, 1990, 1992, 1994) or calculated using SAR (Atkinson(1986, 1987).
Type: Tropospheric ozone creation potential.
The photochemical ozone creation potential (POCP) concept can be
regarded as useful in locations with high NOx concentrations where therate limiting step in ozone creation is the VOC composition and reactivity.
Photochemical Ozone creation potential = 39.9% (Andersson-Skold) or
44.6 (Derwent) relative to ethylene at 100%, with ethanol representing
4.28% of total VOC emissions. POCP figures are considered to be to an
accuracy of +/-5. Ethanol is considered to degrade by reaction with OH
radicals and via carbon/carbon scission.
(4) not assignable(53) (54)Result:Reliability
18.10.2004:

Type
Light source
Light spectrum
Relative intensity

:
:
:
:

Air

Remark:Reliability
18.10.2004:

The estimated half-life of ethanol in the atmosphere ranges
from 4 to 5.9 days, based on a hydroxyl radical
concentration of 800,000 molecules/cubic centimeter.
(4) not assignable

Type:
Light source:
Light spectrum:
Relative intensity:
DIRECT PHOTOLYSIS
Halflife t1/2:
Degradation:
Quantum yield:
INDIRECT PHOTOLYSIS
Sensitizer:
Conc. of sensitizer:
Rate constant:Degradation:
Deg. product:
Method
:
Year
:
GLP
:

82

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Nmbased on intensity of sunlight

(35)
Air
Nm
based on intensity of sunlight= 3 day(s)% after= .0000000000035763 cm³/(molecule*sec)% afterother (calculated)

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS
Test substance:

Remark:Result:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Note: Half life based on 12 hour day.
Calculated value.
SUMMARY (AOP v1.90): HYDROXYL RADICALSHydrogen Abstraction= 3.4363 E-12 cm3/mol-sec
Reaction with N, S and -OH = 0.1400 E-12 cm3/mol-sec
Addition to Triple Bonds = 0.0000 E-12 cm3/mol-sec
Addition to Olefinic Bonds = 0.0000 E-12 cm3/mol-sec
Addition to Aromatic Rings = 0.0000 E-12 cm3/mol-sec
Addition to Fused Rings = 0.0000 E-12 cm3/mol-sec

Reliability
12.11.2004
3.1.2

:

OVERALL OH Rate Constant = 3.5763 E-12 cm3/mol-sec(2) valid with restrictions(45)

STABILITY IN WATERType
t1/2 pH4
t1/2 pH7
t1/2 pH9
Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Remark:Result:Duration (days) of test: Not relevant.
Positive/negative controls: Not relevant.
Analytical procedures used to measure test substance loss:
Not relevant.
Reference is to method. According to Lyman et al. bothalkanes and alcohols are resistant to hydrolysis.
Ethanol is not expected to undergo hydrolysis.

Reliability
19.10.2004:(This is deduced from basics according to Lyman et al.
(1990)).
(2) valid with restrictionsType
t1/2 pH4
t1/2 pH7
t1/2 pH9
Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Remark:Abiotic
at °C
at °C
at °C
other (calculated)
1990
no dataother TS: 100% ethanol(55)
Abiotic
at °C= 2 year at °C
at °C
other
no datano dataEthanol is stable in water and is not hydrolysed. Hydrolysis
as a function of pH is, therefore, an irrelevant endpoint.
Half-life calculated from the rate constant for thereaction with hydroxyl radicals (1.1 x 10sup9) at room
temperature (15 - 25 °C). Actual figure for half-life given
in ENVIROFATE database (Environmental Fate. SilverPlatter,

UNEP PUBLICATIONS

83

OECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

Chem-Bank, June1993).

Reliability
19.10.2004
3.1.3

(56)

STABILITY IN SOIL

TypeRadiolabel
Concentration
Soil temperature
Soil humidity
Soil classification
Year
Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Result:

Test condition:

Reliability
19.10.2004
3.2.1

84

:

Half-lives of between 334 days and 36.6 years have been
calculated for photooxidation in water based on the samerate constant (Handbook of Environmental Degradation Rates(1991) Eds Howard, P.H. et al. Lewis Publishers, Michigan).
(2) valid with restrictions:

Laboratory
32249 mg/kg
25 °C
Other

no data

Addition of ethanol to soil caused an immediate respiration
increase. When 14 mumoles of ethanol were added to soil, 12mumoles of oxygen were consumed before the soil respiration
dropped to the level of untreated soil. 81-92% of CO2
respired from the soil (in excess of that respired fromcontrol soil) was derived from the radiolabelled ethanol
substrate.
Soil tested was Drummer silty clay loam soil. Soil (20 g) ina glass dish was placed in a Warbiurg flask containing 2 ml
sodium hydroxide solution for absorbing carbon dioxide and 2
ml aqueous radiolabelled ethanol solution, which was added
directly to the soil.
Respiratory rate over 4 hr was evaluated by measuring O2
used and CO2 released. The amount of ethanol added waseither 14 or 40 mumoles.
(4) not assignable(57)

MONITORING DATA

Type of measurement
Media
Concentration
Method::::concentration at contaminated site
surface water

Method:Water sampled from Matsubara region of Tatsuno City. Samples divided
into 2, half (1.7l steam distilled with diethyl ether (100ml) for 3 days. Ether
then dried with anyhydrous Na2SO4 then concentrated. Remaining 2l
vacuum distilled at 10-4torr after freezing and receiver cooled at -80C.
Trapped water extracted with diethyl ether (300ml) for 24 hours, thenconcentrated.

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

Concentrates analysed by GC-MS. Column packed with Thermon-1500(5% polyeters, 5% starch, 0.3% phenolic resin on acid washed, DMCS
treated Chromasorb W). Temp 50C for 2 min then ramped to 210C at
4C/min. Injector temp 240C, flow rate (helium carrier) 40ml/min.
Measurements made in the Hayashida River in Tatsumo city, Japan (site ofleather industry). River recognised as one of the most polluted in Japan.
(typical COD 200ppm, BOD 380ppm).
Ethanol was detected (GC-MS) at a concentration of 4020 ppb.
(2) valid with restrictions(58)

Remark:ResultReliability
19.10.2004::

Type of measurement
Media
Concentration
Method::::background concentrationAir
ca. .001 mg/l

Method:Analysis in of the atmosphere of Point Barrow, Alaska.
200ml sample of air taken by gas tight syringe from a windward location. 4
fillings of a 50ml syringe taken over a period of 2 mins and injected into theinlet system of analyser. 25 measurements taken at hourly intervals over a
24 hour period on 2nd/3rd Sept 1967.
Cryogenic condensation trap used to concentrate organic components fromthe air, packed with Carbowax 20M, prior to injection system of GC
analyser (6ft, 20% Carbowax column, 78-82C, He carrier @ 22ml/min).
Trap release using water at 95C. Detector calibrated using acetone.
The authors postulated that as ethanol is a product of fermentation, thelikely source of the measured ethanol is natural.
Ethanol/methanol was detected in 17 of 25 samples. The
average concentration over 24 hours was 0.52 ppb (range 0 1.2 ppb). Using a conversion of 1.9ug/l=1ppb would imply a concentrationof 1.0ug/l.
(2) valid with restrictions(59)

Remark:Result:Reliability
19.10.2004:

Type of measurement
Media
Concentration
Method::::concentration at contaminated site
ground water

Method:Result:

VOC data were obtained from 20 Minnesota Pollution Control Agency
permitted municipal solid waste landfills and two unauthorised dumps. The
following wre taken: 6 leachate samples, 45 groundwater wells at 13
landfills with suspected contamination, 21 groundwater wells at 8 landfills
plus the unauthorised landfills where water quality was believed good.
Samples were collected in 40ml glass vials, no headspace, sealed withteflon lined caps. 40mg sodium thiosulphate added to each sample toremove residual chlorine. Quadruplicate samples used plus field blanks as
controls. Collection methodology described (either direct transfer tosample bottles or stainless steel ladle used.) The authors recognised thepotential for contamination from the plastic materials used in wellconstruction.
Ethanol was found in ground water suspected of leachate contamination(based on levels of inorganics) at 190ug/l at one of 13 sites, and was
detected at 58ug/l in 1 out of 7 landfill ground water samples where
inorganic levels indicated good or unknown water quality. It was found in 2
of 6 landfill leachates at concentrations in the range 23-110mg/l.

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Reliability:(4) not assignable
No data given on analytical methods which makes reliability assignment
not possible.
(60)

Type of measurement
Media
Concentration
Method::::background concentration
drinking water

Result:Reliability
19.10.2004:

Ethanol has been detected (not quantified) in U.S. citywater supplies, hotel drinking water (one of one tested)
and water treatment plants (one of three tested).
(4) not assignable

Type of measurement
Media
Concentration
Method::::background concentration
ground water

Remark:Reliability
19.10.2004:

Sources include animal wastes, plants, insects, forest
fires, microbes, and volcanoes.
(4) not assignable

Type of measurement
Media
Concentration
Method::::background concentration
Food

Result:Values for ethanol concentrations detected in foods are as
follows:
Lima, common, mung and soy beans - 1500 - 7900 ppm (average
4200 ppm)
Split peas - 3600 ppm
Lentils - 4400 ppm
Fried bacon - identified, not quantified as volatile
flavourcomponent

19.10.2004

(35)

(61)

Mountain Beaufort Cheese (French Alps) - identified, notquantified(4) not assignable

Reliability
19.10.2004:

Type of measurement
Media
Concentration
Method::::concentration at contaminated site
Air

Result:Reliability
19.10.2004:

Ethanol was detected (cryogenic trap, GC-FID) atconcentrations of 29 - 57 ppb (air pollution peak in Japan).
(4) not assignable

3.2.2

86

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

(35)

(62)

FIELD STUDIESUNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS
3.3.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS

Type
Media
Air
Water
Soil
Biota
Soil
Method
Year

:
:
:
:
:
:
:
:
:

fugacity model level III

Remark:Fugacity calculations based on the following emissions
scenario:
Air 1000kg/hr
Water 100kg/hr
Soil 10kg/hr.

56.6 % (Fugacity Model Level I)
34.3 % (Fugacity Model Level I)
9.1 % (Fugacity Model Level I)% (Fugacity Model Level II/III)% (Fugacity Model Level II/III)

These are calculated as follows:
TonnesFraction Release
ktpa
air
water
soil
Paint
25
0.8
0.15
0.01
Ink
70
0.95
0.05
0.01
Pharma
35
0.05
0.007
0.0001Food
5
0.01
0.0005 0.001
Flavour
2.5 0.01
0.0005 0.001
Fragrances 2.5 1
00Explosive
50.01 0.0005
0.001
Other
20
0.5
0.01
0
Cosmetic 145
100Detergent25
0.175 0.6
0.002
Deicing40100Cleaning
150.175 0.6
0.002
Other
10
0.175 0.6
0Intermed. 250
0.05
0.007
0.0001
Fuel
180
0.01
0.00050.001
Notes: Emission fractions either from EU technical guidance for risk
assessment or, where not available, by expert judgement. Explosive
emissions deemed similar to fuel use Food/flavour use - assumptions
based on consumption - little emitted - fuel factors used. Fragrances ascosmetics. Split flavour/fragrance tonnage 50/50. To obtain overall
emission rations, total emissions per compartment summed then ration
calculated and rounded to nearest power of 10.
Physicochemical data used:

ResultReliability
Flag
19.10.2004:::

Melting point: -114C
Boiling point: 78.2
LogKow: -0.31
water solubility: 1000g/l
vapour pressure: 59.3mmHg @ 25C
Henry's coefficient: 5E-6 atmm-3mol-1 @ 25C
pKa: 5.9 @ 25C
Total persistence time estimated as 80.3 hrs(2) valid with restrictions
Critical study for SIDS endpoint(63)

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88

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
Media
Air
Water
Soil
Biota
Soil
Method
Year

:
:
:
:
:
:
:
:
:

Volatility
soil - air% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level II/III)% (Fugacity Model Level II/III)

Remark:Reliability
19.10.2004:

Ethanol is relatively volatile and would therefore readily
evaporate from soil at the soil/air interface.
(4) not assignable

Type
Media
Air
Water
Soil
Biota
Soil
Method
Year

:
:
:
:
:
:
:
:
:

Volatility
water – air% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level II/III)% (Fugacity Model Level II/III)

Remark:Reliability
19.10.2004:

The estimated half life for evaporation of ethanol fromwater 1 m deep with a 1 m/sec current and 3 m/sec wind is
6.1 days.
(4) not assignable

Type
Media
Air
Water
Soil
Biota
Soil
Method
Year

:
:
:
:
:
:
:
:
:

Other
water – air% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level II/III)% (Fugacity Model Level II/III)

Remark:Reliability
19.10.2004:

Volatilization from Water:
Henry LC: 5E-006 atm-m3/mole (Henry experimental database)Half-Life from Model River:80.17 hours (3.34 days)
Half-Life from Model Lake :931.5 hours (38.81 days)(4) not assignable

Type
Media
Air
Water
Soil
Biota
Soil
Method
Year

:
:
:
:
:
:
:
:
:

(5)

(5)

(64)
Other
water – air% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level I)% (Fugacity Model Level II/III)% (Fugacity Model Level II/III)

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS
Remark:Reliability
19.10.2004

:

3.3.2

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

When released to the atmosphere, etheanol will photodegradewith a half life ranging from hours in polluted urban areas
to approximately 6 days in the atmosphere. Due to its
solubility in water, rainout may be an important process.
(4) not assignable(35)

DISTRIBUTION

Media
MethodYear:::

air - biota - sediment(s) - soil - waterCalculation according Mackay, Level III
2001

Remark:Adsorption coefficient: Not given.
Desorption: Not given.
Volatility: Not given.
Model used: EQC model of Mackay et al. (1996).
Version 1.01
Date: 1997.
The following input parameters were used:
MWt
Temp.
Water solubility

Result:46.09 g/mol
25 degC
716,000 g/m^3 calculated fromvapour pressure and Henry's law
constant of 5e-06 atm.m^3/mol(Gaffney, 1987)
Vapour Pressure
7870 Pa (59.03 mm)
Log Kow-0.31
Melting point-114 deg C
t1/2 air
203 hr (Graedel, 1978)
t1/2 water
182 hr (from biodegradation data)
t1/2 sediment210 hr (from biodegradation data)
Environmental conditions: left at the default values of themodel.
Air
13.0%
1.60e-8 mol/m^3 (738 ng/m^3)
Water
44.8%
2.75e-5 mol/m^3 (1271 ng/l)
Soil
42.1%
2.88e-4 mol/m^3 (8.3 ng/g)
Sediment
0.039% 9.50e-6 mol/m^3 (0.34 ng/g)
Adsorption coefficient, desorption and volatility not given.
At steady state 67% of additional inputs of ethanol are lost
through reactions and 33% are lost through advection.
(2) valid with restrictions

Reliability
19.10.2004

:

Media
Method
Year

:
:
:

Other

Remark:

PCKOCWIN v1.66 Results(65) (66)First Order Molecular Connectivity Index .......: 1.414
Non-Corrected Log Koc ..........................: 1.3755
Fragment Correction(s):1 Aliphatic Alcohol (-C-OH) .................: -1.5193

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OECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

Corrected Log Koc ..............................: -0.1438
Over Correction Adjustment to Lower Limit Log Koc: 0.0000
Estimated Koc: 1
19.10.2004

(64)

3.4

MODE OF DEGRADATION IN ACTUAL USE

3.5

BIODEGRADATION

Type
Inoculum
Contact time
Degradation
ResultKinetic of testsubst.

:
:
:
:
:
:

Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:

Method:Remark:Aerobic
other: wastewater from domestic sewage= 74 (±) % after 5 day(s)readily biodegradable
5 day(s) = 74 %10 day(s) = 74 %15 day(s) = 95 %20 day(s) = 84 %%other: BOD protocol
1974
no datano data
Biodegradability was measured in fresh water according to "standard
methods for the examination of water and waste water" 13th Ed. American
Public Health Association, New
York, 1971.
Three concentrations tested (3, 7 and 10 mg/l) with at least two in
duplicate. Tested concentrations gave a BOD of 3 to 30mg/l. Dissolved
oxygen measured periodically (5 times in 20 days).
Inoculum: wastewater from domestic sewage (assumed not adapted).

90

Result:Reliability:

Flag
19.10.2004:

Type
Inoculum
Contact time
Degradation
ResultKinetic of testsubst.

:
:
:
:
:
:

Deg. product:

COD measured.
Measured COD was 1.99 mg O2/mg. Theoretical oxygen demand
2.1 mg/mg.
(2) valid with restrictions
Well reported study. Not to GLP and no test substance purity or source
quoted.
Critical study for SIDS endpoint(67)
Aerobic
other: Filtered, settled domestic wastewater as seed in synthetic seawater= 75 (±) % after 20 day(s)readily biodegradable
5 day(s) = 45 %10 day(s) = 68 %15 day(s) = 72 %20 day(s) = 75 %%UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

MethodYear
GLP
Test substance

:
:
:
:

other: BOD protocol
1974
no datano data

Remark:Reliability:

Biodegradability measured in synthetic salt water according
to "standard methods for the examination of water and waste
water" 13th Ed. American Public Health Association, New
York, 1971. Three concentrations tested (3, 7 and 10 mg/l).
COD measured.
Inoculum (other): Seed used was developed in seawater takenfrom Lavaca Bay, Texas. The seed was maintained by adding
small amounts of settled raw wastewater periodically as asource of substrate. No further information available.
Concentration of test chemical, vehicle, pre-acclimation
conditions: 3, 7 and 10 mg/l ethanol was added using 0.1%
stock solution.
Temperature of incubation: Not specified.
Dosing procedure: Not described. Domestic wastewater wasplaced in bottles to which was added aerated dilution watercontaining ethanol.
Sampling frequency: BOD every 5 days; ethanol concentration
was not monitored.
Were appropriate controls and blank system used? Yes.
Analytical method: Cumulative oxygen uptake inethanol-amended and control samples was measured with
dissolved O2 meter.
Method of calculating measured concentrations: Degradation
rate was calculated as the % of theoretical oxygen demand
utilized.
Lag time: Not measured.
Observed inhibition: Not measured.
Excessive biodegradation: Not discussed.
Excessive standard deviation: Not discussed.
Time required for 10% degradation: Not discussed. At 5 days,
74% of ethanol had been degraded
Total degradation at end of test: 84% .
(2) valid with restrictions
Well reported study. Not to GLP and no test substance purity or source
quoted.
(67)

19.10.2004
Type
Inoculum
Concentration:::

Contact time
Degradation
ResultDeg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Remark:Aerobic
activated sludge, domestic
500 mg/l related torelated to= 37 (±) % after 1 day(s)readily biodegradable
other
1966
no dataother TS: Analytical grade
All sludges were capable of oxidizing ethanol as measured by
BOD which was 37.3% of maximum and similar to that for othershort-chain alcohols.
Inoculum (other): Fresh activated sludge: activated sludges

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92

Reliability
12.11.2004:

Type
Inoculum::

Concentration:

Contact time
Degradation
ResultDeg. product
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Method:Remark:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

were obtained from municipal treatment plants in Columbus,
Hilliard and Linworth, Ohio.
Concentration of test chemical, vehicle, pre-acclimation
conditions: 500 mg/l ethanol was added to 125 ml flasks
containing 20 ml blended sludge with a concentration of 2500
mg/l suspended solids.
Temperature of incubation: 20 deg C.
Dosing procedure: see above.
Sampling frequency: BOD 6, 12 and 24 h after inoculation.
Ethanol concentrations were not measured.
Were appropriate controls and blank system used? Yes.
Analytical method: Oxygen uptake measured by Warburg
respirometer.
Method of calculating measured concentrations: Not
discussed.
Lag time: Not discussed.
Observed inhibition: Not measured.
Excessive biodegradation: Not discussed.
Excessive standard deviation: Not discussed.
Time required for 10% degradation: Not discussed. At 6 hours
oxygen demand was 12.9% of theoretical.
Total degradation at end of test: 37.3% at 24 hrs.
(2) valid with restrictions(68)
Anaerobic
other: sediment and groundwater from methanogenic portion of shallow
anoxic aquifer contaminated by landfill leachate
50 mg/l related to Test substance
related to
30 day(s)= 91 (±) % after 30 day(s)readily biodegradable
other
1993
no datano data
Production of methane by ethanol-containing sediment wasmonitored by an automated pressure transducer system.
The acclimation period was 25-30 days.
Inoculum (other): Sediment and groundwater from amethanogenic portion of a shallow anoxic aquifer
contaminated by landfill leachate.
Concentration of test chemical, vehicle, pre-acclimation
conditions: 50 ppm C as ethanol. Ethanol was added toslurries of 50 g sediment and 75 ml groundwater in 160 ml
bottles.
Temperature of incubation: Room temperature.
Dosing procedure: Not described.
Sampling frequency: Not described.
Were appropriate controls and blank system used? Yes,
autoclaved controls.
Methane measurement: GC with flame ionization detector.
Method of calculating measured concentrations: Degradation
rate was calculated as the mean of three tests.
Lag time: Acclimation period was 25-30 days.
Observed inhibition: Not discussed.

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYS

Result:Reliability:

19.10.2004
Type
Inoculum
Contact time
Degradation
ResultKinetic of testsubst.

:
:
:
:
:
:

Control substance
Kinetic::

Deg. product
Method

:
:

Year
GLP
Test substance:::

Method:ETHANOL
ID: 64-17-5
DATE: 19.11.2004

Excessive biodegradation: Not discussed.
Excessive standard deviation: Not discussed.
Time required for 10% degradation: Not discussed. (Ratecalculated as 17.9 ppm C/day.
Total degradation at end of test: 91% of theoretical CH4
production was recovered.
The rapidity and completeness of ethanol biodegradation is
supported by the work of Corseuil et al., 1998 and by Yeh
and Novak (1994) in both of which complete mineralization
was achieved in aerobic or anaerobic conditions.
The rate of biodegradation was calculated to be 17.9 ppm
C/day and total methane recovery was 91% of the theoreticallimit.
(2) valid with restrictions
No detailed results available, only final result reported. Not GLP and no
substance source or purity information.
(69)
Aerobic
activated sludge= 96.8 (±2.4) % after 15 day(s)readily biodegradable
4 day(s) ca. 80 %8 day(s) ca. 88 %11 day(s) ca. 100 %15 day(s) ca. 92 %20 day(s) ca. 98 %Benzoic acid, sodium salt
1 day(s) ca. 10 %5 day(s) ca. 70 %OECD Guide-line 301 B "Ready Biodegradability: Modified Sturm Test
(CO2 evolution)"
1991
no dataother TS
The mineral medium used was adopted from that recommended in the1988 OECD Ring Test of Ready Biodegradability (OECD 301). Stocksolutions based on demineralised water. Stock solution (d) used EDTA asa preservative.
Since only about 1% of cells in activated sludge are active it wasconsidered that a 'cleaner' inoculum of similar activity could be obtainedusing secondary effluent from an activated sludge plant. The level oforganic carbon introduced when using 10% by volume of secondary
effluent to inoculate the test is only about 1-2 mg/litre. The test wastherefore inoculated with secondary effluent from an activated sludge plant
treating domestic sewage. The collected effluent was first passed through acoarse filter to remove gross particulate matter. The level of inorganiccarbon in the inoculum was reduced before use by sparging with carbon
dioxide-free air for about one hour while maintaining the pH at 6.5.
The CO2 produced in the control vessels, using the maximum inoculum
concentration of 10% secondary effluent, was in the range 0.4 - 1.3
mg/litre. Hence, in cases where positive results were obtained less than
10% of the carbon dioxide produced was derived from the control. The
inoculum was not pre-adapted.

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3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

Using suitable volumetric apparatus 100±1 ml of the mineral salts media isdispensed into '125 ml' Hypo-Vial [Pierce Warriner (UK) LtdI. The media isprepared so as to contain 0.5 to 10% by volume of inoculum and 2 to 10
mg/litre of test substance as organic carbon. Controls containing the same
inoculum concentration but no test compound are also prepared. The vials
are sealed with butyl rubber septa and aluminium crimp seals and placedon an orbital shaker in a temperature controlled environment. To follow thecourse of biodegradation and to statistically evaluate the extent of
biodegradation on the final day of the test a minimum of 12 vessels is
required per test substance. This provides for a data point every fourth day
and 4 replicates for the assessment of the final extent of biodegradation on
the 28th day of the test.
A vessel is removed from the shaker as required, a sample of theheadspace gas withdrawn using a gas syringe and the concentration ofcarbon dioxide determined. The seal is then broken and the concentrationof dissolved inorganic carbon (DIG) in the solution is measured
immediately. Similar determinations are made for a control vessel which
does not contain the test substance. The difference in the total inorganiccarbon found in the test and control vessels allows the quantity of carbon
dioxide produced from the test compound to be ascertained.
Positive control: sodium benzoate.
The determination of carbon dioxide in both gaseous and aqueous samples
was performed using a modified lonics 555 TC-T0C Analyser. Carbon
dioxide is released from aqueous samples of carbonate/bicarbonate bydirect injection using a 0-200 ~tl Hamilton constant rate syringe onto aninert support loaded with phosphoric acid. The temperature in the reaction
chamber is controlled at 150"C and pure nitrogen is used as the carriergas. The detection system is a high sensitivity non-dispersive infra-red
analyser. Gaseous samples are injected using a good quality gas-tightsyringe.
The analyser is calibrated for the analysis of gaseous samples by injecting
suitable volumes of a 0.25% v/v mixture of carbon dioxide in nitrogen. For
liquid samples the instrument is calibrated using standard solutions of
sodium hydrogen carbonate in the range 0 - 20 mg/I as DCC.
Remark:Ethanol used as a comparator volatile compound in a study of theapplicability of a modified form of the CO2 production test for assessing
ultimate biodegradability under aerobic conditions.
The test substance in a dilute mineral salts solution is incubated in seatedvessels with appropriate micro-organisms for a period of up to 28 days.
Only about two thirds of the volume of the vessel is filled with liquid. At thetest concentrations used only about 15% of the available oxygen in theheadspace gas is required for the complete oxidation of all test compoundcarbon to carbon dioxide.
Any carbon dioxide produced by the breakdown of the test material is
distributed between the liquid and gaseous phases. Periodically a vessel istaken, a sample of the headspace gas withdrawn using a gas syringe andthe concentration of carbon dioxide in the headspace gas determined. The
seal is then broken and the concentration of dissolved inorganic carbon(DCC) in the solution is measured. Similar determinations are made for acontrol vessel which does not contain the test substance. The difference inthe total inorganic carbon found in the test and control vessels allows thequantity of carbon dioxide produced from the test compound to beascertained. From a knowledge of the quantity of test material added and

94

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3. ENVIRONMENTAL FATE AND PATHWAYS

Result:

Test substance
Reliability::

19.11.2004

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

its carbon content the extent of mineralisation can be calculated.
BOD28 Mean = 96.8%. SD = 2.4
The method is shown to be compatible with existing techniques and isapplicable to the testing of insoluble and volatile compounds.
Reagent grade(2) valid with restrictions
No temperature quoted for study and data only available in graphical form,
otherwise well reported.
(70)

Type
Inoculum
Concentration:::

Contact time
Degradation
Result:::Remark:Reliability
19.10.2004:

Type
Inoculum::

Anaerobic

Remark:The anaerobic pseudo first order rate constants are
published as follows:

Aerobic
predominantly domestic sewage
2 g/l related to Test substance
related to
5 day(s)(±) % after

Ethanol was used as a comparator substance in the description of a new
potential test.
Oxygen utilization was immediate and attained a high rate early in the test
period. After 5 days the O2 uptake had passed the optimum rate and theslope of the O2 utilization curv matched that of the seed bank. Substrateutilization was demonstrated by a decrease in COD and TOC coincident
with the rapid O2 uptake. Bacterial growth reached a peak level of over
10^6 organismsa/ml on the third day of testing. Growth decreased
thereafter to less than 10^4 organisms/ml at the end of the test.
(4) not assignable
(71)

ion
rate constantNO3(-)
0.53
Fe(3+)
0.17
SO4(2-) 0.1
Reliability
19.10.2004:

Remark:Reliability
19.10.2004:

Remark:These are laboratory-derived values.
(4) not assignable(72)

It is shown that 80-100 mg/l degrades aerobically in 5 days
and anaerobically in 10-25 days.
(4) not assignable(73)

Ethanol is biodegraded in aerobic systems using activatedsludge, sewage (including filtered and settled),
wastewater,and soil inocula. Five day theoretical BOD values
range from37-86%. Anaerobic degradation (thermophilic
digestion, 54 °C) of ethanol (5 ml of a 5% aqueous ethanol
solution) produced approximately 1000 ml gas/g sample using
seed whichhad been prepared in a synthetic medium.

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Reliability
19.10.2004:

Remark:Reliability
19.10.2004:

Type
Inoculum
Contact time
Degradation
ResultControl substance
Kinetic

:
:
:
:
:
:
:

Remark:ResultReliability::

The aerobic half-life of ethanol in aqueous systems has been estimated tobe between 6.5 and 26 hours, based upon a river die-away test for one
sample of water from one river. The anaerobic half-life has been estimatedto be between 26 and 104 hours based upon estimated unacclimatedaqueous aerobic biodegradation half-life.
(4) not assignable(35)
Aerobic
activated sludge
89 (±) % after 14 day(s)Aniline%%Concentration of test substance: 100mg/l
Concentration of activated sludge (as the concentration of suspended
solid): 30mg/l
Volume of test solution: 300ml
Temperature: 25C
Initiation time before degradation started: 3 days
(4) not assignable
Results not available in detail and intermediate times only available ingraphical form. No details of test method given although believed to be to
an OECD protocol.
(74)

Deg. product
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:BIOWIN v4.01 Results - predictions::Linear Model: Biodegrades Fast
Non-Linear Model: Biodegrades Fast
Ultimate Biodegradation Timeframe: Days-Weeks
Primary Biodegradation Timeframe: Days
MITI Linear Model: Readily Degradable
MITI Non-Linear Model: Readily Degradable(4) not assignable

3.6

other: modelled data

(64)

BOD5, COD OR BOD5/COD RATIO

BOD5MethodYear
Concentration
BOD5

96

(4) not assignable(35)

19.10.2004

Reliability
19.10.2004

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

:
:
:
:

related to
mg/l

UNEP PUBLICATIONSOECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

GLP
COD
MethodYear
COD
GLP
RATIO BOD5 / COD
BOD5/COD:

no data::::

mg/g substanceno data:ca. .56

Remark:Result:Results from several studies of BODx values as a % of
theoretical oxygen demand (ThOD) are presented.
BOD Values for nonadapted inoculum
BOD5 (4 studies): 37, 44, 45, 74% ThOD (Mean 50%)
BOD
%ThoD (range for 3 studies)
10 days 44-74
15 days 71-95
20 days 75-84
BOD530 days
40 days
50 days%ThOD (1 study)
79
78
77

ThOD = 2.10
BOD5 (20 DegC) = 60% ThOD (acclimated)
COD (5 studies): 90,95,95,97,100% ThOD (Mean 95.4%)

Source:Reliability
22.06.2004:

3.7

BOD5/COD (calculated from above mean values) = 50/95.4 =0.52
Verschueren, K. (1996) Handbook of environmental data on organic
chemicals. 3rd Edition. Van Nostrand Reinhold Company, New York.
(4) not assignable

BIOACCUMULATION

Elimination
MethodYear
GLP
Test substance

:
:
:
:
:

RemarkReliability
19.10.2004

:
:

BCF
Elimination
MethodYear
GLP
Test substance

:
:
:
:
:
:

Remark:otherno data

Not expected to bioconcentrate.
(4) not assignable(5)
3.16
other: modelled

Bcfwin v2.15

UNEP PUBLICATIONS

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OECD SIDS
3. ENVIRONMENTAL FATE AND PATHWAYSETHANOL
ID: 64-17-5
DATE: 19.11.2004

Log Kow (estimated) : -0.14
Log Kow (experimental): -0.31
Log Kow used by BCF estimates: -0.31
Equation Used to Make BCF estimate:
Log BCF = 0.50
Correction(s):Value
Correction Factors Not Used for Log Kow < 1
Reliability
19.10.2004
3.8

Estimated Log BCF = 0.500(4) not assignable(64)

ADDITIONAL REMARKS

Memo

:

Air stripping constantRemarkReliability
19.10.2004::

K = 0.302/day at 2,220 mg/l(4) not assignable

Memo

:

Anaerobic sludge digestion

ResultReliability
29.09.2003::

Inhibited at 1600 mg/l.
(4) not assignable

Memo

:

Impact on biodegradation

Remark:Reliability
29.09.2003:

50% Inhibition of NH3 oxidation by Nitrosomonas at 4,100
mg/l.
(4) not assignable

Memo

:

Waste water treatment

Remark:Anaerobic lagoon:

influent effluent
mg/l
mg/l
13 lb COD/day/1000 cu.ft
80
35
22 lb " " " " " "270
120
48 lb " " " " " "270
130(4) not assignable

Reliability
19.10.2004

98

:

(15)

(15)

(15)

(15)

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY

4.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

ACUTE/PROLONGED TOXICITY TO FISH

Type
Species
Exposure period
Unit
LC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

StaticSalmo gairdneri (Fish, estuary, fresh water)
96 hour(s)mg/l= 13000 measured/nominal

Remark:Biological observations: None described.

No
other
1978
no datano data

Table of cumulative mortality: Not presented.
Lowest concentration causing 100% mortality: Not stated.
Mortality in controls: Not discussed.
Abnormal responses: Not discussed.
Reference substance: None used.
Any observations (e.g. precipitation) that might cause a
difference between measured and nominal values: Not
discussed.
Oncorhynchus mykiss and Salmo gairdneri are synonyms
The Columbia National Fisheries Research Laboratory
conducted aquatic toxicity tests on more than 400 chemicals
during 1965-1978; this is a major research area for the lab.
The lab. also participated in the development of standard
acute toxicity test methodology and only tests meeting
acceptable procedures were included in this compilation.
Test organism: Age fingerlings, Length not stated. Weight
0.8 g.
Loading <=0.8 g/l fish/litre.
Pretreatment: Acclimated for 1-3 day.
Dilution water source: Reconstituted deionized water.
Dilution water chemistry: hardness 40-50 mg/l CaCO3;
Alkalinity 30-35 mg/l CaCO3; pH 7.2-7.5.
Stock solution preparation: Not described.
Flow-through state: Static.
Vehicle, solvent and concentration: Not applicable
Solubility: Not applicable.
Exposure vessel type: 18.9 l wide-mouthed jars containing 15
l test solution, not aerated.
Illumination: not stated.
Replicates: 10 fish per concentration; no. of replicates notstated.
Water chemistry on test: Not described.
Test temperature range : 12 deg C +/- 1 degC.
Method of calculating mean measured concentration: Only
nominal concentrations used.
Reliability:

Statistical method: Litchfield and Wilcoxon (1949).
(2) valid with restrictions
Only nominal concentration measurements are available, However,fugacity data suggests low losses would be expected by evaporation. In
addition there are no water quality data available and no GLP data.

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OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
However, the data is regarded as reliable with restrictions.
Critical study for SIDS endpoint

Flag
17.11.2004:

Type
Species
Exposure period
Unit
LC100
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

flow throughPimephales promelas (Fish, fresh water)
96 hour(s)mg/l= 13480 measured/nominal

MethodRemark::Ethanol concentrations ranged up to 30,000 mg/l
Biological observations: Some fish lost equilibrium.
Table showing cumulative mortality: Not given.
Lowest dose causing 100% mortality: Not stated.
Mortality of controls: Not discussed.
Abnormal response: No abnormal response noted.
Reference substances: None.
Any other observations affecting concentration: None.
These data were collected by the EPA's Environmental
research Lab in Duluth, Minnesota, a lab likely to havesignificant experience in acute toxicity testing of thiskind. These data are rated 'probably reliable'.

(75)

No
other
1974
no dataother TS: Reagent grade

Test organism: Age Juvenile, 4-6 wk; Length 1-3.1 cm; WeightNot stated.
Loading: 20 fish/jar in 2 l test water.
Pretreatment: Acclimated to flowing water for 48 h.
Dilution water source: Lake Superior water.
Dilution water chemistry: Not stated.
Stock solution preparation: Ethanol weighed in to measured
water and whole shaken.
Vehicle, solvent and concentration: Not applicable
Solubility: Not applicable.
Stability: Not measured.
Exposure vessel type: Covered cyclindrical glass battery
jars.
Illumination: 50 ft-c cool, white fluorescent light 16
h/day.
Replicates: 10 fish per concentration, 1 replicate perconcentration.
Water chemistry on test: Not described. DOC kept to 4 mg/l
during test.
Test temperature range : 18-22 deg C.
Method of calculating mean measured concentration:
Concentrations not measured.
Result:Statistical method: Standard graphical.
This LC50 value was within 50% of values previously
reported. LC50 for shorter periods were as follows:
1 h >18,000 mg/l
24 h >18,000 mg/l
48 h =13,480 mg/l
72 h =13,480 mg/l

100

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Reliability
12.11.2004:

Type
Species
Exposure period
Unit
LC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

StaticPimephales promelas (Fish, fresh water)
4 day(s)mg/l> 100 measured/nominal
Yes
No
other
1986
no dataother TS: Reagent grade

Method:Remark:Result:Reliability
17.11.2004:

Juvenile Pimephales promelas fish (4-8 weeks) were exposedto ethanol for 96 hr. EPA method presumed.
Biological observations: Not discussed. Minnows wereconsidered dead if they were motionless and failed torespond to prodding.
Table showing cumulative mortality: Not given.
Lowest dose causing 100% mortality: 100% mortality notachieved at any dose.
Mortality of controls: Not discussed.
Abnormal response: No abnormal response noted.
Reference substances: None although several other substancestested.
Any other observations: None.
Test organism: Age juvenile, not specified; weight 0.2-0.5g.
Loading: <0.5 wet weight/l.
Pretreatment: Acclimated; food with-held 24 h.
Dilution water source: Activated carbon-filtered, dechlorinated andtempered Lake Ontario water.
Dilution water chemistry: hardness 130 mg/l CaCO3;
Alkalinity 93 mg/l CaCO3, pH 7.4, TOC 1.8 mg/l, TSS 180mg/l; salinity 26
mg/l Cl
Stock solution preparation: Not described. Max concentration tested
100mg/l.
Vehicle, solvent and concentration: Not applicable
Solubility: Not applicable.
Exposure vessel type: Unsealed cubic Pyrex chromatograph dishes.
Illumination: 50 ft-c cool, white fluorescent light 16h/day.
Replicates: 10 fish per concentration, 1 replicate per concentration.
Water chemistry on test: Not described. DOC kept below 40% of starting
value.
Test temperature range : 20 deg C +/- 0.1 deg C.
Method of calculating mean measured concentration: Only nominal
concentrations used.
Statistical method: Standard graphical.
The 96 hr LC50 was greater than 100 mg/l, the maximum
concentration tested.
(2) valid with restrictions
(77)

Type
Species
Exposure period
Unit
LC50
Limit test

:
:
:
:
:
:

(2) valid with restrictions(76)

StaticSalmo gairdneri (Fish, estuary, fresh water)
1 day(s)mg/l> 11200 measured/nominal

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4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:

No
other:
1978
no dataother TS

Method:Remark:Fish exposed to ethanol at 6 concentrations up to 30,000
mg/l for 24 hr.
Biological observations:
Table of cumulative mortality: Not presented.
Lowest concentration causing 100% mortality: in static
tests, 25,000 mg/l caused 100% mortality in 3 hr.
Mortality in controls: Not discussed.
Abnormal responses: Not discussed.
Reference substance: None used.
Any observations (e.g. precipitation) that might cause a
difference between measured and nominal values: None.
Test organism: Age fingerlings, 9.2 cm +/1 1.1 cm, weight 9.5 +/- 3.8 g.
Source Caribou Trout ranch, Soda Springs, Ihaho
Loading 1 fish/litre..
Pretreatment: Acclimated for at least 2 wk.
Dilution water source: Dechlorinated city tap water.
Dilution water chemistry: hardness 90 mg/l CaCO3; Conductivity 190
muS/cm, pH 8.0.
Stock solution preparation: Not described.
Vehicle, solvent and concentration: Not applicable
Solubility: Not applicable.
Exposure vessel type: PET-lined 20 l vessels.
Illumination: 12 h light: 12 h dark cycle.
Replicates: 10 fish per concentration, 1 replicate per concentration.
Water chemistry on test: Not described.
Test temperature range : 10 deg C.
Method of calculating mean measured concentration: Only nominal
concentrations used.
Criterion for death: cessation of respiration.

Result:

Test substance
Reliability::

17.11.2004
Type
Species
Exposure period
Unit
LC50
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

102

:
:
:
:
:
:
:
:
:
:
:
:

Statistical method: Litchfield (1949 and APHA (1971).
Nominal concentrations achieved: 0.1, 1.0, 10 and 100 mg/l.
LC50 11,200 mg/l (not achieved).
Reagent grade ethano from Standard Chemicals.
(2) valid with restrictions
This was a screening study for a flow through sub-lethal study. Limited
results are reported. Design was static with no measured concentrations,however short length of study limits impact of these omissions. Study rated
reliable with restrictions.
(78)
Other
Pimephales promelas (Fish, fresh water)
96 hour(s)g/l= 14.2= 14.2Yesother:USEPA methodology
1984
no dataother TS

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Remark:A 96-hour LC50 or 15.3 g/l and a 96-hour EC50 or 12.9 g/lwere recorded in an earlier study under the same condition
but with 95% ethanol supplied by the U.S. Industrial
Chemical Co. Bacterial growth appeared in the exposuretanks. Affected fish were hypoactive and lost equilibrium
prior to death.
fathead minnows were cultureds from brood stock provided by
the USEPA Environmental Research Laboratory (Duluth).
Adults were maintained in flow-through st 25 degC with a 16
-h light/dark photoperiod. Organisms were were fed adultbrine shrimp (Artemia). Fry were fed freshly hatched brine
shrimp nauplii three times daily until 24-h before test
start.
Fish were not fed during exposure to ethanol. 2 replicatesof 25 fish were exposed to each of 5 test concentrations andan untreated control in a flow-through system. The tankvolume was 6.3 l and the volume additions was 6.46 vol/day.
Test fish were 29-30 day-old at start and had a mean lengthof 18.2 +/- 2.22 mm and a mean weight of 0.106 +/- 0.036 g.
The loading rate was 0.421 g/l. Reconstituted of filtered
Lake Superior water was used for control and dilution water.
Water had hardness of 45 mg/l as CaCO3 and an alkalinity of
37.0 mg/l as CaCO3.
Nominal (and range of average measured) concentrations(mg/l) tested were 0 (5.4-8.5), 128 (44-48), 214 (77-79),
356 (130-137), 594 (235-263) and 990 (398-420). Test
temperatures ranged 23.1-25.5 degC. pH ranged 7.3-7.5 SU anddissolved O2 ranged 6.1-7.4 mg/l. Test concentrations in one
replicate were measured daily by GLC.
Mortality and adverse effects were reported at 0, 12, 21,
24, 48, 52, 72 and 96 hr of exposure and the 96-h EC50/LC50
and 95% confidence limits were estimated using the Trimmed
Spearman Karber method.
Test substance was 95% pure ethanol (Aldrich Chemical Co.).
(2) valid with restrictions

Test substance
Reliability
12.11.2004

:
:

Type
Species
Exposure period
Unit
LC0
LC50
LC100
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

StaticLeuciscus idus melanotus (Fish, fresh water)
48 hour(s)mg/l= 7110= 8140= 8690Remark:Reliability
11.11.2004:

Method specified in Deutsche Einheitsverfahren zur Wasser-,Abwasser und Schlamm-Untersuchung L15; Fischtest. Study
duration not specified in reference, but authors refer toMann, H. (1975) Vom Wasser 44, 1 - 3 for details of method.
This specifies 48 hours.
(4) not assignable(79)

no dataother1978
no datano data(80)

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103

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
Species
Exposure period
Unit
LC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

StaticAlburnus alburnus (Fish, estuary)
96 hour(s)mg/l= 11000

Remark:Test condition:

Test substance
Reliability
11.11.2004::

A narcotic effect (loss of equilibrium) was seen in the
fishas an early response, followed by coma and then death.
Fish were tested in filtered brackish water (10/tank) at 10°C in static tanks. At least 6 concentration and one
control were tested. Mortality was recorded and the LC50
was determined by probit analysis.
The test substance was analytical grade.
(4) not assignable

Type
Species
Exposure period
Unit
LC100
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

StaticSemolitus atromaculatus (Fish, fresh water)
24 hour(s)mg/l= 7000 - 9000 measured/nominalYes
no dataother1952
no dataother TS: Commercial grade

Remark:This study was to evaluated fish toxicity as a means ofdetermining the critical range of wastewater contaminationfor 51 different chemicals including ethanol.

no dataotherno dataother TS(81) (82)

Critical range is the concentration range (PPM) above which
all four fish died in 24 hours and below which all 4
survived over the same period.
(4) not assignable

Reliability
11.11.2004:

Type
Species
Exposure period
Unit
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

StaticOryzias latipes (Fish, fresh water)
15 minute(s)mg/l= 1000 - 10000

Remark:Concentration assumed; endpoint is for physiology; EC50
value assumed. Other fish in same battery of tests were

104

(83)

no dataotherno dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Girella punctata, Chasmichthys dolichognathuis and Pagrus
major.
(4) not assignable

Reliability
11.11.2004:

Type
Species
Exposure period
Unit

StaticCarassius auratus (Fish, fresh water)
30 minute(s)g/l=1Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
11.11.2004:

Results expressed as 1% (v/v or w/w not known). Endpoint isbehaviour (temperature selection).
(4) not assignable

4.2

(84)

no dataotherno dataas prescribed by 1.1 - 1.4(85)

ACUTE TOXICITY TO AQUATIC INVERTEBRATESTypeSpecies
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticCeriodaphnia sp. (Crustacea)
48 hour(s)mg/l= 5012 measured/nominal
no dataother: ASTM (see ME)
1987
no dataother TS: Absolute ethanol

Remark:Method was that recommended by the American Society ofTesting and Materials (1980) Standard practice for
conducting acute toxicity tests with fishes,
macroinvertebrates and amphibians. ASTM Standard E729-80.
Philadelphia, Pennsylvania.
Test organism: Cerodaphnia dubia: Source not specified.
Organisms were mass cultured an acclimated to temperature
for at least 10 weeks and maintained in filtered, autoclaved
Lake Huron water. Neonates hatched by isolated gravid
females gathered by sieving.
Age: Neonates.
Control group: Dilution water controls.
Test conditions: Ethanol not discussed.
Test temperature: 24 deg C.
Exposure vessel: Covered vials, not aerated, triplicates foreach concentration.
Dilution water source: See above.
Dilution water chemistry: Hardness 90 mg/l as CaCO3,
Alkalinity 70 mg CaCO3, pH 8.8, TOC 5580 mug/l; TDS 140,000
mug/l; Ca/Mg 2.8. Na/K 4.3.
Lighting: 646 lux +/- 85; 16 hr light, 8 hr dark.
Water chemistry on test: Dissolved Oxygen 8.4-10.3 mg/l, pH

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OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
8.2-8.4.
Endpoint assessment: Assessed microscopically.
Test design: Ten individuals per beaker, 3 replicates perconcentration. Concentrations of ethanol not specified.
Method of calculating mean measured concentration: Not
discussed; Geometric mean LC50's calculated.
Statistical method: Thompson moving averages.
Species: Ceriodaphnia dubia
Biological observations:
Number immobilized as compared to number exposed; Not
discussed.
Concentration response with 95% confidence limits (LC50)
5012 mg/l (4233-6913 mg/l).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
LC50 values ranged from 6325 to 6772 mg/l at 20 degree C and from 3715
to 6076 mg/l at 24 degree C.
Test substance was pure (absolute) ethanol (dehydrated
U.S.P.).
(2) valid with restrictions
These data are regarded as reliable. Fugacity data suggests low losses
would be expected by evaporation, however as no measurements made,only rated reliable with restrictions.
Critical study for SIDS endpoint(86)

Result:

Test substance:

Reliability:

Flag
11.11.2004:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticDaphnia magna (Crustacea)
48 hour(s)mg/l= 12340 measured/nominal
no dataother: ASTM (see ME)
1987
no dataother TS: Absolute ethanol

Remark:Biological observations:
Number immobilized as compared to number exposed; Not
discussed.
Concentration response with 95% confidence limits (LC50)
1,2340 mg/l (11,065-13,948 mg/l).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
LC50 values ranged from 11853 to 13248 mg/l at 20 degree C
and from 9268 to 14221 mg/l at 24 degree C.
Method was that recommended by the American Society ofTesting and Materials (1980) Standard practice for
conducting acute toxicity tests with fishes,
macroinvertebrates and amphibians. ASTM Standard E729-80.

106

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Philadelphia, Pennsylvania.
Test organism: Daphnia magna: Source not specified. Stocks
maintained in adjusted, autoclaved, aerated Lake Huron waterfor 3 years before start of study. Neonates hatched byisolated gravid females gathered by sieving.
Age: Neonates.
Control group: Dilution water controls.
Test conditions: Ethanol not discussed.
Test temperature: 20 deg C.
Exposure vessel: Covered beakers, not aerated, triplicatesfor each concentration.
Dilution water source: See above.
Dilution water chemistry: Hardness 160 mg/l as CaCO3, pH
8.0, TOC 5520 mug/l; TDS 289,550 mug/l; Ca/Mg 5.7. Na/K 4.5.
Lighting: 1916 lux +/- 75; 16 hr light, 8 hr dark.
Water chemistry on test: Dissolved Oxygen 7.6-8.9 mg/l, pH
7.8-8.4.
Endpoint assessment: Assessed microscopically.
Test design: Ten individuals per beaker, 3 replicates perconcentration. Concentrations of ethanol not specified.
Method of calculating mean measured concentration: Not
discussed; Geometric mean LC50's calculated.

Test substance:

Reliability:

11.11.2004
Type
Species
Exposure period
Unit
LC50

Statistical method: Thompson moving averages.
Test substance was pure (absolute) ethanol (dehydrated
U.S.P.).
(2) valid with restrictions
These data are regarded as reliable. Fugacity data suggests low losses
would be expected by evaporation, however as no measurements made,only rated reliable with restrictions.
(87)

Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

other: not specified
Artemia salina (Crustacea)
1 day(s)mg/l= 1833 measured/nominal=
no dataother1994
no datano data

Method:Remark:Artemia salina 24 h nauplius larvae were exposed tounspecified nominal concentrations of ethanol for 24 h.
Biological observations:
Number immobilized as compared to number exposed; Not
discussed.
Concentration response with 95% confidence limits (LC50)
1,834 mg/l (1,324-2,538 mg/l).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Test organism: Artemia salina hatched from dry eggs supplied
by San Francisco Bay Brand hafter hydration in distilled

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OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
water. Cysts were incubated. in synthetic sea water for 24 h
at 25 deg C with continuous side illumination and slightaeration.
Age: 24-h-old nauplius larvae.
Control group: Used but not described.
Test conditions: Synthetic seawater was porepared using 35%
Synthetica sea salt and distilled deionized seawater.
Test temperature: 25 deg C.
Exposure vessel: Plastic 16 mm petri dishes.
Dilution water source: See above.
Dilution water chemistry: Not described.
Lighting: Incubated in the dark.
Water chemistry on test: Not discussed.
Endpoint assessment: Organisms considered dead if they did
not move during 10 sec observation.
Test design: Ten larvae per dish, 3-5 replicates perconcentration; experiment repeated 5 times. Concentrationsof ethanol not specified.
Method of calculating mean measured concentration: Nominal
concentrations only.

Result:Statistical method: Litchfield and Wilcoxon.
Further studies involved older larvae:
48 h LC50 850 mg/l
72 h LC50 695 mg/l
Sensitivity to ethanol was therefore age-related.
(2) valid with restrictions

Reliability
11.11.2004:

Type
Species
Exposure period
Unit
LC50
Limit Test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Staticother: Paramecium caudatum (ciliate Protozoon)
4 hour(s)mg/l= 5840No
no dataother1989
no dataother TS

Remark:Age of test species: 48hrs
Control group: None mentioned. A number of other solvents tested as wellas pesticide compounds.
Test conditions: Stock solutions split-pea medium.
Test temperature range 25 deg C +/- 2 deg.
Exposure vessel: not specified. Test in 100 ml containing 1 ml of culture
containing 1,500-2000 stationary phase organisms/ml.
Dilution water source: not specified .
Dilution water chemistry: Not measured.
Lighting: not specified.
Water chemistry on test: Not measured. Medium was Chalkley's isotonic
inorganic salt (Patterson, 1982)
Endpoint assessment: lethal concentration at which all animals died in 10
mins; death indicated by lack of swimming and or rupture of cell. Also, 4 hr
median lethal concentraton (LC50).
Replicates: 5
Number of concentrations for 4hr LC50 evaluation: 0.1, 0.2, 0.4, 0.8, 1.2, 2

108

(88)

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY

Result
Test substance
Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:::

17.11.2004% v/v (790 to 15,800 mg/l)
10 minute LC50=5% v/v (39,000mg/l)
commercial absolute alcohol(2) valid with restrictions
Method details not comprehensive. Results available for eachconcentration but only in graphical form. However, study sufficiently wellreported to rate reliable.
(89)

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Staticother aquatic crustacea: Palaemonetes kadiakensis
18 hour(s)g/l= 10.1 measured/nominal
No

Method:Remark:Organisms exposed to 4 concentrations of ethanol in therange 1% v/v to 2% v/v. Test conducted at 23 degC.
Biological observations:

Result:Reliability
11.11.2004:

Type
Species
Exposure period

:
:
:

1981
no dataas prescribed by 1.1 - 1.4

Number immobilized as compared to number exposed; mortality
ranged from 0 to 100%. Concentration response with 95%
confidence limits (LC50) 1.28% v/v (1.18-1.38).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Test organism: Palaeomonetes kadlakensis caught in a nearby
lake.
Age: Juveniles.
Control group: None mentioned.
Test conditions: Stock solutions preparation not discussed.
Test temperature range 23 deg C +/- 1 deg.
Exposure vessel: 2 litre beakers containing 100 ml test
medium. Each dilution tested in duplicate.
Dilution water source: Aerated deionized deep well water.
Dilution water chemistry: Not measured.
Lighting: 1 h of typical fluorescent light illumination,
15.5 h 10% normal illumination then 1.5 h typicalillumination.
Water chemistry on test: Not measured.
Endpoint assessment: Organisms considered dead if they did
not respond to light, sound vibration or gentle probing.
Test design: Five organisms per beaker, two beakers perconcentration, at least 5 concentrations of ethanol.
Method of calculating mean measured concentration: Not
described.
Nominal concentration: Range from 1% v/v to 1.5% v/v
according to graph. Measured concentration: Not measured.
Statistical method: Probit.
LC50 quoted as 1.28% v/v (1.18 to 1.38) which is equivalent to 10.1 (9.3 to10.9) g/l. Control response not known.
(2) valid with restrictions(90)
StaticDaphnia pulex (Crustacea)
18 hour(s)UNEP PUBLICATIONS

109OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

g/l
= 12.1 measured/nominal
No

Method:Remark:Organisms exposed to 4 concentrations of ethanol in therange 1% v/v to 2% v/v. Test conducted at 23 degC.
Biological observations:

1981
no dataother TS

Number immobilized as compared to number exposed; mortality
ranged from 0 to 100%. Concentration response with 95%
confidence limits (LC50) 1.53% v/v (1.17-1.80).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Test organism: Daphnia pulex caught from a nearby pond.
Age: less than 24 h old when used.
Control group: None mentioned.
Test conditions: Stock solutions preparation not discussed.
Test temperature range 23 deg C +/- 1 deg.
Exposure vessel: 50 ml culture tubes containing 25 ml test
medium. Tubes loosely capped, not aerated. Each dilution
tested in duplicate.
Dilution water chemistry: Not measured.
Lighting: 1 h of typical fluorescent light illumination,
15.5 h 10% normal illumination then 1.5 h typicalillumination.
Water chemistry on test: Not measured.
Endpoint assessment: Organisms considered dead if they did
not move after being swirled under a light.
Test design: Ten organisms per tube, two tubes perconcentration, at least 4 concentrations of ethanol.
Method of calculating mean measured concentration: Not
described.
Nominal concentration: Range from 1% v/v to 2% v/v according
to graph. Measured concentration: Not measured.
Statistical method: Probit.
LC50 reported as 1.53% v/v (1.17 to 1.80) which is equivalent to 12.1 (9.2
to 14.2) g/l. Control response not known.
USP grade, 95% ethanol(2) valid with restrictions(90)

Result:

Test substance
Reliability
11.11.2004

:
:

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Staticother aquatic crustacea: Hyalella azteca
18 hour(s)g/l= 8.2 measured/nominal
No

Method:Organisms exposed to 4 concentrations of ethanol in therange 1% v/v to 2% v/v. Test conducted at 23 degC.

110

1981
no dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY
RemarkETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Biological observations:
Number immobilized as compared to number exposed; mortality
ranged from 20 to 100%. Concentration response with 95%
confidence limits (LC50) 1.04% v/v (0.761-1.28).
Cumulative immobilization: Not discussed.
Satisfactory control response?: Unknown.
Test organism: Hyalella azteca caught from a nearby slough andmaintained in aquaria with added aerated water and aeration.
Age: Juveniles with 14-16 antenna segments
Control group: None mentioned.
Test conditions: Stock solutions preparation not discussed.
Test temperature range 23 deg C +/- 1 deg.
Exposure vessel: 400 ml beakers containing 100 ml test medium. Beakers
covered with aluminium foil. Each dilution tested in duplicate.
Dilution water source: Aerated deionized deep well water.
Dilution water chemistry: Not measured.
Lighting: 1 h of typical fluorescent light illumination, 15.5 h 10% normal
illumination then 1.5 h typical illumination.
Water chemistry on test: Not measured.
Endpoint assessment: Organisms considered dead if they did not respondto light, sound vibration or gentle probing.
Test design: Ten organisms per beaker, two beakers per concentration, atleast 5 concentrations of ethanol.
Method of calculating mean measured concentration: Not described.
Nominal concentration: Range from 0.8% v/v to 2% v/v according to graph.
Measured concentration: Not measured.
Statistical method: Probit.
LC50 reported as 1.04% v/v (0.761 to 1.28) which is equivalent to 8.2 (6.0
to 10.1) g/l. Control response not known.
(2) valid with restrictions(90)

Result:Reliability
11.11.2004:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticArtemia salina (Crustacea)
1 day(s)mg/l= 23874
no dataother: Artoxkit M
1992
no dataother TS

Method:This test used the Standard Operating Procedures for the ARTOXKIT M
test modified as follows:
The procedures of Vanhaecke (1980) and Vanhaeck & Persoone (1981)
were followed for the hatching of the cysts and collection of the nauplii. For
moulting of instar I to instar II-III larvae the nauplii were transferred after
18-24 h from the start of cyst rehydration to 100 ml Erlenmeyer flasks
containing fresh artificial seawater with continuous aeration and illumination
for a further 24 hr.
LC50s were calculated with the corresponding 95% confidence limits using
the trimmed Spearman Karber method.
Control: Sodium dodecyl sulphate.
An artificial seawater (35 g/l salt) was used:

UNEP PUBLICATIONS

111

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
NaCl 23.9g/l
MgCl2.6H20 10.83g/l
CaCl2 1.15g/l
SrCl2.6H2) 4mg/l
KCl 682mg/l
KBr 9.9mg/l
Na2SO4 9.06mg/l
NaHCO3 200mg/l
NaF 0.3mg/l
H3BO3 2.7mg/l
The EC50 for artemia was 519 mmol/l +/- 29.1 (23.874g/l +/- 1.33).
analytical grade from Sigma Chemical Company.
(2) valid with restrictions

Result
Test substance
Reliability
11.11.2004

:
:
:

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticDaphnia magna (Crustacea)
1 day(s)g/l= 10.7 measured/nominal
no data
OECD Guide-line 202

Method:Result
Test substance::

Reliability
11.11.2004:

1984 version of OECD method. 3 replicates. Statistics: trimmed
Spearman-Karber method. No further data given.
Result quoted as 233mmol/l. No confidence limits given.
No specific data. At least 97% pure, bbut possibly >99% or pharmacopia
purity.
(4) not assignable
(92)

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Staticother: Brachionus calyciflorus
1 day(s)g/l= 29.6 measured/nominal
no dataother: Rotoxkit F

Method:This test used the Standard Operating Procedures for the ROTOXKIT F
test modified as follows:(91) (92)

no dataother TSno dataother TSThe contents of a vial containing B. calyciflorus cysts was emptied into asmall disposable polystyrene Petri dish. 5 ml of EPA water was thenadded after which the whole was covered and incubated at 25C +/-1 in light(19.5uE.m-2) for 18-20hrs. 24 well plates used.
LC50s were calculated with the corresponding 95% confidence limits using
the trimmed Spearman Karber method.
Control: Potassium dichromate
EPA water (<2 weeks old and continuously aerated):

112

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
KCl 4mg/l
NaHCO3 296g/l
MgSO4 60mg/l
CaSO4.2H2O 60mg/l
Result quoted as 644mmol/l (+/-40.6) which is equivalent to 29.6g/l (+/-1.9).
analytical grade from Sigma Chemical Company.
(2) valid with restrictions(91) (92)

Result
Test substance
Reliability
11.11.2004

:
:
:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Staticother: Brachionus plicatilis
1 day(s)g/l= 35.4 measured/nominal
no dataother: Rotox M

Method:This test used the Standard Operating Procedures for the ROTOXKIT M
test modified as follows:

no dataother TSThe contents of a vial containing B. plicatilis cystes , suspended in a saline
medium of 55g/l, were poured into a simall disposable polystyrene Petridish. 5ml of dieonised water was added to bring the salinity of the hatching
medium to 15g/l. The whole was incubated at 25C+/-1 in light (19.5uE.m2) for 24-28 hours. 24 well plates used.
LC50s were calculated with the corresponding 95% confidence limits using
the trimmed Spearman Karber method.
Control: Cupric sulphate.

Result:

Test substance
Reliability
11.11.2004

:
:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

An artificial seawater (15 g/l salt) was used:
NaCl 11.32g/l
MgCl2.6H20 1.97g/l
CaCl2 0.54g/l
KCl 0.36mg/l
MgSO4.7H2O 2.39g/l
NaHCO3 70mg/l
H3BO3 10mg/l
Quoted LC50 was 770 mmol/l (+/- 34.5) which is equivalent to 35.4 (+/- 1.6)
g/l.
analytical grade from Sigma Chemical Company.
(2) valid with restrictions(91) (92)
Staticother: Streptocephalus proboscideus
1 day(s)g/l= 18.8 measured/nominal
no dataother: Streptox F
no dataother TS

UNEP PUBLICATIONS

113

OECD SIDS
4. ECOTOXICITY
MethodETHANOL
ID: 64-17-5
DATE: 19.11.2004:

This test used the Standard Operating Procedures for the STREPTOXKITF test modified as follows:
Hatching of the cysts was initiated 24hrs before the start of the test, incylindrico-conical tube containing 100-125ml of EPA water at 25C +/-1 inlight (19.5uE.m-2) and aerated. As described by Centeno (1992) hatchedlarvae (intstar I) were tranferred afte 16-18hrs from the start of therehydration into Erlenmeyer flasks containing fresh medium and incubatedfor a further 5-7hrs to moult to the intart II-III stage.
Control: Potassium dichromate
LC50s were calculated with the corresponding 95% confidence limits using
the trimmed Spearman Karber method.

Result:

Test substance
Reliability
11.11.2004

:
:

Type
Species
Exposure period
Unit
IC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Result:Reliability
21.08.2003:

Type
Species
Exposure period
Unit
EC50
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

114

EPA water (<2 weeks old and continuously aerated):KCl 4mg/l
NaHCO3 296g/l
MgSO4 60mg/l
CaSO4.2H2O 60mg/l
Quoted LC50 of 409mmol/l (+/- 12.9) which is equivalent to 18.8g/l (+/0.6).
analytical grade from Sigma Chemical Company.
(2) valid with restrictions(91) (92)
Other
1 day(s)mg/l= 13100 - 20900
no dataother1995
no dataas prescribed by 1.1 - 1.4
IC50 is for population growth in this freshwater protozoan,
Tetrahymena pyriformis (Ciliata).
Three incubation periods were evaluated with the following
results:
3 hour 20900 mg/l
6 hour 17700 mg/l
9 hour 13100 mg/l(4) not assignable
(93)
Other
1 day(s)mmol/l= 258= 590
no dataother1999
no dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Remark:EC50 values are for development inthe protozoan Spirostomum
ambiguum.
(4) not assignable

Reliability
21.08.2003:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Reliability
21.08.2003:

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticPalaemonetes pugio (Crustacea)
4 day(s)g/l= 12.07 measured/nominal
no dataother1997
no dataas prescribed by 1.1 - 1.4

Method:Age at start: Adult male and female grass shrimps from local
estuaries, Gulf Breeze, Fla, USA.

(94)
other aquatic crustacea
1 day(s)mg/l= 31700
no dataother1994
no dataas prescribed by 1.1 - 1.4
The species used was the Fairy shrimp, Streptocephalus
rubricaudatus.
(4) not assignable(95)

Acclimation: to 25 Cel and 20 ppt salinity for 2 weeks.
Embryos: Collected for test at embryo cap stage.
Ethanol dosage: increasing concentrations (range not given)
Duration of exposure: 12 days
Result:Conclusion:

Reliability
29.09.2003:

Type
Species
Exposure period
Unit
EC50
Analytical monitoring

:
:
:
:
:
:

Mortality and hatching recorded daily.
Mean LC50 at 4 days: 12.07 g/l
Mean LC50 at 12 days: 3.63 g/l
If ethanol is used as a solvent in this developmental
toxicity test it should not exceed 1 g/l in the test
solution.
(4) not assignable(96)
Daphnia sp. (Crustacea)
1 day(s)mg/l= 12300 - 13400
no data

UNEP PUBLICATIONS

115

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

MethodYear
GLP
Test substance

:
:
:
:

other
1996
no dataas prescribed by 1.1 - 1.4

RemarkReliability
21.08.2003::

The 2 day range was 10100 to 11200 mg/l.
(4) not assignable

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Daphnia pulex (Crustacea)
1 day(s)mmol/l= 251.07
no dataother1995
no dataas prescribed by 1.1 - 1.4

Reliability
21.08.2003:(4) not assignable

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Reliability
11.11.2004:

Type
Species
Exposure period
Unit
LC100
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Daphnia magna (Crustacea)
2 day(s)g/l=1
no dataother1995
no dataas prescribed by 1.1 - 1.4

RemarkReliability
21.08.2003::

Unit of concentration expressed as % (v/v or w/w not known).
(4) not assignable

Type

:

116

(97)

(98)
Daphnia magna (Crustacea)
24 hour(s)mg/l> 10000
no dataother1977
no datano data
Although the figure given in the paper is referred to as an
LC50, it is better defined as an EC50, as the end-point
studied is immobilization.
Twenty-four-hour-old Daphnia exposed to a series ofdilutions of ethanol in tap water. Swimming ability
measured after 24 hours.
(4) not assignable
(99)

(100)
StaticUNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Daphnia magna (Crustacea)
2 day(s)g/l> 100
no dataother1995
no datano data

RemarkReliability
21.08.2003::

Result is expressed in ppm,
(4) not assignable

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Daphnia magna (Crustacea)
1 day(s)mg/l> 10000
no dataother1989
no dataas prescribed by 1.1 - 1.4

RemarkReliability
21.08.2003::

Same value recorded for 2 day exposure.
(4) not assignable

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

RemarkReliability
21.08.2003

:
:

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Reliability
21.08.2003

:

Type

:

(101)

(102)
Daphnia magna (Crustacea)
1 day(s)mg/l= 2500no dataotherno dataas prescribed by 1.1 - 1.4
Effect is on physiology; EC50 (2 day) = 2000 mg/l.
(4) not assignable(103)
Daphnia magna (Crustacea)
1 day(s)mmol/l
= 297.7
no dataotherno dataas prescribed by 1.1 - 1.4(4) not assignable(104)

UNEP PUBLICATIONS

117

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Artemia sp. (Crustacea)
2 day(s)mg/l= 25500
no dataother

Reliability
21.08.2003:(4) not assignable

Type
Species
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Reliability
21.08.2003:

Type
Species
Exposure period
Unit
TLm
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

StaticArtemia salina (Crustacea)
24 hour(s)mg/l> 10000
no dataother

Remark:Reliability
21.08.2003:

Static test carried out in the laboratory at 24.5 C. TLm(concentration causing 50% mortality) determinedgraphicallyfrom measurements at an unspecified number ofconcentrations.
(4) not assignable

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Reliability:

118

no dataas prescribed by 1.1 - 1.4(105)
Artemia sp. (Crustacea)
1 day(s)mg/l= 25500 - 27000 measured/nominal
no dataotherno dataas prescribed by 1.1 - 1.4(4) not assignable(105)

no datano data

(106)
Other
2 day(s)mg/l= 11963
no dataother1990
no dataas prescribed by 1.1 - 1.4
Endpoint is EC50 for growth in the freshwater protozoan
Tetrahymena pyriformis (Ciliata).
(4) not assignable

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

21.08.2003

(107)

Type
Species
Exposure period
Unit
IC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Reliability
21.08.2003

:

4.3

Other
2 day(s)mmol/l= 259.67
no dataother1993
no dataas prescribed by 1.1 - 1.4
IC50 is for population growth in this freshwater protozoan.
Tetrahymena pyriformis (Ciliata).
(4) not assignable(108)

TOXICITY TO AQUATIC PLANTS E.G. ALGAE

Species
Endpoint
Exposure period
Unit
NOEC
LOEC
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Chlorella vulgaris (Algae)
growth rate
4 day(s)mg/l< 500 measured/nominal= 500 measured/nominal= 1000 measured/nominal

Method:Remark:Growth rate measured as chlorophyll content and biomass
accumulation at concentrations of 0.05% (500 mg/l) andhigher (range 500 to 10,000 mg/l).
Cells removed before measurement: cells were not removed.

No
other
1996
no datano data

Biological observations:+Cell density at each flask/each measuring point: Not given.
+Growth curves: Chlorophyll content plotted over time foreach concentration, including control.
Percent biomass/growth rate inhibition per concentration
Observations at 500, 1000, 2000, 5000 and 10,000 mg/l,
the growth inhibition was 37%, 54%, 69%, 86% and 95%
Laboratory culture: isolated from lake Geneva in 1980.
Cultivation method: Cultures were grown in Algal Assay
Procedure (1971) medium in 500 ml flasks containing 250 ml
algal suspension.
Cells were not removed from medium prior to measurement;
cell density not given.
Controls consisted of algal suspensions without solvent ineach experiment.

UNEP PUBLICATIONS

119OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Dilution water source not specified.
Growth/test medium: Algal assay Procedure (1971) medium with
15 mg/l NaHCO3 and 12 mg/l K2HPO4.
Exposure vessel type: 20 x 125 mm test tubes containing
about 20 ml of suspension and ethanol. 3 Tubes per test
concentration were used.
Water chemistry (pH) on test: Not described.
Stock solution preparation: Not described.
Light levels and quality during exposure: 100 microE/m^2-sec
except when reduced to 1.5 microE/m^2-sec 20 min before andduring measurement of chlorophyll content by fluorescence.
Test design: Ethanol was tested three times at eachconcentration (0, 0.05, 0.1, 0.3, 0.5 and 1%).

Result:

Method of calculating mean measured concentrations: Only
nominal concentrations were used.
Growth was inhibited 54% at an ethanol concentration of 1000
mg/l, the value approximating the EC50.
Growth inhibition was 37% at 500 mg/l.
Growth was significantly inhibited (p=0.05) at allconcentrations of ethanol.
Test temperature 21 +/- 1 deg C with continuous illumination
at 100 microE/m^2-sec.
Growth was inhibited 48% at an ethanol concentration of
10,000 mg/l; this approximates the ErC50.
(1) valid without restriction
Critical study for SIDS endpoint

Test condition:

Conclusion:

Reliability
Flag
11.11.2004::

Species
Endpoint
Exposure period
Unit
NOEC
LOEC
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

other aquatic plant: Lemna gibba
growth rate
7 day(s)mg/l= 280 measured/nominal> 280 measured/nominal= 4432 measured/nominal

Method:Remark:Lemna gibba (Fat Duckweed) was exposed to alcohol in waterat nominal concentrations 0, 1.0, 1.7, 2.8, 4.7, 7.8, 13,
21, 36 etc to 21,000 (21 concentrations) for 7 days.
Maintained at 25 degC with 6461 +/- 323 lux continuously;
5382 +/- 89 on test. Grown on Hoagland's with a pH of 4.6 to
5.4. Medium renewed weekly. Acclimation period was 8 weeks.
Cells removed before measurement: Plant fronds counted
visually. Biomass measured by dry weight of plants and

120

(109)

No
EPA OTS 797.1160
1986
no dataother TS: 100% dehydrated

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
frondsBiological observations:+Cell density at each flask/each measuring point: Not
applicable.
+Growth curves: Not shown.
Percent biomass/growth rate inhibition per concentration
Observations: Results were not given for each of the 21
occurrences.%
EPA procedures as described by Holst (1986) and Holst and
Edwanger (1982).
Laboratory culture: Obtained from Smithsonian Institution.
Cultivation method: Cultures were grown in Hoaglands medium
with pH 4.6 to 5.4 medium was renewed weekly.
Acclimation period was 8 weeks.
Plants were not removed from medium prior to measurement;
Plant density not given.
Controls consisted of Lemna cultures without ethanol in eachexperiment.
Dilution water source not specified.
Growth/test medium chemistry: Hoaglands. Water hardness 636
mg/l as CaCO3; Alkalinity 23 mg/l as CaCO3, Conductivity
5000 micromhos/cm, pH 4.5 to 5.1.
Exposure vessel type: 250 ml vessels; Shimadzu closurescovered with paraffin. Each concentration replicated 3 times
Water chemistry (pH) on test: pH ranged 4.6 to 5.1..
Stock solution preparation: Not described.
Light levels and quality during exposure: Mean lux 5382 +/89 during exposure.
Test design: Ethanol was tested three times at eachconcentration (1.0 to 21,000 mg/l, plus control). Only
nominal concentrations were used.
Method of calculating mean measured concentrations: Only
nominal concentrations were used.

Result:Conclusion
Reliability
11.11.2004::

Species
Endpoint
Exposure period:::

Statistical method: EC50 - regression analysis; NOEL:
Dunnett's test.
The EC50 (4432 mg/l) was within the 95% confidence interval
845 to 8018 mg/l for plant growth. The EC50 for biomass(dry weight) was 5987 mg/l (1640 to 10,293) mg/l.
Ethanol was the least toxic of 8 compounds tested.
(1) valid without restriction(110)
other aquatic plant: Lemna minor 6591
growth rate
7 day(s)UNEP PUBLICATIONS

121

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Unit
NOEC
LOEC
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

mg/l
= 778 measured/nominal> 778 measured/nominal= 3690 measured/nominal

Method:Remark:Lemna minor (duckweed) was exposed to alcohol in water at
nominal concentrations 0, 1.0, 1.7, 2.8, 4.7, 7.8, 13, 21,
36 etc to 21,000 for 7 days. Maintained at 25degC with 6461+/- 323 lux continuously; 5382 +/- 89 on test. Grown on
Hoagland's with a pH of 4.6 to 5.4. Medium renewed weekly.
Acclimation period was 8 weeks.
EPA procedures as described by Holst (1986) and Holst and
Edwanger (1982).
Laboratory culture: Obtained from Geobotanischen Institute,
Zurich, Switzerland.

No
EPA OTS 797.1160
1986
no dataother TS: 100% dehydrated

Cultivation method: Cultures were grown in revised Hoaglands
medium with pH 4.6 to 5.4; medium was renewed weekly.
Acclimation period was 8 weeks.
Plants were not removed from medium prior to measurement;
Plant density not given.
Controls consisted of Lemna cultures without ethanol in eachexperiment.
Dilution water source not specified.
Growth/test medium chemistry: Hoaglands. Water hardness 636
mg/l as CaCO3; Alkalinity 23 mg/l as CaCO3, Conductivity
5000 micromhos/cm, pH 4.5 to 5.1.
Exposure vessel type: 250 ml vessels; Shimadzu closurescovered with paraffin. Each concentration replicated 3 times
Water chemistry (pH) on test: pH ranged 4.6 to 5.1..
Stock solution preparation: Not described.
Light levels and quality during exposure: Mean lux 5382 +/89 during exposure.
Test design: Ethanol was tested three times at eachconcentration (1.0 to 21,000 mg/l, plus control). Only
nominal concentrations were used.
Method of calculating mean measured concentrations: Only
nominal concentrations were used.
Statistical method: EC50 - regression analysis; NOEL:
Dunnett's test.
Test highly reliable.

122

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Cells removed before measurement: Unclear. Plant fronds
counted visually. Biomass measured by dry weight of plants
and fronds
Biological observations:+Cell density at each flask/each measuring point: Not
applicable.
+Growth curves: Not shown.
Percent biomass/growth rate inhibition per concentration
Observations: Results were not given for each of the 21
occurrences.%
The EC50 (4690 mg/l) was within the 95% confidence interval
81 to 167,764 mg/l for plant growth. The EC50 for biomass(dry weight) was 6986 mg/l (3155 to 10,817) mg/l. Of threeother clones of Lemna minor, 7120 and 7136 were much moreresistant to ethanol with EC50 values of at least 10,000
mg/l and NOEL values of at least 1000 mg/l.
Ethanol was the least toxic of 8 compounds tested.
(1) valid without restriction

Result:Conclusion
Reliability
11.11.2004::

Species
Endpoint
Exposure period
Unit
NOEC
LOEC
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Selenastrum capricornutum (Algae)
growth rate
4 day(s)mg/l< 500 measured/nominal= 500 measured/nominal= 10000 measured/nominal

Method:Remark:Growth rate measured as chlorophyll content and biomass
accumulation at concentrations of 0.05% (500 mg/l) andhigher (range 500 to 10,000 mg/l).
Cells removed before measurement: cells were not removed.

(110)

No
other
1996
no datano data

Biological observations:+Cell density at each flask/each measuring point: Not given.
+Growth curves: Chlorophyll content plotted over time foreach concentration, including control.
Percent biomass/growth rate inhibition per concentration
Observations at 500, 1000, 2000, 5000 and 10,000 mg/l,
the growth inhibition was 14%, 19%, 26%, 37% and 48%
EPA guidance from 1975 recommends maximum solvent
concentration of 0.05% and 0.01% for acute and chronic tests
and higher concentrations often occur in practice. Ethanol
as solvent in such tests has a significant effect on growthrate of the test alga.
Laboratory culture: Obtained from EPA at Corville, OR.
Cultivation method: Cultures were grown in Algal Assay
Procedure (1971) medium in 500 ml flasks containing 250 ml
algal suspension.

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Cells were not removed from medium prior to measurement;
cell density not given.
Controls consisted of algal suspensions without solvent ineach experiment.
Dilution water source not specified.
Growth/test medium: Algal Assay Procedure (1971) medium with
15 mg/l NaHCO3 and 12 mg/l K2HPO4.
Exposure vessel type: 20 x 125 mm test tubes containing
about 20 ml of suspension and ethanol. 3 Tubes per test
concentration were used.
Water chemistry (pH) on test: Not described.
Stock solution preparation: Not described.
Light levels and quality during exposure: 100 microE/m^2-sec
except when reduced to 1.5 microE/m^2-sec 20 min before andduring measurement of chlorophyll content by fluorescence.
Test design: Ethanol was tested three times at eachconcentration (0%, 0.05%, 0.1%, 0.2%, 0.5% and 1%).

Result:

Method of calculating mean measured concentrations: Only
nominal concentrations were used.
Growth was inhibited 48% at an ethanol concentration of
10,000 mg/l, the value approximating the EC50.
Growth inhibition was 14% at 500 mg/l.
Growth was significantly (p=0.05) inhibited at allconcentrations of ethanol.
(1) valid without restriction

Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
NOEC
LOEC
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Chlamydomonas sp. (Algae)
growth rate
2 day(s)g/l= 7.89 measured/nominal= 19.7 measured/nominal

Method:Ethanol concentrations were 0.5, 1.0, 2.5 and 5.0 %v/v (equivalent to 3.95,
7.89, 19.7 and 39.5g/l respectively) in stocks grown on agar slants at 25
degC with continuous aeration and diurnal light cycle of 12 hr. Before
counting, 5% glutaraldehyde added to test systems.

Remark

:

124

(109)

no dataother1980
no datano dataSpecies: Chlamydomonas eugametos.
Note whether cell removed prior to measurement: 5% glutaraldehyde wasadded to test systems. 1 ml samples
analyzed by haemocytometer or Coulter counter.

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Biological observations: #Cell density: Absolute measurements not given
Growth curves: Not given%Biomass/growth rate inhibition: No inhibition at ethanol concentrations of
0.5 or 1.0%/ At 2.5% cell number was 57% of control. At 5.0%, growth wascompletely inhibited
Observations: None described.
Test organism: Bacteria-free Chlamydomonas eugametos culturecollection No. 9.
Method of cultivation: Stocks grown on agar slants, liquid
cultures made 3-4 days before assay
Liquid cultures grown at 25 degC with continuous aerationand diurnal light cycle of 12 hr.
Controls were used but are not dicussed. Tests for ethanol
and other solvents were controls for tests for herbicides
dissolved in these solvents.
Test conditions: temperature 25 degC.
Growth/Test medium chemistry: Not described. Grown innutrient medium.
Dilution water source: Not described.
Exposure vessel type: 150 ml in 250 Erlenmeyer flasks
aerated. 1x10^6 cells suspended in 20ml nutrient medium in
50 ml flasks not aerated.
Water chemistry in test (pH):Not described.
Stock solutions preparation: Not described.
Light levels and quality: 12 h diurnal at 200 microEm^2/s
PPFD.
Test design: Solvents including ethanol were tersted at 4
concentrations, each concentration was tested at least
twice.
Method of calculating mean measured concentrations: Not
described.
Statistical test: Duncan's Multiple Range.
Cell number was 57% of control at 2.5% (19.7g/l) and there was complete
inhibition at 5% (39.5g/l).
(2) valid with restrictions(111)

Result:Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
NOEC
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Skeletonema costatum (Algae)

Method:Remark:Growth inhibition in Skeletonema costatum evaluated by cellnumber count.
Laboratory culture: Bigelow Lab. for Ocean Sciences, West
Boothbay Harbour, Maine, USA.
Cultured in revised ASP12 medium at 20degC with 14 hr lightat 4304 lux +/- 161/day. Agitated daily and transferred
every 7 days. Acclimated for 4 weeks. Test temperature 19.5
to 20.6 degC.
Controls consisting of Skeletonema in medium without ethanol
were used.

5 day(s)mg/l= 3240 - 5400
= 10943 - 11619
no dataother1989
no dataother TS: 100% dehydrated

UNEP PUBLICATIONS

125OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Temperature range 19.5-20.6 degC.
Growth/Test medium chemistry: Not described.
Dilution water source: Not described.
Exposure vessel type: 100 ml covered with parafilm; eachconcentration tested in triplicate.
Stock solutions prepared with double-distilled sterilewater.
Light levels and quality: Mean lux 4304 +/- 8.2 with a 14 h
light/10 h dark cycle.
Test design: 5 or more concentrations plus control each
replicated 3 times.
Method of calculating mean measured concentrations: Only
nominal concentratiuons used.
Statistical methods: Not described.
Note whether cells removed prior to measurement: Not stated.
Biological observations:#Cell density at each flask at each measuring point:
Not given.
#Growth curves: Not given but growth was stimulated before
inhibition began.
Percent biomass/growth rate inhibition, observations: Not
given.
EC50 for total cell count was 11,619 mg/l (7923 to 15,314)
and for total cell volume, 10943 mg/l (7061 to 14,826) mg/l.
The authors state that, using EPA criteria, ethanol can be
judged 'practically nontoxic, by this test. Ethanol was used
as a carbon source, stimulating growth before inhibition
began.
(2) valid with restrictions

Result:Conclusion:

Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
EC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Chlorella pyrenoidosa (Algae)
growth rate
10 day(s)mg/l= 1180Remark:Alga: Chlorella pyrenoidosa from National Research Councilof Canada.
Culture: Axenic in 250 Erlenmeyer fl;aks containing nitrogen
free medium incubated at 25 degC(112)

no dataother1988
no dataother TS

End point: Growth (biomass) measured by optical density overtime.
Concentration of ethanol: 0.4 to 3.0%
Exposure period: 10 to 14 days (precise duration not
specified).

126

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
EC50 value was the concentration required to cause a 50%
reduction in growth.
Result was converted from an EC50 value of 1.18% v/v.
Stratton also studied the effects of ethanol on 5 further
species of algae, using the same system but withthe concentration of ethanol tested ranging from 0.1 to 8%.
The reported EC50 values were as follows:
Anabaena sp. 6312 mg/l
A. variabilis 10020 mg/l
A. inaequalis 8048 mg/lA. cylindrica 7653 mg/l
Nostoc sp. 22644 mg/l
Test sustance was absolute ethanol.
(2) valid with restrictions

Test substance
Reliability
11.11.2004::

Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Cyclotella sp. (Algae)
Biomass
4 day(s)g/l> 2.37 measured/nominal

Method:Species of marine diatom from the intertidal region of the Gulf of Mexico
were maintained and tested in Guillard's f/2 medium enriched with artificial
sea salt mix.

ResultReliability(113) (114)::

11.11.2004
Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
Method
Year
GLP

:
:
:
:
:
:
:
:
:
:

no dataother1995
no dataas prescribed by 1.1 - 1.4

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was no significant effect on the growth rate.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)
Navicula sp. (Algae)
Biomass
4 day(s)g/l> 2.37 measured/nominal
no dataother1995
no data

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OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test substance:as prescribed by 1.1 - 1.4

Method:Species of marine diatom (Navicula saprophila species) from the intertidal
region of the Gulf of Mexico were maintained and tested in Guillard's f/2
medium enriched with artificial sea salt mix.

Result:Reliability:

11.11.2004

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was an apparent increase in growth rate but no clear dose response
relationship.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)

Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Nitzschia sp. (Algae)
Biomass
4 day(s)g/l> 2.37 measured/nominal

Method:Species of marine diatom (Nitzschia pussilla species) from the intertidal
region of the Gulf of Mexico were maintained and tested in Guillard's f/2
medium enriched with artificial sea salt mix.

ResultReliability

11.11.2004

128

:
:

no dataother1995
no dataas prescribed by 1.1 - 1.4

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was no significant effect on the growth rate.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Nitzschia sp. (Algae)
Biomass
4 day(s)g/l> 2.37 measured/nominal

Method:Species of marine diatom from the intertidal region of the Gulf of Mexico
were maintained and tested in Guillard's f/2 medium enriched with artificial
sea salt mix.

Result:Reliability:

11.11.2004

no dataother1995
no dataas prescribed by 1.1 - 1.4

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was an apparent increase in growth rate but no clear dose response
relationship.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)

Species
Endpoint
Exposure period
Unit
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Nitzschia sp. (Algae)
Biomass
4 day(s)g/lMethod:Species of marine diatom (Nitzschia dissipata species) from the intertidal
region of the Gulf of Mexico were maintained and tested in Guillard's f/2
medium enriched with artificial sea salt mix.

no dataother1995
no dataas prescribed by 1.1 - 1.4

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.

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Result:Reliability:

11.11.2004

There was a dose reponse decrease in growth but with promotion at thelower concentration, the decrease in growth at the higher concentration
was only limited. The results is therefore difficult to interpret.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)

Species
Endpoint
Exposure period
Unit
IC50
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

other algae: Cylindricotheca sp.
Biomass
4 day(s)g/l= 1.97 measured/nominal

Method:Species of marine diatom from the intertidal region of the Gulf of Mexico
were maintained and tested in Guillard's f/2 medium enriched with artificial
sea salt mix.

Result:Reliability:

11.11.2004
Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

130

:
:
:
:
:
:
:
:
:
:
:

no dataother1995
no dataas prescribed by 1.1 - 1.4

A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was a significant dose response decrease with an apparent IC50 ofaround 0.25ml/100ml (equivalent to 1.97g/l). However, taking all resultsfrom the study into account, the authors describe ethanol to be non-toxic.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)
other algae: Thalassiosira weissflgii sp.
Biomass
4 day(s)g/l> 2.37 measured/nominal
no dataother1995
no dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY
MethodETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Result:Reliability:

11.11.2004

Species of marine diatom from the intertidal region of the Gulf of Mexico
were maintained and tested in Guillard's f/2 medium enriched with artificial
sea salt mix.
A 1% (v/v) solution of ethanol was further diluted with test medium atconcentrations of 0.2, 0.25 and 0.3 ml/100ml (1.58, 1.97, 2.37g/l
respectively). Tests were carried out in test tubes containing 25 ml medium
and in triplicate. Each tube was inoculated from exponentially growth cells
at an initial density of 4000 cells/ml. The culture was incubated on a shaker
for 96 h at 30 degC under cool white light producing 100 muEm-2s
irradiation in continuous cycle. Growth was measured
spectrophotometrically at 525nm. Statistical analysis was by Student's ttest with significance at p<0.05.
There was an apparent increase in growth rate but no clear dose response
relationship.
(4) not assignable
Only a very limited range of concentrations was studied which in mostcases produced no effect on growth. No chemical monitoring was carriedon the solutions prepared. Results are only reported in summarized, basic,
graphical form and only as a percentage of control response. There areinsufficient data reported.
(115)

Species
Endpoint
Exposure period
Unit
IC10
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Scenedesmus subspicatus (Algae)
growth rate

Reliability
11.11.2004:(4) not assignable

Species
Endpoint
Exposure period
Unit
IC10
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Scenedesmus subspicatus (Algae)
other: inhibition of protoplast O2 production

Reliability
11.11.2004:(4) not assignable

Species
Endpoint
Exposure period
Unit
EC10
Limit test

:
:
:
:
:
:

mg/l
= 400 measured/nominal
no dataotherno datano data

(116)

mg/l
= 460 measured/nominal
no dataotherno datano data(116)
Chlorococcum sp. (Algae)
Biomass
24 hour(s)mg/l> 1000

UNEP PUBLICATIONS

131OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:

no dataother

MethodReliability
29.09.2003::

In fresh water; method not specified.
(4) not assignable

Species
Endpoint
Exposure period
Unit
EC50
LC100
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dunaliella bioculata (Algae)
Biomass
2 day(s)mg/l= 1000 measured/nominal= .05

Remark:Result:Reliability
11.11.2004:

Two concentrations of ethanol were tested, 0.1% (1000 mg/l)
and 0.05% (500 mg/l).
Growth was reduced 10% at 1000 mg/l. The NOEC and LOEC were
not calculated.
(4) not assignable

Species
Endpoint
Exposure period
Unit
EC0
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Microcystis aeruginosa (Algae, blue, cyanobacteria)
Biomass
8 day(s)mg/l= 1450 measured/nominal

Remark:Reliability
11.11.2004:

Difference in growth rate between ethanol-containing
cultures and controls measured turbidimetrically. Toxic
threshold concentration determined.
(4) not assignable

Species
Endpoint
Exposure period
Unit
LOEC
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

132

(117)

no dataotherno datano data(118)

no dataotherno datano data(119)
Scenedesmus quadricauda (Algae)
growth rate
7 day(s)mg/l= 5000Nootherno datano data

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Remark:Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
Limit test
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

other algae: see method details for species tested
Biomass
96 hour(s)g/lMethod:Algae obtained from University of Texas algal collection. The following
species tested:

Ethanol tested in double-distilled water in a cell
multiplication inhibition test. Endpoint measured was the
toxic threshold.
(4) not assignable(120)

no dataotherno dataother TS

Gleocystis ampla
Scenedesmus obliquus
Nannochloris sp.
Tetraselmis sp.
Chlorella ellipsoidea
Chlorococcum sp.

Result:

Test substance
Reliability::

17.11.2004

4.4

Growth medium prepared according to Bold (1949). pH adjusted to 8 with
HCL or NaOH.
Solution concentrate: 1ml/100ml growth medium
Test concentrations: 0.05, 0.1, 0.2ml solution concentrate in 100ml growth
medium.
Assays carried out in 25ml medium; triplicate samples and triplicate
cultures.
Innoculation: exponentially growing cells at 4x10E3 cells/ml.
Temperature: 30C
Light: cool-white 100uEm-2s in continuous cycle.
Measurement: spectrophotometrically at 525nm on a Fisher
electrophotometer.
Statistical method: student's t test at p<0.05
All algal species stimulated and all concentrations growed faster thancontrol, some almost doubling in rate. No inhibition seen.
Supplied by JT Baker, Phillipsburgh NJ.
(3) invalid
Key details reported. No details of growth medium reported in paper but
referenced elsewhere. No analytical monitoring or details of water used.
Results presented as percentage of control - control value not available
separately. Results only available graphically but this is not a major
limitation bearing in mind result obtained. However, since there isambiguity between the way the test concentrations are described in themethod details versus the way they are reported in the results section, thestudy cannot be considered robust since concentrations cannot be
determined with confidence.
(121)

TOXICITY TO MICROORGANISMS E.G. BACTERIAUNEP PUBLICATIONS

133

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
Species
Exposure period
Unit
LOEC
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Aquatic
Pseudomonas putida (Bacteria)
16 hour(s)mg/l= 6500no dataother: see remarks
1980
no datano data

Remark:Reliability
Flag
11.11.2004::

Stock solutions of the test compound were prepared understerile conditions and diluted to test concentrations (by afactor of 2) with bidistilled water.The inoculated
4-parallel dilution series was prepared in 300-ml Erlenmeyer
flasks, stoppered with cotton-lined plastic caps. The firstflask of each series contained 160 ml of test solution.
Subsequent dilutions from this flask were prepared by adding
80 ml of preliminary pollutant dilution and 80 ml
bidistilled water. Each flask of the dilution series was
inoculated to 100 ml by adding 5 ml of stock solution I, 5
ml of stock solution II, and 10 ml of bacterial cellsuspension from the preliminary culture. Blank controls(not containing inoculum) were prepared by adding 5 ml ofstock solution I, 5 ml of stock solution II, and 10 ml
saline solution (0.50 g NaCl/l sterile, bidistilledwater).Stock solution I: 20.0 g D(+)glucose, 4.240 g NaNO3,
2.40 g K2HPO4, 1.20 g KH2PO4, and 30 ml trace elements
solution dissolved in 500 ml bidistilled water andsterilized for 30 minutes.Stock solution II: 0.20 g
FeSO4o7H20 and 4.00 MgSO4o7H20 dissolved in 1000 ml sterile,bidistilled water. Flasks were cultured at 25 deg. C for 16
hours. The concentration of bacterial cells was measured
turbidimetrically and expressed as the extinction of the
primary light of monochromatic radiation at 436 nm for a 10mm layer. Bacterial cell growth inhibition was graphicallydetermined. Both the highest non-toxic test concentrationand the lowest toxic test concentration were plotted. A 3%
reduction in cell growth was used as the value indicating
the onset of inhibitory action.
(2) valid with restrictions
Critical study for SIDS endpoint

Type
Species
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Aquatic
Photobacterium phosphoreum (Bacteria)
15 minute(s)g/l= 32.1 measured/nominal
no dataother: Microtox
1994
no dataother TS

Method:Test substance:Standard MICROTOX assay procedure (Microbiotics 1989). Unclear
whether exposure time was 5 minutes or 15 minutes. Four replicates.
No specific data. At least 97% pure, bbut possibly >99% or pharmacopia
purity.

134

(122)

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY
Reliability
11.11.2004
4.5.1

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:(4) not assignable(92)

CHRONIC TOXICITY TO FISH

Species
Endpoint
Exposure period
Unit
EC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Pimephales promelas (Fish, fresh water)other: development, histology and mortality
42 day(s)g/l= 1 measured/nominal
no dataother1999
no datano data

Remark:Result:This study assesses the effects of waterborne exposure of
nonylphenol and nonylphenol ethoxylate on the secondary sex
characteristics and gonads of the fathead minnow. Ethanol
was used as a carrier solvent for the test substances. Twocontrols were used in the experiment, one pure water and thesecond ethanol at 0.00001% (v/v or w/w not stated.) This isequivalent to approximately 0.08mg/l (assuming w/w) or
0.1ppm (assuming v/v).Testicular lesions were evaluatedaccording to the severity of relative or absolute Sertoli
cell proliferation and the percentage of seminiferous
tubules affected.
There was no dose dependent trends in mortality of either
males or females exposed to the test substance or controls.
Survival rates of males ranged from a low of 50% to 100%,for females from 60% to 100%.
Survival rates
MALES
Expt1 Expt 2
Water control
100% 50%
Ethanol 0.1ppm 100% 50%
FEMALES
Expt1 Expt 2
Water control
100% 100%
Ethanol 0.1ppm
50% 100%

Reliability:

There was no change in the gross appearance of the fatpad ofmales exposed to ethanol compared to the control. There wasno significant difference in the incidence of histologicallesions observed. There was as difference observed in one of
two experiments in the median size of the tubercules andfatpad. There is no discussion in the paper on therelevance of this difference which was not seen in a second
experiment. There was also no dose response trend for theseend points for the actual test substances in the study,
which varied randomly over a similar range.
(3) invalid
This study was carried out using a single, very low
concentration of ethanol and did not produce any results ofclear significance. It cannot be used to derive a chronic
NOEC for ethanol.

22.09.2003
Species
Endpoint

(123)
:
:

other: Acipenser transmontanum (White sturgeon)other: mortality

UNEP PUBLICATIONS

135

OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Exposure period
Unit
LC0
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

24 day(s)mg/l= 1000 measured/nominal
no dataother1998
no datano data

RemarkReliability::

The authors describe this 24 day test "Acute".
(3) invalid
The primary goal of this study was to investigate thepossibility of using early life stages in both acute andsublethal toxicological testing of forest industry effluents
into the Fraser River. The larval stage was more sensitive
to toxicants than the fry stage and studies wereinconclusive because immature stages did not withstand the
additional handling during determination of body mass. The
investigators had concerns about the reliability of thistesting method and, in any case, ethanol was used only as acarrier solvent control at a concentration of 1 ppm.

04.10.2003
4.5.2

(124)

CHRONIC TOXICITY TO AQUATIC INVERTEBRATESSpecies
Endpoint
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ceriodaphnia sp. (Crustacea)
Mortality
10 day(s)mg/l= 1284 - 2638
no dataother1991

Method:Follows the basic methodology for the three brood test
proposed by Mount and Norberg (Mount, D.J. and Norberg, T.J.
(1984) A seven-day life cycle cladoceran toxicity test.
Environ. Toxicol. Chem. 3, 425 - 434).Analytical methods
used for test substance: no data given.Vehicle used: notrequired. Statistical methods: For LC50: Probit, moving
average and nonlinear interpolation. Calculation of point
estimates and other corresponding 95% confidence intervals
made using a program written by Stephan (1977, Methods forcalculating an LC50, In Mayer FL et al (eds), Aquatic
toxicology and hazard assessment, ASTM STP 634: 65-84).
Calculation of EC50: statistical package SAS GLM (1987,
SAS/STAT guide for personal computers, version 6th ed, SASinstitute Inc, Cary, NC) used to generate regression
equations. NOELs calculated using Dunnett's t-test.Test
organism: Daphnia magna strauss 1820 populations (of British
origin) had been maintained in the Dow Chemical Company
Laboratory since 1982 without drastic changes in population.
Population maintained at 25C for past 3 years and sustainedon Ankistrodesmus convulutus (reared in medium based on
Provasoli and Pintner (1968, Ecological implications of invitro nutritional requirements of algal flagellates", Ann NY

136

as prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Remark:Test condition:

Acad. Sci. 56, 839-851.) and Nitzschia frustulum Kutzing
cultured in ES-I-Si, a medium developed by Provasoli (1968,"Media and prospects for the cultivation of marine algae",in Watanabe A et al, Cultures and collections of algae,
Proceedings of a US-Japan conference, Hakone 1966.) Algal
diet axenic.Test conditions: The. testing conditions
followed the basic tenets of the original three-brood test
proposed by Mount and Norberg (1984) but were revised inthat they emphasize the needs of the animals in terms ofspace and diet. Details of the conditions may be found inin Cowgill and Milazzo (1989). Test vessels were wide mouthclear glass jars graduated in milliliters to contain 150mL.
Into each jar was fitted a glass tube, 3.5 cm diameter.
which had affixed to one end a nytex screen of 243 µm mesh
for C. dubia or 1000 µm mesh for D. magna, These screens
were affixed to the glass tubes with silicone glue. Afterthe screens were glued to the glass tubes, three glass
beads, 8 mm in diameter, were affixed to the undersideequidistant from each other. This was covered with a glass
petal dish 5.5 cm in diameter. The jar containing thescreened tube was filled with double distilled water andautoclaved for 10 mins at a pressure of 124 kPa. This
procedure was repeated three times, renewing the distilledwater each time, before the equipment was used for a test.
This procedure accomplished the complete removal of all
effects of the silicone glue. Only glass vessels were used.
Species: Ceriodaphnia dubia.
Using the USEPA classification scheme, ethanol would beclassified as practically non toxic based on survival.
Based on reproductive parameters, it would be classified asslightly toxic.
Analysis of Lake Huron water used in culturing and testingAl
140
NH3 total
ND(lO)
B
40
Ca
18700
Cr
8
Cu
5
F
80
Fe
17
Pb
(5)
ND (5)
Mg
7800
Mn
(5)
ND (5)
K
1040
Si
3400
Na
4800
S
6000
Zn
8
Total dissolved solids 118000
Total suspended solids NA
Total organic carbon 1600
Test conditions
Test vessel: Capacity 150ml,
Content 100ml
Screen composition: Nytex,
Screen mesh size, 243µm
Light lux: 670 ± 100 lux
Photoperiod, 16 h light, 8 h dark
Temperature 25 ± 2

UNEP PUBLICATIONS

137

OECD SIDS
4. ECOTOXICITY

Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Dissolved oxygen, 8.0 ± 1.5mg/L
pH: 8.2 ± 0.2
Dilution water
Hardness, as mg CaCO3,/L: 90-110
Alkalinity, as mg CaCQ3/L: 55-75
Habitat: Environmental chamber
Habitat changing frequency: Every other day
Diet: algae (A. convolutes, N. frustulum)
Feeding rate (cells/vessel): A. convolutes 9x10^6, N.
frustulum 1.8x10^6Feeding frequency: daily
Age of organisms, <12h (all from fourth brood.)
Number of control broods: 3
Permitted control loss, 20%
Number of organisms/ vessel: 1
Number of organisms/Concentration: 10
Number of organisms!/control: 20
Test length, days: 7-10
Variables monitored Daily: light, temperature, survival,progeny
Variables monitored every second day: w
ater quality variables in renewed solutions
Variables monitored at Test termination: Survival, total
progeny, adult weight
Endpoints: Survival, total progeny, dry adult weight, numberof broods, mean brood size, loss of control limited to 20%(LC50/EC50/NOEC)
Test ended when the control animals had produced threebroods.
Test concentrations used: not specified.
(2) valid with restrictions
This is a well reported study, particularly regarding themethodology of testing. The main weakness is lack of detailon test concentrations used and on the analytical method
used to assess test substance concentration.

11.11.2004(125)

Species
Endpoint
Exposure period
Unit
NOEC
EC50
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Ceriodaphnia sp. (Crustacea)
reproduction rate
10 day(s)
mg/l
= 9.6
26 - 38= 1806no dataother: see free text
1984
no dataas prescribed by 1.1 - 1.4

Method:Follows the basic methodology for the three brood test
proposed by Mount and Norberg (Mount, D.J. and Norberg, T.J.
(1984) A seven-day life cycle cladoceran toxicity test.
Environ. Toxicol. Chem. 3, 425 - 434).Analytical methods
used for test substance: no data given.Vehicle used: notrequired.Statistical methods: For LC50: Probit, moving
average and nonlinear interpolation. Calculation of point
estimates and other corresponding 95% confidence intervals
made using a program written by Stephan (1977, Methods forcalculating an LC50, In Mayer FL et al (eds), Aquatic

138

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:

Test condition:

toxicology and hazard assessment, ASTM STP 634: 65-84).
Calculation of EC50: statistical package SAS GLM (1987,
SAS/STAT guide for personal computers, version 6th ed, SASinstitute Inc, Cary, NC) used to generate regression
equations. NOELs calculated using Dunnett's t-test.Test
organism: Daphnia magna strauss 1820 populations (of British
origin) had been maintained in the Dow Chemical Company
Laboratory since 1982 without drastic changes in population.
Population maintained at 25C for past 3 years and sustainedon Ankistrodesmus convulutus (reared in medium based on
Provasoli and Pintner (1968, Ecological implications of invitro nutritional requirements of algal flagellates", Ann NY
Acad. Sci. 56, 839-851.) and Nitzschia frustulum Kutzing
cultured in ES-I-Si, a medium developed by Provasoli (1968,"Media and prospects for the cultivation of marine algae",in Watanabe A et al, Cultures and collections of algae,
Proceedings of a US-Japan conference, Hakone 1966.) Algal
diet axenic.Test conditions: The. testing conditions
followed the basic tenets of the original three-brood test
proposed by Mount and Norberg (1984) but were revised inthat they emphasize the needs of the animals in terms ofspace and diet. Details of the conditions may be found inin Cowgill and Milazzo (1989). Test vessels were wide mouthclear glass jars graduated in milliliters to contain 150mL.
Into each jar was fitted a glass tube, 3.5 cm diameter.
which had affixed to one end a nytex screen of 243 µm mesh
for C. dubia or 1000 µm mesh for D. magna, These screens
were affixed to the glass tubes with silicone glue. Afterthe screens were glued to the glass tubes, three glass
beads, 8 mm in diameter, were affixed to the undersideequidistant from each other. This was covered with a glass
petal dish 5.5 cm in diameter. The jar containing thescreened tube was filled with double distilled water andautoclaved for 10 mins at a pressure of 124 kPa. This
procedure was repeated three times, renewing the distilledwater each time, before the equipment was used for a test.
This procedure accomplished the complete removal of all
effects of the silicone glue. Only glass vessels were used.
Results based on total progeny
EC50 26mg/l (95% CI 0.5-1443)
NOEL 9.6mg/l
Results based on number of broods
EC50 38mg/l (95% CI 0.6-2554)
NOEL 16mg/l
Results based on mean brood size
EC50 33mg/l (95% CI 0.6-1820)
NOEL 9.6mg/l
Analysis of Lake Huron water used in culturing and testingAl
140
NH3 total
ND(lO)
B
40
Ca
18700
Cr
8
Cu
5
F
80
Fe
17
Pb
ND (5)
Mg
7800
Mn
ND (5)
K
1040

UNEP PUBLICATIONS

139

OECD SIDS
4. ECOTOXICITY

Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

:

Si
3400
Na
4800
S
6000
Zn
8
Total dissolved solids 118000
Total suspended solids NA
Total organic carbon 1600
Test conditions
Test vessel: Capacity 150ml, Content 100ml
Screen composition: Nytex, Screen mesh size 243µm
Light lux: 670 ± 100 lux
Photoperiod, 16 h light, 8 h dark
Temperature 25 ± 2
Dissolved oxygen, 8.0 ± 1.5mg/L
pH: 8.2 ± 0.2
Dilution water
Hardness, as mg CaCO3,/L: 90-110
Alkalinity, as mg CaCO3/L: 55-75
Habitat: Environmental chamber
Habitat changing frequency: Every other day
Diet: algae (A. convolutes, N. frustulum)
Feeding rate (cells/vessel): A. convolutes 9x10^6, N.
frustulum 1.8x10^6Feeding frequency: daily
Age of organisms, <12h (all from fourth brood.)
Number of control broods: 3
Permitted control loss, 20%
Number of organisms/ vessel: 1
Number of organisms/Concentration: 10
Number of organisms!/control: 20Test length, days: 7-10.
Variables monitored Daily: light, temperature, survival,progeny
Variables monitored every second day: water quality
variables in renewed solutions.
Variables monitored at Test termination: Survival, total
progeny, adult weight
Endpoints: Survival, total progeny, dry adult weight, numberof broods, mean brood size, loss of control limited to 20%(LC50/EC50/NOEC)
Test ended when the control animals had produced threebroods.
Test concentrations used: not specified.
(2) valid with restrictions
This is a well reported study, particularly regarding themethodology of testing. The main weakness is lack of detailon test concentrations used and on the analytical method
used to assess test substance concentration

11.11.2004
Species
Endpoint
Exposure period
Unit
NOEC
LC50
Analytical monitoring
MethodYear
GLP
Test substance

140

(125)
:
:
:
:
:
:
:
:
:
:
:

Daphnia magna (Crustacea)
Mortality
11 day(s)
mg/l
= 9.6
= 454
no dataother: see freetext
1990
no dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Method:Remark:Result:

Test condition:

Follows the basic methodology for the three brood test
proposed by Mount and Norberg (Mount, D.J. and Norberg, T.J.
(1984) A seven-day life cycle cladoceran toxicity test.
Environ. Toxicol. Chem. 3, 425 - 434).Analytical methods
used for test substance: no data given.Vehicle used: notrequired.Statistical methods: For LC50: Probit, moving
average and nonlinear interpolation. Calculation of point
estimates and other corresponding 95% confidence intervals
made using a program written by Stephan (1977, Methods forcalculating an LC50, In Mayer FL et al (eds), Aquatic
toxicology and hazard assessment, ASTM STP 634: 65-84).
Calculation of EC50: statistical package SAS GLM (1987,
SAS/STAT guide for personal computers, version 6th ed, SASinstitute Inc, Cary, NC) used to generate regression
equations. NOELs calculated using Dunnett's t-test.Test
organism: Daphnia magna strauss 1820 populations (of British
origin) had been maintained in the Dow Chemical Company
Laboratory since 1982 without drastic changes in population.
Population maintained at 25C for past 3 years and sustainedon Ankistrodesmus convulutus (reared in medium based on
Provasoli and Pintner (1968, Ecological implications of invitro nutritional requirements of algal flagellates", Ann NY
Acad. Sci. 56, 839-851.) and Nitzschia frustulum Kutzing
cultured in ES-I-Si, a medium developed by Provasoli (1968,"Media and prospects for the cultivation of marine algae",in Watanabe A et al, Cultures and collections of algae,
Proceedings of a US-Japan conference, Hakone 1966.) Algal
diet axenic.Test conditions: The. testing conditions
followed the basic tenets of the original three-brood test
proposed by Mount and Norberg (1984) but were revised inthat they emphasize the needs of the animals in terms ofspace and diet. Details of the conditions may be found inin Cowgill and Milazzo (1989). Test vessels were wide mouthclear glass jars graduated in milliliters to contain 150mL.
Into each jar was fitted a glass tube, 3.5 cm diameter.
which had affixed to one end a nytex screen of 243 µm mesh
for C. dubia or 1000 µm mesh for D. magna, These screens
were affixed to the glass tubes with silicone glue. Afterthe screens were glued to the glass tubes, three glass
beads, 8 mm in diameter, were affixed to the undersideequidistant from each other. This was covered with a glass
petal dish 5.5 cm in diameter. The jar containing thescreened tube was filled with double distilled water andautoclaved for 10 mins at a pressure of 124 kPa. This
procedure was repeated three times, renewing the distilledwater each time, before the equipment was used for a test.
This procedure accomplished the complete removal of all
effects of the silicone glue. Only glass vessels were used.
Using the USEPA classification scheme, ethanol would beclassified as practically non toxic based on survival.
Based on reproductive parameters, it would be classified asslightly toxic.
LC50 (48hr) 9248mg/l (95% CI 7560-12600)
LC50 (9 day) 454mg/l (95% CI 232-814)
NOEL (11 day) 9.6 mg/l
Analysis of Lake Huron water used in culturing and testingAl
105
NH3 total
ND(lO)
B
332

UNEP PUBLICATIONS141

OECD SIDS
4. ECOTOXICITY

Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

:

Ca
45050
Cr
ND(5)
Cu
13
F
75
Fe
12
Pb
ND (5)
Mg
7600
Mn
ND (5)
K
2485
Si
4760
Na
5700
S
5585
Zn
15
Total dissolved solids 233500
Total suspended solids 1125
Total organic carbon 1400
Test conditions
Test vessel: Capacity 150ml, Content 100ml
Screen composition: Nytex, Screen mesh size 1000µm
Light lux: 2150 ± 300 lux
Photoperiod, 16 h light, 8 h dark
Temperature 25 ± 2
Dissolved oxygen, 8.0 ± 1.5mg/L
pH: 8.2 ± 0.2
Dilution water Hardness, as mg CaCO3,/L: 160-180
Alkalinity, as mg CaCO3/L: 40-52
Habitat: Environmental chamber
Habitat changing frequency: Every other day
Feeding rate (cells/vessel): A. convolutes 18x10^6, N.
frustulum 3.6 x10^6Feeding frequency: daily
Age of organisms, <12h (all from fourth brood.)
Number of control broods: 3
Permitted control loss, 20%
Number of organisms/ vessel: 1
Number of organisms/Concentration: 10
Number of organisms!/control: 20
Test length, days: 9-11
Variables monitored Daily: light, temperature, survival,progeny
Variables monitored every second day: water quality
variables in renewed solutions
Variables monitored at Test termination: Survival, total
progeny, adult weightEndpoints: Survival, total progeny, dry
adult weight, number of broods, mean brood size, loss ofcontrol limited to 20% (LC50/EC50/NOEC)
Test ended when the control animals had produced threebroods.
Test concentrations used: not specified.
(2) valid with restrictions
This is a well reported study, particularly regarding themethodology of testing. The main weakness is lack of detailon test concentrations used and on the analytical method
used to assess test substance concentration.

11.11.2004
Species
Endpoint
Exposure period
Unit

142

(125)
:
:
:
:

Daphnia magna (Crustacea)
reproduction rate
11 day(s)mg/lUNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

NOEC
EC50
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

= 9.6
14 - 26= 454
no dataother1990
no dataother TS: reagent grade

Method:Remark:Follows the basic methodology for the three brood test
proposed by Mount and Norberg (Mount, D.J. and Norberg, T.J.
(1984) A seven-day life cycle cladoceran toxicity test.
Environ. Toxicol. Chem. 3, 425 - 434).Analytical methods
used for test substance: no data given.Vehicle used: notrequired.Statistical methods: For LC50: Probit, moving
average and nonlinear interpolation. Calculation of point
estimates and other corresponding 95% confidence intervals
made using a program written by Stephan (1977, Methods forcalculating an LC50, In Mayer FL et al (eds), Aquatic
toxicology and hazard assessment, ASTM STP 634: 65-84).
Calculation of EC50: statistical package SAS GLM (1987,
SAS/STAT guide for personal computers, version 6th ed, SASinstitute Inc, Cary, NC) used to generate regression
equations. NOELs calculated using Dunnett's t-test.Test
organism: Daphnia magna strauss 1820 populations (of British
origin) had been maintained in the Dow Chemical Company
Laboratory since 1982 without drastic changes in population.
Population maintained at 25C for past 3 years and sustainedon Ankistrodesmus convulutus (reared in medium based on
Provasoli and Pintner (1968, Ecological implications of invitro nutritional requirements of algal flagellates", Ann NY
Acad. Sci. 56, 839-851.) and Nitzschia frustulum Kutzing
cultured in ES-I-Si, a medium developed by Provasoli (1968,"Media and prospects for the cultivation of marine algae",in Watanabe A et al, Cultures and collections of algae,
Proceedings of a US-Japan conference, Hakone 1966.) Algal
diet axenic.Test conditions: The. testing conditions
followed the basic tenets of the original three-brood test
proposed by Mount and Norberg (1984) but were revised inthat they emphasize the needs of the animals in terms ofspace and diet. Details of the conditions may be found inin Cowgill and Milazzo (1989). Test vessels were wide mouthclear glass jars graduated in milliliters to contain 150mL.
Into each jar was fitted a glass tube, 3.5 cm diameter.
which had affixed to one end a nytex screen of 243 µm mesh
for C. dubia or 1000 µm mesh for D. magna, These screens
were affixed to the glass tubes with silicone glue. Afterthe screens were glued to the glass tubes, three glass
beads, 8 mm in diameter, were affixed to the undersideequidistant from each other. This was covered with a glass
petal dish 5.5 cm in diameter. The jar containing thescreened tube was filled with double distilled water andautoclaved for 10 mins at a pressure of 124 kPa. This
procedure was repeated three times, renewing the distilledwater each time, before the equipment was used for a test.
This procedure accomplished the complete removal of all
effects of the silicone glue. Only glass vessels were used.
Using the USEPA classification scheme, ethanol would beclassified as practically non toxic based on survival.
Based on reproductive parameters, it would be classified as

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Result:

Test condition:

144

slightly toxic.
Results based on total progeny
EC50 14mg/l (95% CI 0.8-274)
NOEL 9.6mg/l
Results based on number of broods
EC50 26mg/l (95% CI 1-640)
NOEL 16mg/l
Results based on mean brood size
EC50 15mg/l (95% CI 0.9-278)
NOEL 9.6mg/l
Analysis of Lake Huron water used in culturing and testingAl
105
NH3 total
ND(lO)
B
332
Ca
45050
Cr
ND(5)
Cu
13
F
75
Fe
12
Pb
ND (5)
Mg
7600
Mn
ND (5)
K
2485
Si
4760
Na
5700
S
5585
Zn
15
Total dissolved solids 233500
Total suspended solids 1125
Total organic carbon 1400
Test conditions
Test vessel: Capacity 150ml, Content 100ml
Screen composition: Nytex, Screen mesh size 1000µm
Light lux: 2150 ± 300 lux
Photoperiod, 16 h light, 8 h dark
Temperature 25 ± 2
Dissolved oxygen, 8.0 ± 1.5mg/L
pH: 8.2 ± 0.2
Dilution water Hardness, as mg CaCO3,/L: 160-180
Alkalinity, as mg CaCO3/L: 40-52
Habitat: Environmental chamber
Habitat changing frequency: Every other day
Feeding rate (cells/vessel): A. convolutes 18x10^6, N.
frustulum 3.6 x10^6Feeding frequency: daily
Age of organisms, <12h (all from fourth brood.)
Number of control broods: 3
Permitted control loss, 20%
Number of organisms/ vessel: 1
Number of organisms/Concentration: 10
Number of organisms!/control: 20
Test length, days: 9-11
Variables monitored Daily: light, temperature, survival,progeny
Variables monitored every second day: water quality
variables in renewed solutions
Variables monitored at Test termination: Survival, total
progeny, adult weightEndpoints: Survival, total progeny, dry
adult weight, number of broods, mean brood size, loss ofcontrol limited to 20% (LC50/EC50/NOEC)UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Test ended when the control animals had produced threebroods.
Test concentrations used: not specified.
(2) valid with restrictions

Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Palaemonetes pugio (Crustacea)
other: embryo aute toxicity
12 day(s)g/l= 3 - 4.5
no dataother: see freetext
1995
no dataas prescribed by 1.1 - 1.4

Method:Remark:Adult male and female grass shrimp were collected by dip net
during spring and autumn 1995 from relatively uncontaminatedlocal estuaries near the U. S. Environmental Protection
Agency. Gulf Ecology Division. Gulf Breeze, Florida. The
grass shrimp were placed in ice chests with water from thecollection site, transported to the laboratory . The grass
shrimp were acclimated slowly to 251 and 20-ppt salinity
over a 4-h period and were maintained communally in glass
aquaria with flow-through seawater at 250C and 20-ppt
salinity. Adult grass shrintp were held in the aquaria forat least two weeks before use.For each test, gravid female
grass shrimp were examined with a dissecting microscope forpresence of embryos in the tissue cap stage (2-3 d after
oviposition. Female grass shrimp with embryos at thetissue-cap stage were placed under a dissecting microscope
and the embryos gently removed from female pleopods andseparated from other embryos using forceps and fine-tip
probes. Separated embryos were washed three times infiltered 2O-ppt seawater.SEATOX, a 4-d test protocol, wasused as a screening test that extended from 2-3 d prior tohatch through the hatching period (Rayburn 1996,
Characterisation of grass shrimp embryotoxicity test using
the water soluble fraction of no 2 fuel oil. Mar Poll Bull,
32(12) 860-8). Embryos at the tissue cap stage (3 d after
Oviposition were collected and placed individually into
wells of 24-well plastic tissue culture plates. These wereplaced into an incubator at 27C and gently rotated at 60rpm.
Six days later (9d after oviposition) the embryos were
removed and re-examined microscopically. Normal embryos
exhibited well developed eyes, a beating heart and visible
limbs. Dead or abnormal embryos were discarded and replacedwith normal embryos from excess plates kept in the same
conditions. Plates of embryos were then randomly selected
for a given exposure concentration and the seawater was
removed and replaced with 2 ml of test solution. Plates werereturned to the 27C incubators and examined daily formortalities and hatching. The test was terminated after a
4-d exposure, 13d after oviposition. Three tests were
performed with 5-6 solvent different concentrations (nofurther data).Statistical analysis: LC50s with confidence
intervals were calculated by Litchfield-Wilcoxon probit
analysis.
The overall control mortality was 7.4% (16/216).

(125)

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DATE: 19.11.2004

Result:Reliability
29.09.2003:

Species
Endpoint
Exposure period
Unit
NOEC
LOEC
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Palaemonetes pugio (Crustacea)
other: embryo teratogenesis
12 day(s)g/l> .079= .39
no dataother: see freetext
1995
no dataas prescribed by 1.1 - 1.4

Method:Adult male and female grass shrimp were collected by dip net
during spring and autumn 1995 from relatively uncontaminatedlocal estuaries near the U. S. Environmental Protection
Agency. Gulf Ecology Division. Gulf Breeze, Florida. The
grass shrimp were placed in ice chests with water from thecollection site, transported to the laboratory . The grass
shrimp were acclimated slowly to 251 and 20-ppt salinity
over a 4-h period and were maintained communally in glass
aquaria with flow-through seawater at 250C and 20-ppt
salinity. Adult grass shrimp were held in the aquaria for atleast two weeks before use.For each test, gravid female
grass shrimp were examined with a dissecting microscope forpresence of embryos in the tissue cap stage (2-3 d after
oviposition. Female grass shrimp with embryos at thetissue-cap stage were placed under a dissecting microscope
and the embryos gently removed from female pleopods andseparated from other embryos using forceps and fine-tip
probes. Separated embryos were washed three times infiltered 2O-ppt seawater.Three 12-d tests were performed
according to Rayburn (Rayburn 1996, Characterisation ofgrass shrimp embryotoxicity test using the water soluble
fraction of no 2 fuel oil. Mar Poll Bull, 32(12) 860-8)
using 24-well plastic tissue culture plates for each solvent
tested. Each well contained a single embryo and 2 ml of test
solution (static). Each treatment dilution or control was
conducted within a single 24-well tissue culture plate N =24). Plates were placed on rotary shakers (60 rpm) in
incubators in the dark and kept at 27 ± IC. Plates were
removed from the incubator daily and each embryo was
examined for abnormalities of the eye, yolk, heart, head,
hepatopancreas, and telson. Both the number and type ofabnormality were recorded. Survival was determined bystructural integrity of the embryo during the first fourdays of the test, and thereafter by the presence or absenceof heartbeat. Mortalities and hatching were recorded daily.
Exposures were terminated after 12 d(14--l5 d after
oviposition). Three tests were conducted. LC50 values with
95% confidence intervals and coefficients of variation werc

146

LC50 (4 day) average 12.07g/l (of three replicates, values
12.39, 12.15, 11.6). The mortality curve was extremely
sharp and demonstrated a linear concentration response
curve.LC50 (12 day) average 3.63g/l (of three replicates,
values 4.5, 3,31, 3.00). The range of concentrations overwhich mortality occured was much broader that with the 4 day
test.
(2) valid with restrictions(126)

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
calculated from total mortalities.Treatment concentrations:
0.079, 0.39, 0.79, 1.97, 3.95, 7.9 g/l.Statistical analysis:
LC50s with confidence intervals were calculated by
Litchfield-Wilcoxon probit analysis.
The overall; control mortality was 7.4% (16/216).
Developmental abnormalities were not noted at the 0.079g/l concentration
but were seen at 0.39 and 0.79g/l and resulted in delayed hatching. Twoembryos failed to hatch properly and had swimming difficulties as larvae.
Malformations were detected in 21.7% of the embryos exposed to the 3
lowest concentrations and most of the malformed embryos died before theend of the assay. Developmental delay was detected 6 days after
oviposition at the three highers concentrations. By 9 days nearly all theembryos exposed to the highest concentration died before the end of theassay and all embryos that died were malformed before the died. Only the
1.97g/l treatments had larvae that survived with malformations.
(2) valid with restrictions(126)

RemarkResult::Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
LC50
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Palaemonetes pugio (Crustacea)
Mortality
12 day(s)mg/l= 2000 - 9100
no dataother1997
no dataas prescribed by 1.1 - 1.4

Method:Adult male and female grass shrimps were collected using push nets froma site in Escambia Bay, Pensacola, Florida. Collections were made in themonths of Sept. 1995, March 1996, and May 1996. For the Sept 1995
collection, 70 gravid females and 60 males were collected during high tide.
Water temperature was 28.3C and salinity was 20 ppt. For the March 1995
collection, 180 non-gravid females and 150 males were collected during
low tide. Water temperature was 16.80 C and salinity was 8 ppt. For the
final collection in May 1996, 200 gravid females and 200 males werecollected during high tide. Water temperature was 28.1C and salinity was 3
ppt. Shrimp were placed into coolers filled with site water, transported to
the Gulf Ecology Division Laboratory, Gulf Breeze, FL, and identified as
Palaemonetes pugio (Williams 1984). Approximately 60 female and 40
male shrimp from each of three collections were transferred into flowthrough aquaria (80 L) and maintained on a flow rate of- 24L per hour,salinity of 19-22 ppt, and temperature of 19-25C for approximately a sixmonth period. Protective habitats were not furnished in the aquaria. Under
these laboratory conditions, the shrimp were able to reproduce and supply
adequate numbers of embryos for experiments described here andelsewhere (Little 1968). Shrimp were fed - 2.5 grams of flake food
(Tetramin R) daily and twice a week with 25 ml of concentrated brine
shrimp (Artemia saline) nauplii.
Experiments were conducted with over a 9 month period using embryosfrom shrimp that had been maintained in aquaria for different periods oftime, from 1-160 days. A single gravid female shrimp with a clutch ofembryos 3-d (after oviposition) was selected for testing. Embryos were
removed from the female, placed in disposable 24-well flat bottom plastic
culture plates and individually exposed to 2 ml of ethanol at five different
dilutions ( 0.05, 0.10, 0.50, 1.0 and 2.0% v/v%) based on 12-d LC5O
values from Rayburn and Fisher (1996). Dilutions were made withhistological grade 100% EtOH and 20 ppt 0.22 pm filtered natural sea

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water. Filtered sea water was also used as control. Embryos were placedon rotators (Model G2, New Brunswick Scientific Co.) in an incubator
maintained at 2700±1 for 12 d. Rotators were set at 60 rpm to provide agentle agitation of the embryos in the test wells. After a 12-d exposure,embryos were examined for mortality. 12-d LC5O values and 95%
confidence intervals (Cl) were calculated using the trimmed SpearmanKarber method (Hamilton et al. 1977). Average of mean 1 2-d LC5O values
and coefficients of variation (CV) were calculated according to Steel and
Torrie (1980).
Five LC50s were determined from the Sept collection over a period from30-160 days from collection, Four LC50s from the March collection (2-60days) and two LC50s from the May collection (1-30 days.)
11 tests performed in total in saltwater solutions containing 0.37% to 1.10%v/v ethanol. Average control mortality 11.7% with a standard error of 3.2%.
Three results showed mortalities over 16.7%. One results produced a very
low value (order of magnitude lower. High control mortality was observed
and it was not possible to calculate an LC50. The average value from theremainder was an LC50 value 0.53% (4.18g/l) SD=0.20% (1.58g/l).
Embryos oviposited in the field had greater sensitivity (2-10x) than those
ovipositied in the laboratory.
(2) valid with restrictions(127)

Result:Reliability
11.11.2004:

Species
Endpoint
Exposure period
Unit
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

other aquatic crustacea
Mortality
28 day(s)mmol/l= 25 - 150
no dataother

Remark:Reliability
05.10.2003:

Behaviour = 25 mmol; Growth = 150 mmol; Mortality = 75-150mmol.
(4) not assignable

Species
Endpoint
Exposure period
Unit
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

other aquatic crustacea
Mortality
56 day(s)Reliability:(4) not assignable
This otherwise robust study of the acute and chronic
toxicity of pentachlorophenol in 95% ethanol referred to a'solvent control' but the concentration of ethanol was not
given, nor was a median lethal concentration determined forthe solvent control. Mortality rate in controls was highfor Calamoecia lucasi sublethal tests because of the
difficulties experienced in their culture.

no dataas prescribed by 1.1 - 1.4(128)

no dataotherno dataas prescribed by 1.1 - 1.4

05.10.2003

148

(129)

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4. ECOTOXICITY

Species
Endpoint
Exposure period
Unit

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
other aquatic mollusc: Littorina littorea
other: morphological changes (imposex)

Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:Result:Reliability
22.09.2003:

The study was primarily on the effects of tributyl tin, Pb
and Sn. Ethanol only controls received 50 or 100 ng/l.
Ethanol significantly increased length of penis confounding
the interpretation of the effects of heavy metal compounds.
(4) not assignable

Species
Endpoint
Exposure period
Unit
NOEC
Analytical monitoring
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Daphnia magna (Crustacea)
Reproduction rate
21 day(s)
mg/l
> 10
= .1
no dataother1995
no dataas prescribed by 1.1 - 1.4

Remark:Result:Reliability:

This study was designed to identify possible physiologicaland biochemical target sites for the oestrogenic effect ofdiethylstilbestrol on the freshwater crustacean daphnia
magna. Ethanol was used as the solvent for the DES. The
controls used in the experiment a water and solvent , thelatter at 0.001% ethanol (w/w or v/vnot specified but would
approximate to 10mg/l).
Exposure to 0.001% ethanol had no adverse effect on survivalof the test species.
(3) invalid
This study only used a single ethanol concentration and did
not establish a LOEL. It cannot be considered valid to use
to asses the chronic toxicity of ethanol to invertebrates.

=
no dataotherno dataas prescribed by 1.1 - 1.4(130)05.10.2003

(131)

4.6.1

TOXICITY TO SEDIMENT DWELLING ORGANISMS

4.6.2

TOXICITY TO TERRESTRIAL PLANTS

Species
Endpoint
Exposure period
Unit
EC50
Method

:
:
:
:
:
:

other terrestrial plant: Allium cepa (onion)
Growth6 day(s)mg/l= 7890 - 15780 measured/nominal
other

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Year
GLP
Test substance

:
:
:

1984
no datano data

Method:Winter-rested onions (Allium cepa), 1.5 to 2.0 cm diameter, were fitted into
10 cm x 1.5 cm (dia) test tubes after skinning and exposure of root
primordia. Test liquids were added and the whole incubated at about 20
degC, protected against direct sunlight. The original form of the Allium test
had root growth initiated in distilled water and a modified test involved
placing onions directly into test solutions.
Root tips were examined microscopically following e.g. Feulgen or orcein
staining.
Root lengths were measured and compared with controls (%).

Result:Reliability:

12.11.2004

For compounds other than ethanol the test was compared with othermutagen tests.
Ethanol concentration Root length (cm) %Control
2%
2.88
33.8
1%
5.74
67.3
0.1%
7.34
86.1
Control
8.52
100
Density of ethanol = 789 g/l(2) valid with restrictions
This study is primarily an evaluation of this test methodology as a standard
short-term test in environmental monitoring. For ethanol, only the root
growth inhibition end-point is reported. Given the detail presented in thistest development and evaluation this is considered to be valid withrestrictions.
(132)

Species
Endpoint
Exposure period
Unit
EC50
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Lactuca sativa (Dicotyledon)
Emergence
3 day(s)mg/l= 5382 measured/nominal
other
1977
no datano data

Method:Result:Lettuce fruits (Lactuca sativa cv. Great Lakes) were germinated at 30 degCin the absence or presence of concentrations of a wide variety of aliphaticorganic compounds including ethanol. The inhibitory activity of thecompounds was expressed as the millimolar concentration of compound
producing 50% inhibition at 30 deg C.
Effect of Ethanol on Lactuca sativa Reproduction

Reliability

150:

ENDPOINT: 72 h EC50 of = 117 mM (equivalent 5382 mg/l)
Measurement: Germination.
(2) valid with restrictions
This seems to be a well conducted study. There is some concern regarding
the specific germination temperatures of different lettuce cultivars which
may affect reproducibility. This study is considered to be valid withrestrictions; see also Meyer, H. 1971.

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

12.11.2004(133)

Species
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

other terrestrial plant: Lactuca sativa (Lettuce) and Pisum sativum (Pea)
Emergence
2 day(s)Method:Result:Peas (Pisum sativum cv. Alaska) and lettuce seeds (Lactuca sativa cv.
Grand Rapids) were treated for 24 and 44 hours with absolute ethanol orwater. Lettuce was germinated in light and peas in the dark.
Percent germination-------------------

other
1971
no dataas prescribed by 1.1 - 1.4

24 h
44 h
--------------Lettuce
Treatment:
None
65
65
Abs Ethanol 20
16____________________________
Peas
Treatment:

Reliability:

12.11.2004

None
80
80
Abs Ethanol(2) valid with restrictions
Although this study was instituted to investigate solvents as carriers of
substances into seeds it shows that ethanol inhibits germination in lettuce
but not in peas. This is confirmatory to the findings of other studies. This
study, although deficient in detail, is regarded valid with restrictions; seealso Reynolds T., 1977.
(134)

Species
Endpoint
Exposure period
Unit
inhibition respiration
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

other terrestrial plant: Zea mays mays (Corn)other: coleoptile and root size and respiration
1 day(s)mg/l= 3000 - 4000 measured/nominal
other
1958
no datano data

Method:Maize (Purdue hybrid) tissues were established in Warburg reaction
vessels for 9 h before total growth and respiration were determined.
Seedlings were raised in the dark at 30 degC in tap water and sections 6
mm long were cut 2-3 mm from the tips of coleoptiles. Primary leaves were
removed. CO2 was determined by absorption onto KOH soaked filter
paper.
All studies were carried out at 25 degC. Tips were grown with and/or
without ethanol (0, 0.05%, 0.1%, 0.2%, 0.3% and ranging up to 1.5% vol)UNEP PUBLICATIONS

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and indole-3-acetic acid.

ResultReliability::

12.11.2004

Respiratory Quotient (RQ) was also determined.
The threshold concentration for the inhibitory effect on growth andrespiration was approx. 0.1% and the degree of inhibition was increased
markedly through 0.2 to 0.3% ethanol.
The growth of coleoptiles was more sensitive to ethanol than that of theroots. Oxygen uptake was less sensitive to added ethanol in coleoptiles.
Respiratory quotients were depressed by addition of ethanol. The inhibitiveeffect of ethanol was 0.4%.
(2) valid with restrictions
This report gives experimental detail and clear evidence of an effect of
ethanol. Although old, the study can be regarded to be valid withrestrictions.
(135)

Species
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

other terrestrial plant: Solanum tuberosum (Potato)other: tuber respiration
1 day(s)Method:Result:Irish Cobbler potatoes were exposed to concentrations of a number ofchemical species and rates of respiration were determined and expressed
and total CO2 per 100 g in 48 h.
Respiration expressed as total carbon dioxide per 100 g in48 h:Reliability:

12.11.2004

other
1935
no datano data

Controls:
76.9
Ethanol 0.063 mmol/l: 50.3 (2.9 mg/l)
Ethanol 0.25 mmol/l: 60.9
Ethanol 1 mmol/l: 164.7 (46 mg/l)
Ethanol 2 mmol/l: 105.2
Ethanol 8 mmol/l:
44.1 (368 mg/l)(2) valid with restrictions
This old study is well controlled and yields a respiration rate end point inwhich low and high dosages of ethanol are shown to inhibit respiration, witha significant increase in respiration at 1 mmol/l. There are data from a later
study by Rychter (1979) supporting this observation together withcorresponding changes in respiratory enzyme activities. This study istherefore as regarded valid with restrictions.
(136)

Species
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Avena sativa (Monocotyledon)
Growth7 day(s)Method:15 Replicates per treatment were studied in an experiment in which oat
plants were pretreated in darkness with or without 5% carbon dioxide, withor without ethanol (0.2%, 0.3%) for 3 days from the time of planting.

152

other
1958
no datano data

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4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:Reliability:

12.11.2004

The growth of both mesocotyls and coleoptiles was depressedby each factor during pretreatment. Between 3 and 7 days indarkness, both treatments promoted mesocotyl by incrementsof 33.6 and 38.8 mm. Both carbon dioxide and ethanol are
therefore seen to prolong the meristematic phase.
(2) valid with restrictions
Ethanol is seen to stimulate growth in oat seedlings in this study withcomprehensive controls. The observed effect may be considered a positive
finding and the endpoint to be of uncertain toxicological significance.
(137)

Species
Endpoint
Exposure period
Unitrespiratory inhibition
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

other terrestrial plant: Solanum tuberosum (Potato)other: Tuber respiration
2 day(s)?g/l> 1578 measured/nominal
other
1979
no datano data

Method:Result:Potato (cv. Norchip) were preconditioned for 2 wk following harvest. 6
Whole tubers weighing ca 100 g were placed in 2L kjars and ventilated withdifferent gas mixtures at a rate of 400 ml/min containing ethanol,
acetaldehyde or acetic acid vapours. Respiration and respiratoryintermediates were evaluated.
An increase in the concentration of ethanol from 500 to 5000 mul/l led to a
progressive increase in respiration.
Higher concentrations (20,000 mul/l) reduced respiratory upsurge.

Reliability:

12.11.2004

Ethanol induced a decline in the level of 2-phosphoglyceric acid andphosphoenolpyruvate while leading to the accumulation of tricarboxylic acidcycle intermediates including isocitrate and alpha-ketoglutarate. This was
similar to but independent of the action of ethylene.
(2) valid with restrictions
This study is more recent than Miller (1935) and gives similar results
together with supporting respiratory enzyme values. Although the endpoint
is of uncertain toxicological significance the study is regarded as valid withrestrictions because of the complementary findings in the earlier Miller
study.
(138)

Species
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

other terrestrial plant: Helianthus tuberosus (Girasole)other: cell enzyme activity and pigmentation
3 day(s)Method:Slices (1 mm thick) of Jerusalem artichoke were incubated in a dark roomat room temperature in 2 L Erlenmeyer flasks with 1.5 ml distilled water or
distllled water containing the test compound. Solvents were avoided asthese affected induction patterns.

other
1979
no datano data

The incubation medium was vigorously bubbled with a stream of filtered,hydrated air. Microsomal fractions were prepared and tri-cinnamic acid 4-

UNEP PUBLICATIONS

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OECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
hydroxylase activity determined. Cytochrome P-450 and microsomal
proteins were estimated. These were shown to reach maximum activity 24
h after slicing.

Remark:Result:Ethanol, butanol, isopropanol, methanol, phenobarbital, Mn and Fe were
included in medium for up to 72 hr.
This study shows that cytochrome P450 is inducible in plant cells by
ethanol.
Effect of Ethanol on Helianthus tuberosus Enzyme(s)Concentration / Dose 300.00mM
ENDPOINT:
hydroxylase activity
72h
15% +/- 9%P450 content

72h

85 +/- 25 pmol/mg protein

Concentration / Dose 0.00mM (WATER)
ENDPOINT:
hydroxylase activity
72h
13% +/- 1%
Reliability:

P450 content
72h
35 +/- 2 pmol/mg protein(2) valid with restrictions
The endpoint is of uncertain toxicological significance but may haveinterpretative value.

12.11.2004(139)

Species
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

other terrestrial plant: Saccharum officinarum (Sugarcane)other: root abundance
28 day(s)Method:Result:Reliability:

Sugar cane sets were immersed in ethyl alcohol solutions of strength 1, 2,
4, 6, 8 and 10% and root primordia and production was monitored.
A very marked increase in root production was noted, especially at
marginal temperatures(2) valid with restrictions
Ethanol stimulated root production in sugar cane sets which is considereda positive response. The endpoint is of uncertain toxicological significance.
(140)12.11.2004

other
1940
no datano dataSpecies
Endpoint
Exposure period
Unit
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

other terrestrial plant: Solanum tuberosum (Potato)other: shoot size and abundance (15 days); tuber enzymes (5 days)
15 day(s)Method:Irish Cobler potatoes were dipped in water or ethanol
solutions (20, 40 and 80 ml/l) for 24 hours and thenevaluated after 5 days for peroxidase and catalase
activities in their juice.

other
1931
no datano data

After 15 days the number of sprouts on 12 pieces were

154

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
counted and their total length recorded.

Result:Reliability:

12.11.2004

Other compounds evaluated included acetaldehyde, ethylene
chlorohydrin, potassium thiocyanate and thiourea.
Conc. ml/l Peroxidase Catalase Sprouts Total length
0 (water)
2.18
10.0
2
0.2
20 ml/l
1.94
11.9
7
1
40 ml/l
2.36
14.5
12
3
80 ml/l
2.13
17.5
12
7
(2) valid with restrictions
Ethanol stimulated potato tubers into growth from dormancy, an effect that
may be regarded either a positive or negative effect. There were control
groups in this old but apparently well conducted study and the dosage is asingle immersion. However, the endpoint is of uncertain toxicologicalsignificance.
(141)

Species
Endpoint
Exposure period
Unit
EC50
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

other terrestrial plant: Daucus carota (Wild carrot)
Growth28 day(s)mg/l= 395 measured/nominal
other
1978
no datano dataResult:Cultures of carrot cells exposed to 2% ethanol showedarrested growth and soybean cell growth could be decreased
by about 50% by 0.05% ethanol. Concentrations of 2% could be
reduced to nontoxic levels after 20-25 days byvolatilization.

Reliability:

12.11.2004
4.6.3(Density of ethanol = 789 g/l; 0.05% is approximately 395
mg/l).
(3) invalid
This study evaluated the possible effects of compounds used as herbicide
solvents on plant cell growth and ultrastructure. The results show ethanol
to have cytotoxic activity in carrot and soybean cells in vitro, with wide
variation in cytotoxic dose between the two species. The endpoint is of
uncertain toxicological significance and relevance. Overall, this study istherefore regarded as invalid.
(142)

TOXICITY TO SOIL DWELLING ORGANISMS

Type
Species
Endpoint
Exposure period
Unit
LC50
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

filter paper
Eisenia fetida (Worm (Annelida), soil dwelling)
Mortality
48 hour(s)other: microgram/cm^2 filter paper= 100 - 1000 calculated
other
1984
no datano data

UNEP PUBLICATIONS

155

OECD SIDS
4. ECOTOXICITY
MethodETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Eisenia foetida earthworms were obtained from Bert's Bait Farm, Irvine KY.
They were housed in Nalgene boxes filled with moist peat moss and rabbit
manure and stored at 18 degC. Cornmeal was added as an additional foodsource and calcium carbonate was added to prevent pH falling below 5.5.
Mature worms with developed clitellum, weighing 370 to 450 mg wereselected for exposure to test substances.
Worms were exposed individually to deposits of test subtances on filter
paper for 48 hr, in the dark, and mortality was recorded.
Glass shell vials (22 mm x 85 mm) were lined with Whatman No. 1 filter
paper strips (9.5 x 6.8 cm = 65cm2) and placed in cardboard scintillation
vial trays. Chemical concentrations were expressed in microg/cm^2.
For volatile solvents, microlitre quantities were added to pre-moistened
vials when high concentrations were used (as for ethanol)
After inserting earthworms, the vials were capped and stored in the dark in
a horizontal position for 48 hr.
Death was ascertained by gentle prodding of the anterior end. A minimum
100 earthworms were used to determine each LC50 in a replicated tests
involving a geometric series of 5 to 7 exposure concentrations.

Remark:Test substance:Conclusion:

Reliability:

LC50 was determined by the Litchfield-Wilcoxon log dose-effect probit
transformation method.
This study could be regarded as an 'inhalation' exposure study. It is
possible to calculate the vapour loading in the test vessels from theirdimensions and the dose of ethanol used. This equates to 2002000mg/litre.
All chemicals tested in this study were of either technicalor analytical grade.
Regarded as moderately toxic (LC50 <1000 mg/square cm) inthis test(2) valid with restrictions
This experimentally robust study involving 90 chemical
species demonstrated the unpredictability of chemical
toxicity to different animal species. The 'dosage',presented interms of concentration in a given area of
substrate, is difficult to interpret.

12.11.2004

(143)

Type
Species
Endpoint
Exposure period
Unit
LC50
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

other: aquatic
Lumbricus sp. (Worm (Annelida), soil dwelling)
Mortality
96 hour(s)other: mg/l
> 100 measured/nominal
other
1986
no datano data

Method:Result:

Methodology is proposed for simultaneous multispecies
testing by static immersion of test organisms in 20 l oftest solution.
Asellus intermedius (pillbug), Daphnia magna (water flea),
Dugesia tigrina (flatworm), Gammarus fasciatus(Sideswimmer), Helisoma trivolvis (snail), Lumbricus
variegatus (segmented worm) and Pimephales promelas (fathead

156

UNEP PUBLICATIONSOECD SIDS
4. ECOTOXICITY

Reliability
10.08.2003

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

minnow) all gave a 96 h LC50 of >100 mg/l.
(3) invalid
(144)

4.6.4

TOX. TO OTHER NON MAMM. TERR. SPECIES

4.7

BIOLOGICAL EFFECTS MONITORING

4.8

BIOTRANSFORMATION AND KINETICS

4.9

ADDITIONAL REMARKSUNEP PUBLICATIONS

157

OECD SIDS
5. TOXICITY

5.0

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

TOXICOKINETICS, METABOLISM AND DISTRIBUTIONIn Vitro/in vivo
: In vivo
Type: Metabolism
Species: HumanNumber of animals
Males
: 12
Females : 12
Doses
Males
: 150 mg/cu m; 750 mg/cu m and 1500 mg/cu m; and exceeding MAK
Females : 150 mg/cu m; 750 mg/cu m and 1500 mg/cu m and exceeding MAK
Vehicle: other: airRoute of administration: inhalation
Exposure time
: 4 hour(s)Product type guidance:
Decision on results on acute tox. Tests :Adverse effects on prolonged exposure : no significant effects
Half-lives: 1st:
2nd:
3rd:
Toxic behaviour:
Deg. product:
Method: OtherYear: 1994
GLP: no data
Test substance: no data
Method:Remark:Result:SourceReliability

158

:
:

12 Male and 12 female healthy volunteers carried out a day of training andthen 3 experiments at intervals of about 4 days between each. Inhalation
exposures to ethanol were at 80 ppm (150 mg/m^3), (smell threshold); 400ppm (750 mg/m^3) and 800 ppm (1500 mg/m^3) for 4 hours. In a second
series 8 males and 8 females were exposed to concentrations above the
MAK upper limit, 1.e. 1000 ppm (1900 mg/m^3) and at hourly changing
levels to a maximum 3610 mg/m^3. Blood alcohol levels were measured
and subjects were evaluated by the Swedish Performance Evaluation
System.
Statistical tests included analysis of variance, F-test and correlation
coefficients.
The highest dose was below the current MAK value of 1900 mg/cu m (=1000 ppm) and it is concluded that the maximum blood alcohol level willremain below 0.001% both in men and in women. Regression analysis of
the data shows that the blood ethanol concentration (BEC) can bemodelled using the following equation:[BEC] = [exposure (ppm)] × 0.0029 (with a 7% error for 95% confidence).
Blood alcohol levels were between 0.00023 and 0.0021 mg/ml in the firstseries and 0.00066 and 0.0056 mg/ml in the second (Units of concentration
not clear in results). There was good correlation between inhalation
exposure concentrations and resultant blood ethanol concentrations.
In both experiments there were no significant exposure-related effects inthe psychological performance variables in both men and women. In thesecond experiments where concentrations varied about the MAK there
were no significant effects at and below the MAK but at concentrations
above the MAK, exposure was troublesome.
CEFIC Ethyl Alcohol Group(2) valid with restrictions
This appears to be a well-run study evaluating a broad range of air

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
concentrations in relation to the MAK value. It is not clear from the original
paper and its translation whether the blood alcohol levels expressed as'promille' are mg/l or parts per million, former most likely, although this does
not materially affect the conclusions.
Critical study for SIDS endpoint
(145)

Flag
24.06.2004:

In Vitro/in vivo
Type
SpeciesNumber of animals
Males
Females
Doses
Males
Females
VehicleRoute of administration:::

In vivo
Absorption
Human

:
:

11

Method::::other: inspired air: other: oral drink in orange juice followed by prolonged
inhalation
Exposure time
: 1 hour(s)Product type guidance:
Decision on results on acute tox. Tests :Adverse effects on prolonged exposure :Half-lives: 1st:
2nd:
3rd:
Toxic behaviour:
Deg. product:Result:SourceConclusion::

Reliability

:

Flag

:

Expired and inspired air and blood was collected andanalysed for ethanol following ingestion of 0.7 g/kg
bodyweight of ethanol followed by prolonged (more than 1 hr)
inhalation exposure to ethanol in air (14 mg/l).
All experiments were performed on 11 normal, adult, malehuman subjects. Concentrations of ethanol in atmospheric air
were determined by quadrupole mass spectrometer. Air was
sampled into 20 ml syringes which were then injected with 4
ml of freshly boiled hot water and shaken for 8 minutes totransfer ethanol from air to water. Whole blood aliquots
were precipitated with 0.33 M perchloric acid (1:8).
Supernatants were taken for enymatoc analysis or gas
chromatographic analyses.
About 55% of ethanol in air was absorbed by adult volunteers
and fractional absorption was not affected by variations in
tidal volume (0.7 to 2.1 litres), nor by the presence ofsystemic blood alcohol levels up to 50 times higher thanthat of inspired air. Absorption fractions were about 0.55
and the concentration in end expiratory air did not fall
below some 30% of the of the inspired air.
CEFIC Ethyl Alcohol Group
Inspired ethanol is a significant contributor to elevationsof blood alcohol concentration.
(2) valid with restrictions
This well-designed study with intention for use in forensic
studies gave consistent respiratory uptake values andfractional uptake values across all 11 volunteers and values
close to previously published data. There was robust support
for the conclusion that inspired ethanol can be considered
as a source of elevation of blood alcohol concentration.
Critical study for SIDS endpoint

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

24.06.2004(146)

In Vitro/in vivo
: In vivoType: Excretion
Species: HumanNumber of animals
Males
: 976
Females : 114
Doses
Males
:
Females :Vehicle:Route of administration: other: drinking
Exposure time:
Product type guidance:
Decision on results on acute tox. Tests :Adverse effects on prolonged exposure :Half-lives: 1st:
2nd:rd
3 :Toxic behaviour:
Deg. product:
Method: OtherYear: 1996
GLP: no data
Test substance: other TS: alcoholic beverage
Method:Result:SourceReliability
Flag
24.06.2004:::

In Vitro/in vivo
Type
Species

:
:
:

160

150 Volunteers (20-60 year-old) received neat whisky (40%v/v) or an ethanol cocktail (15-20% v/v) at a dose of 0.35,
0.51, 0.68, 0.85 or 1.05 g ethanol/kg bodyweight. To
evaluate possible interference by gastric acid secretion, afurther 12 volunteers also received cimetidine, ranitidine
or omeprazole to suppress acid secretion. Additionally, 16male and 4 female chronic acoholics with blood alcohol
levels greater that 250 mg/dl were recruited. Twoconsecutive blood samples were taken from each of 188
suspected drunk drivers and for subjects with falling blood
alcohol concentration (176), the frequency distribution of
apparenmt rates of disappearance were plotted.
The mean blood alcohol levels for driving under the
influence (DUI) suspects was 1.88 +/- 0.748 mg/ml in males
and 1.86 +/- 0.702 for females. The overall mean rate ofalcohol elimination from DUI suspects was 0.191 +/- 0.049
mg/ml/hr with 95% limits spanning from 0.09 to 0.29 mg/ml/h.
The slowest rate of ethanol disappearance was in a healthy
male who ingested 0.68 g ethanol per kg bodyweight after an
8 hr fast; the beta-slope was 9 mg/dL/h. The fastest rateof disappearance was observed in a male chronic alcoholicduring detoxification; the beta-slope was 36 mg/dL/h. This
4-fold observed difference should be considered when thepharmacokinetics of ethanol become an issue in drinking anddriving trials, for example, when retrograde estimations areattempted.
CEFIC Ethyl Alcohol Group(2) valid with restrictions
Critical study for SIDS endpoint(147)
In vivo
Metabolism
Human

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY
Number of animals
Males
Females
Doses
Males
Females
VehicleMethodYear
GLP
Test substance

ETHANOL
ID: 64-17-5
DATE: 19.11.2004
:
:
:
:
:
:
:
:
:

Remark:Result:SourceConclusion::

Reliability
24.06.2004:

In Vitro/in vivo
Type
SpeciesNumber of animals
Males
Females
Doses
Males
Females
Vehicle:::Remark:Reliability:

The oxidation of ethanol in mammals, in partucular humans
and the regulation of the rate of ethanol oxidation by
enzymes, is reviewed.
Ethanol is metabolized by conversion to acetaldehyde which
can then be further oxidized to acetate which is thenconverted to CoA ester before entering the general pathways
for fat oxidation. Ethanol is oxidized by either cytoplasmic
dehydrogenase (ADH) or microsomal enzymes (the microsomal
ethanol oxidation system, MEOS) to acetaldehyde. Genetic andenvironmental factors that can alter the rates of ethanoloxidation are discussed.
CEFIC Ethyl Alcohol Group
During the oxidation of ethanol at blood levels of about 10
mM, acetaldehyde is present at about below 2 microM whichindicates that the removal of acetaldehyde is via an enzymeof very low Km.
Alcohol dehydrogenase is responsible for the oxidation of
approximately 90% of ingested ethanol.
(4) not assignable(148)
In vivo
Metabolism
Human

:
:
:
:
:

12.11.2004
In Vitro/in vivo
Type
SpeciesOther1987
no data:::In the major pathway for alcohol metabolism, ethanol is metabolised toacetaldehyde mainly by alcohol dehydrogenase (ADH) which is then further
metabolised by aldehyde dehydrogenase (ALDH) to acetic acid. ADH and
ALDH can exist in multiple isoenzymes and ADH polymorphism leads toindividual differences in alcohol sensitivity and metabolism. Deficiency of
the ALDH2 isozyme (ALDH2*2) can lead to flushing syndrome due to
deficient metabolism of the aldehyde intermediate species. Such a genetic
deficiency is only found prevalently amongst people of Mongoloid origin(including Japanese and Chinese.)(4) not assignable
Review article.
(149)
In vivo
Metabolism
Human

UNEP PUBLICATIONS

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Number of animals
Males
: 3
Females :
Doses
Males
: 0.1, 0.4g/kg 16% solution for oral, 7% solution for i.v.
Females :Vehicle: Water
Route of administration: other: intravenous and peroral
Exposure time
: 6 hour(s)Product type guidance:
Decision on results on acute tox. Tests :Adverse effects on prolonged exposure :Half-lives: 1st:nd
2 :3rd:
Toxic behaviour:
Deg. product:
Method:Year:
GLP: no data
Test substance:
MethodResult::Subject ages 29, 33, 34. One carried the low Km isozyme of aldehyde
dehydrogenase (ALDH) and received both low and high doses of ethanol.
Peroral administration was at the rate of 0.1g/kg per 2.5mins and i.v. at
0.1g/kg per 10 mins. Dosing was either after 12hrs fasting or 15 minutes
after a light meal (100g bread, boiled egg, 200ml milk.)
Blood samples were collected from the mean cubital vein anddeproteinized by the PCA method (Okada 1982, Eriksson 1982).
Laboratory temperature 24-26C. Ethanol and acetaldehyde determined bygas chromatography.
i.v. concentrations - fasted
ALDH type Dose Peak EtOH Peak Acetald.
--------------- ------- --------------- -----------------normal
0.4
20mM
9uM
deficient
0.4
16mM
25uM
normal
0.1
5.5mM
<2uM
deficient
0.1
3.5mM
14uM
p.o. concentrations - fasted
ALDH type Dose Peak EtOH Peak Acetald.
--------------- ------- --------------- ----------------normal
0.4
3.5mM
8uM
deficient
0.4
11mM
21uM
normal
0.1
3.5mM
<2uM
deficient
0.1
2.5mM
14uM
0.1g/kg dose - after meal
ALDH type Route Peak EtOH Peak Acetald.
--------------- -------- --------------- -----------------normal
p.o
1.5mM
<2uM
deficient
p.o
1.0mM
28uM
normal
i.v
5.5mM
<2uM

162

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Conclusion:

Reliability:

12.11.2004

deficient i.v
3.0mM
21uM
Peak blood ethanol concentrations lower in peroral than i.v. dosing. Whilst
there were differences in peak concentrations there was no marked
difference in elimination rate between the normal and deficient ALDH types(both 0.12-0.13mg/ml/hr)(2) valid with restrictions
No analytical information provided. Some data only available in graphicalform. Otherwise reasonably well reported.
(150)

In Vitro/in vivo
: In vivo
Type: Metabolism
Species: HumanNumber of animals
Males
: 30
Females :
Doses
Males
: 0.4g/kg
Females :Vehicle:Route of administration
Exposure time
Product type guidance
Decision on results on acute tox. Tests
Adverse effects on prolonged exposure
Half-lives: 1st:
2nd:
3rd:
Toxic behaviour:
Deg. product:
MethodResult

:

:

:
:
:
:
:

oral unspecified

Volunteers split equally between those with normal aldehyde
dehydrogenase (ALDH) and those with the low Km isozyme of ALDH, as
determined from their hair roots by an isoelectric technique (Harada 1978).
Mean ages and body weights were not significantly different between thegroups. Blood samples were collected from the mean cubital vein at 0.5, 1,
2, 3 and 4 hrs after start of ethanol intake and deproteinized by the PCA
method (Okada 1982, Eriksson 1982). Laboratory temperature 24-26C.
Ethanol and acetaldehyde determined by gas chromatography.
Widmark's beta60 value in each subject was calculated from the slope ofthe elimination curve by linear least squares regression of the psuedo liner
portion. The C0 value was calculated from the the regression line yintercept. the r value was derived from the dose divided by C0. Ethanol
elimination rate ER was calculated from beta60 and r. Computer
calculations were used to derive the Michaelis Menton parameters.
Widmark model
normal ALDH------------

Deficient ALDH--------------

beta 60 0.16 ±0.033
C0
0.61 ±0.095
r0.67 ±0.11
ER104.91±10.55

0.14 ±0.017 mg/ml/hr
0.56 ±0.053 mg/ml
0.72 ±0.067 l/kg
96.88±11.73 mg/kg/hr

Michaelis Menton model
normal ALDH--------------

Deficient ALDH----------------UNEP PUBLICATIONS

163

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Conclusion:

Reliability:

12.11.2004
5.1.1

Vmax 0.18 ±0.032
0.15 ±0.026 mg/ml/hr
Km
0.047 ±0.053 0.037 ±0.039 mg/ml
k120.087 ±0.064 0.060 ±0.037
k12
0.093 ±0.071 0.073 ±0.049
Vd
0.61 ±0.91
0.66 ±0.068 l/kg
According to the data presented, there is a slight but not a significant
difference in the ethanol elimination rates between those with normal ALDH
and those with deficient ALDH.
(2) valid with restrictions
There are some apparent discrepancies between the conclusions in the
text and the data. Insufficient information is available to check the statistics
provided. Little analytical information is presented. Assuming the data to becorrect, the study is considered reasonably well reported and reliable.
(150)

ACUTE ORAL TOXICITY

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 9.8 - 11.6 ml/kg bw
Mouse
NMRI
male/female
5physiol. saline
3 doses lying between LD16 and LD84
other
1976
no dataother TS: Analytical grade

Remark:Method: Ethanol administered to SPF NMRI mice (5 per sex per group) by
gavage, diluted as necessary with physiological saline (0.9% NaCl).
Volume administered was 20 ml/kg, or where necessary, 30 ml/kg.
Quantities of solvent were varied so that at least three mortality values
between 16 and 84% were obtained.
All deaths occurred within 24 h. No signs or necropsy findings described.

Result:

LD50 calculated by probit analysis and is for both sexes combined.
Time of death: All occurred within 24 hr. Individual times not given.
Description, severity, time of onset and duration of clinical signs at eachdose level: Not described.
Necropsy findings: Not done.
Potential target organs: Not discussed.
Sex comparison: Not given; LD50 is for both sexes combined. Values citedare 95% confidence limits.
Values cited are 95% confidence limits. Average LD50=10.5ml/kg, whichis equivalent to 8300mg/kg.
LD50's for i.v. and i.p. routes also determined:-

Test condition

164:

i.v. 2.8
i.p. 4.0
Age of animals: not given. Animals (5 of each sex) were housed inpolycarbonate cages in air-conditioned rooms at a temperature of 22 deg.
C and relative humidity of 55%.
Food (Ssniff Standard diet R from Intermast GmbH, Bockum-Hovel) andwater were available ad lib.
Doses: Not stated. At least 3 doses between LD16 and LD84 were used.

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Doses per time period: One.
Volume administered or concentration: 20 ml/kg total volume.
Post dose observation period: 7 days.
Exposure duration: Not applicable.
Test substance was analytical grade.
(2) valid with restrictions
Critical study for SIDS endpoint

Test substance
Reliability
Flag
12.11.2004

:
:
:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 15010 mg/kg bw
Rat

Remark:19 ml/kg converts to 15.01 g/kg bw.
Method used female rats only, 8 per dose and 7 doselevels.

(151)

Female8other: gavaged after 5% gum acacia
16,17,18,19,20,21 ml/kg
other
1992
no dataother TS

Ethanol gavage was preceeded by gum acacia gavage,
intended to reduce local irritation in stomach.
Post dose observation period was 24 h.

Result:

Test condition:

Test substance
Reliability
Flag
12.11.2004

:
:
:

Type
Value

:
:

Potential target organs, male-female comparison, necropsy
findings not reported.
Time of death: Individual times not given.
Description, severity, time of onset and duration ofclinical signs at each dose level: Inebriation to gait
disturbance, dose-related decrease inresponse to painfulstimuli, respiratory depression and coma.
Necropsy findings: Diffuse congestion of the gastric mucosa
without gross haemorrhage or ulceration.
Potential target organs: Not discussed.
Sex comparison: Not applicable.
Clinical observations ranged from inebriation to gait
disturbance and dose-related decrease in response to painfulstimuli, repiratory depression and coma. Deaths were due tocardiorespiratory failure.
Age of animals: Adults, 180 g. Animals were housed at atemperature of 22-26 degC with 12 hr light-12 hr dark cycle.
Food and water were available ad lib.
Doses: 16, 17, 18, 20, 21 and 22 ml/kg.
Doses per time period: One.
Volume administered or concentration: See above.
Post dose observation period: 24 hrs.
Exposure duration: Not applicable.
Test substance was 99.8% ethanol and 0.1% methanol.
(2) valid with restrictions
Critical study for SIDS endpoint
(152)
LD50
= 7000 - 11000 mg/kg bw

UNEP PUBLICATIONS

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Rat
WistarMale
10
no data
six to 8 dose levels
other
1970
no datano data

Method:Remark:Rats were about 100 days old in one experiment, 10-12 months
old in another. Six to eight dose levels with a doseinterval of 1.1 used. Ethanol given as a 40% w/v solution,
Results range of values for old rats to young rats.
Time of death: All deaths occurred within 24 hr. Individual
times not given.
Description, severity, time of onset and duration ofclinical signs at each dose level: Not described.
Necropsy findings: Not conducted.
Potential target organs: Cause of death was respiratory
failure.
Sex comparison: Not applicable.
Age of animals: About 100 days or 10-12 mth. Food and water
were available ad lib.
Doses: 6-8 dose levels, not described.
Doses per time period: One.
Volume administered or concentration: As a 40% w/v solution.
Post dose observation period: 24 hrs.
Exposure duration: Not applicable.
Old rats were considerably more sensitive than young rats.
(2) valid with restrictions
Critical study for SIDS endpoint

Test condition:

Conclusion
Reliability
Flag
12.11.2004

:
:
:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 14.6 ml/kg bw
Rat
Sprague-DawleyMale6other: none

Method:ResultReliability::

Age at start of treatment: Older rats (300-470 g). Dosing by straight needle
in undiluted form in non-fasted animals. LD50 determined by method of
Litchfield and Wilcoxon.
95% confidence limits of result: 12800-16700mg/kg.
(4) not assignable
Very little method descrption was given.
(154)

(153)

12.11.2004
Type
Value
Species
Strain

166

:
:
:
:

other
1971
no dataother TS: analytical grade

LD50= 7800 mg/kg bw
Rat
Sprague-Dawley

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Sex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

male/female

Method:ResultReliability::

Age at start of treatment: 14 day old rats (16-50 g). Dosing by straightneedle in undiluted form in non-fasted animals. LD50 determined bymethod of Litchfield and Wilcoxon.
Number of animals: 6-12
95% confidence limits of result: 6300-9700mg/kg.
(4) not assignable
Very little detail provided of test method

other: none
other
1971
no dataother TS: analytical grade

12.11.2004

(155)

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 11500 mg/kg bw
Rat
Sprague-DawleyMale6other: none

Method:ResultReliability::

Age at start of treatment: young adult rats (80-160 g). Dosing by straightneedle in undiluted form in non-fasted animals. LD50 determined bymethod of Litchfield and Wilcoxon.
95% confidence limits of result: 18800-2700mg/kg.
(4) not assignable
Very little detail given of method used.
(155)

12.11.2004

other
1971
no datano dataType
ValueSpecies
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
11170 - 16710 mg/kg bw
Rat

RemarkReliability
12.11.2004::

Method:(4) not assignable

Type
Value
Species
Strain
Sex

:
:
:
:
:

other
no datano data(156)
LD50= 7060 mg/kg bw
Rat

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167

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

other

Reliability
12.11.2004:(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
ca. 11850 mg/kg bw
Rat

Remark:Reliability
12.11.2004:

BASF Test.
LD50 value was between 11850 and 12640mg/kg.
test substance was ethanol at 90%,70% 50% and 30%.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 9500 mg/kg bw
Mouse

Test substance
Reliability
12.11.2004::

Test substance was 95% ethanol.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 3450 mg/kg bw
Mouse

Remark:Method not specified.

168

no datano data

(157)

other
No
other TS

(158)

other
no dataother TS

(159)

other
no datano data

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
> 790 mg/kg bw
Mouse

RemarkReliability
12.11.2004::

Converted from 10ml/kg.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
5060 - 7850 mg/kg bw
RabbitRemarkReliability
12.11.2004::

Value = 6300 mg/kg body weight(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LDLo
= 7000 mg/kg bw
RabbitRemarkReliability
12.11.2004::

Method not specified.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
Vehicle

:
:
:
:
:
:
:

(4) not assignable
(160)

other
1972
No
no data

(161)

other
no datano data

(162)

other
no datano data

(163)
Other
= 9900 mg/kg bw
RabbitUNEP PUBLICATIONS

169

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Doses
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LDLo
= 6000 mg/kg bw
Cat

RemarkReliability
12.11.2004::

Method not specified.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Other
5500 - 6500 mg/kg bw
Dog

Remark:Reliability
12.11.2004:

Time of death reported to be "12 to 14 hours".
Value is reported as the lethal dose.
(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Other

Remark

:30% aqueous ethanol.

170

other
no datano data
Although the value was reported as an LD50 value, in alater publication (Munch J.C. 1972. Ind. med. Surg. 41, 31)
it is said to be the minimum lethal dose.
(4) not assignable
(164)

other
1936
no datano data

(165)

other
1875
no datano data

(166)

Dog

no dataother TS

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Dogs were dosed by gavage with 4ml/kg (ca. 3160mg/kg) or
8mls/kg (ca. 6320mg/kg) of 33% aqueous ethanol solution.
(4ml/kg dosed within 1 hour, 8ml/kg dosed within 2 hours).
Liver function was tested by use of Bromosulphothalein(BSP) tests. The 8ml/kg ethanol dosed group gave an mean
increased BSP retention time of ca. 10% that of the 4ml/kg
group.
(4) not assignable

Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LD50
= 5560 mg/kg bw
guinea pig

Reliability
12.11.2004:(4) not assignable

5.1.2

(167)

other
no datano data(168)

ACUTE INHALATION TOXICITY

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Result:LC50> 60000 ppm
Mouse
CD-1
male/female
640,000, 50,000 and 60,000 ppm
60 minute(s)
other
1985
no dataother TS: 95% USP
The sexes of the animals were not specified and the numbers given are
estimates as 12 animals per exposure concentration were used.
Time of death: Not applicable, no deaths.
Description, severity, time of onset and duration ofclinical signs at each dose level: Slight to moderate ataxia
occurred and recovery time was more than 4 hours at all
exposure levels.
Necropsy findings: Not applicable
Potential target organs: Not applicable.
Sex comparison: Not applicable
No LC50 was determined as no deaths occurred at any of the exposure
concentrations.
Slight to moderate ataxia was observed and recovery from this exceeded 4
hours at all exposure levels.

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171

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test condition:

Reliability:

Flag
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:172

Necropsy and target organ study not applicable.
Age of animals: Not stated but weighed 25-30 g. Animals were caged withwood-chip bedding in a room at a temperature of 22-24 deg. C and 121 hrlight/12 hr dark cycle.
Doses: 40,000, 50,000 and 60,000 ppm for different exposure duration.
Doses per time period: One exposure pereiod per exposure level.
Volume administered or concentration: Not applicable.
Post dose observation period: 72 days.
Exposure duration: 60, 30 and 10 minutes.
(2) valid with restrictions
The study is reasonably well reported but there are the following deviations
from an ideal protocol.
Exposure period only 60 minutes.
Species mouse rather than preferred rat.
Observations reported for only 3 days rather than 14.
Volume of chamber 29 litres (above 20 litres)
No detailed observations of effects.
No pathology
No detailed reporting of findings down to individual animal.
Critical study for SIDS endpoint
(169)
LCLo
> 29.43 mg/l
Rat
Other
no data12
saturated air
7 hour(s)
other
1981
No
no data
12 rats were exposed to a saturated vapour concentration of
the test substance at a temperature of 20 degrees C. There
were no deaths. The mean atmospheric concentration of test
substance was 29.43mg/l.
BASF Test(4) not assignable
(170)
Other
Mouse

other
1982
no datano data
13,300 ppm for 1.33 hours caused ataxia.
23,940 ppm for 1.25 hours caused narcosis.
29,300 ppm for 7 hours caused narcosis and deaths.

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
31,900 ppm for 0.33 hours caused ataxia.
(4) not assignable

Reliability
18.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

6 hour(s)
other
1980
No
no data

RemarkReliability
12.11.2004::

BASF test(4) not assignable

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

4 hour(s)
other
1980
No
no data

RemarkReliability
12.11.2004::

BASF test(4) not assignable

(171)
Other
Rat

7 hour(s)
other
1982
No
other TS
12 rats were exposed to a saturated atmosphere of the test
substance at 20 degrees C for seven hours. No deaths
resulted.
50% ethanol in water
BASF test.
(4) not assignable
(172)
LC50
= 5.9 mg/l
Rat

(173)
LC50
= 124.7 mg/l
Rat

(174)

UNEP PUBLICATIONS173

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

SourceReliability
29.09.2003

:
:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

:

174

LC0
= 16000 ppm
Rat

8 hour(s)otherno datano data
Patty's Toxicology of INdiustrial Chemicals(4) not assignable
(175)
Other
Rat

8 hour(s)otherno datano data
No details of method. Some deaths occurred at 16000 ppm
andat 32000 ppm.
(4) not assignable
(176)
Other
Rat

other
1918
No
no data
Duration of exposure varied from 0.5 to 21.75 hours. No
effects at 3260 ppm for 6 hours but drowsiness by 8 hours.
Incoordination at 5660 ppm for 1.75 hours and lightnarcosis at 6400 ppm for 12 hours. At 12,400 to 12,700 ppm
there was deep narcosis by 8.5 hours and deaths by 21.75
hours. Deep narcosis and death occurred at 44,000 ppm by
6.5 hours.
(4) not assignable(177)

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Method:Other
guinea pig

other
No
no data
Duration of exposure varied from 3.75 to 24 hours.
No overt effects at 6400 ppm for 8 hours or 9080 ppm for5.25 hours. Light narcosis and incoordination at 12,850 to
13,300 ppm for 8.75 to 24 hours. No effects at 19,260 ppm
for 3.75 hours, but 20,000 ppm for 6.5 hours caused
incoordination, and 21,900 ppm for 9.8 hours caused deep
narcosis and death.
(4) not assignable
(177)
LC50
= 39 mg/l
Mouse

4 hour(s)otherno datano data(4) not assignable
(178)
Other
guinea pig
HartleyMaleother: 0.9% saline
concentration 31, 62,5, 125, 250mM
other
1994
no datano data
Animals with tracheas cannulated with polyethylene tubes, artificiallyventilated (tidal vol 10ml/kg). Changes to resistance to inflation measured
by pressure required to overinflate by 2x tidal volume for 2 breaths.
Animals subjected to 15mins saline aerosol, followed by 15second burstsof ethanol containing aerosol, with a 5 minute gap before next burst of

UNEP PUBLICATIONS

175

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
ethanol aerosol, with increasing concentrations used. 46.4% of aerosol
measured as deposited in lungs by radiolabel technique.
Study designed to assess if ethanol in aerosol form causes
bronchoconstriction.
Ethanol did not cause bronchorestriction.
(4) not assignable(179)

Remark:ResultReliability
18.11.2004

:
:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time

:
:
:
:
:
:
:
:
:

other: volunteer study

Method:Result:Reliability
18.11.2004:

Volunteers: Healthy; 2 atopic, 5 non-atopic; 4 women, 2 men; 5 nonsmokers, 1 regular smoker; age 28-45yrs.
Inhalation via the mouth of an aerosol, particle size 0.5-4.0um. 5 day
interval between exposures.
Lung function assessed by recording partial and maximum expiratory flow
volume at time zero and repeatedly during the 4 hrs after exposure (1 sec
forced expiratory volume and flow rate at 40% of the forced vital capacity.)
Mean of three repeats used.
Statistical analysis: student's t test.
Ethanol concentrations were measured using Draeger tubes in inhaled(measured in breathing tube) and expired air (5 & 30 mins after exposure.)
Subjects reported coughing at start of exposure and 3 reported chest
tightness at end. None reported signs of intoxication normally associatedwith ethanol ingestion. No symptoms were experienced with the saline
control.
Ethanol decreased the maximum expiratory flow rates for the whole of the
4 hour period after exposure (8-37% statistically significant reduction for
the first 90 minutes after exposure.) There was no significant effect on the
one second forced expiratory volume.
The ethanol concentration in inspired air was 0.18-0.2% (1800-2000ppm)
and in exhaled air for 30 minutes post exposure 0.06-0.1% (600-1000ppm)(4) not assignable
(180)

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Method

:

176

Human
male/female
6other: saline
0, 25% in aerosol form
30 minute(s)

Other
Mouse
C57BL
Femalesingle group exposed to 25-38mg/l
24 hour(s)
other
1986
no data

Animals 12-24 weeks old.
Caged mice placed in perspex inhalation chambers.
Feed: CRM pellets (K&K Greefe), freely available during exposure.
Aged matched control mice used
Blood obtained in heparinised syringes (cardiac puncture) then mixed with

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:Reliability
18.11.2004

:

5.1.3

EDTA. Hb, RBC and WBC determined by Coulter counter. PVC measured
using microhaematocrit tubes. Platelet count determined (after 100x
dilution in formal citrate) using a Neubauer counting chamber.
Reticulocytes counted on unfixed smears of supervitally -stained blood.
Blood films stained by the May-Grunwald-Giemsa method and differential
leucocyte counts performed on 500 consecutive nucleated cells.
Femoral marrow expekked into heparinised Hank's solution, dispersed intoa single cell suspension and used for determination of marrow cellularity or
deoxyuridine suppression values.
Quantification of granulocyte-macrophage progenitor cells: Femoral
marrow expelled into MEM alpha medium. Samples dispersed into single
cell medium, washed twice, assays of CFU-GM performed on each marrowcell suspension in triplicate. Details of procedure given in reference.
Many mice showed locomotor depression and ataxia. Blood ethanol levels
were in the range 150-560mg/dl. Ethanol exposed mice developed
leucopenia, neutropenia, lymphopenia, monocytopenia, thrombocytopenia
but not anaemia or macrocytosis. There was no effect on deoxyuridine
suppression values or number of granulocyte-macrophage progenitor cells.
There was a slight reduction in the number of megakaryocytes.
(4) not assignable(181)

ACUTE DERMAL TOXICITY

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

LDLo
= 20000 mg/kg bw
Rabbitno datano data
4

Remark:Reliability:

Dosage translated from 200 Proof. Reported that dose used killed 1 out offour animals.
(4) not assignableNo details of method reported therefore not possible to assess compliant
with relevant testing protocol. No reference source quoted for reported
data.
(182)

12.11.2004
5.1.4

no dataother1968
no datano data

ACUTE TOXICITY, OTHER ROUTES

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Route of admin.
Exposure time
Method

:
:
:
:
:
:
:
:
:
:
:

LC50
= 9450 - 9710 mg/kg bw
Mouseother: HS
male/female
10physiol. Saline
6, 8 and 10 g/kg in 20% w/v solutioni.p.
24 hour(s)UNEP PUBLICATIONS

177OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Year
GLP
Test substance

:
:
:

1995
no datano data

Remark:Result:

Test condition:

Reliability
12.11.2004:

Time of death: All deaths occurred within 30 min. Individual
data were not given.
Description, severity, time of onset and duration ofclinical signs at each dose level: Not described.
Necropsy findings, included doses affected, severity andnumber of animals affected: Not done.
Potential target organs: Not discussed.
Sex comparison: LD50 males; 9.71 g/kg. LD50 females; 9.45
g/kg.
The LD50 vales were 838 to 1127 (9.71 g/kg) in males and8.45 to 1049 g/kg (9.45 g/kg) in females.
Age of animals: 25-30 days. Animals were housed in
Plexiglass cages with aspen shavings in a climate-controlled
room with 12 hr light and 12 hr dark cycle. Food and water
were available ad lib.
Doses: 6, 8 and 10 g/kg.
Doses per time period: One.
Volume administered or concentration: 10 ml/kg total volume
as a 20% w/v solution.
Post dose observation period 24 hr.
Exposure duration: Not applicable.
(2) valid with restrictions

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Route of admin.
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:(183)

Result:

Test condition

:

178

LD50
= 9000 mg/kg bw
Mouse
Swiss Webster
Male
8
Water
6 ranging 5000 to 11000 mg/kg bw
i.p.
Other
1979
no dataas prescribed by 1.1 - 1.4
The LD50 was calculated using the Lichfield-Wilcoxon method.
Time to death, clinical signs, necropsy findings andpotential target organs not reported.
Time of death: Not reported.
Description, severity, time of onset and duration ofclinical signs at each dose level: Not reported.
Necropsy findings, included doses affected, severity andnumber of animals affected: Not reported.
Potential target organs: Not discussed.
Sex comparison: Not applicable.
The LD50 in male mice was 9.2 g/kg bodyweight with a 95%
confidence interval of 8.9 to 9.4 mg/kg.
Age of animals: Not stated but weighed 25-30 g. Animals werehoused in plastic cages in a climate-controlled room with 12
hr light and 12 hr dark cycle. Food and water were available
ad lib.
Doses: Not stated but at least 6 doses between 5.0 and 11.0

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Reliability
12.11.2004

:

Type
Value
Species
StrainSex
Number of animals
VehicleDoses
Route of admin.
Exposure time
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Method:Remark:(184)

SourceTest condition::

Reliability
12.11.2004

:

5.2.1

g/kg.
Doses per time period: One.
Volume administered or concentration: 0.2 to 0.25 ml using a
20% w/v solution.
Post dose observation period 7 days.
Exposure duration: Not applicable.
(2) valid with restrictions
LD50= 5100 - 6710 mg/kg bw
Rat
WistarMale
10
Six to eight dose levels
i.p.
Other
1970
no datano data
Rats were about 100 days old in one experiment, 10-12 months
old in another.
Results are values for old rats to young rats.
Time of death: All within 24 hr, individual times notreported.
Description, severity, time of onset and duration ofclinical signs at each dose level: Not reported.
Necropsy findings, included doses affected, severity andnumber of animals affected: Not conducted.
Potential target organs: Cause of death was respiratory
failure.
Sex comparison: Not applicable.
EU Existing Chemicals Programme HEDSETAge of animals: About 100 days or 10-12 mth. Food and water
were available ad lib.
Doses: Not stated but 6 to 8 doses with interval 1.05.
Doses per time period: One.
Volume administered or concentration: A 15% w/v solution.
Post dose observation period 24 hrs.
Exposure duration: Not applicable.
(2) valid with restrictions(153)

SKIN IRRITATION

Species
Concentration
Exposure
Exposure time
Number of animals
VehiclePDII
ResultClassification
Method

:
:
:
:
:
:
:
:
:
:

RabbitUndiluted
Occlusive
4 hour(s)6
not irritating
not irritating
OECD Guide-line 404 "Acute Dermal Irritation/Corrosion"

UNEP PUBLICATIONS179OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Year
GLP
Test substance

:
:
:

1981
no dataas prescribed by 1.1 - 1.4

Remark:Reliability
Flag
17.11.2004::

Ethanol was applied to six shaved New Zealand white albino
rabbits for 4 hours under exposure chamber of 6 cm2.
Draize scoring criteria.
Mean score for erythema was 1.0 after 1 and 24 hours.
Scores for erythema and oedema were 0.0 at all other time
points.
(2) valid with restrictions
Critical study for SIDS endpoint

Species
Concentration
Exposure
Exposure time
Number of animals
VehiclePDII
ResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Test substance
Reliability
12.11.2004::

Species
Concentration
Exposure
Exposure time
Number of animals
VehiclePDII
ResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

RemarkReliability
12.11.2004::

Species
Concentration
Exposure
Exposure time
Number of animals

:
:
:
:
:

180

(185)
Rabbit95 %4
Waterslightly irritating
not irritating
other
1971
no dataother TSClassification according to Directive 67/548/eec is notpossible from the data presented in this paper.
Method was a modified Draize test employing groups of 4
rabbits and 24-hour covered application.
The average score was 0.5 out of a possible 8 (scores of
0.62, 0.62 and 0.25 were recorded for 3 repetitions).
Test compound was 95% ethanol.
(2) valid with restrictions(186)
Rabbitnot irritating
other
1979
No
as prescribed by 1.1 - 1.4
After Fed Reg vol 38 No 187 27-05-1973 1500.41(4) not assignable(187)
RabbitUNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

VehiclePDII
ResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

RemarkReliability
12.11.2004::

Species
Concentration
Exposure
Exposure time
Number of animals
VehiclePDII
ResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Human
Undiluted
Occlusive
4 hour(s)31Method:Result:Reliability
17.11.2004:

Application of 0.2ml ethanol on a 25mm plain Hill Top chamber containing
a Webril pad to the skin of human volunteers for 4 hours. Full details given
in York (1996)) and Basketter (1997). Treatment sites assessed for
irritation on a four point scale at 24, 48 and 72hrs after pad removal. Any
weakly positive reaction (mild erythema or dryness across most of contactsite) considered a positive reaction. Interpretation of results in terms of EUclassification done by statistical comparison with a concurrent positive
control (20% sodium dodecyl sulphate).
One out of 31 subjects produced a positive result. Positive control
produced a reaction in 15 out of 31 subjects Ethanol therefore considered
non-irritant.
(4) not assignable
(189)

5.2.2

not irritating
Draize Test
1978
No
other TSethanol 96%(4) not assignable(188)

not irritating
not irritating
other
2004
no datano dataEYE IRRITATION

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark

:RabbitUndiluted
100 other: microlitre

moderately irritating
Irritating
OECD Guide-line 405 "Acute Eye Irritation/Corrosion"
1987
no dataas prescribed by 1.1 - 1.4
Method: six New Zealand white rabbits, application of 100
microlitre into the lower conjunctival sac. Draize scoring

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181

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004:

criteria.
Average scores

Test substance
Reliability
Flag
12.11.2004:::

24hr 48hr 72hr------- ---Conjunctivitis
2.50 2.61 2.06
Chemosis1.67 1.17 0.83
Iritis0.50 0.33 0.00
Corneal Opacity 1.00 1.50 1.00
Test substance was neat ethanol.
(2) valid with restrictions
Critical study for SIDS endpoint

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Method:Result:Result(190)

Test substance
Reliability
Flag
12.11.2004:::

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification

:
:
:
:
:
:
:
:
:

182

Rabbit
100 % active substance3
Nonemoderately irritating
not irritating
OECD Guide-line 405 "Acute Eye Irritation/Corrosion"
1998
Yesother TS: 100% ethanol
The method was fundamentally OECD Guideline 405 withinstillation of 0.1 ml, observation for 7 days and standard
grading scales for lesions. However, a Modified Maximum
Average Score (MMAS) was derived by averaging the individual
animal weighted scores at each time of observation and thenselecting the highest (maximum) of these averages. This isa preferred result for this end point as it is a recent
study carried out to a recognized protocol that is reported
in detail.
Average scores
Day 1 Day 2 Day 3------------ ----Corneal opacity
1.33 1.33 0.66
Iritis0.33 0.66 0.33
Conjunctival redness 2.66 2.00 1.66
Chemosis1.66 1.66 0.66
Individual animal observations reported. Full reversal of
all symptoms in animals within 14 days. Most persistenteffect conjunctival redness, still present, grade 1, at 7days (last observation time before 14 day observation.)
Concentration undiluted/100%(2) valid with restrictions
Critical study for SIDS endpoint(191)
other: human, rabbit

other: review of published work

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

MethodYear
GLP
Test substance

:
:
:
:

Remark:Test substanceConclusion::

Reliability
Flag
12.11.2004::

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

not irritating
not irritating
OECD Guide-line 405 "Acute Eye Irritation/Corrosion"
1983
Noother TS

RemarkReliability
12.11.2004::

Pure ethanol(4) not assignable

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

1986
other TS
Routes include direct contact with and without anesthesia,
vapour exposure, injection into orbit, acute and chronic
alcohol intoxication and fetal alcohol syndrome.
spirits, toiltery solutions; various concentrations alcohol
1. Splashes of alcoholic spirit (20-50% alcohol causes
stinging discomfort and reflex closure with no lastic
effects.
2. On rabbit cornea, 50% alcohol causes mild reaction
graded 20 on a scale of 100.
3. repeated applicatuon of 7 drops of 40 to 80% alcoholcaused loss of corneal epithelium and endothelium followedby haemorrhage into conjunctiva, inflitration andvascularization of corneal stroma.
4. Shaving lotions etc may contain up to 90% alcohol;
severe reactions with slow recovery may occur, possibly due
to other components.
5. High vapour concentrations may cause stinging andwatering of eyes above 0.25%.
By other routes - ocular effects are not irritation.
(4) not assignable
Critical study for SIDS endpoint(192)
Rabbit(193)
Rabbitnot irritating
other
1979
No
as prescribed by 1.1 - 1.4

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OECD SIDS
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DATE: 19.11.2004

RemarkReliability
12.11.2004::

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

RemarkReliability
12.11.2004::

Species
Concentration
Dose
Exposure time
Comment
Number of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:Test substance
Reliability
12.11.2004

:
:

184

After Fed Reg vol 38 no. 187 27-09-1973(4) not assignable(194)
Rabbitmoderately irritating
Draize Test
1978
No
other TS
96% ethanol.
(4) not assignable(195)
Rabbithighly irritating
other
1982
no dataother TSClassification of the results according to Directive
67/548/EEC was not possible based on the data presented in
the paper.
Method apparently complied with the main requirements of
the OECD protocol (1979). Groups of six rabbits each had
100 microulitre test substance instilled into the lower
conjunctival sac of one eye. In one group the eyes were notrinsed out, while intwo further groups, rinsing was
performed after 4 and 30 seconds respectively. Scoring was
carried out based on AFNOR recommendations (Association
Francaise de Normalisation, 1982).
When classified according to AFNOR recommendations, ethanol
was severely irritating if not rinsed out of the eye, andvery irritating if rinsed out of the eye after 4 or 30
seconds. Reclassification of the results based only on
readings taken (without rinsing) at the observation times
specified in the OECD protocol, gave rise to aclassification of "severely irritant".
Test substance was ethanol 90 %?.
(3) invalid(196)

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY
5.3

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

SENSITIZATION

Type
SpeciesNumber of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Mouse ear swelling test
Mouse
23

Remark:Age at start of treatment: 6-8 weeks
Acclimation: 7 days

not sensitizing
not sensitizing
other
1988
no datano data

On day 0, mice (9 males and 10 females) were injected s.c.
with 0.05 ml of the test substance in complete Freund's
adjuvant (scapular region) and the test substance was also
applied topically to the abdomen (amount not specified).
On days 3, 5, 7, 10, 12 and 14 they received topicalapplications to the shaved abdomen, and a second scapulars.c. injection of 0.05 ml in CFA was given on day 7. On
day 26, the thickness of the left ear was measured using amobile disk caliper with an accuracy of 0.01 mm immediately
prior to application of the test substance to both sides ofthe ear. Left ear thickness was measured again on days 27
and 28 (i.e. 24 and 48 hours after challenge).
No increase in ear thickness following challenge
application of ethanol.
Measurement of 94 untreated mice showed an ear thickness of
0.214 mm with a typical variation of 0.002 mm. There was no
statistically significant increase in ear thickness
following challenge application of ethanol. Average ear
thickness before: 21.66 +- 1.85 mm.
Average thickness after: 21.69 +/- 1.91 (Swelling 0.1%).
Known moderate and strong sensitizers applied as controls
produced significant swelling in this study.
Test substance was 95% ethanol.
(2) valid with restrictions
Critical study for SIDS endpoint

Result:

Test substance
Reliability
Flag
12.11.2004

:
:
:

Type
SpeciesNumber of animals
VehicleResultClassification
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Guinea pig maximization test
guinea pig
10

Remark:Effective concentrations of ethanol used in the induction
phase were:(197)

not sensitizing
not sensitizing
other
1984
Yes
other TS

Subst. 1; intradermal: 25%
Subst. 1; topical: 37.5%
Subst. 2 intradermal and topical: 37.5%

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185

OECD SIDS
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DATE: 19.11.2004
Effective concentratiuons of ethanol used in the challenge
phase:
Subst. 1 1st and 2nd challenges: 25%, 47.5%
Subst. 2 1st Challenge: 37.5%, 60% and 71.25%
Subst. 2 2nd Challenge: 22.5%, 60% and 71.25%
Method: Test procedure was based on that of Magnusson andKligman (1969) J. Invest. Derm. , 52, 269.
Test animals were femnale Dunkin-Hartley albino guinea pigs;
10 test and 10 controls.

Result:No skin reactions were evoked at challenge with thepolyalkalene glycol in 75% ethanol in either test or
control group animals.

Reliability:

Flag
12.11.2004:

5.4

Although this study was not primarily carried out to assess
ethanol, it can be reliably concluded that ethanol did notshow any signs of sensitizing propert(2) valid with restrictions
There is no detailed information provided on method usedother than reference to a second source document. No
positive control was used.
Critical study for SIDS endpoint(198)

REPEATED DOSE TOXICITY

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
NOAEL
LOAEL
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronicRatmale/female
Sprague-Dawleyoral feed
90 days
Daily

Method:Age at study start: 43 days
No. of animals per sex per dose : 10
Ethanol supplied in nutritionally balanced liquid diet.
Controls received diet without ethanol.
Parameters recorded: Bodyweights weekly, food consumption
daily.
Blood aspartate aminotransferase and alanine
aminotransferase levels determined at termination.
Liver and kidneys were examined macroscopically andmicroscopically at necropsy and the spleen was weighed.

RemarkResult::186

1,2,3,4,5%,10%w/w ethanol in liquid diet
= 2%
= 3%
other
1986
no dataother TS: pureNo statistical tests for significance were used.
2% dose calculated to be equivalent to 2400mg/kg/day
Bodyweight: All groups gained weight though final weights decreased with

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
dose.
Food/water consumption: Consumption in the 10% group was reduced
relative to controls (182 ml diet/kg-d versus 195 ml diet/kg-d).
Clinical signs: No adverse signs were observed
Ophthalmology, haematology: Not examined.
Clinical biochemistry: Serum liver enzymes were not affected by treatment
and kidney findings were minimal.
Mortality and time to death: No deaths occurred.
Fross pathology: Liver yellowing, dosage-related.
Histopathology: Hepatic centrilobular steatosis increased in severity withdose as did the frequency and severity of Mallory bodies (hyaline) andacidophilic degeneration andnecrosis. Most liver findings were absent or mild at 2% w/w ethanol but
became more significant at 3% and higher dose.
Reticulo-endothelial cell proliferation was slight at 1 and 2%. A few kidney
casts were noted in animals from the 1-3% dose groups and there were a
few calcifications in the 3-5% groups. Slight tubular fatty change occurredin all groups.
(2) valid with restrictions
Critical study for SIDS endpoint(199)

Reliability
Flag
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
NOAEL
LOAEL
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Mouse
Male
B6C3F1drinking water
90 days
7 days/week ad libitum

Method:Remark:Result:Age at study start: 43-46 days
Number of animals per sex per group: 10
Ethanol was diluted in deionized water.
No satellite animals were included.
Parameters observed were bodyweights, water consumption and clinicalobservations weekly. Sperm motility was assessed at termination.
Complete necropsies were performed.
Statistical tests were t-tests and F-tests.
The 5% dose was calculated as equivalent to 7300-9400 mg/kg bodyweight over the various urethane dose groups, based on average bodyweight and drinking water consumption.
LOAEL dose was much greater than 5% w/v for observed body and organ
weight increases and decreased sperm count.

5% w/v in deionized water
yes, concurrent vehicle
< 5%
= 5%
other: NTP 13-wk toxicity protocol
1996
Yes
as prescribed by 1.1 - 1.4

No premature deaths occurred.
Bodyweight-relative liver weight was increased and there were increases inabsolute heart, liver, kidney and lung weight
Minimal nephropathy occurred in 30% of treated animals and in 10%
controls. Sperm count in the cauda epididymis was decreased.

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DATE: 19.11.2004

Source:Reliability:

12.11.2004

U.S. Environment Protection Agency High Production Volume, Chemical
Right to Know Program.
(2) valid with restrictions
Single dose used did not allow a NOAEL to be determined so therefore
only reliable with restrictions.
(200)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
NOAEL
LOAEL
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Mouse
FemaleB6C3F1drinking water
90 days
7 days/week ad libitum

Method:Remark:Age at study start: 43-46 days
Number of animals per sex per group: 10
Ethanol was diluted in deionized water.
Satellite animals were not included.
Parameters observed were bodyweights, water consumption andclinical observations weekly. Vaginal cytology was assessed
before termination.
Complete necropsies were performed.
Statistical tests were t-tests and F-tests.
Oestrus cycle length was determined.
Bodyweight: Unaffected by treatment
Food/water consumption: water consumption lowered in ethanol group.
Clinical signs: None noted
Ophthalmological, haematological and blood chemistry findings: Not
examined.
Mortality and time of death: No premature deaths occurred.
Gross pathology: Time spent in dioestrus and pro-oestrus was increased.
Organ weight changes: Ethanol treatment did not affect organ weights.
Histopathology: Non-neoplastic lesions did not significantly differ fromcontrols

ResultReliability::

12.11.2004
Type
Species
Sex
Strain

188

:
:
:
:

5% w/v in deionized water
yes, concurrent vehicle
= 5%
> 5%
other: NTP 13-wk toxicity protocol
1996
Yes
as prescribed by 1.1 - 1.4

The 5% dose was calculated as equivalent to 17000-24000 mg/kg bodyweight over the various urethane dose groups, based on average bodyweight and drinking water consumption.
NOAEL effects were body and organ weights and oestrous cycle length.
The only treatment-related change in female mice was the time spent indioestrus and pro-oestrus but it was unclear whether this was significant.
Cycle length was unchanged.
(1) valid without restriction
Highly reliable. Single dose but sufficient to determine a NOAEL.
(201)
Sub-chronic
Rat
FemaleFischer 344

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Route of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
NOAEL
LOAEL
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

drinking water
90 days
7 days/week ad libitum

Method:Remark:Result:Age at study start: 43-46 days
Number of animals per sex per group: 10
Ethanol was diluted in deionized water.
Satellite animals were included for haematological and clinical chemistry
examination at 3 and 23 days.
Parameters observed were bodyweights, water consumption and clinicalobservatons weekly. Haematology, blood chemistry and vaginal cytology
was assessed before study termination.
Complete necropsies were performed.
Statistical tests were t-tests and F-tests.
The 5% dose was calculated as equivalent to 4800-5600 mg/kg bodyweight over the various urethane dose groups, based on average bodyweight and drinking water consumption.
Body and organ weights were unaffected by treatment while alanine
aminotransferase was decreased and serum bile acids were increased at
week 13. NOAEL and LOAEL were not achieved at this dosage.

5% w/v in deionized water
yes, concurrent vehicle
< 5%
= 5%
other: NTP 13-wk toxicity protocol
1996
Yes
as prescribed by 1.1 - 1.4

No clinical signs, ophthalmological, haematological, or organ weightchanges were observed.
No premature deaths occurred.

Reliability:

12.11.2004

Minimal nephropathy occurred in 40% test animals and in 0% of controls.
No liver lesions were found in controls but hepatodiaphragmatic nodules
were observed in ethanol-exposed animals.
(2) valid with restrictions
Single dose used did not allow a NOAEL to be determined so therefore
only reliable with restrictions.
(200)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
NOAEL
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Rat
Male
Fischer 344
drinking water
90 days
7 days/week ad libitum

Method:Age at study start: 43-46 days
Number of animals per sex per group: 10

5% w/v in deionized water
yes, concurrent vehicle> 5%other: NTP 13-week toxicity protocol
1996
Yes
as prescribed by 1.1 - 1.4

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ID: 64-17-5
DATE: 19.11.2004

Remark:Result:Reliability:

12.11.2004

Ethanol was diluted in deionized water.
Satellite animals were included for haematological and clinical chemistry
examination at 3 and 23 days.
Parameters observed were bodyweights, water consumption and clinicalobservations weekly. Haematology, blood chemistry and sperm motility
was assessed at termination.
Complete necropsies were performed.
Statistical tests were t-tests and F-tests.
The 5% dose was calculated as equivalent to 2800-4100 mg/kg bodyweight over the various urethane dose groups, based on average bodyweight and drinking water consumption.
There was a 20% decrease in thymus weight relative to controls.
Reticulocyte count was increased and serum bile acid concentration
increased. Some other blood biochemical parameters differed
inconsistently from control values at day 3 or 23. Reproductive tissues andsperm counts were not affected by treatment.
(1) valid without restriction
Highly reliable. Single dose but sufficient to determine a NOAEL.
(201)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronicRat
no datano dataoral feed
Up to 36 weeks
ContinuousNone
2 ml ethanol/rat/day given to 6 rats
yes, concurrent no treatment
other

Remark:Reliability
12.11.2004:

Body weight gain decreased in treated rats. Haematologicaleffects evident at all time points (p <0.05) included
reductions in erythrocytes, haematocrit and haemoglobin
concentration, while erythrocyte sedimentation rate, MCV
and MCH were increased. A significant fall in total whitebloodcells was also seen in treated rats (p <0.001) at alltime points. Lymphocytes were reduced while neutrophils
were increased. Monocytes were increased at 10 and 14
weeks only.
Six rats given alcohol equivalent to approximately 8 g/kg
body weight/day (based on body weight mid-way through thestudy). Six control rats given isocaloric amount ofsucrose. Body weights monitored. Haematologicalparameters measured after 10, 14, 18 and 22 weeks.
(4) not assignable

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:
:

190

No
other TS(202)
Sub-chronicRatmale/female
Other
Inhalation
90 days
Continuous
no data
86 mg/m3, 15 rats exposed

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Yes
other
1970
no dataother TS

Remark:Result:

Test substance
Reliability::

Both Long-Evans and Sprague-Dawley rats were used in the
study.
In this study, 15 male and female Princeton-derived
guinea-pigs, 3 male New Zealand albino rabbits, 3 malesquirrel monkeys and 2 male beagle dogs were exposed at thesame time as the rats. No deaths, signs of toxicity or
histopathological effects were reported in these species
either.
One control group (123 rats) used for experiments with 5
chemicals. Blood samples taken before and after exposure
to determine haemoglobin concentration, haematocrit andtotal
leukocyte counts. Sections of heart, lung, liver, kidneyand spleen retained for histopathological examination fromabout half of the rats. Limited number of biochemical andhistochemical determinations carried out.
None of the treated rats died and no signs of toxicity were
evident. Histopathological examination revealed
non-specific circulatory and inflammatory changes that were
not considered to be chemically induced.
Test substance was absolute ethanol (USP grade).
(3) invalid
Insufficient detail reported. Single low dose limits valueof study.

12.11.2004

(203)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Rat
Male
Wistardrinking water
8 monthContinuous
Noneca. 7.7 g/kg body weight/day given to 6 rats.
Yes
other

Remark:Result:Reliability
12.11.2004:

Ethanol was given as a 10% solution in drinking water for 8
months after which a range of haematological parameters wasstudied. Control rats received pure drinking water.
Body weight gain was unaffected in the treated rats.
Following ethanol exposure the osmotic fragility of theerythrocytes was increased. There were no statisticallysignificant effects on haematocrit, haemoglobin
concentration, erythrocyte count, reticulocyte count, MCV,MCH or MCHC.
(4) not assignable

Type
Species::no dataother TS

(204)
Chronic
Other

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OECD SIDS
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ID: 64-17-5
DATE: 19.11.2004

Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:

Test substance
Reliability
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:192

male/female
Other
Other
up to 5 years
Continuous
None
Ethanol added to diet and drinking water of a group of 4 baboons inincrements to reach 25 g/kg body weight/day after 1 year.
Yes
other
no dataother TS
Ethanol was administered in a semi-liquid diet and also inthe drinking water to a treated group of 3 male and 1
femaleanimal and a control group of 1 male and 1 female
animals. The baboons were weighed and given blood tests
regularly. Liver biopsies were performed every 3 months
for 2 years, then every 6 months. The treated baboons werestudied for 9, 18, 48 and 60 months.
Species: Baboon
Strain: Papio
The ethanol-containing diet had no effect on body weightgain. Moderate fatty change was seen in the livers of theanimals treated for 18 and 48 months, while the livers of
those treated for 9 and 60 months were normal. No
cirrhosis was evident at post-mortem.
Test substance was absolute alcohol.
(4) not assignable(205)
Chronic
Other
no dataOtheroral feed
up to 22 months
Continuous
None
Ethanol added to diet of a group of 9 baboons to provide 50% of their total
calorific intake as alcohol.
Yes
other
no datano data
A figure of 80 mg/kg body weight day is given in the paper,
which may relate to the total volume of liquid dietconsumedor to the mean intake of ethanol (which would equate
with 63g ethanol/kg body weight/day).
An additional group of 6 treated baboons and their pair-fed
controls had been given a solid diet with either ethanol orcarbohydrates in the drinking water for periods of from 17
to 34 months. They were then changed to the liquid dietforan average of 17 months. When the ethanol was given in asolid diet, no lesions more severe than fatty liver wereseen, while with the liquid diet, one baboon developed
hepatitis (after 29 months on the solid and 19 months ontheliquid diet), two developed incomplete cirrhosis (after

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
30 months on the solid and 15 months on the liquid diet)and one developed complete cirrhosis (after 34 months on
the solid and 19 months on the liquid diet).
Ethanol was administered in a liquid diet to 9 treatedbaboons and 9 pair-fed controls which were given isocaloric
carbohydrate. Liver biopsies were performed at regularintervals. The baboons were exposed for from 8 to 22
months,the average exposure being 15 months.
Species: Baboon
Strain: Papio hamadryas or olive and yellow
The ethanol-containing diet reduced body weight gain, andinebriation was observed. Fatty liver developed in alltreated baboons, and the liver triglyceride content
increased progressively. Mild inflammation, cellulardegeneration and and some fibrosis were noted in the liver,and ultrastructural changes were seen in the mitochondria
and endoplasmic reticulum.
Three baboons fed ethanol
for 9 months, and one treated for 12 months developed
alcoholic hepatitis. When two of these animals were
biopsied at 20 months, cirrhosis was found.
Serum cholesterol and gutamic-oxaloacetic transaminase
activity were increased in the treated animals, while
haematocrit and haemoglobin values tended to be lower.
(4) not assignable

Result:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Rat
Male
WistarOther
Up to 85 days
ContinuousNone
80 ml ethanol/kg body weight/day given to 16 rats
Yes
other
1985
no datano data

Remark:Body weight gain decreased in treated rats. Haematologicaleffects evident at all time points (p <0.05) included
reductions in erythrocytes, haematocrit and haemoglobin
concentration, while erythrocyte sedimentation rate, MCV
andMCH were increased. A significant fall in total whitebloodcells was also seen in treated rats (p <0.001) at alltime points. Lymphocytes were reduced while neutrophils
were increased. Monocytes were increased at 10 and 14
weeks only.
Treated rats were fitted with two gastrostomy canulae. A
low-fat liquid diet was infused through one, and ethanol
solution through the other. Controls were given the dietand glucose solution (which was isocaloric with theadministered ethanol) through one canula. An initial doseof 8 g ethanol/kg body weight/day was gradually increased
to12 g ethanol/kg body weight/day over 85 days. Blood
sampleswere taken at 2, 4, 6 and 12 weeks. The rats were
killed at15 days (n=2), 30 days (n=7), 45 days (n=2) and 85
days (n=5). Their livers were removed and subjected tomicroscopy and lipid analysis.

(206)

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DATE: 19.11.2004

Result:Reliability:

Body weight gain was unaffected. Fatty degeneration of theliver was seen in all exposed rats. The degree of severity
varied, but correlated with the duration of intoxication,
and with blood alcohol levels (p <0.001). Mild, focal
mononuclear cell infiltration and necrosis of hepatocytes
was mainly found in rats with severe fatty degeneration.
Liver triglycerides were elevated (p <0.001). No similareffects were seen in the controls.
(3) invalid
Study inadequately reported with regard to dosing regime.

12.11.2004

(207)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-acuteRat
Male
Sprague-Dawleyoral feed
4 weekContinuous
None
Ethanol given in the diet at a concentration of 6% v/v to 30 rats.
Yes
other

Remark:Result:Reliability
17.11.2004:

Cell proliferation (crypt cell production rate) was examinedin thegastrointestinal tract (oesophagus, stomach, duodenum, ileum, proximal
colon and rectum) of 30 ethanol-treated rats and their pair-fed controls
given an isocaloric liquid diet containing 36% total calories as
either ethanol or carbohydrate. Blood samples were taken at the time ofdeath.
The crypt cell production rate was increased 2.1 fold in
the rectal mucosa of treated rats (p <0.005) but not in theother gut tissues examined. The proliferative compartmentof the crypt was also expanded towards the colonic lumen inethanol-treated rats (p <0.001). Serum gastrin
concentrations were significantly increased (p <0.01). All
tissues of the gastrointestinal tract were normal by lightmicroscopy.
(4) not assignable
(208)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-acuteguinea pig
no datano dataInhalation
10.5 weeks
Every day for first 2 weeks; thereafter 4 hours/day, 6 days/week.
no data
3000 ppm
yes, concurrent no treatment
other

Remark

:3000 ppm is equivalent to 5643 mg/m3.
Guinea-pigs were exposed to ethanol vapour on 64 occasions.

194

no datano dataNoother TS

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:

Test substance:

Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Precise number exposed not specified but at least 3. Body
weights recorded weekly, blood counts and urinalysis
carriedout every 2 weeks.
No untoward effects other than slight transient albuminuria
in one animal.
Test substance used was "denaturing formular 2b",containing0.5% benzene.
(4) not assignable(209)
Sub-acuteRat
Male
Sprague-Dawley
drinking water
12 week
ContinuousNone3.26 M (equivalent to 10.2 g/kg body weight/day by the end of the study)
given to 12 rats.
yes, concurrent vehicleotherno datano data
No overt signs of toxicity. Body weight gain was reduced
inthe treated rats. No effects on liver function (asmeasuredby GOT, GPT and serum albumin) or haematologicalindices. One treated rat had increased serum creatinine
but there were no microscopic effects on the kidneys.
Organ weights were unaffected and there was no grosspathology at autopsy.Microscopic examination of the liver
revealed fatty degeneration in 10/12 treated rats, aneffect not seen in the controls.
Two groups of 12 rats given either pure water or drinking
water containing ethanol. Rats weighed weekly. Blood
samples collected at the end of study for determination of
haemoglobin, leukocytes, differential white cell count,serum creatinine, GPT, GOT and serum protein. Liver,kidneys, heart and spleen weighed and samples of liver,pancreas, kidney and heart taken for microscopic
examination.
(4) not assignable(210)
Chronic
Monkeymale/female
Other
oral feed
up to 48 months
Continuous
None
Ethanol given as 40% of total ingested calories to a group of 4 monkeys.
Yes
other
1983
no datano data

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OECD SIDS
5. TOXICITYETHANOL
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DATE: 19.11.2004

Remark:Result:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-chronic
Monkey
Male
Other
Other
3 monthtwice daily
None
Ethanol comprised 40% of the ingested calories in a group of 14 animals.
Yes
other

Remark:Result:Reliability
12.11.2004:

A dose level in g/kg body weight can not be derived fromtheinformation presented in the report of this study.
Fourteen rhesus monkeys were given a nutritionally adequate
liquid diet containing 40% of the total calories as
alcohol,twice a day by gavage for 3 months. A control groupof 12 monkeys received the same diet, with the ethanol
replaced bycarbohydrate. At the end of the study, acomplete necropsy was performed, and an unspecified range oftissues was examined microscopically.
The treatment had no effect on body weight gain. Marked
accumulation of triglycerides, cholesterol andphospholipids occurred in the serum and liver. Althoughgeneralized fatty change was evident in the liver, cirrhosis
did not develop. The relative heart weight was increased andmicroscopic effects on the heart were observed (fattychange in the myocardium, focal myocytolysis, atrophy ofmuscle bundles and early fibrosis). Triglyceride andcholesterol ester levels were increased in the heart. ECGs
were normal.
There were no effects on haematological parameters or grossor microscopic effects on the pancreas, kidneys, spleen or
lungs.
(4) not assignable

Type
Species
Sex
StrainRoute of admin.

:
:
:
:
:

196

Eight monkeys were given a nutritionally adequate liquid
diet which provided 50% of their total calorific intake asethanol. Four control animals were given isocaloric
amountsof carbohydrate in the same diet. Liver biopsies
were taken at 3, 12 and 24 months, and at 40 or 48 months
when the monkeys were killed. Detailed description of
methods not provided.
Strain: Macaca radiata
There were no effects on body weight gain or on relative
liver weights. Control liver biopsies were normal. Fatty
infiltration was seen in biopsies from the treated monkeys
at all time points, but no necrosis, inflammation, fibrosis
or effects on hepatic collagen were found.
(3) invalid(211)

no datano data(212)
Sub-acuteRat
Male
Sprague-Dawley
Gavage

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Up to 10 weeks
Daily
None
5 g/kg body weight/day given to groups of 4 or 6 rats.
Yes
other

Remark:Groups of 4 rats were treated with ethanol daily for 0, 1,
2, 5 or 10 weeks, and then given a final dose of 5 gethanol/kg body weight 3 hours prior to being killed. The
livers were removed and the mitochondria and microsomes
wereexamined for evidence of diene conjugation (a method for
detecting lipid peroxidation).

no datano data

Further groups of six rats were treated daily for 0, 1, 2,
3, 5 or 7 weeks prior to being killed. The livers werethenremoved and enzyme activities determined.
Control rats were given isocaloric sucrose in both cases.
Mitochondrial lipid peroxidation was increased in 2 of 4
rats at week 0 (i.e. after an acute dose of ethanol but
withno pre-treatment), 3 of 4 rats after 1 week of ethanol
treatment, and in all treated rats from week 2 onwards.
Microsomal peroxidation was not seen at week 0, but wasevident in treated rats from 2 weeks onwards.
The activities of hepatic glutathione peroxidase andglutathione reductase were increased by ca. 45 and 14%
respectively at all time points.
(4) not assignable

Result:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-acuteRat
Male
Sprague-Dawley
Inhalation
14 days
Continuous

Remark:Result:Study designed to assess the effects of ethanol on immune andhematopoietic systems. Full method details provided in reference.
Ethanol blood levels measured at 169+/-14mg%. No weight changes wereobserved.
A decrease in cellularity was found in the spleen, thymus and bonemarrow. Red and white blood cell counts and haemoglobin concentration
were not affected. Ethanol treatment did alter the relative proportions oflymphocytes and polymorphonuclear leukocytes in the peripheral blood. In
the bone marrow, granulocyte macrophage progenitor cells were notaffected but there was a decline in erythroid progenitor cells. The
proliferation ability of splenic lymphocytes when stimulated by mitogens
was unaffected.

(213)

10mg/l for 3 days then 25mg/l
yes, concurrent no treatment
other
1988
no dataother TS

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test substance
Reliability
18.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-acuteRat
Male
Sprague-Dawley
Inhalation
3, 6, 9, 26 day groups
Continuous

Method:Remark:Result:Reliability
17.11.2004:

Full details provided in reference.
Additional set of animals treated sub-cutaneously daily with pyrazole and acontrol set treated with saline.
Ethanol levels assayed twice daily and in triplicate.
Blood ethanol levels measured in duplicate by gas chromatography details of method provided in reference.
Also measured:- plasma retention of sodium sulfobromopthalein- Plasma activity of glutamic pyruvic transaminase and - glutamic
oxalacetic transaminase- Liver triglycerides- Phagocytic function- Liver and spleen histopathology.
Statistical analysis by student's t test.
Whilst a well reported study, the results are of limited value in assessing
the toxic properties of ethanol relevant to its use as a chemical substance.
Ethanol exposure produced a small but noticeable retardation inbodyweight gain.
Initial exposure produced lethargy, ataxia and intoxication but animals
adapted and appeared normal at the end of the study.
Blood ethanol levels in the ethanol-saline group peaked on day 9 at 126+/40mg/100ml and declined by day 26. In the ethanol-pyrazole group theypeaked at 219mg/100ml (+/- 35). No blood ethanol was measured in either
the air-saline or air-pyrazole controls..
Liver triglycerides were raised (doubled) for the ethanol groups at theearlier time points but were the same as the controls by day 26. Plasma
triglycerides showed no consistent pattern.
Plasma glutamic pyruvic transaminase levels were raised by 20% in theethanol-saline group compared to the control.
Liver samples from the ethanol-saline group exhibited mild vacuolisation for
the early time periods, but this was not seen at 26 days.
No other parameters were significantly effected between the ethanol-saline
and air-saline groups.
(2) valid with restrictions
(215)

Type
Species
Sex
StrainRoute of admin.
Exposure period

:
:
:
:
:
:

198

95% ethanol.
(4) not assignable(214)

20mg/l
Other
other
1979
no datano data

Sub-acuteRat
Male
Sprague-Dawley
Inhalation
35 days

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Continuous

Method:Result:Conclusion:

Reliability
17.11.2004:

Detailed method provided in reference.
Ethanol vapour concentration adjusted to maintain a blood ethanol level of
200-300mg/dl (sufficient to produce signs of intoxication and ataxia levels
2-3 as defined by Majchrowijz (1975).
Rats sacrificed within 1 hour of exposure, lungs and livers removed,
weighed and snap frozen in liquid nitrogen.
Blood ethanol levels determined (method provided in reference).
Liver and lung glutathione and malonaldehyde levels measured (method
details provided.) Enzyme assays also carried out and vitamin E levels
measured.
Ethanol exposed rats showed retarded weight gain. Lung and liver weights
not affected but a moderate decrease in hepatic total protein and a small
decrease in pulmonary soluble protein observed with ethanol exposure.
Ethanol did not affect levels of glutathione of vitamin E in the lung but
levels were significantly diminished in the liver. However, this change
disappeared if levels expressed per gram of protein. Ethanol had no effecton malondialdehyde levels in either tissue..
Of the enzymes, catalase and superoxide dismutase levels weresignificantly increased in the lung by ethanol exposure. Other enzyme
levels (glutathione peroxidase and reductase) were not affected. There
was no effect on anti-oxidant enzyme levels in the liver.
Ethanol exposure does not produce a significant degree in oxidative stressin rat lung.
(4) not assignable
(216)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:see method details
yes, concurrent no treatment
other
1990
no datano data

Sub-acute
Rat
Male
Other
oral feed
3 to 4 weeks (not further defined)
Continuous
no dataethanol consumed by groups of 12 rats equivalent to 12.1 to 16.9 g/kg
body weight/day
Yes
other
no datano dataIn the first experiment, rats received 36% of their total
calories as either ethanol or isocaloric dextrin maltose.
In the second experiment, they received 36% of theircalorific intake as ethanol or isocaloric fat. The ethanol
was administered in a liquid diet at 5 g/100 ml diet. At
the end of the study, the small intestine was removed for
examination.
Strain: CD
Reduced body weight gain was seen in the ethanol-treatedrats regardless of whether ethanol was substituted for

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OECD SIDS
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carbohydrate or fat (p <0.01). There was no effect on
smallintestine weight. The intestinal villi were shorterand contained fewer cells in the treated rats (p <0.001),
but showed no haemorrhagic erosions. Effects on thecrypts indicative of cellular proliferation were seen
(increased epithelial cell count and mitotic index,
increased thymidine kinase activity and higher
incorporation of tritiated thymidine into intestinal DNA).
(4) not assignable

Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Sub-acute
Mouse
Female
C57BL
Inhalation
20-43 days
Continously

Method:Result:Reliability
18.11.2004:

Animals 12-24 weeks old.
Caged mice placed in perspex inhalation chambers.
Feed: CRM pellets (K&K Greefe), freely available during exposure.
Aged matched control mice used
Blood obtained in heparinised syringes (cardiac puncture) then mixed with
EDTA. Hb, RBC and WBC determined by Coulter counter. PVC measured
using microhaematocrit tubes. Platelet count determined (after 100x
dilution in formal citrate) using a Neubauer counting chamber.
Reticulocytes counted on unfixed smears of supervitally -stained blood.
Blood films stained by the May-Grunwald-Giemsa method and differential
leucocyte counts performed on 500 consecutive nucleated cells.
Femoral marrow expekked into heparinised Hank's solution, dispersed intoa single cell suspension and used for determination of marrow cellularity or
deoxyuridine suppression values.
Quantification of granulocyte-macrophage progenitor cells: Femoral
marrow expelled into MEM alpha medium. Samples dispersed into single
cell medium, washed twice, assays of CFU-GM performed on each marrowcell suspension in triplicate. Details of procedure given in reference.
Mice exposed to ethanol developed thrombocytopenia only and none of themore extensive effects seen following short exposure to higher
concentrations. There was no effect on bone marrow. Effects are only
believed to occur following exposures >22-24mg/l(4) not assignable
(181)

5.5

10-25mg/l
yes, concurrent no treatment
other
1986
no data

GENETIC TOXICITY ‘IN VITRO‘

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
Method

200

(217)

:
:
:
:
:
:
:

Ames test
TA97, 98, 100, 104 and 1535
1, 3, 10, 33, 100, 333, 1,000, 3,333, 10,000 microgram/plateNot determinedwith and without
Negative
Other

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Year
GLP
Test substance

:
:
:

1992
no dataother TS: 91% pure

Method:Remark:Result:No. of replicates: 5 + complete repeat of experiment.
Frequency of dosing: Once, including pre-incubation.
Positive and negative controls: Positive controls were
included.
No. of metaphases analyzed: Not applicable.
Solvent used: Not applicable.
Follow-up: Not applicable.
Criteria for evaluating results: Combination of magnitude ofincrease in number of his+ revertants and shape of
dose-response curve.
Positive controls (-S9): sodium azide (for TA1535, TA100), 9-aminoacridine(TA97), 4-nitro-o-phenylenediamine (TA98), methylmethane sulphonate(TA104).
Positive control (+S9): 2-aminoanthracene.
Solvent control: water.
A preincubation assay. This test is considered to be highly
reliable in view of inclusion in NTP mutagenicity testing
program, conducted in 5 strains over a wide range ofconcentrations, with and without two metabilic induction
systems in two concentrations.
Negative.
Test-specific confounding factors: None.
Dose-effected related observations: Ethanol at any dose did
not produce a 2-fold increase in his+ revertants in theabsence or presence of rat or hamster liver extracts.
Mitotic index: Not applicable.
Ethanol failed to induce reversions in any S. typhimurium
tester strain with or without metabolic activation over awide range of doses up to 10 mg/plate.
(1) valid without restriction
Critical study for SIDS endpoint

Conclusion:

Reliability
Flag
12.11.2004

:
:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ames test
Salmonella typhimurium/microsome test
100 microlitre
Not determinedwith and without
Negative
Other
1982
no datano data

Method:Test design: Salmonella/microsomal assays were carried out by making
post-mitochondrial preparations from livers of male Sprague-Dawley rats
induced with Aroclor 1254. Reversion of all strains by 5 microgram/plate of
the promutagen 2-aminoanthracene was included in each assay system.

(218)

Ethanol was one of 25 chemicals examined by spot testing with 5
microgram with and without S9 mix. Compounds
positive in the spot test were then subject to plate incorporation testing (notnecessary for ethanol). Ethanol was actually being used as an inert
solvent.
Positive and negative controls: Both negative controls and positive (2-UNEP PUBLICATIONS

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aminoanthracene, 4-nitro-o-phenylene diamine [frameshift mutagen], 9aminoacridine [frameshift mutagen]and sodium azide [base-pair
substitutions]) were used.

Result:Reliability:

12.11.2004

The Salmonella histidine auxotrophs hisTA98, hisTA100, hisTA1535,
hisTA1537 and hisTA1538 were used.
No evidence of mutagencity was observed for ethanol with and without S9
mix. 9 Of 25 chemicals demonstrated potential mutagenicity in the spot
test but only one of these, DEA laureth sulphate, gave a positive test in theplate incorporation test.
(2) valid with restrictions
Spot testing with confirmatory incorporation testing with 4 positive controls
indicates a valid study. However, there is no mention of Good Laboratory
Practice.
(219)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ames test
Salmonella/microsome
10, 100, 500, 1000 microgram/plate

Method:Ethanol was one of 300 chemicals tested in the standard
Salmonella/microsome Ames test using human or rat liver S9 mix. The test
method is given in detail in Ames, McCann and Yamasaki (1975) Mutat
Res. and is reviewed in McCann & Ames (1975) Ann N.Y. Acad Sci.

With
Negative
Other
1975
no dataas prescribed by 1.1 - 1.4

Remark:ResultReliability::

Salmonella typhimurium strains TA1535, TA1537, TA100 and TA98 wereused.
It is noted that there was a high correlation between carcinogenicity andmutagenicity (90%; 156 carcinogens in 174 mutagens) whereas few
noncarcinogens showed any degree of mutagenicity.
There were <70 revertents per 10,000.
(1) valid without restriction
The methodology of this test is now accepted and repeatedly used as astandard in vitro test for mutagenicity. It is considered robust for detecting
environmental carcinogens. This study is regarded as valid without
restrictions.
(220)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ames test
Salmonella typhimurium LT2 strains
100 microlitre
not presented
with and without
Negative
Other
1983
no datano data

Method:Different concentrations of 9delta-tetrahydrocannabinol and positive
mutagen controls were added in 0.1 ml proportions together with 0.1 ml ofa 16 hr nutrient broth culture of the bacterial test strain or 0.1 ml of theculture and 0.5 ml of S-9 mix. These were then poured onto minimal
glucose agar plates to form an even layer across the agar.

12.11.2004

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Remark:Result:Reliability:

12.11.2004
Type
System of testing::

Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Method:Duplicate plates were made for each strain and plates were incubated for
48 hr at 37 degC. Colonies were counted on a Quebec colony counter.
Background lawn and unreverted bacteria were evaluated by microscopy.
Appropriate control combinations and growth study plates were prepared.
Salmonella typhimurium strains TA1538, TA1537, TA1535, TA100 and
TA98.
This study incorporated ethanol as a negative control in an evaluation of
the mutagenicity of delta9-tetrahydrocannabinol and other mutagens.
Absolute ethanol was used as the solvent for 9-aminoacridine, one of thepositive mutagen controls.
No evidence of mutagencity was observed in the absence or presence of
S9 mix.
(2) valid with restrictions
The study gave the results expected for positive controls and ethanol
evaluated as a solvent to a positive control gave negative results with andwithout S-9 mix. However, there is no mention of Good Laboratory
Practice.
(221)
other: Ames reversion test and DNA repair test in E. coli
Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, TA1538
and E.coli strains
Tested to "toxicity limit" (not defined).
with and without
Ambiguous
Other
1984
no dataother TS
Ethanol tested in revised plate incorporation test as described in Maron,
D.M. & Ames, B.N. (1983) Revised methods for the Salmonellamutagenicity test, Mutation Res.113;173-215.
The Ames reversion test was conducted with his- S. typhimurium strains
TA1535, TA1537, TA1538, TA98, TA100 and, in part, TA97.
S9 Mix contained 10% liver S9 fractions from Sprague-Dawley rats pretreated with Aroclor 1254.
Mutagenic potency was expressed by dividing the number of revertants inexcess of controls by the corresponding amount of ethanol in nmoles.
The genotoxic activity of Escherichia coli was assessed using strains WP2(repair proficient), WP67 and CM871.

Result

Test substance
Reliability:::

A ratio of more than 2 between the MIC's in repair proficient (rep+) and deficient (rep-) strains was considered to be sufficient.
All strains of Salmonella typhimurium showed no reversion in the presenceof ethanol with potency (revertants/nmole of <0.00006.
In the DNA repair test there was equivocal activity in the 2 hr preincubation
assay in the presence of S9, otherwise, ethanol was inactive in theabsence of S9 and in the spot test.
Reagent grade(2) valid with restrictions
Consistency of results in the two tests for 71% of the substances tested
together with an overall predictive accuracy of 64.5% for the reversion test
and 72.4% for the DNA-repair test in 75 compounds classified for their

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carcinogenic activity, demonstrated the validity of this study oncomparability of methods. TRhis study is considered to be valid withrestrictions.

12.11.2004

(222)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ames test
Salmonella typhimurium strains TA97 and TA102

Method:Plate incorporation test (as described in Maron and Ames - 1983. Muta
Res 113, 173.) S9 mix contained 10% liver S9 from SD rats pretreatedwith Aroclor 1254.

Result:with and without
negative1984
no dataas prescribed by 1.1 - 1.4

Mutagenic potential expressed by dividing number of revertants in excess
of controls by corresponding amount of compound in nmoles.
Ethanol was negative for mutagenic activity in the Ames reversion test
using strain TA97 but showed a reproducible increase in revertants overcontrols in TA102 but this was less than two-fold increase which is notnormally considered to be biologically significant in the Ames test. It washowever repeatable.
The authors tentatively classified it as an uncertain or questionablemutagen. However, considering the response against the high dose used
(160mg/plate) suggests that this is an excessively conservative conclusion
and that the balance of evidence points to a negative result.
Source from Carlo Erba
(2) valid with restrictions(223)

Test substance
Reliability
12.11.2004

:
:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Ames test

Remark:Reliability:

Ethanol has been reported as being compatible with the
Salmonella/microsome test at 200ul/plate in the plate incorporation assay
and up to 100 ul/plate in the pre-incubation assay (Maron et al., 1981). At
Safepharm Laboratories, ethanol has been used as one of the validated
vehicle controls for more than 10 years. The typical dose volume used is
100ul/plate, which is equivalent to 79mg/plate, or approximately 16 times
the normal maximum recommended dose level of 5mg/plate used inregulatory mutagenicity tests. In 1998 it was used as the vehicle for 18 test
materials, which was approximately 5% of the total number of studies
completed in that year. The mean, minimum and maximum frequencies ofrevertant colonies for the ethanol vehicle control plates were allcomparable to the 1998 vehicle control history profile for all vehicle controls
used at Safepharm Laboratories in 1998.
(2) valid with restrictions

204

with and without
1997
yes

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

29.06.2004

(20)

Type
System of testing::

Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

Method:DNA damage and repair assay
343/636 (genotype uvrB+/recA+/lac-) and DNA repair deficient 343/591(uvrB-/recA-/lac+)
Up to 1720 mmol/l>1720 mmol/lwith and without
negativeother1992
no dataother TS
Differential DNA repair test as described by Mohn, G.R. et al. (1984)
Methodologies for the direct and animal mediated determination of various
genetic effects in derivatives of strain 343/113 of E. coli K-12, in: B.J.
Kilbey et al. (Eds.) Handbook of Mutagenicity Test Procedures, 2nd edn.,
Elsevier, Amsterdam, pp. 189 - 215.
For each concentration of test compound 100 ul of test compound or thesolvent, 100 ul of bacterial mix and 500 ul S9 mix (where used) were madeup to 1 ml with buffered saline. The mixture was incubated at 37 degC in
the dark before seeding NR agar plates.
The relative survival of DNA repair deficient and proficient bacteria were
calculated.

Remark:Result:

Test substance:

Reliability:

12.11.2004
Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
Method
Year
GLP

:
:
:
:
:
:
:
:
:

Solvent: not specified, but since ethanol was a solvent used for othercompounds and a high concentration was used, it is likely no solvent wasused.
Controls: The positive control was 4-nitroquinoline-N-oxide without S9 mix.
No positive control was used for the S9 mix as this had been previously
validated.
Statistical methods: confidence interval determined according to thevariance of each strain, determined from an experiment with 100 untreatedsamples. A reduction in number of colonies by 2 standard deviations taken
as significant.
This study was a screening test of 61 compounds, giving a mixture of
positive and negative results.
In both the absence and presence of S9 mix, the high dose of 1720
mmole/l ethanol gave a negative result in DNA repair deficient strain of E.
coli.
Test substance was of the highest purity obtainable from commercial
sources.
(2) valid with restrictions
Although conducted to a standard published method this paper does notpresent method detail in full. There was an overall concordance of 80%
between this and the results from Ames tests on the 51 chemicals studied .
The study is therefore considered to be valid with restrictions.
(224)
Bacterial forward mutation assay
Escherichia coli RK+ (replicative killing competent strain CHY832)
11 to 23% v/v
17% v/v
without
positiveother1985
no data

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Test substance:other TS: see TS

Method:Test design: The test strain carries a lethal gene (RK+) that is repressed
below 39 degC and derepressed above this temperature. After treatment
with ethanol at 30 degC cells were plated and cultured at 42 degC to detect
RK- mutants.

RemarkResult::No. of replicates: 3 per concentration.
Frequency of dosing: exposure to ethanol for 10 min before plating.
Positive and negative controls: Negative controls were used.
No. of metaphases analyzed: Not relevant.
Solvent: with and without DMSO.
Evaluation criteria: Positive result when mutation index twice that of control.
The authors suggest that there is a threshold concentration below which
ethanol is not genotoxic. This concentration appears to be the upper limit
for cellular tolerance to ethanol.
Whilst positive, the massive concentration at which this result was seen
can be extrapolated to conclude that the result would be negative at moreconventional test concentrations.
The 5 ethanol preparations showed similar dose-response curves for
induction of RK- mutants with thresholds of 18-19% v/v. Addition ofdimethylsulfoxide lowered the thresholds by around 5% to 13-15%.
Test-specific confounding factors: None.
Dose-effect related observations: All ethanol preparations induced RKmutants with mutation indices of 2 or more. Steep dose-reponse curves
showed threshold at 18-19% v/v.
Frequency of reversions etc: All preparations gave mutation indices of up
to 50 at the highest dose tested.
Mitotic idex: Not relevant.
Synthetic anhydrous 100%, synthetic 95%, 95% grain alcohol,
96.6% grain alcohol and dehydrated absolute 100% grain
alcohol.
The positive result could be due to trace contaminants inethanol, a bacterial metabolite, direct mutagenic effect of
ethanol and indirect effect of ethanol.
(2) valid with restrictions

Test substance:Conclusion:

Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Bacterial reverse mutation assay
Escherichia coli
140 or 180 mg/ml

Method:Result:

The escherichia coli selector strain CHY832 deleted forbio-uvr-chlA was used with and without S9 activation to
examine the mutagenic potential of 48 environmental
chemicals including ethanol. The study was run parallel with
the McCann and Ames Mutatest.
In the Mutatest, ethanol was negative for mutagenicity at 10000 microg/ml
with S9. In the RK test, ethanol was positive for mutagenicity at 180000
microg/ml without S9.

206

(225)

with and without
positiveother1984
no dataas prescribed by 1.1 - 1.4

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Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Chromosomal aberration test
Human peripheral lymphocyte
1% v/v
Not recorded
without
negativeother1985
no dataother TS: analytical grade

Method:Remark:Result:Reliability:

Flag
12.11.2004:

No of replicates: 2 (as solvent control to two other substances).
Duration of treatment: 24 hours.
Number of metaphases analyzed: 100 or 200.
In vitro actication, chromosomal aberrations in blood cultures (without andwith S9 mix delivered via an improvised dialysis bag); sister chromatid
exchange and C-mitotic effects and polyploidies in blood cultures werestudied.
All compounds produced C-mitoses, polyploidies and micronuclei, the latterinterpreted as resulting from errors in the anaphase distribution ofchromosomes by spindle disturbances rather than from structural
chromosome aberration.
Ethanol produced 3 aberrations in 100 metaphases in one study and 4
aberrations in 200 metaphases in another study.
(2) valid with restrictions
This study appears to be well conducted with appropriate controls. The
study is regarded as valid with restrictions.
Critical study for SIDS endpoint
(227)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Chromosomal aberration test

Remark:Reliability
30.06.2004:

During the 1990's, ethanol was used as a vehicle in a sufficient number ofstudies at Safepharm Laboratories to demonstrate that it is not clastogenic
to either human lymphocytes or to Chinese hamster lung cells (CHL). As ingene mutation assays, the dose volume of 1% (100ul/10ml) exceeds the
maximum recommended dose levels suggested by the OECD test
guideline. In the data for 1997, in both cell types, the mean values forethanol controls were slightly higher than the overall control means but the
maximum values were similar in all cases.
(2) valid with restrictions
(20)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result

:
:
:
:
:
:

(4) not assignable(226)

with and without
1997
yes

Chromosomal aberration test
Chinese hamster ovary cell
5%>5%no datanegative

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MethodYear
GLP
Test substance

:
:
:
:

other
1989
no dataother TS

Method:This study examined the potentiating effect of ethanol on other clastogens
in Chinese hamster ovary cells in vitro.

Result:

Test substance
Reliability::

12.11.2004

Plating rate: 3x10^5 cells per petri dish.
Number of replicates: not given.
Frequency of dosing: single dose for 3 hrs.
Positive controls: Methyl methanesulphonate, bleomycin, mitomycin.
Number of metaphases analysed: 100-200 per treatment.
Information cited on aberrant metaphases, chromatid breaks andexchanges, chromosome types (break or ring/dicentric).
Solvent: double distilled water.
Statistical methods: chi-square analysis to assess if effects between twotreatments give statistically significant differences.
Treatment with ethanol alone (5% for 3 hours) had no clastogenic activity
as demonstrated in lack of induction of chromosome breaks and chromatid
exchanges. Tabulated data is reported for 0 and 4% ethanol. Ethanol was
found to potentiate the clastogenicity of known clastogens (those used as
positive controls) with a clear dose response relationship.
Absolute ethanol (ex Merck)(2) valid with restrictions
Key data that would be required to assess compliance with the OECD
protocol are not reported in this study. However, it does appear to beotherwise reliable.
(228)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
human lymphoblastoid cells
1% and 2%
Not statedno datanegativeother1992
no datano data

Method:This study involved ethanol (1% or 2%) alone as a control in an interaction
study to evaluate the effect of ethanol on lobeline sulfate and bleomycin.
Two cell lines evaluated, one derived from a female with multiple primarymalignancies and the second from a patient with cutaneous melanoma.

Result:Reliability:

12.11.2004

208

Number of replicates: 3
Frequency of dosing: no data
Controls: negative and positive (bleomycin)
Number of metaphases analysed: 100.
Solvent: no data, presumed water.
Statistical method: student t test. Results quoted as number of chromatid
breaks per cell, with comparison made with cultures with no treatment.
Ethanol (1%) alone showed breakage rates not significantly different fromcontrols.
(4) not assignable
Study reasonably well reported. Study not designed primarily to assess
clastogenicity of ethanol and some details, required to assess compliancewith OECD protocol, are not reported. Appears to be reliable withrestrictions.
(229)

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
mouse embryo
22, 65, 220 and 650 mM plus control

Method:Result:Reliability
12.11.2004:

This study investigated whether acetaldehyde, the primarymetabolite of ethanol, is responsible for evoking theobserved embryotoxicity, embryolethality,
chromosome-breaking activity and induction of sister
chromatid exchange in mouse embryos in vitro.
4-Methylpyrazole was used to inhibit alcohol dehydrogenase.
It is shown that mouse oocytes as well as morulae andblastocysts are able to xidise ethanol in the presence ofNAD+
Embryotoxicity in pre-implantation embryos was due toacetaldehyde.
(4) not assignable

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
human lymphoid cells
2%, 4%, 6%, 8% and 10%
8% and 10%
no datapositiveother1991
no datano data

Method:Result:Reliability
12.11.2004:

This study was part of a cocarcinogen evaluation involving cigarette smoke
condensates in vitro. Concentrations of ethanol were included as controls.
Ethanol alone showed no demonstrable clastogenic activity as measured
by the frequency of chromatid breaks per cell. At relatively high doses(below cytotoxic doses of 8% and 10%)ethanol inhibited DNA and chromosome repair systems. At 4% there waspronounced uncoiling of chromatids and at 6% the uncoiling was difficult to
identify mitotic figures.
(4) not assignable
(231)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Remark:no datanegativeother1991
no datano data(230)

Cytogenetic assay
Human lymphocytes
0.8 and 1% ethanol (equivalent to 6.31 and 7.89 mg/ml respectively)
without
negativeother1984
no datano data
Peripheral lymphocytes were exposed to ethanol for 24 hours and 100
metaphases were analyzed per treatment.

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Result:Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
Human lymphocyte and Chinese Hamster Ovary cells
0.5 to 10 mg/ml

Method:RemarkResult::Reliability
12.11.2004:

Human lymphocytes and Chinese hamster ovary cells were grown for 12 to48 hors in the presence of 0.5 to 10 mg/ml in the absence or presence of
S9 liver homogenate.
This paper is only an abstract.
CHO cells in SP metabolised ethanol to acetaldehyde. Acetaldehyde
produced a dose-dependent increase in chromosome damage below 5
mg/ml in the same study.
(4) not assignable
(233)

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
Chinese hamster ovary cells
160 mmol/l (equivalent to 7.37 mg/ml)

Remark:Result:Cells were incubated with ethanol for 30 minutes at 30°C. 100 cells werescored for chromosome aberrations. Controls contained 3% DMSO.
No increase in chromosome aberrations was seen in the absence of ametabolic activation system.

There was no increase in either chromatid breaks or isochromatic lesions
at 0.8 or 1% ethanol.
(4) not assignable(232)with and without
ambiguousotherno dataother TS

without
negativeother1987
no datano data

The S2 fraction from Zea mays induced chromosome aberrations (gaps,
breaks and exchanges) when tested on the cells in the absence of ethanol.
In the presence of ethanol and S2 fraction, the aberration rate wasincreased.
(4) not assignable(234)

Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Chromosomal aberration test
human lymphocytes
25, 150 and 500 mg/100 ml

Remark:Blood obtained fron 6 men and 4 women and serum was cultured in the210

without
negativeother1973
no datano data

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
presence of 25 mg, 150 mg or 500 mg/100 ml ethanol for 3 days. Cells
from each culture were examined for chromosome gaps, breakages, rearrangements and aneuploidy. Blastic transformation was studied byThomas' method.
Ethanol had no effect on chromosomes in vitro.
(4) not assignable(235)

ResultReliability
12.11.2004

:
:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
Human lymphocytes
1.16, 2.32, 3.48 mg/ml

Remark:Reliability
12.11.2004:

A significant dose-related increase (p <0.05) in chromosome aberrations(particularly chromatid and chromosome gaps and breaks) was seen at alldose levels.
Cells from 5 donors incubated with ethanol for 50 hours. 100
metaphases/donor screened for chromosome aberrations.
(4) not assignable
(236)

Type
System of testing
Test concentration:::

Cycotoxic concentr.

:

Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:

Method:Remark

:without
positiveother1977
no datano data

Mouse lymphoma assay
Mouse lymphoma L5178Y cells, TK +/0.092, 0.184, 0.369, 0.553, 0.738 mol/l without activation; 0.414, 0.465 and
0.517 mol/l with activation
Maximum concentration with metabolic activation caused <10% fall ingrowthwith and without
negative
other: Clive et al. (1979)1988
no datano data
Test design: mouse lymphoma cell TK +/- forward mutation
assay with and without metabolic activation.
No. of replicates: 3 per dose level but 6 for negative
control.
Frequency of dosing: One 4 h exposure.
Positive and negative controls: Negative (no ethanol) only.
Number of metaphases analyzed: Not relevant.
Solvent/vehicle: Not discussed.
Follow up: Not relevant.
Criteria for evaluating results: 2-fold or greater increasein mutation frequency at 10% or greater total growth cf.
controls. Statistical test 2-tailed Student's t-test.
Results are supported by those of Amacher, D., et al. (1980)
Mutat. Res. 72:447-474.
Test specific confounding factors: None.
Dose-effect related observations: No clear-cut dose-effectrelated observations were seen.
Frequency of reversions etc.: Without activation, mutationindex values from lowest to highest dose were 1.3, 1.1, 1.2,

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1.1 and 1.6. With metabolic activation these values were1.1, 1.3 and 1.8.

Result:Mitotic index: Not strictly applicable. Total growth cf.
controls were 88, 84, 53, 34 and 17% from lowest to highest
concentrations in the absence of activation. With
activation, total growth was 43, 24 and 6% from lowest to
highest concentration.
Only at the maximum concentration, with metabolic activation
was total growth <10% control.
Without activation, the lowest and highest concentrations of
ethanol produced statistically significant increases inmutation frequency.
See Remarks.
Ethanol is judged not to have significant mutagenic activity
in this system.
(2) valid with restrictions

Conclusion:

Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Mouse lymphoma assay
Mouse lymphoma L5178Y cells, TK +/Up to 7.79 x 10-1 M (equivalent to ca. 35.9 mg/ml)
More than 2%
without
negativeother1980
no dataother TS

Method:Cells determined free of mycoplasma before use. Stock cells treatedweekly with THMG mixture to reduce spontaneous mutant levels.
No. of replicates: 3 Plates each and two controls
Frequency of dosing: Once.
Positive and negative controls: Positive controls were
included. 10 Noncarcinogens and 13 putative animal
carcinogens were tested.
Solvent used: none.
Criteria for evaluating results: Gene mutation at thethymidine kinase (TK) locus in trifluorothymidine-resistantL5178Y mouse lymphoma cells.

(237)

Cytotoxicity test: 6x10E5 cells/ml suspension, 5 log range of concentrations
used as range finder. Estimated ID50 used as median dose for main
study. Protocol: 3 hours treatment followed by cell washing. Cell counts at
24 and 48 hours.

Remark:Result:212

Mutagenicity test protocol: As cytotoxicity test then split with cells
resuspended in soft agar cloning medium, with or without
trifluorothymidine.
Mouse lymphoma thymidine kinase assay, as described in Amacher, D.E.
et al. (1979) Point mutations at the thymidenekinase locus in L5178Y
mouse lymphoma cells. I. Application to genetic toxicology testing.
Mutation Res., 64, 391 - 406.
Concentration Cell survival Mutants/10E4 survivors------------------------- ---------------------0
100%
0.73
0.173
91%
0.69
0.26
82%
0.77

UNEP PUBLICATIONSOECD SIDS
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0.346
0.433
0.52
6.06
6.93
7.79

81%
75%
63%
52%
36%
3%

0.81
0.74
0.92
0.68
0.60
0.72

No increase in mutants at clearly cytoxic concentrations.
Purity not specified(2) valid with restrictions

Test substance
Reliability
12.11.2004

:
:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Mammalian cell gene mutation assay
S49 mouse lymphoma cells
1%

Method:This study evaluated ethanol as the solvent control in a study of theinduction of dexamethasone, 6-thioguanine and ouabain resistance.

(238)

with
negativeother1983
no datano data

Materials:
Dulbecco's modified Eagle's medium with 4.5g glucose/l, heat inactivatedhorse serum and fetal calf serum, Bacto Agar, ICR 191 and ethanol ex
Sigma. S49.1 ML cells from P Coffino, San Francisco, originally isolatedby Horibata and Harris (1970).
Cells grown in stationary medium without antibiotics at 37C in a humidified
CO2 incubator. Freshly cloned cultures frozen at -80C. Stock cultures
frequently discarded and repaced with thawed frozen ones to prevent build
up of mutants. Cells stained with trypan blue for counting.
Mutagenic treatment and selection:
4 hour treatment. Cells (6x10E6/dose) centrifuged and re-suspended in
medium containing S9. Maximum solvent concentration 1%. Positive
control used.

Result:Reliability
12.11.2004:

Type
System of testing
Test concentration
Cycotoxic concentr.

:
:
:
:

Criteria for positive result: survival >=40%, factor by which frequency
elevated compared to control >3, frequency of 6-TG mutants elevatedagainst controls.
The dexamethasone resistance marker was induced at the highest
frequency and was expressed within 3 days after mutagenesis. Ethanol
had no effect on the mutagenesis of
this marker.
Surviving fraction: 100%
Mutant frequency: 0
Mean and standard deviation of control: 104 +/- 9
Elevation of mutant freq. compared to control: 0(2) valid with restrictions(239)
Micronucleus test in vitro
Male Chinese hamster lung fibroblast (V79) cells
50 microlitre/ml

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Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:

without
negativeother1984
no dataother TS

Method:This study evaluated the micronucleus assay by comparison with sister
chromatid exchange results for known mutagens/carcinogens. Ethanol,
methanol, butanol and propanol were also examined.
Micronucleus induction was studied in vitro in cells treated with ethanol (50microliter/ml) for 1 hour.
No significant induction of micronuclei was evoked by ethanol in V79
Chinese hamsetr cells.
Absolute ethanol.
(4) not assignable(240)

Result:

Test substance
Reliability
12.11.2004

:
:

Type
System of testing
Test concentration
Cycotoxic concentr.
Metabolic activation
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

Sister chromatid exchange assay
Chinese hamster ovary cells

Method:Result:Methanol, ethanol (0.1% w/v), propanol, butanol and acetaldehyde(0.0005% and 0.001% w/v) were evaluated for effect on SCE in CHO cells
in vitro.
SCE in ethanol treated cells occurred at 4.83 SCE/mitosis versus 4.52 incontrols.

Conclusion:

Reliability
12.11.2004

:

5.6

1977
no dataas prescribed by 1.1 - 1.4

In acetaldehyde treated cells, there were 13.56 SCE/mitosis at the lowest
concentration and 28.25 SCE/mitosis at the highest concentration versus
4.69 SCE/mitosis in controls.
It is acetaldehyde rather than ethanol responsible for an increase in SCE in
CHO cells.
(4) not assignable(241)

GENETIC TOXICITY ‘IN VIVO‘

TypeSpecies
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance214

negative

:
:
:
:
:
:
:
:
:
:
:
:

Micronucleus assay
ratmaleother: BD6
drinking water
10-30 days
5% or 10%
negativeother1993
no dataother TS

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY
MethodETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Remark:Result:No of rats: 51 in 3 separate experiments.
Weight: 180-200 g
Diet and drinking water: standard rodent dioet; water ad libitum alone orwith added ethanol.
Exposure period: 10-30 days.
Investigations: At end of exposure animals were killed and pulmonary
alveolar macrophages and bone marrow erythroblasts were harvested.
Both cytotoxic and cytogenetic effects were examined.
Assuming that rat drinking water consumption is 100ml/kg/day, 5% ethanol
in drinking water would be equivalent to 5000mg/kg, well above the normal
upper limit stated in OECD 474)
No effect on micronucleus incidence was observed.
Polynucleated PAM were enhanced.

Test substance:

Reliability
Flag
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses

:
:
:
:
:
:
:

Result
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:

10% dose was cytotoxic to bone marrow
5% or 10% in drinking water as part of a co- clastogenicity study withtobacco smoke.
(2) valid with restrictions
Critical study for SIDS endpoint(242)

Reliability
12.11.2004:

Type
Species::

Micronucleus assay
ratmale
Wistardrinking water
3 or 6 week
10% or 20% ethanol in the drinking water given to one to four rats per doseand exposure period
other
1980
no dataother TSAge of animals at start: Adult.
No. of animals per dose: 1, 2 or 4.
Dosage: 10% or 20% v/v
Vehicle Control: Tap water.
Duration of test: 3 or 6 weeks.
Frequency of treatment: Daily
Sampling: Hepatocytes, bone-marrow cells and blood lymphocytes for
micronuclei, micronuclei in polychromatic erythrocytes and for sister
chromatid exchanges and chromosomal aberrations.
Group sizes in this study small.
Negative. Drinking ethanol did not affect the incidence of micronuclei inbone marrow cells or hepatocytes at either ofthe two dose levels.
Also, drinking ethanol did not affect the incidence of chromosome
aberrations in bone marrow cells or cultured lymphocytes at either of thetwo dose levels. Frequencies of sister chromatid exchanges in blood
lymphocytes are significantly enhanced in rats exposed to either dose andat the higher dose, ethanol increased the frequency of micronuclei andchromosomal aberration in polychromatic erythrocytes.
(2) valid with restrictions(243)
Micronucleus assay
mouse

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Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

male
Swiss
drinking water
27 days
up to ca. 65 g/kg body weight/day
negativeother1977
no dataother TS: see TS

Method:Remark:Result:Age at study start: 72-75 days.
No. of animals per dose: 3 in negative control, 5 in ethanol groups and 6 inpositive control.
Vehicle: Water.
Duration of test: 27 days.
Frequency of treatment: Ethanol ad libitum; for positive control, ethyl
methanesulfonate by injection 30 and 6 h before sacrifice.
sampling times: Sacrificed on 27th day and 4 slides of stained bonemarrow taken from each mouse.
Controls: see above.
Parameters observed: Bodyweight
Organs/tissues at necropsy: Bone marrow smears only.
Criteria for evaluating results: An average of 4000 polychromaticerythrocytes and corresponding normochromic cells were counted for eachanimal. The % of cells with micronuclei and groups means were calculated.
Criteria for selecting MTD: Not discussed. 2 animals receiving 40% overthe last 2 wk died.
Age of animals at start: 72-75 days
No. of animals per dose: 3 or 5
Dosage: Two groups of mice. Group 1 given 10% alcohol in the drinking
water for 6 days, then 20% for 7 days followed by 30% for 14 days. Group
2 given 10% for 6 days, 30% for 7 days, then 40% for 14 days. Control
group was untreated.
Duration of test: Total 26 days.
Controls: Ethyl methyl sulfonate opr dimethylsulfoxide.
Investigations: Bone marrow preparations were made and examined forpolychromatic and normochromatic erythrocytes.
This investigation suffers from the limitation of a relatively short period of
alcohol ingestion. However, these data were considered sufficiently
reliable by US EPAS for inclusion in the GeneTox Program report and forthis reason have been assigned a reliability score of 2.
Time weighted average concentrations of ethanol were 23% and 33%.
Actual intakes were not determined.
40% level in drinking water is equivalent to an intake of approximately 65
g/kg body weight/day.

Test substance216:

The P/N ratio was not affected by ethanol but was significantly increased in
the positive(ethylmethylsulfonate) control. The incidence of micronuclei wassignificantly increased in the positive control group but not by ethanol.
Mortality at each dose level: 2 aniumals receiving 40%
ethanol died possibly of dehydration. 2 positive control
animals and 0 negative control animals died.
Mutations etc observed: The %PCE's with micronuclei innegative control, low dose, high dose and positive control
groups were 0.37, 0.26, 0.24 and 0.88 respectively.
Clinical signs: Not discussed.
Body weight changes: Not affected by treatment.
Food/water consumption: Not discussed.
Test substance: "distilled ethanol".

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DATE: 19.11.2004

Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Cytogenetic assay
ratmaleother: CDoral feed
6 weeks
36% of dietary energy
positiveother1981
no dataother TS

Method:Age of animals at start: Weanling fed until 130-150 g weight.
No. of animals per dose: 16 males per pair-fed group
Rat strains: CD
Dosage: 12 to 16 g/kg bodyweight/day representing 36% of total energy
intake.
Duration of test: 6 weeks
Frequency of treatment: Fed ad libitum in pair fed groups with group fed
diet without alcohol.
Controls: Untreated only.

RemarkResult(2) valid with restrictions
(244)

:

:

Examinations: Blood samples examined for frequency of micronuclei,
polychmomasia, orthochromasia.
Blood ethanol concentrations were measured at 9am by tail tip excision.
Statistics: student t test.
Dose of 6g/kg given by gavage the day before sacrifice produced nochange in bone marrow cell population, mitotic index or percentage of cellswith micronuclei.
In the authors' opinion, the decreased proportion of nucleated cells most
likely reflects hypoplasia, conceivably from cytogenetic damage of stem
cells.
Blood ethanol concentrations 149mg/100ml +/- 20mg.
Despite pair feeding equal consumption of calories, ethanol fed animals
had significantly lower bodyweight.
Ethanol treatment significantly (P<0.001) decreased the number ofnucleated cells per rat relative to pair-fed controls (4929 ± 774 versus 7996
± 708) and significantly (P<0.05) increased the percentage of nucleatedcells undergoing mitosis to 2.43 ± 0.47 from 1.48 ± 0.38.
Ethanol significantly (P<0.001) increased the number of erythrocytes per
rat from 4789 ± 525 to 7595 ± 390 with significant increases in bothpolychromatic and orthochromatic components. The number oferythrocytes with micronuclei, per rat, was increased significantly (P<0.01)
from 35 ± 7 to 71 ± 12 of which both polychromatic and orthochromaticcomponents were equally affected. This was also reflected in thepercentage of erythrocytes with micronuclei although significant only withrespect to polychromatics.

Reliability:

Overall, the percentage of erythrocytes with micronuclei increased from0.74 ± 0.13 ti 0.94 ± 0.16 (p<0.03) with the PCEs increaseing from 0.95 ±0.13 to 1.30 ± 0.20 (p<0.05) and the OCEs increasing from 0.67 ± 0.12 to
0.84 ± 0.16 (not significant.)(2) valid with restrictions

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12.11.2004

(245)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses

:
:
:
:
:
:
:

Result
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:Reliability:

12.11.2004
Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
Method
Year
GLP

218

:
:
:
:
:
:
:
:
:
:
:

Micronucleus assay
mouse
male/female
other
i.p.
2 day
0.3 ml of 20, 40 or 60% ethanol injected into groups of 4 males and 4
femalesother1977
no datano data
Age at start of treatment: 10 weeks.
Strain: Parkes.
Dosages: Equivalent to 0.62, 1.24 and 1.86 g/kg body weight.
Route of administration: i.p.
Period of treatment: 2 consecutive days.
Controls: no treatment.
Examinations: Femur bone marrow was collected 6 hours after the lastdose and erythrocytes were examined for micronuclei and chromatin
masses.
Positive. Statistically significant increases (p <0.05) inthe occurrence of micronuclei were seen at 0.62 and 1.24
g/kg body weight. At 1.86 g/kg body weight, the occurrence
was increased, but statistical significance was not
attained. No sex difference was evident.
(3) invalid
The data show several inconsistencies that indicate that they may not bevalid. The numbers of observed micronuclei are presented in both tabularand graphical form but the data appear to be inconsistent between the twoforms of presentation. Also, the tabulated data report a control group mean
micronucleus frequency of 4.63% in NCEs and 5.6% in PCEs. These
values are 20-50 times the typical frequency seen in laboratory mice usedin standard regulatory tests. This difference indicates that either the miceused in this study were grossly abnormal in this respect or that there may
have been a problem with the technical quality of the slides or the slide
scoring method used. The data also indicate similar frequencies of
micronuclei in NCEs and PCEs. This cannot have occurred as a result of
exposure to ethanol because the kinetics of the development of NCEs from
PCEs does not allow for the observation of equivalent numbers of
micronucleated erythrocytes of the two types at the single time-point of 30
hours that was used. Because of the apparent errors in this report, and thedeviations from internationally recognized procedures, it is considered thata reliable conclusion cannot be drawn from this data.
(246)
Cytogenetic assay
Chinese hamster
male/female
oral feed
9 weeks
10% v/v
negativeother1979
no data

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test substance:no data

Method:Age at start of study:10-20 weeks.
Dosages: Ethanol (10%) in liquid feed (11 animals); salt water control (36animals), cyclophosphamide (6 animals), ethanol + cyclophosphamide (8animals); patulin (6 animals); ethanol + patulin (7 animals); aflatoxin B (10animals) and ethanol + aflatoxin B (7 animals).

ResultReliability
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses

:
:
:
:
:
:
:

Result
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:Reliability
12.11.2004:

Type
Species
Sex:::

Investigation: Bone marrow examined after 9 weeks for chromatid breaks,
isochromatid breaks, chromatid translations and mitosis with multiple
aberrations.
Ethanol alone had no effect on bone marrow chromosomes in either sex.
(2) valid with restrictions(247)
Cytogenetic assay
hamstermale/female
otherdrinking water
12 week
10% in the drinking water during week 1, 15% in weeks 2 - 3, 20% in
weeks 4 - 12.
negativeother1981
no dataother TS: Absolute, extra pure
Age at study start: 10-20 weeks.
No. animals/dose: Ethanol treated 8 females, 9 males.
Controls 9 females, 7 males.
Vehicle: Water.
Duration of test: 12 weeks.
Frequency of treatment: Drinking water ad libitum. Controls received plain
water.
Diet: (Altromin 7024)
Dosage: 10% v/v in the first week; 15% during the second and third weekand 20% from the 4th to the 12th week. Fluid intake approximately 5.2
ml/hamster/day. Maximum intake therefore up to 26 g/kg body weight/day
in males and 33 g/kg body weight/day in females.
Investigations: Some animals in each group were exposed to cigarettesmoke during the last 4 weeks. Bone marrow was examined forchromosomal aberrations.
Ethanol also failed to induce chromosome aberrations inbone marrow cells in a similar study by the same group inwhich hamsters were given 10% v/v ethanol in the diet for 9
weeks (Korte, A. et al. (1979). The influence of ethanol
treatment on cytogenetic effects in bone marrow cells ofChinese hamsters by cyclophosphamide, aflatoxin B1 andpatulin. Toxicology 12, 53-61.
2.3% aberrant metaphases detected in controls, 3.7 % in treated animals.
No sex difference evident. Mitotic index significantly elevated in smoke
treated group (P <0.001) but not in ethanol controls.
(2) valid with restrictions(248)
Cytogenetic assay
hamstermale/female

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StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

other
drinking water
46 week
10 % v/v ethanol in the drinking water given to 5 females and 2 males
negativeother1981
no dataother TS

Method:Remark:Result:Age at study start: 15 mth. Animals housed individually.
No. animals/dose: Controls 3 males, 2 females. Ethanol 2 males, 5
females.
Vehicle: Water.
Doses were given as 10% v/v (180 g/kg/day).
Duration of test: 46 wk.
Frequency of treatment: Drinking water ad libitum. Controls received plain
water.
Sampling: Blood taken in the 47th week. Two samples per animal
analyzed.
Clinical observations: None.
Organs examined at necropsy: None.
Criteria for examining results: Chromosomal aberrations in lymphocytes
included chromatid breaks, isochromatid breaks and chromatidtranslocations. An aberrant metaphase cell contained at least one
aberration.
Criteria for selecting MTD: None.
Statistics: Chi square test.
Group size in this study was small and ingested dose was uncertain.
Hamsters are reported to have ingested about 1.4 ml/g body weight/week(157g/kg body weight/week). This figure may have been erroneous as the
hamsters appear to have consumed ca. 45 ml fluid/week, corresponding toan approximate intake of 17 g ethanol/kg body weight/day.
The rate of aberrant metaphases was higher inthe ethanol-treated group than in the control group (10.8
versus 7.7%) but the difference was not statisticallysignificant (p >0.25).
Mortality at each dose level: None.
Clinical signs: None described.
Body weight changes: Did not change significantly.
Food/water consumption changes: Animals consuming ethanol
ate 30% less food than controls.
Test substance was ethanol absolute, extra pure, Merck.
(2) valid with restrictions

Test substance
Reliability
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
male
other
gavage5 day
10 or 40% ethanol in water (dose volume 2 ml/kg) to groups of 15 mice
ambiguous
other
1982
no datano data

Remark:Age at start of treatment: 10-12 weeks

220

(249)

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Result:Conclusion:

Reliability:

12.11.2004

Strain: CFPL or Alderly Park.
Treatment: Mice dosed with ethanol in distilled water on 5 consecutive
days.
Dosages: equivalent to 0.25 of the MTD (10% ethanol) and the MTD (40%ethanol). Actual doses administered were 0.16 and 0.63 g/kg bodyweight/day by oral gavage.
Controls: treated with distilled water only.
Mating: Immediately after completion of dose schedule, each male wascaged sequentially with 2 undosed females each week for 8 consecutive
weeks. All females were killed and examined 18 days after first being
caged with males.
Implantation sites and dead implants/female were recorded.
Replicates: In 3 different laboratories.
No effect on pregnancy rate. Occasional positive results with regard to
preimplantation loss during weeks 7 and 8 (reduction in the number of
implants/male). It was suggested that in most cases this was due to alower number of implants in the corresponding control groups. There werealso occasional increases in the number of postimplantation deaths/male
although the majority of the post implantation results were not significant.
Ethanol is unlikely to be a dominant lethal mutagen, at least up to the
maximum tolerated dose.
(1) valid without restriction
This is a highly reliable study that was well reported and compliant with
OECD protocols
(250)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
male/female
Swissother: i.p. (acute) and drinking water (chronic)
3 days (acute) and 11 weeks (chronic)
1.26 g/kg/day and 1.04 g/mouse/day
positiveother1994
no datano data

Method:Remark:Result:Age at study start: 12-16 weeks (25-30 g).
Strains: Inbred Swiss, C57Bl6 and CBA
No. animals/dose: Controls 3 males, 2 females. Ethanol 2 males, 5
females.
Dosage: 0.1 ml 40% alcohol i.p. (acute study). 5% in drinking water
increased by 5% every week to 40% and then at 40% for 4 weeks.
Equivalent dose 0.13 g/mouse/day at 5%; 1.04 g/mouse/day at 40%.
Period of treatment: 3 days (acute), 11 weeks (chronic).
Vehicle: Water.
Replicates: 2 or 3
Mating: 4-day schedule post last treatment.
Investigations: Uterine contents, deciduomas, post-implantation losses.
This study was designed to reproduce the results of Badr using i.p.
injection rather than intubation, but it was unable to do. The authors
concluded that ethanol did not have a significant dominant lethal effect but
caused some pre-implantation loss, which might be due to an effect on thefertilization capacity of sperm.
In Swiss mice, the mutagenic index based on both pre- and postimplantation lethality was consistently positive.
There was a marked reduction (34% and 30%) in the number of pregnant
females at the first two mating times in the treated group and a significant

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decrease in total and live implants in the second mating. There was noincrease in dead implants from the first two matings and only a small
increase at the third mating (P<0.05).
(2) valid with restrictions(251)

Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
male/female
C3H
oral feed
4 weeks in males, then mated with untreated females
0%, 20% and 30% of ethanol derived calories
negativeother1982
no dataother TS: 95%

Method:Result:Conclusion:

Reliability
12.11.2004:

Age at start of treatment: 10 weeks.
Strain: C3H/HE mice.
Feeding: ad libitum.
Environment: temperture and humidity controlled constant with diurnal
daylight/dark rhythm.
Dosage: 0%, 20% or 30% of isocaloric diet made up of ethanol-derived
calories.
Replicates: pair-fed regimen.
Sampling: Weekly blood for blood alcohol concentration.
Investigations; Implantation sites, dead, resorptions and live foetuses
counted.
Statistical analysis: Undernutrition and gender factors considered.
No differences were found between the litters of alcohol-treated males andcontrols in terms of number of implantation sites, prenatal mortality, foetal
weight, sex ratio or frequency of soft tissue malformations.
Paternal alcohol consumption does not grossly alter foetal growth anddevelopment in C3H mice.
(2) valid with restrictions
(252)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
male/female
other: CF1
oral feed
5 weeks
5% v/v liquid diet (28% ethanol-derived calories)
positiveother1991
no dataother TS: 95% USP

Remark:Age of animals at start: 8-9 weeks; 30.1 g
No. of animals per dose: 10 per group
Dosage: 5% v/v in liquid diet representing 28% of total energy intake.
Duration of test: 5 weeks
Frequency of treatment: Fed ad libitum in pair fed groups with group fed
diet with alcohol replaced by sucrose.
Mating: 3 Females were housed with each male and examined daily forpresence of vaginal plug to a maximum 6 days.
Examinations: Tail blood samples examined for haematocrit.

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Females were housed until day 14 when ovaries and uteri were scored fordominant lethal mutations. A mutation index (MI) was calculated: MI/100 =(no of corpora lutea + dead foetuses - total foetuses)/no of corpora lutea.
This study was part of a co-mutagencity study involving delta9tetrahydrocannabinol and Trenimon.
Ethanol caused minimal impairment of fertility at this dosage, but increased
the frequency of dominant lethal mutations.
(2) valid with restrictions(253)

Result:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
ratmaleSprague-Dawleyoral feed
5 week
6% ethanol in the diet for 1 week, then 10% given to 6 rats
positiveother1976
no dataother TS

Method:Result:Animals: weight at start of treatment: 318 g (males); 259 g (females). No ofanimals: 12 males; 25 females.
Environment: Individually housed, 22 degC, RH 45% +/- 10 with diurnal
light cycle (nocturnal 20:00 to 8:00hrs.)
Dosage: Six treated rats were given a 6% v/v ethanol-containing liquid diet(providing 35% of calories.) This was increased to 10% v/v after 7 days
exposure (58% of dietary calories.
Controls: Six controls given an isocaloric amount of sucrose.
Diet 'Metrecal' (chocolate or vanilla.)
Duration of treatment: 15 days (males). females on lab chow and water.
Investigations: After treatment each male was placed in a cage with 2
females every night. No food or water was provided during this time. The
experiment was continued for 5 weeks, with the males still receiving
alcohol during the day. The males were killed on day 36. Blood samples
were collected.
Pregnancies were terminated on day 20 of gestation and litter size andfoetal mortality was assessed.
Treated males showed signs of intoxication and considerable weight gain
compared to controls. Treated animals were much less succesful atmating; the numbers of successful matings were 6/12 in the treated groupand 13/13 in the controls. The number of offspring/litter was greater in thecontrols (p <0.01). A higher incidence of early resorption was seen in thetreated group (p <0.01).

Test substance
Reliability::

12.11.2004
Type
Species
Sex
StrainRoute of admin.

:
:
:
:
:

From graphical data presented in the reference, it is possible to estimate
ethanol consumption as being in the range 7.2 to 14.4 g/kg/day.
Test substance was 95% v/v ethanol.
(2) valid with restrictions
Only six pregnancies examined in treatment group and males alsochronically treated with ethanol such that the quality of the study is
reduced.
(254)
Dominant lethal assay
ratmaleLong-Evansdrinking water

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Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

60 day20% v/v ethanol solution given to 10 rats
positiveother1982
no dataother TS

Method:RemarkResult::Age at Study start: Not stated. Animals 200-300 g and were acclimated for
2 wk before mating.
No of animals/group: 10.
Dosage: Level of alcohol in the drinking water equivalent to a daily dose of
15.7 g/kg body weight.
Vehicle: Distilled water.
Duration of Test: Males treated for 60 days then mated with 3 females overthree weeks.
Frequency of treatment: Ad libitum for 60 days.
Sampling: Testicular tissue examined after the third mating.
Uterine contents examined on gestation day 20.
Controls: Untreated males.
Clinical observations: male bodyweights before and after 60 day exposureand at sacrifice.
Histopathology: Testicular tissue.
Criteria for evaluation: Dominant lethal index calculated as 100%x(1-litter
size in treated group/litter size in control group).
Criteria for selection of MTD: Not discussed.
Diluted to 20% v/v in distilled water.
Both the number of resorptions and the percentage of litters withresorptions were increased. The index of dominant lethal mutations
declined from 16.4 to 7.8 over three successive matings.
Absolute and bodyweight relative testicular weights were decreased byethanol treatment (20%) and seminiferous tubule diameters weredecreased together with an increase in the number containing cellulardebris.
Mortality at each dose: None.
Mutations etc. Not relevant.
Clinical signs: No adverse signs were observed.
Body weights: Male bodyweights were unaffected by ethanol treatment.
Food/water consumption: Not presented.
Test substance was USP alcohol, 200 proof.
(2) valid with restrictions(255)

Test substance
Reliability
12.11.2004

:
:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses

:
:
:
:
:
:
:

Result
MethodYear
GLP
Test substance

:
:
:
:
:

Dominant lethal assay
ratmale
Wistardrinking water
up to 35 days
up to 30% alcohol in the drinking water given to an unspecified number ofrats
negativeother1980
no datano data

Remark:Age at start of treatment: 6-7 weeks.

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Treatment: Three groups of rats (numbers unspecified) were treated asfollows:group I, 30% ethanol for 4 days; group II, 15% ethanol for 5 days, then
20% for 30 days; group III, 15% for 5 days, 20% for a further 5 days, 25%
for 10 days and then 30% ethanol for the final 15 days. One control group
was untreated, while a positive control group was exposed to x-rays (200
R) prior to mating. After treatment, each male was paired with 2-3 females
per week for 8 consecutive weeks. The females were killed 10 - 11 days
after removal from the males and examined for live and dead
implantations.
Investigations: Dead implantations, reduction of live implantations and total
implantations were enumerated.
There were no significant differences in the numbers of dead, live and total
implantations at the pre-or postimplantation levels in the control (untreated)or ethanolic groups. In the positive control group there was a highincidence of dead implants and a reduction in the number of live implants.
The pregnancy rate was lower in group II, but this was not thought to betreatment-related, as the effect was not seen at 30%.
(2) valid with restrictions(256)

Result:Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
femaleno dataoral unspecified
single dose
5 ml/kg body weight given to 31 micenegativeother1975
no datano data

Remark:Result:Reliability
12.11.2004:

Females in pro-oestrus given a single dose of ethanol and mated on sameday (2 females to 1 male). Pregnant females then dissected. Control
group of 33 mice.
Route of administration: presumably gavage.
Ethanol produced no dominant lethal effects at a daily dosage of 5 ml/kg
p.o.
(4) not assignable
(257)

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
femaleC3H
gavagesingle dose 1, 1.5 or 2 hr after mating
1 ml of 12.5% ethanol or distilled water

Remark:Age at study start:10-12 weeks.
Strain: ( C3H x C58BL)F1
No. animals/dose: 47 ethanol treated vs. 43 controls matedat 1 hr; 24 ethanol treated vs. 24 controls mated at 1.5
hr; Three further experiments involved mating 1 hr (29 vs.

otherno datano data

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ResultETHANOL
ID: 64-17-5
DATE: 19.11.2004:

25 controls) or 2 hours (118 vs. 110) after ethanol
treatment, with uterine analysis.
Vehicle: Distilled water.
Duration of test: 17 days
Investigations: Number of implantations per pregnant female,number of living embryos per pregnant female and % dead
implants.
Sampling: Eggs taken and examined microscopically;first-cleavage embryos taken for chomosome analysis.
Clinical observations: None.
Organs examined at necropsy: None.
Treatment with ethanol at 1 or 1.5 hr after mating did not affect the numberof implantations per pregnant female, the number of living embryos perpregnant female and the % dead implants. However, the pooled data (118ethanol-treated vs. 110 controls) for rats treated at 2 hr post mating
showed a significant increase in late (post 11 days) deaths (P = 0.002).
Ethanol treatment was associated with a higher number of abnormal cells(P=0.039), ie trisomy possibly the result of clastogenicity or lagging ofchromosomes in M-II.
(4) not assignable(258)

Reliability
12.11.2004:

Type
Species
Sex
StrainRoute of admin.
Exposure period
Doses
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Dominant lethal assay
mouse
male
CBA
gavage3 day
0.1 ml of 40 or 60% ethanol (1.24 and 1.88 g/g bodyweight)
positiveother1975
no datano data

Remark:Age of animals at start: About 7-10 weeks.
No. of animals per dose: 13 at lowest dose; 6 at the highest dose.
Mouse strains: CBA/Fa Cam and CBA/Jackson
Dosage: 0.1 ml 40% ethanol solution (1.24 g/g bodyweight) daily by oral
gavage, both strains; and 0.1 ml 60% (1.88 g/g) in CBA/J
Vehicle: Distilled water.
Duration of test: Mated with untreaed females about every 4 days for 7
weeks.
Frequency of treatment: Gavaged once daily on 3 consecutive days.
Controls: Untreated only.
Replicates: 2 in CBA/Jackson mice
Mating: Each mouse was mated to 2 females and to further females after 3days,and then new females 7 more times at 4-day intervals.
Sampling: Pregnant females were sacrificed 13-15 days after conception.
Pregnant CBA/Fa females were allowed to produce their first litters, andthe size and sex ratios were recorded. Young were examined forabnormalities.
Pregnant CBA/Jackson females were killed and dissected 13 - 15 days
after conception. Dead and live implants were recorded.
Doses of ethanol administered equivalent to 1.24 and 1.88 g/kg bodyweight.
Identical results are reported in another paper (Badr, F.M. et al. (1977)
Adv. exp. Med. Biol. 85A, 25 - 46), but the number of mice exposed to 40%
ethanol is given as 25 and there are said to be 20 controls.

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ResultReliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004::

12.11.2004
5.7

Dead implants increased and live implants decreased significantly (P<0.01)
compared to controls. The dominant lethal mutation index increased to a
maximum of 46% in low
dose ltters and 67% in high dose litters.
It was evident that late spermatids were most affected by ethanol
treatment.
(3) invalid
No examination of the uterine contents was undertaken yet this was taken
as evidence of post-implantation loss due to dominant lethal mutations.
The method that was used to calculate the dominant lethal index is only
appropriate for very potent mutagens. The study did not take into accountthe fact that genetic effects can result in a reduction in mating frequency,
fertilization frequency or implantation frequency. This study cannot be
considered reliable.
(259)

CARCINOGENICITY

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

rat
male/female
Sprague-Dawley
drinking water
179 weeksad libitumnone
0, 10% in drinking water

Method:Result:Drinking water: tap water. Replace daily after measuring consumed
volume.
Animals: Treatment started at 39 weeks old (breeders), 7 days before
mating or from embryo life (offspring) and continued until death. 100
animals per dose level.
Animals housed in groups of 5, markrolon cages, stainless steel wire top,
white wood shavings for bedding.
Temperature: 23+/-2C, relative humidity 50-60%
Animals weighed weekly for 13 weeks, then biweekly to 104 weeks thenevery 8 weeks. Status and behaviour recorded 3x daily and clinicalexamination every 2 weeks.
Tissue necropsy: Skin and subcutaneous tissue, brain, pituitary gland,
Zymbal glands, parotid glands, submaxilliary glands, Harderian glands,
cranium (oral/nasal cavities, ear ducts), 5 head sections, tongue, thyroid,parathyroid, pharynx, larynx, thymus, mediastinal lymph nodes, trachea,
lung, mainstem bronchi, heart, diaphragm, liver, spleen, pancreas, kidneys,adrenal glands, esophagus, stomach (fore/glandular), intestine (4 levels),
bladder, prostate, gonads, interscapular fat pad, subcutaneous andmesenteric lymph nodes and any other organs with visible lesions. All
slides examined by the same group of pathologists
Statistical analysis: chi-squared test.
Intake of food and water was reduced but there was no significant
difference in bodyweight or survival, except females in the 104-152 week
age range showed higher mortality. No effects were visible from grossinspection.

yes, concurrent vehicle
other
1986
yes
other TS

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Increased tumour incidences reported.

Test substance:

Reliability:

18.11.2004

total malignant tumours (M+F)
Total malignant mammary tumours (F)Head and neck carcinomas, especially of oral cavity, lips and tongue (M+F)
Squamous cell carcinoma of forestomach (M+F)
Interstitial adenomas of the testes (M)Sertoli cell tumours (ovary) (F)Adenocarcinoma of uterus
Pheochromoblastoma (M+F)
Osteosarcoma of the head and other sites (M+F)
ethanol >99.8% supplied by Carlo Erba, Milan. Material replaced every 3months.
(4) not assignableNo detail other than histopathology reported. Animals were allowed to live
out their natural lives. No mortality or historical incidence data available.
Statistical analysis and basis of conclusions unclear. Study is severly
limited by the single high dose used with no intermediate concentrations.
(260)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

mouse
male/female
B6C3F1drinking water
2 years
ad libitum daily

Method:Groups of 48 male and 48 female mice were exposed to 2.5% or 5%ethanol in drinking water ad libitum for 2 years. This study was designed
and conducted to determine the long-term toxicity and carcinogenicity ofurethane in ethanol with control groups consuming drinking water alone orcontaining ethanol as specified above.

Result:3300 mg/day (males) 80 and 5000 mg/day (females) approx.
ambiguous
yes, concurrent vehicle
EPA OPPTS 870.4300
2002
yes
as prescribed by 1.1 - 1.4

Draft report issued 2002.
Ethanol caused a marginal exposure-related increase insurvival in males but had no effect on the survival of
females. There was evidence of an ethanol-induced reduction
in water consumption that was more marked in males than infemales.
There was equivocal evidence of carcinogenic activity of
ethanol in MALE based on increased incidences ofhepatocellular neoplasms.
There was no evidence of carcinogenic activity of ethanol in
FEMALE mice exposed to either concentration of ethanol.

Reliability

228:

Overall, the findings were insufficient to establish adefinitive effect of ethanol on the carcinogenicity ofurethane in this strain of mouse.
(2) valid with restrictions
Well reported and reliable study from a reputable toxicology laboratory.
However, only a single and very high dose used limit ultimate value, hence

UNEP PUBLICATIONSOECD SIDS
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ID: 64-17-5
DATE: 19.11.2004
only reliable with restrictions.

12.11.2004(261)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Remark:1999

Ethanol or the metabolites may be important factors for the humancarcinogenic effect of alcoholic drinks. This was concluded as a result of
the observation that all kinds of alcoholic drinks, if taken in sufficientquantity, can lead to an increased incidence of certain types of tumour (Blot
1992; IARC 1988). The mechanisms of ethanol-induced carcinogenisis
have not been clearly settled. Carcinogenisis of the mouth, throat, larynx
and possibly also gullet, as a result of the local effects of the ethanol, on
the one hand, must be distinguished from the carcinogenic effects in theliver and breast, which arise from the systemic availability of ethanol or its
metabolites, on the other. A local mechanism, among others, has been
postulated because the frequency of cancer in the mouth and throat wasincreased in persons who used, but as a rule did not swallow, mouthwashes with an ethanol content of at least 25% (Blot 1992).
Mechanistically, because of its physical-chemical properties, ethanol couldchange the barrier function of the cell membrane thereby making thepenetration of carcinogenic substances into the cell easier. This hypothesis
is supported by the synergistic effect of ethanol and tobacco smoke, withits many carcinogenic constituents, in relation to a carcinogenic effect in
the region of the mouth and throat. In addition, the influence of ethanol on
the expression of enzymes that metabolise particular foreign substances,from which an increased activation of the carcinogens might result, could
be responsible (Blot 1992; IARC 1988). An increase in the incidence oftumours induced by various carcinogens when ethanol is administeredsimultaneously has been demonstrated in several animal experiments. For
example, ethanol increases the carcinogenic effect of certain nitrosominesin the upper gastrointestinal and respiratory tracts or the vinyl-chlorideinduced hepatocarcinogenisis (Anderson et al. 1995; Blot 1992; IARC
1988; Mufti et al. 1997; Seitz et al. 1992).
To date, it has not been convincingly shown that ethanol acts as acomplete liver carcinogen. It is possible that the formation of liver cirrhosis,
which is as a precancerous lesion independent of the initiating factors,
plays a causal role. The induction of cytochrome-P450-2E1 and theresulting increased formation of reactive oxygen species, and also theincreased lipid peroxidation, has been assigned an important role inalcohol-induced liver damage (Brooks 1997). A modulating effect of ethanol
on hepatocellular carcinogenisis by other initiating factors, such as infection
with hepatitis B or C viruses, has also been discussed. Further, theinduction of various enzymes which metabolise foreign substances and thechanging of the membrane properties of the endoplasmic reticulum and thecell by ethanol could alter the toxicokinetics and bio-availability of particular
carcinogens or pre-carcinogens and thus promote the formation ofhepatocellular or extrahepatic carcinomas (Anderson et al. 1995; Farber

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1996; Seitz et al. 1992).
The increased rate of breast cancer which has been linked to theconsumption of alcoholic drinks, may possibly result from the demonstrated
influence of ethanol upon the hormone system. However, the in-vitro
animal investigations available at present offer no conclusive mechanistic
explanation for the epidemiological findings (Blot 1992; Longnecker 1995;
Singletary 1997). In how far the weak genotoxic properties of ethanol or
acetaldehyde and the reactive oxygen species formed during ethanol
metabolism are involved in the carcinogenisis has not been clarified. With
alcoholics and persons with a genetically determined reduced ALDH
activity, at least, increased acetaldehyde concentrations have been foundin the peripheral venous blood (Eriksson and Fukunaga 1993). Indications
as to the critical role of acetaldehyde in the aetiology of tumours in theupper digestive tract caused by ethanol have been observed in Japanesewith the inactive ALDH2 genotype which leads to increased acetaldehyde
concentrations after consumption of ethanol: 40 alcohol-dependent and 29
non-dependent patients with squamous cell carcinoma of the oesophagus
and groups of 55 and 28 control persons respectively were examined withrespect to their ALDH2 genotype. In the case of the alcoholics, and also
the non-alcoholics with an ALDH*2~ allelomorph which codes for an
inactive ALDH2, a clearly increased risk of gullet cancer was found. The
OR was 7.6 95% -CI; 2.8 - 20.7) for alcoholics and 12.1(95%-Cl; 3.4-42.8)
for non-alcoholics. The alcoholics consumed ca. 120 g ethanol per day, thenon-alcoholics ca. 55g. Perturbing factors such as smoking and diet could
not be taken into account because of the small size of the groups(Yokoyama et al. 1996a). Supporting this, a relationship between thefrequency of primary tumours in the oesophagus and the ALDI-12
genotype was observed in 33 Japanese male alcoholics with squamouscell carcinoma of the oesophagus. Of 17 patients with the genotype for theinactive ALDH2, 13 had multiple primary tumours as opposed to only S
from 16 with the active ALDH2. The difference was statistically significant(p '~ 0.01). The age and drinking and smoking habits of the patients withsingle and multiple carcinomas were not a distinguishing feature. The
prevalence of further tumours in the upper regions of the respiratory
passages and digestive tract was also higher; 29.4% for the patients withinactive ALDH2 ascompared with 6.3% of the patients with active ALDH2. Since the cells of
the oesophagus epithelium show no ALDH2
activity, the authors assume that the observed differences between thepatients with active and those with inactive ALDH2 are the result of anincreased systemic availability of acetaldehyde in the latter group(Yokoyama et al. l996).
Investigations of a possible carcinogenic effect due to occupational
exposure to ethanol inhalation are not available to date.
It can be proved that the consumption of alcoholic drinks leads to anincrease of the incidence of tumours in various locations. The
corresponding epidemiological investigations have been documented indetail by the IARC. In summary, many retrospective and some prospective
cohort studies with alcoholics, or brewery workers with a high permitted
consumption of alcohol during working hours, showed a clear connection
between the consumption of alcoholic drinks and the appearance oftumours of the mouth, throat, larynx, oesophagus and liver. Many casecontrol studies have confirmed the connections and given indications as tothe dose-effect relationships that are, however, subject to large
uncertainties. According to these studies, the relative risks of tumours ofthe mouth, throat or larynx are, in part, already significantly increased by
the daily consumption of ca. 10 g of ethanol. Mostly, the effects wereindependent of the preferred type of alcoholic drink (IARC 1988). In themeantime, a connection between the taking of alcoholic drinks and an

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increased risk of breast cancer may also be assumed. Similarly, there areindications of a relationship between the consumption of alcoholic drinks
and the increased incidence of colorectal tumours (Blot 1992; Longnecker
1995; Singletary 1997).
There are no investigations of the carcinogenic effect of ethanol under aninhalative condition relevant to the work-place.
The available animal experiments with chronic oral dosing with alcohol or
alcoholic drinks are described in detail elsewhere (IARC 1988). For mouse,
rat and hamster there is in the majority no indication concerning treatmentrelated increased incidence of tumours. But, hardly any of theinvestigations met the requirements of a valid carcinogenicity study
because, for example, of the absence of suitable control groups, because
the number of animals was too small or because of an inadequate
histopathalogical assessment (LARC 1988).
Studies which essentially fulfill the required criteria are discussed in moredetail in the following. In an
investigation on the influence of ethanol on vinylchloride-induced
hepatocarcinogenisis, a control group of 80 male Sprague-Dawley rats
received drinking water with 5% ethanol (v/v) for 30 months. If an averagedrinking water consumption of 15 ml/day and a body weight of 300 g isassumed, then from this a mean dosage of 2 g ethanol/kg BW per day canbe estimated. A further control group of the same size received pure water.
Thus, there was no compensation of the increased calorie intake which
may be assumed for the ethanol group. The survival rate after 18 months
was comparable in the two groups; 73% (ethanol treated) and 70%. Up
until the end of the study, there were 8 hepatocellular carcinomas and 29
hyperplastic nodules in the ethanol group and 1 corresponding carcinoma
and 10 nodules in the untreated group. 57 cases of tumours of theendocrine system were observed in the ethanol-treated group; in thecontrol group only 8. The incidences were 26/79 compared with 8/80 fortumours of the hypophysis, 14/79 and 0/80 for tumours of the adrenal
gland, 14/79 and 0/80 for tumours of the pancreas (not further specified)and 3/79 and 0/80 for testicular tumours. In total, 91 tumours werediagnosed in the alcohol group, 44% of which were classified as malign. In
the control group there were 16 tumours, of these 5 were malign (Radike etal. 1981).
Sprague-Dawley rats (50 animals per dose and sex) received a semisynthetic liquid feed with 1 or 3% ethanol for 2 years. A dosage of ca. 1
and 3 g ethanol/kg BW per day can be estimated from the foodconsumption. In place of ethanol, the control groups received in their dietan equi-calorific quantity of glucose. From week 104 until the end of theexperiment (week 120) all animals received a normal liquid diet. The
survival rates in the ethanol-treated groups were not reduced incomparison with the corresponding control values. The body weights in thehigher dosage group were significantly reduced in comparison with thecorresponding control values; for the males from the 13th week and for thefemales from the 69th week. The maximum body-weight reduction was ca.
15%. The weights of the kidneys and livers of the ethanol-treated animals
did not deviate significantly from the corresponding control values. In thehistological assessment, various nonneoplastic findings arose significantly
more often in the ethanol-treated group than in the corresponding control
group. In the male animals of both dosage groups these were cystic andfocal degeneration of hepatocytes and chronic inflammation in thepancreas. In addition, the higher dosage group showed increased fibrosisin the bile duct, and the lower dosage group hyperplasia in the pancreas,
C-cell hyperplasia of the thyroid gland and demyelination of the peripheral
nerves. In the female animals of both dosage groups focal hyperplasia in
the adrenal cortex was noticeable; in the higher dosage group only, C-cell

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hyperplasia of the thyroid gland, inflammation of the clitoral gland,
pigmentation of the mandibular lymph nodes and also demyelination of theperipheral nerves. The analysis of the neoplastic findings gave no
indication of a significant ethanol-induced change in the tumour spectrum
or tumour frequency. In the female animals of the lower dosage group,
fibroma, fibroadenoma and adenoma of the mammary gland were
statistically significantly increased. In contrast, tumours of the islet cells in
the pancreas were reduced. In the higher dosage group, the number of (notmore exactly specified) neoplasia of the hypophysis was increased, thenumber of adenoma of the adrenal cortex reduced. Overall, the total
number of tumours in the higher dosage group was significantly reduced.
The authors concluded from their results that ethanol itself has no
carcinogenic effect (Holmberg and Ekstrom 1995).

Reliability:

12.11.2004
Species
Sex
StrainRoute of admin.

232

:
:
:
:

The influence of a life-long ethanol consumption on life-expectancy wasinvestigated on male CS7BL-mice. At first the animals received 3.5%
ethanol (v/v) in their drinking water from the 14th to the 19th week of life.
After that they were divided into three groups which received 3.5, 7.5 or
12% ethanol in their drinking water until the end of their lives. Each group
comprised 100 animals which were caged separately. Two control groups,
also with 100 animals each, were maintained. In one group each animal
was individually caged while in the other the animals were held 5 to a cage.
From the consumption of liquid the authors calculated mean ethanol doses
for the three exposed groups as 2.8, 7.3 and 11.1 g/kg BW per day. The
blood-ethanol concentrations were measured for the first time in the 13th
week of the experiment and additionally at three further points in time 3
months apart and at different times of the day. The values averaged over
all measurements were, with rising dose, 66, 142 and 268 mg/kg blood(approximately equal to mg/l blood). The development of body weight in the
5 groups was not different. The average life in the middle dosage groupwas statistically significantly raised with respect to that of the control groupwith individual caging. In the other groups there was no difference from thecontrol. On the natural death of the animals, or after killing in extremis, adetailed histological examination was carried out. Of the animals caged
individually, between 72 and 89 animals per group were examined; ofthose caged in groups only 55 on account of cannibalism. There were no
significant differences between the frequency, type and severity of the nonneoplastic liver damage observed in the 5 groups. No damage attributable
to the treatment with ethanol could be detected in the other organs (brain,lungs, heart, pancreas, spleen, kidneys, small intestine, testes) which werealso examined histologically. The type and frequency of the neoplasia in
the individual groups gave no indication of a significant, ethanol-relatedchange in the tumour spectrum or incidence (Schmidt et al. 1987). Thus,
overall, the results of those animal experiments carried out with aconvincing methodology are contradictory. The incidences of tumours inmale Sprague-Dawley rats following chronic consumption of ethanol in theirdrinking water were raised, but not, however, in the case of male andfemale animals of the same strain to which ethanol was administered in aliquid feed. A drinking water study with mice also gave negative results.
Since the MTD was not reached in all three studies, the strength of theirevidence in the assessment of the carcinogenic potential of ethanol is inany event limited.
(2) valid with restrictions
Whilst this is a review it is from a reputable body (German MAK
commission).
(262)
Mouse
Other
drinking water

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:

ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:

Result:Reliability:

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period

:
:
:
:
:
:
:

Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:

Method:Life (up to 160 weeks)
Continuous
None
100 males/group given 3.5 to 15% solutions for 5 weeks followed by 3.5,
7.5 or 12% v/v ethanol in water for life.
Yes
other
1987
no datano dataTreatment had no adverse effects on survival. There were increased
incidences of liver sarcomas (probably lymphomas) at 7.5% (13/87) and
12% (10/72) compared with 4/79 in controls and 6/77 at 3.5% ethanol. No
increase in liver carcinomas was found.
(3) invalid
Study design inadequate: not all mice were necropsied, as many wereautolysed, histopathology was inadequately reported and no female mice
were used.
(263)
Rat
male/female
Long-Evans
Other
Days 11 to 21 of pregnancy inclusive
Daily in two divided doses
All rats exposed in utero followed until natural death (up to 2 years 6months).
7 g/kg body weight/day to 20 dams in two divided doses
Other
other
1987
no datano data
One male and one female pup from each of 20 ethanol-treated litters wereselected for lifetime study at age 6 months.
Route of administration: rat litters exposed in utero. Dams gavaged withethanol on days 11 to 21 of gestation. Litters were transferred to untreatedsurrogate mothers within 24 hours of birth.

Result:Reliability:

12.11.2004
Species:

Control groups consisted of litters from pair-fed (isocaloric sucrose
solution) an ad lib fed (lab. feed) mothers.
Minimal detail reported in this study which was part of an investigation into
the effect of in utero ethanol exposure on longevity. Six unspecified
tumours were found in 6/20 males and 6/20 females in each group except
female pair-fed controls which had 8/20 animals with tumours. The authors
concluded that there was no significant difference in tumourincidence.
Alcohol-exposed females had significantly shorter lifespans (about 2
weeks) than pair fed and ad lib. fed controls (p <0.02). A similar but lessmarked effect on longevity was seen in males.
(3) invalid
Small group sizes (n=20) and absence of tumour histopathology make thisstudy inadequate.
(264)
Mouse

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Sex
StrainRoute of admin.
Exposure period

:
:
:
:

Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Remark:Result:Reliability:

12.11.2004

Male
C3H
Other
mothers treated either during pregnancy or for 1 week beginning when thepups were 1 week old
Continuous
15 months
0.5 or 5% in the drinking water
Other
other
no datano data
Method: moths given ethanol in their drinking water either during
pregnancy or when pups 1 week old. Control group not clearly described.
Route of exposure: transplacental and neonatal.
In offspring treated during embryogenesis, there were liver
tumours (diagnosed grossly and described as hepatomas) in
3/25 exposed to 0.5% and 1/10 exposed to 5%. In offspringtreated via the milk, liver tumours developed in 5/31
exposed to 0.5% and 5/45 exposed to 5%.
The incidence in pooled control males was 27/62,
significantly higher (p <0.005) than in the (pooled)exposedgroups.
(3) invalid
Study design inadequate: short treatment duration, females not examined,tumours not examined histopathologically. No information on initial groupsizes.
(265)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

HamsterMaleother: outbred Syrian golden
oral feed
29 weeks in total over a 33 week period
Daily
up to a further 19 months
19 males given a liquid diet containing 6% ethanol.

Remark:Reliability:

Nineteen males were maintained on a liquid diet containing 6% ethanol(w/v) from age 9 to 29 weeks, then left untreatedfor 4 weeks, then treatedwith 6% ethanol in the diet for a further 9 weeks. It is unclear how thecontrol group of 21 males was treated.
(3) invalid
Study design inadequate: small group sizes, no females used, short
exposure duration.
(266)

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period

234

:
:
:
:
:

Yes
other
no datano dataRat
MaleFischer 344
oral feed
26 weeks

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

Remark:ResultReliability::

12.11.2004

63 weeks
26 rats given 6% ethanol in liquid diet
yes, concurrent vehicleotherno datano data
Method: Groups of 26 nine-week old rats were given a liquid diet containing
0 or 6% ethanol for 26 weeks and then fed a normal laboratory diet for afurther 63 weeks prior to sacrifice.
No statistically significant difference in tumour incidence was seen.
(3) invalid
Study design inadequate: short exposure period, no females used, groupsizes too small. A dietary level of 6% ethanol corresponded to 35% total
calories.
(267)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
Male
other: Holtzman
oral feed
up to 370 days
Continuous
no data
15 males given 5 g/kg bw/day

Remark:ResultReliability::

Method: Treated rats given a diet containing alcohol equivalent to 35% of
total calories. Control rats pair fed.
No liver tumours seen in either group.
(3) invalid
Study design inadequate: no female rats used, small numbers of animals
used, study duration too short, pathological examination limited to the liver.
(268)

12.11.2004

yes, concurrent vehicleotherno datano dataSpecies
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Mouse
male/female
C57BL
Gavage50 weeks
twice per week
no data
0.2 ml of 40% ethanol in water to 36 males and 32 females

ResultReliability::

No treatment-related tumours.
(3) invalid
Study quality inadequate: limited dose of ethanol administered, short

No
other
no datano data

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duration and no control group.

12.11.2004(269)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
male/female
other: BDVI
Gavage78 weeks
twice per week
unclear - see remarks
25 rats per sex given unspecified amounts of 40% ethanol.

Remark:ResultReliability::

Method: Observation period unclear, but survivors killed when 120 weeks
old.
No obvious effects on survival. No treatment-related tumours.
(3) invalid
Study design inadequate: small group sizes, unspecified doses, short
exposure period.
(270)

12.11.2004

yes, concurrent no treatment
otherno datano dataSpecies
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat

RemarkResult::Reliability:

Method: 10 control rats were treated with saline.
Average lifespans were 500 and 396 days for the low and highdose
groups. No lifespans given for the controls. No oesophageal, stomach or
hepatic tumours were seen in any group.
(3) invalid
Study inadequately reported. Limited pathological examination carried out.
(271)

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
Method
Year

236

:
:
:
:
:
:
:
:
:
:
:
:

Sprague-Dawley
Gavage
Life
Daily
None
40 rats/group given 0.5 ml/day of 30 or 50% ethanol
Yes
other
no datano data

HamsterMaleother: outbred Syrian golden
drinking water
29 weeks
Continuous
47 weeks
groups of 27 males given 7.4 or 18.5% aqueous ethanol
yes, concurrent vehicle
other

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

GLP
Test substance::no datano data

RemarkResultReliability:::

Method: control group of 27 males given tap water.
No statistically significant differences in tumour incidence were observed.
(3) invalid
Study design inadequate: short treatment duration, no females used, small
group sizes.
(272)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Hamster

ResultReliability::

No tumours of pancreas, common duct or gall-bladder observed.
(3) invalid
Study design inadequate: no tissues examined other than pancreas,
common duct and gall bladder. Group sizes small, exposure period short,no controls.
(273)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Hamstermale/female
Otherdrinking water
up to 807 days
5 days/weekNone
7 males and 3 females given 19.5% aqueous ethanol

ResultReliability::

No tumours reported.
(3) invalid
Study inadequately reported. Group sizes were small and no controls wereused.
(274) (275)

:
:
:
:
:
:
:
:

Mouse
Femaleother: C3H/St
drinking water
80 weeks
Continuous
no data
12% in water to 15 females

12.11.2004

12.11.2004

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

Other
drinking water
up to 46 weeks
Continuous
None
20 hamsters of each sex given 5% (w/v) aqueous ethanol
no data specified
otherno datano dataNootherno datano data

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DATE: 19.11.2004

ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:

Result:Reliability:

12.11.2004

yes, concurrent vehicleotherno datano data
Mammary tumours developed in 8/11 treated and 22/27 control mice.
Though the incidences did not differ, median time to tumour appearance
was significantly (p <0.001) shorter in the treated group (8 months versus
14.2 months of age).
(3) invalid
Study design inadequate: small group sizes, no males used, no
histopathological examination of tumours.
(276)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
Male
Wistardrinking water
up to 40 weeks
Continuous
None
10 rats given 10% aqueous ethanol

ResultReliability::

No tumours observed in either group.
(3) invalid
Study inadequate to assess carcinogenicity: group sizes too small, only
male rats used, duration of exposure short.
(277)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
Male
Sprague-Dawley
drinking water
up to 30 months
Continuous
no data
80 males given 5% v/v in water

Result:Reliability:

At 18 months survival was about 70% in both groups. Hepatocellular
carcinomas were found in 1/80 controls and 8/79 treated rats (p = 0.016).
Hyperplastic nodules occurred in the liver of 10 controls and 29 treatedrats. In the treated group, 57/59 had endocrine tumours compared with
8/80 controls. Of these, the treated group showed 26 pituitary (P=0.00004), 14 adrenal, 14 pancreatic and three testicular tumours. All eightcontrol group tumours were pituitary tumours.
(3) invalid
Isocaloric and isonutrient diets not used, no females were included in the12.11.2004

238

yes, concurrent vehicleotherno datano datayes, concurrent vehicleotherno datano data

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
study. Some histopathology inadequately described.

12.11.2004(278)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:

ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:

Result:Reliability:

12.11.2004

Ratmale/female
Wistardrinking water
up to 23 months
each day or on alternate days
no data
15% aqueous ethanol daily or 55% aqueous ethanol on alternate days,
each to groups of 20-25 rats of each sex.
yes, concurrent vehicleotherno datano data
Survivors were killed at 23 months. No excess of tumours ofany kind wasfound in either treated group.
(3) invalid
Study design inadequate due to small group sizes. Ethanol intakes at thehigh concentration were lower.
(279)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses
ResultControl group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat

RemarkResult::Reliability:

Method: Control group consisted of 48 rats.
No adverse effects on survival. No statistically significant increase intumour incidence reported.
(3) invalid
Data inadequately reported.

Sprague-Dawley
drinking waterLife
5 days/week
None
25% ethanol in water to 48 rats
yes, concurrent no treatment
otherno datano data

12.11.2004(280)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Post exposure period
Doses

:
:
:
:
:
:
:
:

ResultControl group
Method
Year

:
:
:
:

Mouse
male/female
other: C57BL and CF1
Dermalup to 802 (CF1) or 830 days (C57BL)
three times/week
unspecified amounts of 50% aqueous ethanol applied to 31 female and 33
male C57BL mice and to 57 male CF1 mice.
No
other

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

GLP
Test substance::no datano data

RemarkResultReliability:::

Method: no data.
Only one skin papilloma was seen in a C57BL mouse.
(3) invalid
Study inadequately designed and reported.

12.11.2004(281)

Remark:Reliability
12.11.2004

:

5.8.1

In addition to studies in which ethanol alone was investigated, a number ofcarcinogencity studies have been carried out where ethanol has been coadministered with other compounds (IARC, 1988). Such studies havegenerally involved the use of ethanol as a vehicle for another test materialof interest or have been carried out where mechanistic or other reasons
have required the deliberate study of the co-exposure of ethanol withanother test material e.g. the use of ethanol as an experimental
hepatotoxin in carcinogenicity studies of vinyl chloride. These studies are
generally inadequate in one or more aspects: short duration, small groupsizes, inadequate reporting, inconsistent reporting and inadequate
pathological examination. They are consequently inadequate fordetermining the effects of ethanol.
(4) not assignable(282)

TOXICITY TO FERTILITYType: Two generation study
Species: MouseSex: male/female
Strain: CD-1
Route of admin.
: drinking water
Exposure period: 105 weeks
Frequency of treatm.
: ad libitum
Premating exposure period
Male: Parental 7 days; F1 74 days
Female : Parental 7 days; F1 74 days
Duration of test:
No. of generation: 2
studies
Doses: 5, 10 and 15% v/v in water
Control group: yes, concurrent no treatment
NOAEL parental: = 15 %NOAEL F1 offspring: = 10 %NOAEL F2 offspring: < 15 %Result: No observed effect on fertility
Method: other: NTP protocol
Year: 1985
GLP: no data
Test substance: other TS: 92%
Method:Age at onset: P animals 6 weeks at receipt, 11 weeks at first exposure.
No. of animals per sex per group: 20 also 20 F1 animals at the high dosemated at 74 days old.
Ethanol administered in deionized, filtered water.
P generation dosed for 7 days pre-mating and then for 98 days. F1
animals continued on dosing until mating.
Animals mated in cohabiting pairs; litters were proof of pregnancy.

240

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5. TOXICITY

ResultETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Litters were not standardized.
Clinical signs, oestrous length etc. were not evaluated.
Epididymal and vas sperm were evaluated for concentration,
motility and morphology in F1 males only.
High dose F1 animals had liver, kidney/adrenal and male sexorgans weighed at termination.
F2 data were for litter sizes etc. Only.
Parental/F1 data: Ethanol treatment had no effect on bodyweights and on
the proportion of breeding pairs producing at least 1 litter during thecontinuous breeding phase or the number of litters per pair. Fertility indiceswere 97, 100, 100 and 94% in the controls and 5%, 10%, 15% ethanolgroups respectively.
Offspring data: F1 offspring of the 15% ethanol pairs had fewer live pups
per litter. their F2 offspring weighed less as pups than control pups, males,
females or both sexes. Fertility indices in F1 matings were 85% and 85% in
the controls and 15% ethanol groups respectively. Other reproductive
performance indices e.g. gestation index, changes in lactation and changes
in oestrous cycles were not studied
Effects on sperm and male reproductive organs: In the F1, 15% ethanolgroup there was a significantly decreased %motile sperm but no changes
in sperm concentration, %abnormal sperm or %tailless sperm. There was asignificant decrease in testis, epididymis and seminal vesicle weight
Gestation index, changes in lactation and changes in oestrus cycles were
not studied.
Haematological, clinical biochemical, gross pathological andhistopathological changes were not studied.
Mortality in P animals is reported but not discussed. F1 males from the
15% group at adulthood had decreased bodyweight and and decreased
weight of testis and epididymides and seminal vesicles. In F2 females,
relative liver and kidney/adrenal weights were increased.

Conclusion:

Reliability:

Flag
12.11.2004:

Type
Species
Sex:::

Offspring toxicity:
Litter size and weights: Not given.
Sex ratios: Not influenced by treatment
Viability index: Not reported. Litters born to P at 15%
ethanol had reduced number of live pups per litter.
Post natal survival until weaning: Not reported.
Postanatal growth: Pups born in final F1 generation of animals exposed to
15% ethanol pre- and post-natally weighed less than controls at birth anddays 21 and 74.
Vaginal opening or preputial separation: Not studied.
Anogenital distance: Not measured.
Organ weights: Described above.
Gross pathology: Not examined.
Overall, ethanol in drinking water at concentrations up to 15% (equivalent
to 20.7 g/kg/day) had no demonstrable effect on fertility in this twogeneration study.
(1) valid without restriction
Well reported study but not to a standard protocol. Very high doses usedand no NOAEL identified for all endpoints.
Critical study for SIDS endpoint(283)
One generation study
Mouse
Male

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241OECD SIDS
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ID: 64-17-5
DATE: 19.11.2004

Strain: Swiss Webster
Route of admin.
: oral feed
Exposure period: 49 days
Frequency of treatm.
: ad libitum
Premating exposure periodMale: Sequential matings through 7 weeks of exposure
Female : None
Duration of test
: 7 weeksNo. of generation: 1
studies
Doses: 10% and 25% of ethanol-derived calories
Control group: Yes
NOAEL parental: = 10 %NOAEL F1 offspring: = 25 %Result: Fertility not affected
Method: otherYear: 1989GLP: no data
Test substance: no data
Method:Number/age of animals: 20 males per group, 75 days old at start oftreatment.
Vehicle etc: Ethanol providing 10 or 20% of calories in a nutritionallybalanced liquid diet. Two control groups consisting diets compensated or
not for ethanol calories.
Dosing schedules: Males were given ethanol or control treatments for 7
weeks prior to mating with untreated females.
Mating procedure: 2 females per male for 4 hours; vaginal plugs weretreated as evidence of pregnancy. Females were allowed to give birth andoffspring were counted, weighed, culled and re-weighed at 21 days. Litterswere standardized by culling at birth to 8 per dam.
Parameters assessed: Vital and functional observations were maintainedon P and F1 generations.

Remark:Sperm quality, anogenital distance and organs at necropsy were notevaluated.
Parental data
Bodyweight: Paternal bodyweights were less at 25% ethanol-derived
calories than at 10 or 0%. Offspring bodyweights were not affeceted bytreatment.
Food/water consumption: High-dose males consumed less diet. (NB pairfed controls).
Clinical signs: None reported.
Fertility index: At least 80% for each ethanol concentration at each time
point. Feretility was at least as great as in pair-fed controls.
Precoital interval: Not measured.
Duration of gestation: To term.
Gestation index: Not given.
Changes in lactation, oestrus cycles, sperm, haematology, clinicalchemistry, gross pathology, no. of implantations, no.of corpora lutea,
ovarian primordial follicle count, organ weight change and histopathology:
Not studied.
Mortality: Not reported.
Offspring toxicity:
No dose-related observations were made.
Litter sizes and weights: Not affecetd by paternal exposure.
Sex and sex ratios: Not affected by paternal, exposure.
Viability index: Not measured.
Post natal survival: No mortality reported.

242

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5. TOXICITY

ResultETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Effects on offspring: Not studied.
Postnatal growth: Not affected by day 21.
Vaginal opening: Not studied.
Anogenital distance, organ weights and gross pathology: Not studied.
No toxic responses were noted in treated males other than decreased
bodyweight gain at 25% ethanol-derived calories in diet. Fertility over 7
weeks of treatment was not affected.
No adverse effects on offspring were noted as a function of either level ofpaternal ethanol treatment or duration of treatment.

Reliability:

12.11.2004

Fertility was at least as great as in pair-fed or standard controls.
(2) valid with restrictions
Well reported study but not to a standard protocol. Very high doses used.
(284)

Type
: One generation study
Species: Rat
Sex: Female
Strain: other: Holtzmann
Route of admin.
: oral feed
Exposure period: 8 or 16 weeks before mating
Frequency of treatm.
: ad libitum dailyPremating exposure periodMale: no treatment
Female : 3 or 6 weeks
Duration of test:
No. of generation:
studies
Doses: 5% in liquid feed
Control group: Yes
NOAEL parental: < 5%NOAEL F1 offspring: <= 5 %Result: Fertility not affected although oestrous cycle length was prolonged andirregular
Method: otherYear: 1982
GLP: no data
Test substance: no data
Method:Remark:No. and age of animals: 10 per group, age 20 days. No F2 generation.
Ethanol was supplied in a liquid diet for 16 weeks prior to mating or for 8
weeks followed by 8 weeks on standard diet. Dosing ended after mating.
Two Control groups were used, one receiving standard diet the other pairfed with 5% ethanol in diet.
Mating procedure was by 1:1 cohabitation with a fertile male for 14 hr.
proof of pregnancy was a sperm-positive vaginal smear. Study ended withdelivery of F1 pups.
Oestrous cycle length and pattern was recorded.
Growth performance in F1 pups was followed.
Statistical test was one-way ANOVA.
Parental data:
Effect of duration of exposure, not dose, was assessed.
Administration of 5% for 16 weeks, not 8 weeks, increased oestrus cycle
length and irregularity. Age to vaginal patency was increased by bothregimen. No abnormalities ofovaries or uteri were found.
Bodyweight: Maternal bodyweights were measured but not reported.
Offspring bodyweights were not affected by treatment.
Food/water consumption: Not reported but must have been recorded.
Clinical signs: None reported.

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Fertility index: At least 80% for each ethanol concentration at each time
point. Fertility was at least as great as in pair-fed controls.
Precoital interval: Not measured.
Duration of gestation: Not reported.
Gestation index: All females delivered live litters.
Changes in lactation, oestrus cycles, sperm, haematology, clinicalchemistry, gross pathology, no. of implantations, no.of corpora lutea,
ovarian primordial follicle count, and organ weight change: Not studied.
Histopathology: All ovaries and uteri examined were normal.
Mortality: None reported.

ResultReliability::

12.11.2004

Offspring toxicity:
No dose-related observations were made.
Litter sizes and weights: Not affecetd by maternal exposure.
Sex and sex ratios: Not given.
Viability index: Not measured.
Post natal survival: No mortality reported.
Effects on offspring: Not studied.
Postnatal growth: Not studied.
Vaginal opening: Average age of vaginal patency was 72-77 days in bothgroups and significantly older than in control groups (41-58 days).
Anogenital distance, organ weights and gross pathology: Not studied.
No adverse effect on fertility, litter size or neonatal bodyweight was
detected. Irregular cycles and longer oestrous cycles were noted in rats fed
for 16 weeks but not after 8 weeks with 8 weeks recovery period.
Average age of vaginal patency was 72-77 days in both groups of ethanoltreated rats versus 41-58 days in controls.
(2) valid with restrictions
Well reported study but not to a standard protocol. Very high doses usedand no NOAEL identified.
(285)

Type
: One generation study
Species: Rat
Sex: Female
Strain: other: Holtzmann
Route of admin.
: oral feed
Exposure period: 55 days
Frequency of treatm.
: ad libitum dailyPremating exposure period
Male: NoneFemale : 50-55 days
Duration of test:
No. of generation:
studiesDoses: 2.5% and 5% in feed, estimated 8-12 g/kg/day and 12-14 g/kg/day
Control group: no data specified
NOAEL parental: = 2%
Result: Ovarian function was suppressed at the high dose
Method: otherYear: 1982
GLP: no data
Test substance: no data
Method244:

No. and age of animals: 8-11 per group aged 20 days at start.
Vehicle etc: Ethanol was supplied in a liquid diet.
Dosing schedule; Pair-fed controls were used at each dose for P animals
with ethanol in diet ad lib for 50-55 days.
No matings attempted so no F1 and F2 animals.
Litter standardization: Not applicable.

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Remark:Result:Reliability:

12.11.2004

Animals were weighed weekly and examined daily for vaginal patency.
Once patent, vaginal lavages were made daily.
Oestrous cycle length and pattern: - length but not pattern determined.
Sperm examination: Not applicable.
F1 and F2 observations: Not applicable.
This study was to assess ovarian function as a possible factor in fertility
studies following ethanol exposure. It is an adjunct to a study by the sameauthors demonstrating absence of effect on fertility but evidence ofdisturbed ovarian function.
Parental and offspring data are not applicable in this study.
Ovarian function was supressed in rats that achieved blood alcohol levelsof 250 mg/100 ml.
Animals receiving 5% ethanol(but not 2.5%) in liquid diet had longer time tovaginal patency, failed to begin oestrus cycles, showed decreased
bodyweight gain, had ovaries containing only a single generation of
corporea lutea, had infantile vaginal and uterine epithelium and decreased
uterine and ovarian weight.
(2) valid with restrictions
Well reported study but not to a standard protocol. Very high doses used.
(286)

Type
: One generation study
Species: Rat
Sex: Female
Strain: Wistar
Route of admin.
: oral feed
Exposure period: 49 days (animal age 28-77 days)
Frequency of treatm.
: Daily
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 5% in feed, estimated 12,000 to 14,000 mg/kg/day
Control group: other: See method details
Method: otherYear:
GLP: no data
Test substance: no data
Method:Number of animals: 100, ex Charles River Breeding Labs, individually
caged.
Ethanol fed animals received a liquid diet with ethanol accounting for 35%
of total calories. Pair fed isocaloric controls (using dextrimaltose asalternative to ethanol.)Controls: ad libitum intact and ad libitum oophorectomized (Feed: Lab Blox
F4 ex Best Feeds, Oakdale, Pa)
Gross and microscopic anatomy: Animals sacrificed by exsanguination, 67mls of blood kept for analysis. Liver, ovaries, uterus and fallopian tubes
removed and weighed, preserved in Bouin's solution. Histological sections
stained with hematoxylin and eosin. Liver assessed for degree of fat,necrosis and inflammation. Ovaries assessed for corpus lutea, corpus
hemorrhagica, and numbers/development of Graafian follicles.
Uterus/fallopian tubes assessed for thickness of endometrium, type of andsecretory activity of the epithelial lining cells and thickness of the muscle
wall of each organ. Cervix/vagina examined for thickness of epithelial celllining and signs of estrogen stimulation.
Plasma steroids and gonadotropins. All assessed by radioimmunoassays

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Remark:Result:with following levels of detection:
Progesterone 10pg, Estradiol 0.1pg, estrone 0.2pg, corticosterone 20pg,
gonadotropin 4ng/ml. All measurements in duplicate.
Liver: Enzyme function assessed by measuring serum alkaline
phosphatase, gamma glutamyl transpeptidase, glutamic pyruvic andglutamic oxalacetic transaminase activities.
Blood ethanol: measured using blood samples obtained 1 day before
sacrifice between 9-11am and before feeding.
Statistical analysis: Student t test. "Probable significance" at p<0.05 and"significance" at p<0.01
The reported blood ethanol level was relatively low (110±9mg/dL) but thetiming of the sample (taken 09.00 - 11.00hours) was probably inappropriate
to detect the peak likely at the usual time of feeding during the previous
evening.
Growth (body weight at sacrifice): ethanol fed: 138 +/-5.3g; isocaloric
controls: 161.6+/-3.8g. Significant difference (p<0.01).
Anatomy: Livers of alcohol fed animals significantly larger than controls(p<0.01. Treated animals 10.6g+/-0.5, isocaloric controls 6.4g+/-0.3, ad
libitum controls 8.3g+/-0.2). Ethanol treated animal livers markedly more
fatty in appearance.
Ovaries: Marked weight loss in treated animals (30.6+/-2.2mg versus
75.5+/-3.9mg for isocaloric controls and 91.4mg+/-0.2mg for ad libitum
controls). Weight loss significant even when corrected for body weight.
Reduction in ovarian mass due to absence of developing follicles, corpus
lutea and corpus hemorraghagica.
Histology: Differences in appearance of uterus, cervix and vagina between
treated and untreated animals.
Uterus/fallopian tube: Marked weight loss in treated animals (39.0+/-4.1mg
versus 180.5+/-18.7mg for isocaloric controls and 306mg+/-15.5mg for ad
libitum controls). Weight loss significant even when corrected for bodyweight.
Plasma steroid and gonadotropin levels (n=25):
Plasma estradiol reduction seen in treated animals (ethanol fed: 27.5+/1.2pg/ml; isocaloric controls: 33.3+/-1.5pg/ml; ad libitum intact controls:
48.0+/-1.4pg/ml; p<0.01). However no statistically significant reduction
relative to oophorectomized control (29.8+/-1.6pg/ml).
Plasma progesterone reduction seen (ethanol fed: 23.3+/-4.3pg/ml;
isocaloric controls: 54.3+/-7.3pg/ml; ad libitum intact controls: 41.7+/6.7pg/ml; p<0.01). However no statistically significant reduction relative tooophorectomized control (18.0+/-0.6pg/ml).
Plasma estrone increase seen (ethanol fed: 156.0+/-26.7pg/ml; isocaloric
controls: 114.9+/-13.9pg/ml; ad libitum intact controls: 80.5+/-6.3pg/ml;
p<0.01); oophorectomized control (48.0+/-5.2pg/ml).
Plasma corticosterone levels increased (ethanol fed: 74.0+/-9.0ug/dl; ad
libitum controls: 48.0+/-6.0ug/dl; p<0.05). However no statisticallysignificant reduction relative to pair fed controls (78.0+/-9.0ug/dl).
Plasma lutenizing hormone levels increased (ethanol fed: 68.7+/-5.7ng/ml;
ad libitum controls: 43.5+/-7.0ng/ml; p<0.01). However no statisticallysignificant reduction relative to pair fed controls (79.4.0+/-6.8ng/ml). All
significantly less (p<0.01) than oophorectomized ad libitum control.
Plasma follicle stimulating hormone levels not statistically different but all
significantly less (p<0.01) than oophorectomized ad libitum control.
Biochemical liver function:
Serum glutamic oxalo-acetic-acid-transaminase and serum glutamic
pyruvic transaminase levels increased in treated animals (2.5x control
levels). Alkaline phosphatase 50% greater in treated animals (all p<0.01).
Gamma glutamyl transpeptidase activity not detected in any controls but
reproducibly measured (2.3+/-IU/ml) in ethanol fed animals.

246

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Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

12.11.2004

Blood ethanol levels: 110+/-9.0mg/l. Not detected in any controls.
(2) valid with restrictions
Well reported study but not to a standard protocol. Very high doses usedand no LOAEL identified.
(287)

Type
: Fertility
Species: Rat
Sex: MaleStrain: Sprague-Dawley
Route of admin.
: Gavage
Exposure period: 9 weeks
Frequency of treatm.
: twice per day
Premating exposure periodMale: 9 weeks
Female :Duration of test:
No. of generation:
studies
Doses: 0, 2, 3 g/kg
Control group: other: concurrent vehicle and concurrent no treatment
Method: otherYear:
GLP: no data
Test substance: other TS
Method:Remark:Result:Animals: Supplied by Charles River, 2 Months old at acquisition;acclimated for 2 weeks.
Environment: 22 +/- 1 degree C; relative humidity 45 +/- 5%; 12 hr:12 hrlight:dark cycle.
Feed: Laboratory feed and water ad libitum.
Treatment: After acclimation, groups of 20 male rats were intubated at
0900 and 1600 hrs with 3g/kg ethanol (15% w/v in distilled water), 2g/kg
ethanol or vehicle (distilled water) only. Groups receiving 2 g/kg or 0 g/kg
were paired fed with those receiving 3 g/kg and a fourth group received nogavage treatment. This continued for 9 weeks before breeding with 70 to
90-day-old females.
Controls: Distilled water by gavage (Group 3) or no gavage treatment(Group 4). Offspring of these groups were compared to evaluate thepotential for handling stress.
Evaluation of pups: After birth, pups were examined and weighed andlitters were culled to 10 per female. Culled pups were subjected to brain
and adrenal gland weight measurements. Offspring were weighed at 7, 14
and 21 days. At 7 days old, three males and 3 females from each of 6
litters were culled and their brain and adrenal gland weights determined.
At 21 days, this was repeated with inclusion of more organ weights. Blood
alcohol levels were determined after breeding was determined at 1,2,3 & 5hours after dosing.
Statistical analysis: ANOVA and Duncan's Multiple Range tests onparametric data; Chi-square on non-parametric data.
Paternal alcohol exposure did not influence litter size, average birth weight
per pup or postnatal bodyweights in offspring. A study of runts suggested
an influence of ethanol on individual sperm rather than on entire sperm
production. The small but significant effect on male:female ratio (53 to45%) was unexpected and is without explanation. An apparent effect on
adrenal gland weight at birth is difficult to interpret, as this did not persist
through offspring growth and development. An effect on spleen and heart
weight indicates that paternal alcohol exposure may produce grosschanges in offspring as well as functional changes.
6 Males in the top dose group and 1 ad lib control rate died due to illness
prior to breeding. Peak blood alcohol level was 338 +/- 15 mg% in the top

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dose group and 132 +/- 5 mg% in the lower dose group. There were no
adverse effects on male reproductive performance and female fecundity
was no affected. Litter sizes and birth weights were not affected. Litter
sizes and birth weights were not affected by paternal ethanol intake ateither dose. Ethanol treatment in fathers had no effect on offspring growth
rate.
There was a significantly higher number of female runts (bodyweight <5.5g) in the groups sired by rats exposed to ethanol. There was also asignificantly higher number of male runts in the groups sired by rats
exposed to ethanol but only in comparison to the intubated control; thedifference with the non-intubated control was not significant.

Conclusion:

Reliability:

12.11.2004

The % of males in litters sired by ethanol treated rats was significantly
lower than the % sired by vehicle-treated fathers (p<0.04) although thedifference with the non-intubated control was not significant. Ethanol
treatment at both levels resulted in a significant increase in absoluteadrenal gland weight but not in brain/bodyweight ratios. Organ weights(both absolute and bodyweight relative) were unaffected at 7 days but
significant reductions of spleen and heart weight were noted at 21 days atthe 3g/kg dose level (NOEL=2g/kg).
There was no effect on fertility in a group of 20 male rats given 3 g or 2
g/kg ethanol by oral intubation daily for nine weeks, achieving BAL's of
338±15 and 132±5mg/dL, respectively. Although fertility was unaffected,
this study did reveal higher incidences of runted pups in the resulting
offspring especially at the highest exposure level (3g/kg).
(2) valid with restrictions
Study well enough reported for a valid with restrictions rating
(288)

Type
: Fertility
Species: MouseSex: MaleStrain: C57BL
Route of admin.
: oral feed
Exposure period: 35 or 70 days
Frequency of treatm.
: Daily
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studiesDoses: 5 or 6% in feed calculated to yield 12,000 to 14,000 mg/kg/day
Control group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Method248:

Animals: supplied by Jackson Laboratories (Bar Harbor, ME).
Age: 60 days.
Environment: 22 ± 1 degree C; relative humidity 45 ± 5%; 14 hr:10- hrlight:dark cycle. Animals housed individually.
Feed: Laboratory feed and water ad libitum.
Treatment: Vitamin supplemented (3g/l) chocolate flavoured Carnation
Slender containing sucrose in amounts isocaloric to either 5% (v/v) or 6%(v/v) ethanol for 3 days after which on half of the animals in each group
received diets containing 5% or 6% ethanol. All mice initially treated for 3
days on contro followed by 70 days treatement for the 5% ethanol groupand 35 days for the 6% ethanol group. 48 Hr after cessation of treatment,

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ID: 64-17-5
DATE: 19.11.2004

Remark:Result:Source:Conclusion:

mice were anaestheticsed then hemicastrated and the right testis andassociated structures weight recorded. Spermatozoal function (number,
motility, forward progression, in vitro fertilisation of mouse oocytes) andblood alcohol levels (BAL) were determined from 25ul aliquots of tail blood(head space sampling technique used). After castration, the incision wasclosed and the mice allowed free access to lab chow and water (notreatment) for a further 70 days. All animals then sacrificed.
Group sizes consisted 5 to 12 individuals (treatment and control groups
respectively).
Controls: Vehicle (Carnation Slender containing sucrose as above).
Statistical analysis: Wilcoxon tests; parametric data analysed following
appropriate transformations and Newman-Keuls multiple range test.
Hemicastration was used to evaluate the reversibility of ethanol's effects onmale reproductive function. Hemi-castrated pair fed controls were used tominimize the effect of hemicastration on the data (as it is known to produce
compensatory effects on the remaining testis).
Except for germ cell desquamation, all effects seen at the 5% ethanol dietwere reversible. The authors speculated that Sertoli cells rather than
Leydig cells are involved in reproductive failure of abstinent alcoholics.
BAC peaked at 166± 38mg% (1660mg/l, n=15) for the 5% dose and 260±
35mg% (2600mg/l) for the 6% dose, both at day 34/35.
After treatment with either 5% ethanol or 6% ethanol, testicle weightdecreased by 24% and 28%, but the effect was reversible and there wasno significant difference between the control and recovery animals (10weeks no treatment, P>0.1).
Seminal vesicle/prostate weights decreased by 20% for those on the 6%
diet but the effect was reversible and there was no significant difference
between the control and recovery animals (10 weeks no treatment, P>0.1).
There were significant increased frequencies of germ cell desquamation(480% in the 5% treatment group, 400% in the 6% treatment group) and ofinactive seminiferous tubules (186% in the 5% treatment group, 567% in
the 6% treatment group) Improvement in both parameters was noted in thecontralateral organs after 10 weeks alcohol abstinence but all remained
significantly elevated except for the % inactive tubules which returned tocontrol group levels in the 5% treatment group. (Note: Germ cell
desquamation in 5% treatment recovery group - 95% confidence limit
range: 1.2-3.2% lumina showing desquamation versus control level of 0.31.0%.)
Quality of spermatogenesis was significantly poorer in testes from both
treatment groups compared to their respective controls. After 19 weeks
abstinence, some pathology persisted in animals that had been exposed toethanol although the differences were not significant.
Caudal epididymal sperm content was not significantly affected bytreatment with the 5% ethanol diet but was 6% lower in the animals
receiving the 6% diet (p<0.01). This difference disappeared following 10
weeks without treatment.
Sperm motility was not significantly affected by treatment with the 5%ethanol diet in the animals receiving the 6% diet molility was reduced by
85%. This difference disappeared following 10 weeks without treatment.
Forward progression was reduced in both treatment groups (apparently
more so in the lower treatment group). The difference disappeared in therecovery group that had been receiving 5% ethanol but persisted in the 6%group.
In vitro fertilization of mouse oocytes by epididymal spermatozoa was
reduced by 20% in the 5% treatment group and 63% in the 5% treatmentgroup but these differences disappeared in both treatment groups following
10 weeks abstinence.
IARC monographs on the evaluation of carcinogenic risk to humans. Vol
44, Alcohol Drinking. IARC, Lyon, France, 1988.
No NOAEL established (<5% ethanol diet, <166mg% BAC). However, for
persistent effects the NOAEL would appear to be close to 5% ethanol diet.

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Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:(2) valid with restrictions
Not a standard study protocol but appears to be well conducted andreported. Limited and very high doses does not allow a NOAEL to beestablished.

12.11.2004

(437)

Type
: Fertility
Species: Rat
Sex: MaleStrain: Sprague-Dawley
Route of admin.
: Inhalation
Exposure period: 3 to 4 weeks
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 22, 23, 25, 27 mg/l (approx 11500, 12000, 13000, 14000ppm)
Control group: yes, concurrent no treatment
NOAEL parental: = 23 mg/l
LOEL parental: = 25 mg/l
Method: otherYear: 1983
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Method:Remark:Result:250

Animals: adults, 330-350 g; acclimated for 3-5 days. Animals housed
together in airtight chambers.
Controls: Housed in similar chambers to exposed animals without ethanol
vapours present.
Environment: Not described.
Feed: Laboratory feed and water ad libitum during exposure. Feedsupplemented with vegetables and peanut bars.
Treatment: During acclimation, all treated rats were exposed to 22 mg/l
ethanol vapour in air. After acclimation, groups of 10-12 rats were exposedto ethanol at 23, 25 or 37 mg/l for up to 3 to 4 weeks. Five exposure wererun as separate experiments a concurrent controls. After treatment,
animals were immediately weighed then sacrificed and trunk bloodcollected over ice, the plasma separated and frozen for later analysis. Sex
organs (testes, seminal vesicles, prostate) were removed and weighed. In
one experiment, animals were injected with saline or gonadotropin
releasing hormone (GnRH) and sacrificed 10 minutes later.
Blood samples were withdrawn periodically from a tail vein for blood
alcohol concentration (BAC) determination.
Plasma testosterone and lutenising hormone (LH) were recorded.
Statistical analysis: Dunnett's and Duncan's tests after analysis of variance.
This data supports the premise that fertitily effects observed following highdoses of ethanol may well be confounded by malnutrition stress.
BACs of 94-127 mg/100ml resulted in smaller increase in bodyweight gain
than observed in control animals but was not associated with significant
changes in plasma testosterone levels or weights of sex organs. The
maximum BAC observed was 187mg/100ml. A BAC of 180 mg/100 ml was
associated with an inhibition of testosterone secretion only in animals that
had failed to grow but this was not seen in a second experiment with asimilar BAC.
In the experiment where GnRH was injected, a BAC of 163mg/100ml was
associated with a marked weight loss and reduction of plasma testosterone
levels. Basal plasma LH levels were comparable in the control and ethanol
treated rats and intravenous administration of GnRH produced comparable

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
elevations in LH level in the controls and ethanol treated rats.

Conclusion:

Reliability:

12.11.2004

NOAEL (male fertility) = 127mg/100ml BAC
LOAEL (male fertility) = 163mg/l00ml BAC
Authors' conclusion is that data indicates that in growing animals
testosterone secretion appears to be directly related to changes in bodyweight but not the degree of alcohol exposure. Adequate function of the
hypothalamic-pituitary-testicular axis provided normal growth is maintained.
Alcohol does not appear to lower pituitary gland sensitivity to GnRH.
(2) valid with restrictions
Not a standard protocol but reasonably well reported and a no effect levelestablished by a relevant route of exposure.
(289)

Type
: One generation study
Species: Rat
Sex: MaleStrain: Sprague-Dawley
Route of admin.
: Gavage
Exposure period:
Frequency of treatm.
: Daily
Premating exposure periodMale: 3 and 9 weeks
Female : not treatedDuration of test:
No. of generation: 1
studiesDoses: 2.5 and 5.0g/kg
Control group: yes, concurrent vehicle
Method: otherYear: 1995
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Method:Remark:Animals: Supplied by Charles River, Portage, MI but test animals bred inlaboratory.
Age: 70 days.
Number of animals: 18-26 per dose group.
Environment: 22 ± 1 degree C; relative humidity 45 ± 5%; 12 hr:12 hrlight:dark cycle.
Feed: Laboratory feed and water ad libitum.
Vehicle: distilled water.
Controls: Vehicle (distilled water without ethanol) and non-intubatedcontrols to evaluate the effects of intubation stress. Animals in the low andcontrol treated groups were pair-fed to the high dose group animals.
Treatment: Males were bred in once after 3 weeks exposure and twice
after 9 weeks of exposure to 75-90-day-old females for 2 weeks or until
sperm plugs were observed. Males were intubated throughout the mating
period. Females with plugs were separated and housed individually. At 20days gestation, females from the 3 week exposure breeding and one fromthe 9 week exposure breeding were killed and their foetuses counted,
sexed and weighed. The second female from the 9 week breeding wasallowed to deliver its litter, which was similarly assessed.
Blood alcohol levels (BALs) were determined in males after 15 weeks
exposure. Animals were sacrificed 1 hour after intubation (previously
determined to co-incide with peak BAL) and trunk blood measured. BAL
was assessed by reduction of NAD by alcohol dehydrogenase using a
Beckman automated analyser.
Statistical analysis: Chi-square, ANOVA and Duncan Multiple Range Tests
were used.
The 3 week exposure period was intended to assess the effect on

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Result:Conclusion:

Reliability:

12.11.2004

postmeotic spermatids and spermatogonia whilst the 9 week exposure
peroid was intended to assess the effect on germ cells throughout theirmaturation prior to as well as meiosis.
Male fertility and litter size was not affected by treatment. Fecundity was
not reduced in individual breedings but was significantly reduced at the
5g/kg dose level if the breeding periods were combined. There was nodifferences between the two control groups.
At the first breeding (3 weeks) the 2.5g/kg and concurrent control males
weighed significantly more than ad lib controls. After 9 weeks, only the
concurrent control animals weighed significantly more than the ad lib
controls.
There did not appear to be any treatment related effects on resorptions andlitter size.
The mean BACs for the two ethanol dose groups was 248±14mg% (5g/kg)
and 155±9mg% (2.5g/kg).
There was a treatment related increase in the number of male foetuses inthe 3 week breeding at 5g/kg, but the effect was not repeated at the 9 week
breedings.
At the 3 and 9 week breedings there was a significant dose related
increase in fetal weights. There was also a significant increase in placental
weights, but only at the 9 week breedings.
There were no treatement related effects on newborns sired by alcohol
treated males.
There was no significant effect on male fertility. Female fecundity was
reduced at the high dose level when all breedings were combined but litter
size was not decreased nor were resorptions significantly increased. The
main finding was an increase in fetal weights of offspring sired by alcohol
treated males.
(2) valid with restrictions
Not a standard protocol but reasonably well reported. Dose levels very
high and no clear no effect level established.
(290)

Type
: One generation study
Species: rat
Sex: maleStrain: Sprague-Dawley
Route of admin.
: oral feed
Exposure period: 5 weeks
Frequency of treatm.
: continuous
Premating exposure periodMale: 15 days
Female : no treatment
Duration of test
: 36 days
No. of generation:
studiesDoses: 6 and 10% in diet; calculated to be much greater than 12,000 mg/kg
Control group: no data specified
NOAEL parental: < 6 - 10 %Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Method252:

Animals: weight at start of treatment: 318 g (males); 259 g (females). No ofanimals: 12 males; 25 females.
Environment: Individually housed, 22 degC, RH 45% +/- 10 with diurnal
light cycle (nocturnal 20:00 to 8:00hrs.)
Dosage: Six treated rats were given a 6% v/v ethanol-containing liquid diet(providing 35% of calories.) This was increased to 10% v/v after 7 days
exposure (58% of dietary calories.

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5. TOXICITY

Result

Test substance
Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:::

12.11.2004

Controls: Six controls given an isocaloric amount of sucrose.
Diet 'Metrecal' (chocolate or vanilla.)
Duration of treatment: 15 days (males). females on lab chow and water.
Investigations: After treatment each male was placed in a cage with 2
females every night. No food or water was provided during this time. The
experiment was continued for 5 weeks, with the males still receiving
alcohol during the day. The males were killed on day 36. Blood samples
were collected.
Pregnancies were terminated on day 20 of gestation and litter size andfoetal mortality was assessed.
Treated males showed signs of intoxication and considerable weight gain
compared to controls. Treated animals were much less succesful atmating; the numbers of successful matings were 6/12 in the treated groupand 13/13 in the controls. The number of offspring/litter was greater in thecontrols (p <0.01). A higher incidence of early resorption was seen in thetreated group (p <0.01). Adverse effects on the testes, seminal vesicles,
ductules and in serum testosterone levels were noted.
From graphical data presented in the reference, it is possible to estimate
ethanol consumption as being in the range 7.2 to 14.4 g/kg/day.
Test substance was 95% v/v ethanol.
(2) valid with restrictions
Only six pregnancies examined in treatment group and males alsochronically treated with ethanol such that the quality of the study is
reduced. General toxicity such as ataxia, lethargy and weight loss during
the experiment may confound the results.
(254)

Type
: Fertility
Species: Rat
Sex: male/female
Strain: Sprague-Dawley
Route of admin.
: Inhalation
Exposure period: 7 hours
Frequency of treatm.
: Daily
Premating exposure periodMale: 6 weeks
Female : None
Duration of test
: see method details
No. of generation:
studies
Doses: 0, 10000, 16000ppm
Control group: Yes
other: NOAEL paternal : > 16000 ppm
Result: Negative
Method: otherYear: 1985
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Method:Exposure was conducted in 0.5m3 chambers with dynamic air flow (one airchange per minute.)
Animals: 18/group. Starting weights 400-500g.
Temperature: 73 +/- 3F. Humidity: average 40-50%.
Exposure period: 2 day non exposure period before mating. Mating period
5 days. Mating confirmed by presence of sperm plugs under cages orvaginal smears. Females housed individually.
Analytical monitoring: Yes (IR analyser - exposures found to be within 11%
of nominal). Independently cross-checked with charcoal adsorption tubes.
Parameters measured: weights (daily), food and water intake.

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Result:Reliability:

12.11.2004

No effect on weight gain, feed or water intake. No effect on fertility or litter
sizes.
Previous studies quoted as showing exposures to 10000 and 16000ppm
ethanol typically give rise to blood ethanol concentrations of 30 and
500mg/l ethanol. Authors calculate that for rats exposures in excess of
11000ppm are required to begin accumulating ethanol in the blood and thatethanol is no more toxic by the inhalation route than by other routes.
(2) valid with restrictions
Well reported study but not to a standard protocol but no pathology on
males. Study incomplete as a comprehensive assessment of effects onmale fertility. Route of exposure highly relevant.
(291) (292) (293)

Type: Fertility
Species: Rat
Sex: Female
Strain: CD-1
Route of admin.
: s.c.
Exposure period: OnceFrequency of treatm.
:
Premating exposure period
Male:Female :Duration of test
: OnceNo. of generation:
studies
Doses: 7900 mg/kg bodyweightControl group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
ResultReliability
12.11.2004::

Ovulatory surges of LH and therefore ovulation were blocked by ethanol.
(4) not assignable
(294)

Type
: Fertility
Species: Rat
Sex: MaleStrain: Sprague-Dawley
Route of admin.
: Inhalation
Exposure period: 1 week
Frequency of treatm.
: 6 hours/day
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 0, 1000ppm
Control group:
Method:Year: 1985
GLP: no data
Test substance: other TS
Method254:

Full method details provided in White (1983).
Equilibrium concentrations achieved after 30 minutes.
Temperature 20-22C, relative humidity 40-60%
Controls: air only.

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Remark:Result:

Test substanceConclusion::

Reliability:

17.11.2004

No food or water during treatment.
Statistical methods: student's t test, significant p<0.05
Animals sacrificed immediately after exposure or after an 18hr rest period.
Measurement of hormones was by radioimmunoassay.
Study designed to assess the effect on male reproductive function by
quantifying serum concentrations of testosterone and luteinizing hormone.
No significant effect on luteinising hormone or corticosterone. A significant
depression of testosterone occurred after the first exposure, but the level
returned to normal after 18hrs recovery and was absent at the end of thestudy.
99% ethanol, supplied by Merck.
The relevance of this transient effect was not considered significant withrespect to the ability of the testes to produce testosterone.
(2) valid with restrictions
Very restricted range of parameters assessed and exposure short, but
within these restrictions, results appear reliable.
(295)

Type
: Fertility
Species: Monkey
Sex: Female
Strain: Macaca Fascicularis
Route of admin.
: i.v.
Exposure period: 3-6.5 months
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 2900-4400 mg/kg bodyweightControl group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Result:Reliability
12.11.2004:

Amennorrhea, atrophy of the uterus, decreased ovarian mass andsignificant lowering of LH levels were observed.
(4) not assignable
(296)

Type
: One generation study
Species: Rat
Sex: male/female
Strain: Wistar
Route of admin.
: oral feed
Exposure period: gestation day 12 to ten days postpartum
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test
: in utero and as neonates
No. of generation:
studies
Doses: 36% of calorie intake; estimated to be greater than 12,000 mg/kg/day
Control group: no data specified
Method: otherYear:
GLP: no data

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Test substance:as prescribed by 1.1 - 1.4

Remark:Result:Reliability
12.11.2004:

The authors concluded that there was less phenotypic masculinization atbirth in the treated offspring.
In male progeny there was decreased anogenital distance; the weights ofthe testes and seminal vesicles/prostate were decreased 55 and 110 days
postpartum; serum testosterone and luteinizing hormone levels weredecreased on day 55 but not on day 110; and sexual motivation andperformance were reduced.
(4) not assignable
(297)

Type
: One generation study
Species: Rat
Sex: Female
Strain: Wistar
Route of admin.
: oral feed
Exposure period: while in utero and as neonates
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 36% of calorie intake
Control group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Result:Reliability
12.11.2004:

Male offspring gonadal growth and sexual performance was adversely
affected.
(4) not assignable
(298)

Type
: One generation study
Species: Rat
Sex: Female
Strain: Wistar
Route of admin.
: drinking water
Exposure period: Before mating, through gestation and lactation
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 12% in drinking water
Control group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
ResultReliability
12.11.2004

256

:
:

No effect on reproductive performance was noted.
(4) not assignable(299)

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Type: One generation study
Species: Rat
Sex: MaleStrain: Sprague-Dawley
Route of admin.
: i.p.
Exposure period: OnceFrequency of treatm.
: OncePremating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 2500 mg/kg bodyweightControl group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
Result:Reliability
12.11.2004:

There was a significant decrease in the level of LH and testosterone withmarked attenuation of testicular
steroidogenesis.
(4) not assignable
(300)

Type
: One generation study
Species: Rat
Sex: Female
Strain: Wistar
Route of admin.
: i.p.
Exposure period:
Frequency of treatm.
:
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses:
Control group:
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
RemarkResultReliability
12.11.2004:::

Information taken from secondary source - no dosage information given.
Ethanol increased prolactin secretion.
(4) not assignable
(301)

Type
: One generation study
Species: Rat
Sex: male/female
Strain: Sprague-Dawley
Route of admin.
: Other
Exposure period:
Frequency of treatm.
:
Premating exposure period
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Duration of test
No. of generation
studies
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:

11600 mg/kg bodyweight
no data specified
other

Result:Reliability
12.11.2004:

Patterns of LH secretion in both male and female offspring,as adults, indicated an effect on the central mechanisms
controling secretion of pituitary LH.
(4) not assignable

no dataas prescribed by 1.1 - 1.4

(302)

Type
: One generation study
Species: MouseSex: Female
Strain: C57BL
Route of admin.
: drinking water
Exposure period: before mating, through gestation and lactation
Frequency of treatm.
: Daily
Premating exposure period
Male:Female :Duration of test:
No. of generation:
studies
Doses: 10% in drinking water
Control group: no data specified
Method: otherYear:
GLP: no data
Test substance: as prescribed by 1.1 - 1.4
ResultReliability
12.11.2004
5.8.2

No significant effect on reproductive capacity.
(4) not assignable(303)

DEVELOPMENTAL TOXICITY/TERATOGENICITY

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Teratogenicity
Method
Year
GLP

258

:
:

:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleSprague-Dawley
Inhalation
17 hours per day
DailyDays 1-19 of gestation
10,000, 16,000 or 20,000 ppm
no data specified= 16000 ppm> 20000 ppm= 20000 ppm

:
:
:
:

>= 20000 ppm
otherno data

UNEP PUBLICATIONSOECD SIDS
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Test substance:as prescribed by 1.1 - 1.4

Remark:Age at study start - not stated - 176-200 g.
Number of animals per dose per sex: not explicitly stated but approximately
16.
Doses are calculated to be equivalent to 17, 29 and 28 g/kg bodyweight.
Vehicle used - not applicable
Mating conditions: Virgin females were housed with males and vaginal
smears were taken.
Foetuses were examined externally and internally for malformations;
implants and resorptions were recorded as was litter weight.

Result:No maternal organs were examined and fetuses were examined forexternal and visceral malformations.
Maternal LOAEL effect was narcosis and lowered food consumption.
Development LOAEL effect - none seen
Development NOAEL effect - visceral or skeletal malformations or variants.
Statistical tests were t-tests or chi-square tests; p=<0.05 regarded
significant.
This was considered in the IARC Monograph 1988.
No mortality occurred.
Maternal data are not given for the following:
Number aborting
Number of corporea lutea(Duration of pregnancy not relevant)Bodyweights
Haematology and blood chemistry findings
Gross pathology in dams
Organ weight changes
Histopathology incidence and severity.
Maternal data are given for the following:
The number of pregnant per dose level were 15/15, 15/16 and 14/16 in thelow, medium and high dosage groups.
The numbers of resorptions were not affected by ethanol inhalation.
Number of implantations were 14-16/litter in all ethanol-treated groups.
Number of corpora lutea were 14-16/litterFood consumption was lowered in the high-dose group.
Clinical signs: the highest dose induced complete narcosis (described assevere toxicity); lower doses did not cause narcosis but caused
hyperactivity after exposure.
Maternal weight gains were not affected by treatment.
Blood alcohol levels ranged 0.02 to 0.04 mg/ml at 10000ppm, 0.43 to 0.53
mg/ml at 16000ppm and 1.48 to 1.93 mg/ml at 20000ppm. Measurements
were made on non-pregnant rats and represent the ranges of the averagevalues measured at days 1, 10 and 19.
Foetal data are not given for the following:
Litter size (but are deduced to average 6.0 to 7.1
foetuses/litter across the groups).
Number viable
Postnatal growth (not applicable)
Postnatal survival (not applicable)

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Foetal data are given for the following:
Litter weights were not significantly affected by ethanol treatments.
Sex ratio did not differ significantly from controls.
Grossly visible abnormalities are given in detail but the frequency of eachdid not differ significantly between groups. More litters contained abnormal
foetuses in the 20,000 ppm group compared to controls.
No definite evidence of malformations due to ethanol exposure were seen
although the incidence of abnormal changes at the highest concentration
was of borderline significance.
(2) valid with restrictions
Well reported study which established a NOAEL
Critical study for SIDS endpoint(293)

Conclusion:

Reliability:

Flag
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
male/female
WistarGavagethroughout gestation and gestation + lactation
Daily

Method:Age at study start 3 mth (females 180 g)

1 g/kg/day (12.5% v/v in distilled water)
yes, concurrent vehicle
other
1998
no dataas prescribed by 1.1 - 1.4

Number of animals: not explicitly stated for parental stock but 252 offspring
subjected to further treatments and tests.
Treatment: Ethanol was administered as a 12.5% v/v solution in distilledwater by gavage to yield a dosage of 1 g/kg/day to dams throughout
gestation alone or through gestation and lactation.
Controls: Sucrose was added to distilled water vehicle to provide isocaloric
control.
Mating conditions: Females were housed singly with male for 4 days.
Offspring were appropriately cross-fostered and divided into those that had
not experienced exposure to ethanol, those that had experienced ethanol
in utero and those that had experienced etanol exposure in utero andthroughout lactation.
Dams were weighed during gestation. Offspring were weighed on days 3,
10, 20, 30, 45 and 63.
Offspring were subjected to assessments in Two-Way Active Avoidance(Shuttle-Box) Tests at 9 and 12 weeks and at 5 months.
Blood alcohol levels were determined on gestation day 14 and post natal
day 14.
Remark260:

Statistical analysis was by ANOVA and Chi-squared analysis.
This study is included as a neurotoxicity study in Chapter
5.9.

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY
ResultETHANOL
ID: 64-17-5
DATE: 19.11.2004:

Mortality was significantly increased (32% versus 7% in controls) inoffspring exposed to ethanol during pregnancy with continued postnatal
exposure having no significant further effect.
Offspring cross-fostered to dams that had been exposed to ethanol only
during pregnancy showed even greater mortality (59%). Growth inoffspring was delayed during the first 9
weeks. Learning was impaired in rats of both gender at 9 weeks relative tocontrols. This remained evident in males, but not females, at 5 months.
No visible malformations observed.
In offspring treated both pre- and post-natally with ethanol, 60% were poor
learners compared with 33% in sucrose controls.

Conclusion:

Reliability:

12.11.2004

Blood ethanol levels were 35.0 +/- 5.8mg/100ml.
Ethanol at a dose of 1 g/kg/day administered to dam rats during gestationand lactation produce growth and behavioural changes in the offspring.
(2) valid with restrictions
Single dose used and no NOAEL established. High levels of mortality alsoa concern; high levels of cannibalism (possibly due to aggressive
behaviour linked to alcohol withdrawal) offered as a possible explanation
by authors.
(304)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Teratogenicity
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleC57BL
oral feed

:
:
:
:
:

= 25 %
other1979
no datano data

Remark:Result:Age at study start 4-5 months
Dosage: NOAEL and LOAEL doses given as % ethanol-derived calories.
NOAEL (teratogenicity) effect is malformed foetuses and litter weight.
Doses are calculated to be equivalent to 17, 29 and 28 g/kg bodyweight
Ethanol or sucrose was added to provide calories.
Number of animals per dose per sex: not explicitly stated but approximately
16.
Mating conditions: Females were housed singly with proven studs until
vaginal plugs were found.
Dams were weighed on days 0, 4, 10 and 18 (at sacrifice).
Foetuses were examined externally and internally for malformations;
implants and resorptions were recorded as was litter weight.
No maternal organs were examined and foetuses were examined forexternal and visceral malformations.
Statistical tests were t-tests or chi-square tests; p=<0.05 regarded sign
No mortality occurred.

ad lib17%, 25% and 30% ethanol-derived calories
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Maternal data are not given for the following:
Number pregnant per dose level
Number aborting
Pre- and post-implantation losses
Number of corporea lutea(Duration of pregnancy not relevant)Haematology and blood chemistry findings
Gross pathology in dams
Organ weight changes
Histopathology incidence and severity.
Maternal data are given for the following:
The numbers of resorptions were one per litter at the twolower doses and 2/litter at the higher dose.
Number of implantations were 7.3/litter in all
ethanol-treated groups.
Maternal weight gains were not affected by treatment.
Rates of diet consumption were 12.02 ml/d, 12.86 ml/d and10.31 ml/d in the 3 ethanol dosed groups.
Clinical signs included slight tremulousness at the high-dose group when
ethanol-containing diet was removed.
Blood alcohol levels ranged 3 mg% to 384mg% across the 3 treatmentgroups.
Foetal data are not given for the following:
Litter size
Number viable
Sex ratio
Postnatal growth
Postnatal survival
Foetal data are given for the following:
Litter weights were not significantly affected by ethanol treatments.
Grossly visible abnormalities affected limb, eye, brain, heart, uro-genitaltract and abdomen. Litter weight was not affected by ethanol-containing
diets but malformations were significantly increased by maternal diets
containing 25% or more of ethanol-derived calories.

Conclusion
Reliability::

Grossly visible abnormalities, external, soft tissue and skeletal
abnormalities affected the limb, eye, brain, heart, urogenital tract andabdomen of foetuses.
The teratogenicity of ethanol is demonstrated.
(2) valid with restrictions
Well reported study which established a NOAEL.

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.

262

(305)
:
:
:
:
:
:
:
:
:
:

mouse
male
Swiss Webster
oral feed
28 daysad libitum
28 days
6.3% ethanol in liquid diet (32% ethanol-derived calories)
yes< 32 %UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
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DATE: 19.11.2004

NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Teratogenicity
MethodYear
GLP
Test substance::< 32 %= 32 %:::::= 32 %
other1981
no dataas prescribed by 1.1 - 1.4

Method:Age at study start: 190 days.
Number of animals per group is not stated.
Ethanol was added to diet and control diet contained an
isocaloric amount of sucrose.
Bodyweights were measured every two days. Blood samples
were taken for blood alcohol levels.
Mating: 48 h after ethanol or sucrose diets were removed
males were mated with nulliparous females by 5-days
cohabitation in the same cage. If no vaginal plugs were
found, new females were offered. Mating lasted until 11 days
after the last ethanol treatment.
Only pregnancy rate and resorptions were recorded for dams.
Corpora lutea were counted although data are not presented.
Males were not examined.

Remark:Result:Foetal examination included crown-rump length, viability,
sex ratio, litter size and weight and grossly visible
visceral and skeletal abnormalities.
These are PATERNAL NOAEL and LOAEL, not maternal.
Fertilizaton rate was decreased 1/9 among matings 3-5 days after
treatment.
Crown rump length was reduced in the one litter produced by mating 3-5days after paternal ethanol treatment.
Nine females became pregnant per dose level. There were no abortions
although pregnancy rates were reduced. Resorption rates varied 0-27%
across mating intervals but were unaffecetd by ethanol treatment.
Implantations, implantation losses and numbers of corpora lutea were notreported.
Paternal bodyweights were unaffected by treatment.
Clinical signs, haematological and blood biochemical parameters were notreported.
Blood alcohol levels reached 296 +/- 19 mg%.
Gross pathology, organ weight changes and histopathological incidenceswere not studied.
Foetal changes:

Conclusion:

Reliability:

Litter size and weights, percentage of live foetuses and sex ratios were
unaffecetd by ethanol treatment.
Only 2 abnormalities (undescended testes and body asymmetry) occurredin 95 pups sired by treated males.
Skeletons were not examined.
The role of paternal alcohol intake on anomalies seen in foetal alcohol
syndrome was not conclusively established.
(2) valid with restrictions
Well reported study but only a single high dose used, which did not allow ano effect level to be established.

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DATE: 19.11.2004

12.11.2004(306)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Teratogenicity
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleCBAoral feed-31 to 17 gestationad libitum

:
:
:
:
:

= 15 %
other1977
no datano data

Remark:No early deaths were reported,
No. pregnant per dose level: 8-10
All implants were resobed at the highest concentration of ethanol in diet.
Resorption rates were 2% and 0% in controls and 57%, 72% and 100% in
respective treatment groups.
Numbers of implantations were 4.8 and 5.6 in controls and 4.0, 5.5, 5.2 and
0 in the treatment groups.
Pre- and post-implantation losses were not specified.
No. of corpora lutea were not counted.
Clinical signs were not discussed although dams were described as'alcoholic'.

15, 20, 25 and 30% ethanol-derived calories
Yes< 15 %< 15 %= 15 %

Blood alcohol levels before mating werer 0 and 0 mg/dl in controls and 73,
121, 174 and 315 mg/dl in treatment groups.

Result:Liver weight relative to body weight measured in 3 females per group
before mating were not affected by treatment and there were no
histopathological effects seen in liver tissue.
Foetal data:
Litter size is not given. Foetal weights were depressed by treatment withmeans of 0.97 and 0.95 g in controls and 0.64, 0.33 and 0.51 g in the 3
lowest ethanol dose groups.
Defects included skeletal abnormalities at 100% incidence in all 3 ethanol
treated groups. These effects were primarily of the occipital bone but also
affected the sternum and ribs.

Test condition:

Visceral abnormalites affected 0% of foetuses in either control group and
36%, 100% and 100% of foetuses in the 3 ethanol treated groups. Dilated
brain ventricles were the most frequent anomaly but open eyelids,
exencephaly, gastroschisis and heart defects also occurred in the higher
dose groups.
Age at study start: 60-100 days.
Number of animals per group: at least 8 per group..
Ethanol was added to diet and control diet contained an
isocaloric amount of sucrose. Females received ethanol indiet for 10 days before dose graduated to next higher

264

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concentration until there were 10 females in each dietgroup. Depending on dose group, animals received ethanol
for 30 to 80 days before mating.
Bodyweights were measured every two days. Blood samples
were taken for blood alcohol levels.
Mating: 48 h after ethanol or sucrose diets were removed
males were mated with nulliparous females by 5-days
cohabitation in the same cage. If no vaginal plugs were
found, new females were offered. Mating lasted until 11 days
after the last ethanol treatment.
Only pregnancy rate and resorptions were recorded for dams.
Corpora lutea were counted although data are not presented.
Males were not examined.

Reliability:

Foetal examination included crown-rump length, viability,
sex ratio, litter size and weight and grossly visible
visceral and skeletal abnormalitie(2) valid with restrictions
Not a standard study protocol but appears to be well conducted andreported. Limited and very high dose does not allow a NOAEL to beestablished.

12.11.2004(307)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Teratogenicity
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleC3H
oral feed-31 to day 17 gestation
Ad libitum

:
:
:
:
:

= 20 %
other1977
no datano data

Remark:Maternal data:
No early deaths were reported.
No. pregnant per dose level: 8-10
All implants were resorbed at the highest concentration of ethanol in diet.
Resorption rates were 7% and 0% in controls and 0% 30%, 72% and 100%in respective treatment groups. Early and late resorptions were notdistinguished.
Numbers of implantations were 11 and 7.3 in controls and 6.8, 6.5, 6.1 and
0 in the treatment groups. Pre- and post-implantation losses were notspecified.
No. of corpora lutea were not counted.
Clinical signs were not discussed. Animals were descriubed as 'alcoholic'.
Bodyweight: Not given.
Food/water consumption: Calorific intake was similar across all groups.
Haematological findings: Not measured.
Blood alcohol levels were 103, 160, 289 and 398 mg/dl in the treatment

20, 25, 30 and 35% ethanol-derived calories
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groups.
Clinical biochemistry and gross pathology: Not examined.
Organ weight changes: Liver weight relative to bodyweight was notaffeceted by treatment.
Histopathology: No liver pathology was detected.
Foetal data:
Litter size and weights: Not given. Foetal weights in controls weredepressed (1.14 and 1.27 g in controls and 0.77, 0.50 and 0.58 g in theethanol dosed groups).
Number viable and sex ratio: Not reported.
Postnatal growth and postnatal survival: Not applicable.
Foetuses showed high rates of skeletal and visceral anomalies at all doses
yielding foetuses.
Blood alcohol levels before mating werer 0 and 0 mg/dl in controls and 73,
121, 174 and 315 mg/dl in treatment groups.

Result:Liver weight relative to body weight measured in 3 females per group
before mating were not affected by treatment and there were no
histopathological effects seen in livertissue.
These data and test conditions were taken from Source
document which did not present the author and reference.
Foetal data:
Litter size is not given. Foetal weights were depressed by treatment withmeans of 0.97 and 0.95 g in controls and 0.64, 0.33 and 0.51 g in the 3
lowest ethanol dose groups.
Defects included skeletal abnormalities at 100% incidence in all 3 ethanol
treated groups. These effects were primarily of the occipital bone but also
affected the sternum andribs.

Test condition:

Visceral abnormalites affected 0% of foetuses in either control group and
36%, 100% and 100% of foetuses in the 3 ethanol treated groups. Dilated
brain ventricles were the most frequent anomaly but open eyelids,
exencephaly, gastroschisis and heart defects also occurred in the higher
dose groups.
Age at study start: 60-100 days.
Number of animals per group: at least 8 per group.
Ethanol was added to diet and control diet contained an isocaloric amountof sucrose. Females received ethanol in diet for 10 days before dose
graduated to next higher concentration until there were 10 females in eachdiet group. Depending on dose group, animals received ethanol for 30 to
80 days before mating.
Bodyweights were measured every two days. Blood samples were taken
for blood alcohol levels.
Mating: mated in pairs during 1.5 hr periods. Copulation plugs indicative ofmating.
Parameters assessed: Blood ethanol levels and relative liver weights
before mating and fetal weights and abnormalities.
Organs examined at necropsy: Adult livers. Foetuses were examined for

266

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
abnormalities of the skeleton and internal organs.

Reliability:

12.11.2004

Foetal examination included crown-rump length, viability, sex ratio, litter
size and weight and grossly visible visceral and skeletal abnormalities.
(2) valid with restrictions
Well reported study but not to a standard protocol. Very high doses usedand no NOAEL identified.
(308)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
NOAEL teratogen.
LOAEL Maternal
Toxicity
LOAEL Embryotoxicity
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleCD-1
Gavage8-14 gestation
once per day

:
:
:
:
:

> 6400 mg/kg bw
other
1987
no dataother TS: 200 proof

Method:Age at start: 8-10 wks
Number of animals per dose pewr sex: 6 confirmed pregnant/group.
Ethanol was administered by gavage in distilled water in 10 ml bolus
doses.
Clinical observations performed were physical examination and weightrecording 6 times through pregnancy. Viability checked twice daily.
Females were paired 1:1 with males and vaginal plugs were indicative ofpregnancy.
Parameters assessed were maternal bodyweights, numbers of implantationsites, resorptions, live and dead foetuses, external abnormalities.

2,200, 3,600, 5,000 and 7800 mg/kg bw
Yes= 2200 mg/kg bw>= 6400 mg/kg bw= 3600 mg/kg bw

No organs were examined at necropsy.

Remark:Statistical tests were Bartlett's for homogeneity of variance, one-way
ANOVA, Dunnett's, Duncan, Kruskal-Wallis, Dunn's and nested ANOVA.
Maternal data:
Mortality and day of detah: No control animals died.
Mortality rates in treated groups were 0/6, 1/6, 4/6, 5/6 and 6/6. Day ofdeath not reported.
Number pregnant per dose level: 6
Number aborting: Not reported. Possibly 2 litters were aborted at 5000
mg/kg. A surviving dam at 6400 mg/kg delivered a litter.
Number of resorptions: Not distinguised. Resorptions per litter did not
differ from control below 5000 mg/kg.
Mean implantations ranged from 10.5 in controls to 13.83 but no significant
difference noted.
Pre- and post-implantation loss: Not reported.
No. of corpora lutea: Not measured.
Duration of pregnancy: dams killed on gestation day 18.
Bodyeight: Not affected by treatment.
Food consumption: not reported.
Clinical signs: Timing and duration not reported. At 3600 mg/kg and above

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
dams were lethargic with staggering gait and or laboured breathing.
Haematological findings: Not measured.
Clinical biochemistry: Not measured.
Gross pathology: Not reported.
Organ weight changes: Not measured.
Histopathology: Not reported.
Foetal data:

Result:Litter size and weights: Not reported. Group means were not significantly
different from controls.
Number viable: Mean number of dead foetuses per litter did not varysignificantly with dose and ranged 0 to 0.5. Number of live foetuses per
litter differed significantly from controls at 5000 mg/kg.
Sex ratio: Not reported.
Postnatal growth: Not applicable.
Postnatal survival: Not applicable.
Grossly viable abnormalities etc: No externally malformed foetuses were
found in treated groups. Other types of abnormality were not sought.
No maternal mortality occurred at 2200 mg/kg but 1/6 dams died at 3600
mg/kg rising to 6/6 at 7700 mg/kg. At doses of at least 3600 mg/kg, dams
were lethargic and showedstaggered gait and/or laboured breathing.
At 5000 mg/kg, resorption of litter were increased and live foetuses/litterswere decreased. This was not apparent in the one litter at 6400 mg/kg. No
other fetal effects were seen.
Foetal data:
Group mean litter weights ranged from 1.33 g (controls) to 0.99 g and did
not vary with statistical significance. Mean number of dead foetuses per
litter was not dose related and ranged from 0 to 0.5.

Conclusion:

Reliability:

12.11.2004

No externally visible malformations were found in foetuses from treatedanimals.
No dose-related adverse effects on foetuses were observed at doses close
to those causing acute maternal toxicity.
(2) valid with restrictions
Well reported study but not to a standard protocol.
(309)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL teratogen.
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleSprague-Dawley
drinking water
Days 6-15 gestation
Dailyad-libitum
15% ethanol in water
yes, concurrent vehicle> 15 %
not developmentally toxic
other
1978
no dataas prescribed by 1.1 - 1.4

Method:Animal details: supplied by Spartan Research Animals, Haslett, Michigan;weight 250-270 g
Acclimation: 2-3 weeks at 72 degF and 45% RH;light cycle 12 hr dark, 12

268

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY

ResultETHANOL
ID: 64-17-5
DATE: 19.11.2004:

hr dark.
Dosing: Liquid provided via a ball tipped waterer to minimise evaporation.
Food: Purina lab chow.
Day 0 taken when sperm observed in a vaginal smear.
Blood alcohol determinations in non pregnant animals (using an alcohol
dehydrogenase method): 10 am, 2 pm, 10 pm, 2 am and 6 am on day 4 oftreatment.
Sacrifice: day 21
Observations:-Weights (dams daily and offspring)- Live, dead and resorbed fetuses.
- Crown-rump length- Number of each sex- External examination and check for cleft palate.
- One third of fetuses examined histopathologically for soft tissue damage;
heads preserved in Bouins solution and examined for soft tissue damage
but not skeletal alterations.
- All fetuses preserved in alcohol and subsequently processed for skeletal
alterations.
Statistical evaluation: Litter used as experimental unit. Wilcoxon test as
modified by Hasseman and Hoel to evaluate incidence of fetal alterations
and resorptions. Analysis of variance used for maternal and fetal
bodyweights. Level of significance p<0.05.
Mean consumption of food and liquid by rats given ethanol was significantly
less that that of control rats during the experimental period. As a result,mean gain in body weight of the exposed rats was also significantly lessbetween days 6 and 16 of gestation. Ethanol ingestion did not affect fetal
survival adversely, but mean fetal body weight was significantly less thanthat of the control litters.
No malformed fetuses were found in the experimental litters. No external orsoft tissue alterations were observed among the fetuses of the control orexperimental litters. Skeletal malformations were not detected in theexperimental group but skeletal variants consisting of unfused bones of theskull and cervical vertebra with missing centra occurred in the ethanol
litters at an incidence significantly greater than in the control litters.
Reduced fetal body weight (5.41g+/-0.25; control 5.70+/-0.35)
Non fused sternebrae (100 in 18 litters; control 45 in 8)
Vertebrae-missing centra (117 in 18 litters; control 15 in 8)
Wavy ribs (13 in 5 litters; control 3 in 3)
Note: total skeletal fetuses examined 223 in 29 litters
Overall there were no definite teratogenic abnormalities.

Test substance
Reliability::

12.11.2004
Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL teratogen.

:
:
:
:
:
:
:
:
:
:

Peak blood alcohol levels: 40 mg/100ml blood at 6am.
Reagent grade ethyl alcohol 190 proof from US Industrial Chemicals Co.
(2) valid with restrictions
Well reported study but only single dose used.
(310)
Mouse
FemaleCD-1
drinking waterdays 6-15 of gestation
Daily
ad libitum
15% ethanol in drinking water
yes, concurrent vehicle> 15 %UNEP PUBLICATIONS

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result
MethodYear
GLP
Test substance

:
:
:
:
:

no developmental toxicity observed.
other

Method:Result:Animals: Virgin mice from Carworth Animals, Portage, Michigan, weight 2225 g.
Acclimation: 2-3 weeks at 72 degF and 45% RH;light cycle 12 hr dark, 12
hr dark.
Dosing method: Liquid provided via a ball tipped waterer to minimise
evaporation.
Food: Purina lab chow.
Day 0 taken when vaginal plug observed in mice.
Blood alcohol determinations in non pregnant animals (using an alcohol
dehydrogenase method): 10 am, 2 pm, 10 pm, 2 am and 6 am on day 4 oftreatment.
Sacrifice: day 18
Observations:-Weights (dams daily and offspring)- Live, dead and resorbed fetuses.
- Crown-rump length- Number of each sex- External examination and check for cleft palate.
- One third of fetuses examined histopathologically for soft tissue damage;
heads preserved in Bouins solution and examined for soft tissue damage
but not skeletal alterations.
- All fetuses preserved in alcohol and subsequently processed for skeletal
alterations.
Statistical evaluation: Litter used as experimental unit. Wilcoxon test as
modified by Hasseman and Hoel to evaluate incidence of fetal alterations
and resorptions. Analysis of variance used for maternal and fetal
bodyweights. Level of significance p<0.05.
Mice receiving ethanol consumed significantly less food and liquid thancontrol mice. Consumption returned to control levels within two days after
removal of the ethanol. Maternal body weight gain reflected the decreased
consumption of food and liquid. The number of live fetuses/litter was not
significantly affected but fetal weight and length were significantly
decreased upon comparison to control values. Other than one fetus withcleft palate and two fetuses from different litters that had exencephaly with
open eye, no external or soft tissue alterations were noted among theoffspring of dams given ethanol. The incidence of exencephaly, open eye,and cleft palate did not differ significantly from control values. Skeletal
malformations were not detected but the incidence of several minor
skeletal variants e.g. delayed ossification of the centra of cervical vertebra,
non-fused sternebrae and delayed ossification of sternebrae (less than
90% ossified), was significantly increased among the litters of miceingesting ethanol.

no dataas prescribed by 1.1 - 1.4

Significant effects in mice:
reduced fetal body weight (0.95g+/-0.12; control 1.11+/-0.11)
Reduced crown rump length (22.2+/-1.0mm; control 23.5+/-1.2mm)
Non fused sternebrae (52 in 18 litters; control 26 in 12)
Delayed ossification (59 in 17 litters; control 7 in 4)
Note: total skeletal fetuses examined 239 in 21 litters
Overall there were no significant teratogenic abnormalities.
Test substance
Reliability

270

:
:

Peak blood alcohol levels were 204 mg/100ml at 2am.
Reagent grade ethyl alcohol 190 proof from US Industrial Chemicals Co.
(2) valid with restrictions

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Well reported study but only single dose level used.

12.11.2004(311)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL teratogen.
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

RabbitFemaleNew Zealand whitedrinking water
Days 6-18 of gestation
Daily
ad libitum
15% ethanol in drinking water
yes, concurrent vehicle> 15 %
not developmentally toxic
otherMethod:Result:Animals: supplied by Langshaws Rabbitry, Augusta, Michigan. Initial
animal weights not specified.
Acclimation: 2-3 weeks at 72 degF and 45% RH;light cycle 12 hr dark, 12
hr dark.
Dosing: Liquid provided via a ball tipped waterer to minimise evaporation.
Food: Purina lab chow.
Day 0 taken when mating observed.
Blood alcohol determinations in non pregnant animals (using an alcohol
dehydrogenase method): 10 am, 2 pm, 10 pm, 2 am and 6 am on day 4 oftreatment.
Sacrifice: day 29
Observations:-Weights (dams daily and offspring)- Live, dead and resorbed fetuses.
- Crown-rump length- Number of each sex- External examination and check for cleft palate.
- One third of fetuses examined histopathologically for soft tissue damage;
heads preserved in Bouins solution and examined for soft tissue damage
but not skeletal alterations.
- All fetuses preserved in alcohol and subsequently processed for skeletal
alterations.
Statistical evaluation: Litter used as experimental unit. Wilcoxon test as
modified by Hasseman and Hoel to evaluate incidence of fetal alterations
and resorptions. Analysis of variance used for maternal and fetal
bodyweights. Level of significance p<0.05.
Liquid consumption of animals receiving ethanol was significantly less thanthat of controls as was mean body weight (latter difference as statisticallysignificant on days 12 and 18 of gestation.)
The incidence of resorptions among litters of rabbits given ethanol wasapproximately twice that observed in the control litters; this increase wasdue primarily to the complete resorption of two litters in the ethanol group.
Fetal body measurements and the number of malformed fetuses werecomparable between the control and experimental litters. No alterations
were observed at an incidence that was significantly increased in theethanol group compared to the control group. Minor vascular alterations
observed have all been found to occur spontaneously in control groups ofthis strain of rabbit.

no dataother TS

Overall there were no teratogenic abnormalities
Peak blood alcohol level was 24mg/100ml at 6am.

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OECD SIDS
5. TOXICITY
Test substance
Reliability

ETHANOL
ID: 64-17-5
DATE: 19.11.2004::

Reagent grade ethyl alcohol 190 proof from US Industrial Chemicals Co.
(2) valid with restrictions
A well reported study but only a single dose used.
(311)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
NOAEL maternal tox.
other: NOAEL
behavioural
teratogenicity
other: NOAEL fertility
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Rat
male/female
Sprague-Dawley
inhalation
7 hours
Daily
see method details
0, 10000, 16000ppm
Yes> 16000 ppm> 16000 ppm

:
:
:
:
:
:

> 16000 ppm
Negative
other: see method details
1985
no dataas prescribed by 1.1 - 1.4

Method:Behavioural testing procedures:
Behavioral testing was from days 10 - 90. Female and male pups wereselected randomly. For each test, one female and one male were usedfrom each litter. Testers were not aware of the treatment groups to which
subjects belonged. Tests used:
1. Rotorod, 9cm in diameter and 10 cm long, and the surface was roughwith sand. Rotation speed increased until the animals had fiveunsuccessful trials.
2. The open field was 1 m in diameter, with an enclosure wall 0.5 m high.
Animals were tested for 3 min.
3. Optical digital animal activity monitor. The animal test area was a
40x40x20 cm Plexiglas cage which had 30 photodiodes per side. Activity
scores were summed over the 3 days of testing at each age.
4. Running wheel activity over a 24 hr period, separated into day and night
activity scores.
5. Two shuttle boxes in sound-attenuated chambers, with 4 cm center
partitions. Metal grid floors to which electrical shocks could be applied. 5
sec warning stimulus.
hr/day until they no longer responded sufficiently to receive reinforcement.

02.06.2004

Exposure was conducted in 0.5m3 chambers with dynamic air flow (one airchange per minute.)
Animals: 18/group. Starting weights 400-500g.
Temperature: 73 +/- 3F. Humidity: average 40-50%.
Exposure period: Males: 6 weeks; 2 day non exposure period before
mating. Females: GD 1-20. Mating period 5 days. 7 hours per day.
Mating confirmed by presence of sperm plugs under cages or vaginal
smears. Females housed individually.
Analytical monitoring: Yes (IR analyser - exposures found to be within 11%
of nominal). Independently cross-checked with charcoal adsorption tubes.
Parameters measured: weights (daily), food and water intake as measuresof maternal toxicity.
Parturition: All litters culled to 4 pups of each sex and fostered to untreateddams which had delivered within past 2 days. On PND10 pups individually
identified by ear punch and randomly assigned to behavioural study

272

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5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:Reliability:

12.11.2004

groups.
No effect on weight gain, feed or water intake. No effect on fertility or litter
sizes.
Behavioural testing showed no difference from controls in any of the
behavioural tests.
Previous studies quoted as showing exposures to 10000 and 16000ppm
ethanol typically give rise to blood ethanol concentrations of 30 and
500mg/l ethanol. Authors calculate that for rats exposures in excess of
11000ppm are required to begin accumulating ethanol in the blood and thatethanol is no more toxic by the inhalation route than by other routes.
(2) valid with restrictions
Well reported study but not to a standard protocol. Route of exposure
highly relevant.
(291) (292)Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
male/female
Sprague-Dawley
inhalation
7 hours
Daily
see method details
0, 10000, 16000pm
Yesother: see method details
1988
no dataas prescribed by 1.1 - 1.4

Method:Behavioural testing procedures:
Behavioral testing was from days 10 - 90. Female and male pups wereselected randomly. For each test, one female and one male were usedfrom each litter. Testers were not aware of the treatment groups to which
subjects belonged. Tests used:
1. Rotorod, 9cm in diameter and 10 cm long, and the surface was roughwith sand. Rotation speed increased until the animals had fiveunsuccessful trials.
2. The open field was 1 m in diameter, with an enclosure wall 0.5 m high.
Animals were tested for 3 min.
3. Optical digital animal activity monitor. The animal test area was a
40x40x20 cm Plexiglas cage which had 30 photodiodes per side. Activity
scores were summed over the 3 days of testing at each age.
4. Running wheel activity over a 24 hr period, separated into day and night
activity scores.
5. Two shuttle boxes in sound-attenuated chambers, with 4 cm center
partitions. Metal grid floors to which electrical shocks could be applied. 5
sec warning stimulus.
hr/day until they no longer responded sufficiently to receive reinforcement.
Exposure was conducted in 0.5m3 chambers with dynamic air flow (one airchange per minute.) Dosing method described in detail.
Animals: Starting weights: females, 15 per group, 176-200g; males, 18 pergroup >390g.
Temperature: 24+/-2C. Humidity: ~40%. 12hr light/dark cycle.
Exposure period: Males: 6 weeks; 2 day non exposure period before
mating. Females: GD 1-20. Mating (1:1 male:female) period 5 days.
Mating confirmed by presence of sperm plugs under cages or vaginal
smears. Females housed individually. Stock males used as sires forcontrols.
Analytical monitoring: Yes (IR analyser - exposures found to be +/-200ppm
of nominal). Independently cross-checked with charcoal adsorption tubes

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OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
analysed by gas chromatography.
Parameters measured weekly: weights food and water intake as measuresof maternal toxicity.
Parturition: All litters weighed within 16 hrs. Litters less than 3 pups per
sex discarded. Offspring weighed on PND 7 and checked for abnormalities
Neurochemical analysis: One female and one male (untested) from 5 litterssacrificed PND 21 for analysis of concentrations of protein and theneurotransmitters acetylcholine. dopamine, norepinephrine, 5hydroxytryptamine, substance P. beta-endorphin, and Met-enkephalin. Pup
brains were separated into the four general brain regions of cerebrum,
cerebellum, brainstem meduila-pons), and midbrain and frozen until
assayed by sonic homogenization in 8 ml of 0. 1 N HCl.

Remark:Result:Reliability:

12.11.2004

Statistical Analyses: Behavioral data were analyzed using multivariate
analysis of variance or an m-ranking procedure. Repeated measures
analyses were conducted where appropriate to p<0.05. Neurochemical
data were analyzed using Analysis of Variance followed by Duncan'sMultiple Range post-hoc tests where a significance was found.
Authors concluded that industrial inhalation exposure to ethanol may not be
expected to produce alarming blood ethanol levels and that inhalation
exposures to ethanol which produce narcosis in maternal rats are notteratogenic.
Males: weight gain retarded during 1st week but normal thereafter.
Females: no effect on weight gain. Feed intake retarded during 1st week
but normal thereafter at 16000ppm.
No effects on litter size, still births, length of pregnancy, offspring survival.
No effect observed in behavioural study tests.
No effect on dopamine, substance P, beta-endorphin and acetylcholine
levels.
Significant effects on norepinephrine, 5-hydroxytryptamine but magnitude
and direction of changes not correlated with dose. Level of Met-enkephalinaffected at lower but not higher dose.
(2) valid with restrictions
Well reported study but not to a standard protocol. Route of exposure
highly relevant.
(291)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

MonkeyRemark:Result:Reliability:

There was some correlation with cognitive performance and this study isfurther considered in Neurotoxicity.
Standardised craniofacial cephalograms of 18 macaques exposed weekly
to ethanol or sucrose solution in utero were measured at ages 1, 6, 12 and24 mths showed that there may be a critical period for induction of alcoholinduced craniofacial alterations. These were most prominent at 6 mths anddiminished thereafter as underlying changes inskeletal structure caused disappearance of the thin upper lip and smoothphiltrum characteristic of feat alcohol syndrome.
(4) not assignable274

Macaca Fascicularis
other: in utero expusure
throughout gestation
Daily

other
1999
no datano data

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

12.11.2004(312)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Human

Remark:Reliability
12.11.2004:

The large and growing evidence of the effects of parental alcohol
consumption on offspring was assessed. Study of clinical cases support
the conclusion that chronic heavy maternal drinking consistent with alcohol
dependence or alcoholism is an aetiological factor in foetal alcohol
syndrome. The effects of this can extend beyond puberty with cognitive
and neurobehavioural problems continuing and with the most deleterious ofoutcomes. Animal studies have demonstrated a dosage relationship in theabsence of the confounding factors prevalent in humans (e.g. poormaternal health and tobacco smoke) and that the effects of alcohol aremost pronounced on the brain. Individual differences in maternal
metabolism and genetic factors may explain why the infants of someproblem-drinking mothers are more affected than others but there is also
some evidence of a threshold of drinking below which adverse effects
cannot be detected. This may be around 30 to 40 g per day (4-5 UK drinks
or 2.5 to 3.5 US drinks per day). This level is well above the levels definedas moderate drinking for non-pregnant women.
This is a literature review presented as a book chapter.
(4) not assignable(313)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses

:
:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Result:Reliability
12.11.2004:

SpeciesSex::oral feedother:literature review
1999

Rat
Femaledrinking water
4 weeks and throughout gestation
20% alcohol before mating and 30% through gestation; 3500 mg/kg/day
throughout treatment.
otherno dataas prescribed by 1.1 - 1.4
Offspring of female Sprague-Dawley rats that were administered ethanol inwater as the drinking fluid at a concentration of 20% v/v for four weeks
before mating and at a concentration of 30% v/v during gestation were
physically and developmentally retarded and failed to catch up with control
offspring during the first four weeks postpartum.
(4) not assignable
(314)
Rat
Female

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275

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Sprague-Dawley
drinking water
4 weeks before mating and on days 1 to 20 of gestation.

Result:Reliability
12.11.2004:

The offspring of Sprague-Dawley rats given 20% ethanol in the drinkingwater four weeks before mating and 30% ethanol in drinking-water until
gestation day 20 had retarded skeletal development and decreased bodyweight but no gross malformation.
(4) not assignable(315)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses

:
:
:
:
:
:
:
:

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

Remark:Result:Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

RemarkResult::Reliability
12.11.2004:

Study was designed to assess ethanol effects on sexual differentiation.
Gestation was prolonged and offspring of each sex showed decreased
anogenital distances at birth. Pups nursed by ethanol-drinking mothers
had a significantly earlier preputial separation, but there was no effect on
adult masculine sex behaviour, plasma testosterone or weights ofaccessory sex glands.
(4) not assignable(317)

Species:276

20-30% in water; estimated 4000 mg/kg/day.
no data specified
otherno dataas prescribed by 1.1 - 1.4

Rat
FemaleLong-Evansdrinking water
10 or 90 days before gestation and during gestation
9220 mg/kg/day during 10 day pre-exposure; 14500 mg/kg/day in 90 day
preexposure and 11,300 mg/kg/day during gestation
no data specified
otherno dataas prescribed by 1.1 - 1.4
This is an important study in that it confirms that 5% v/v in feed delivers adosage of 12,000 to 14,000 mg/kg.
A decrease in foetal bodyweight was noted in each dosage regimen. There
were no gross abnormalities or skeletal defects noted.
(4) not assignable(316)

Long-Evansdrinking water
GD 7 to delivery
10%

Mouse

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:

Femaleother: not specified
drinking waterfrom 11 weeks before mating and throughout gestation

Result:Reliability
12.11.2004:

Retardation of muscle growth was seen in offspring killed at 12 weeks ofage of inbred mice given 10-20% ethanol in the drinking-water for 11
weeks before mating and 30% ethanol after breeding until delivery.
Prenatally, there was suppression of hyperplasia of muscle fibres during
myogenesis; postnatally, there was suppression of normal hypertrophy ofindividual muscle fibres.
(4) not assignable(318)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleC57BL
drinking water
Before mating, throughout gestation and lactation

Result:Reliability
12.11.2004:

Fetotoxicity and structural teratology:
When female C57Bl/Crgl mice were given 10% ethanol (v/v) in water as thedrinking fluid before mating, throughout gestation and lactation, no
significant effect on pup development or behaviour was seen.
(4) not assignable(303)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Dog
FemaleBeagleGavageMethod:Result:Dogs were administered 1.8 g/kg bw ethanol as a 25% solution by gavage
twice daily and were given either a normal-proteino r low-protein dietthroughout gestation.
Ethanol consumption and low dietary protein intake, independently of each

10-20% in drinking water; Estimated 3000 to 5000 mg/kg/day
no data specified
otherno dataas prescribed by 1.1 - 1.4

as above
10% (v/v)
no data specified
otherno dataas prescribed by 1.1 - 1.4

twice daily
throughout gestation
1800 mg/kg/day
no data specified
otherno dataas prescribed by 1.1 - 1.4

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DATE: 19.11.2004
other, significantly decreased maternal weight gain as well as the weight of
the neonates(4) not assignable(319)

Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Dog

Result:Reliability
12.11.2004:

There were no gross or histological abnormality, a slight decrease in thenumber of offspring per litter and in pup weight, and an increase in thenumber of still births. Blood ethanol concentrations were 1.3-1.75 g/l,(4) not assignable(320)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

MonkeyFemaleMacaca Fascicularis
GavageStarting before day 10 or on day 40 of gestation
once per weekthroughout gestation as above
0.3 - 4.1 g/kg bw
no data specified
other

Result:Reliability
12.11.2004:

Spontaneous abortion frequency increased at peak plasma
ethanol concentrations above 2 g/l. Developmental alterations wereobserved consistently in offspring of monkeys with blood levels greaterthan 1.5 g/l when treatment was initiated at the start of gestation; infants
exposed only after gestation day 40 were less consistently abnormal
despite higher maternal blood ethanol levels (5.5g/l). There weredevelopmental alterations and an increase in spontaneous abortions at
peak plasma ethanol levels above 200 mg/100 ml. Developmental
alterations in offspring were consistent at blood alcohol levels in excess of
150 mg/100 ml.
(4) not assignable(321) (322)Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

278

Gavagethroughout gestation
3 or 3.6g/kg bodyweight

no dataas prescribed by 1.1 - 1.4

Ratmale/female
oral feedfrom 6 weeks before mating
Estimated much greater than 12,000 mg/kg/day
no data specified
otherno dataas prescribed by 1.1 - 1.4

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Method:Result:Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleSprague-Dawleyoral feed
Days 1-21 of gestation

Result:Reliability
12.11.2004:

When Sprague-Dawley rats were given ethanol as 35% of total calories ina liquid diet on gestation days 1-21, offspring had abnormal distribution of
nerve fibres in the temporal regions of the hippocampus, which persisted tomaturity.
(4) not assignable(324)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleWistaroral feed
Days 6-19 of gestation

Result:Reliability
12.11.2004:

Gestation length, litter survival or birth weight. Pup weight was not affectedby treatment from birth to 28 days. Offspring motor activity wassignificantly increased at 16 days of age.
(4) not assignable(325)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
Method

:
:
:
:
:
:
:
:
:
:

Male Sprague-Dawley rats maintained for six weeks on a liquid dietcontaining 10% ethanol were paired with untreated females.
There was body weight loss and central nervous system impairment, andonly half of the treated animals had successful matings, compared to all of
the controls. There was a decrease in litter size and an increase inprenatal mortality among the litters.
(4) not assignable(323)

35% of calorie intake; estimated 12150 mg/kg/dayotherno dataas prescribed by 1.1 - 1.4

5% in diet; average dose 11,200 mg/kg (range 12,000-14,000)
no data specified
otherno dataas prescribed by 1.1 - 1.4

Rat
FemaleLong-Evansoral feed
Days 6-20 gestation
35% of calorie intake, estimated 12,150 mg/kg/day
no data specified
other

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279

OECD SIDS
5. TOXICITYETHANOL
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DATE: 19.11.2004

Year
GLP
Test substance:::

Result:Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleLong-Evansoral feed
Days 6-20 of gestation

Result:Reliability
12.11.2004:

Males showed feminization; females showed masculinization indicating
prenatal hormonal disruption.
(4) not assignable(327)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleWistaroral feed
From day 12 of gestation to 10 days post partum

Result:Reliability
12.11.2004:

Offspring showed decreased ano-genital distance; decreased male
reproductive organ weights 55 and 110 days post partum; lowered sex
hormones (LH and testosterone), sexual motivation and performance;
lowered phenotypic masculinization.
(4) not assignable(328)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
Method
Year
GLP

:
:
:
:
:
:
:
:
:
:
:
:

280

no dataas prescribed by 1.1 - 1.4
Offspring showed lower maximal suckling pressure as well as suckling
pattern changes.
(4) not assignable(326)

35% of calorie intake. Estimated 12,150 mg/kg/day.
no data specified
otherno dataas prescribed by 1.1 - 1.4

Estimated 12,150 mg/kg/day; 36% of calorie intake.
no data specified
otherno dataas prescribed by 1.1 - 1.4

Rat
Femaleoral feed
16 weeks
16 weeks
5% ethanol in liquid feed
no data specified
otherno data

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Test substance:as prescribed by 1.1 - 1.4

Result:Reliability
12.11.2004:

Mating of female Holtzman rats fed a liquid diet containing 5% ethanol for
16 weeks with untreated males resulted in no adverse effect on litter size or
neonatal body weight.
(4) not assignable(285)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleWistaroral feed
Before mating, throughout gestation and lactation

Result:Reliability
12.11.2004:

When female Wistar rats were given 20-25% of the calories consumed as
12% ethanol in a sucrose solution as the drinking fluid before mating andthroughout gestation and lactation, there was no effect on development ofoffspring
There was no effect on development or offspring.
(4) not assignable(329)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleMethod:Remark:Result:To study the severity and degree of in utero alcohol effects in relation to therate of maternal alcohol damage, multiparous 1-year alcohol-fed rats wereused, with an appropriate pair-fed control group. During pregnancy,
alcoholic dams showed relatively high acetaldehyde levels (41 +/- 19
mumol/l) and blood alcohol levels of 22.8 +/- 14 mmol/l.
The stage of maternal alcohol illness, as indicated mainly by the extent ofliver damage, may play an important role in the frequency and severity of inutero alcohol effects in the rat.
Dams showed marked histological alterations in liver as well as high serum
aspartate-aminotransferase, alanine- aminotransferase, alkaline
phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase
activities. The increase in serum enzyme levels did not correlate with anincrease in hepatic enzyme levels since only glutamate dehydrogenase
was enhanced in liver after 1 year of alcohol intake. In addition, except for
an increase in low Km aldehyde dehydrogenase activity, there were nochanges in liver alcohol metabolizing enzymes in chronic alcohol vs. pairfed females.

as above
12% ethanol in sucrose solution (20-25% calorie intake)no data specified
otherno dataas prescribed by 1.1 - 1.4

oral feed
1 year
Daily
Blood alcohol level of 22.8 mmol/lyes, concurrent no treatment
otherno dataas prescribed by 1.1 - 1.4

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DATE: 19.11.2004
Alcoholic rats showed a high incidence of damage in their progeny(resorptions, immature fetuses, decrease in fetal weight, etc.), and rats withthe highest serum levels of theabove enzymes (especially glutamate
dehydrogenase and gamma-glutamyl transferase) had severely affected
progeny. Rats with minimal histological liver damage, in contrast, did notshow resorptions.
(4) not assignable(330)

Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

Absence of sexual dimorphism (saccharin preference and maze learning)
was seen among offspring, suggesting disrupted perinatal androgen status.
(4) not assignable(331)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

Offspring males showed feminized behaviour and females showed
masculinized behaviour, suggesting disruption of the hormonal
environment prenatally.
(4) not assignable

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

There was evidence of behavioural deficits, which persisted until
adulthood. Female offspring showed a variety of deficits in maternal
behaviour when adult, which may have been related to prenatal hormonal
alterations.
(4) not assignable(333)

Species
Sex
Strain:::282

Sprague-Dawleyoral feed
GD 7 to parturition
liquid diet containing ethanol as 35% of calories

Long-Evansoral feed
GD 6-20
Liquid diet containing 35% ethanol derived calories(332)

Long-Evansoral feed
GD 6-20liquid diet containing 35% ethanol derived calories

Mouse
FemaleC3H

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Route of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:

oral feed
Days 0-17 of gestation

Method:Result:Reliability
12.11.2004:

Groups of C3H mice were given a liquid diet or a fortified liquid diet, eacheither alone or with 4.1% w/v ethanol, from days 0-17 of pregnancy; afurther group was given an amount of liquid diet equal to that consumed by
the group given liquid diet plus ethanol.
Ethanol consumption inhibited foetal growth and development but did notaffect litter size irrespective of the diet used.
(4) not assignable(334)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Mouse
Femaleother: RAP
i.v.

Method:Result:Reliability
12.11.2004:

Preimplantation effects in a study on the effects of preimplantation
exposure: mice were given ethanol intravenously on days 3 and 4 ofpregnancy and offspring were examined on day 19 of pregnancy.
On day 19 of gestation the mean foetal and placental weightswere
significantly lowered but there was no effect on skeletal development.
(4) not assignable

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

MonkeyFemaleother: rhesus and cynomolgus
i.v.
1-2 minutes

Result:Reliability
12.11.2004:

In monkeys given 3g/kg bw ethanol intravenously over 1-2 min on gestation
days 120-147, transient but marked collapse of umbilical vasculature wasobserved within 15 min. This resulted in severe hypoxia and acidosis in
the fetus, but recovery occurred during the succeeding hour.
(4) not assignable(336)

Species
Sex::

Days 0-17 gestation
4.1% w/v in liquid dietno data specified
otherno dataas prescribed by 1.1 - 1.4

once on day 3 or 4 of gestation
not specified
no data specified
otherno dataas prescribed by 1.1 - 1.4(335)

Gestation days 120-147
3g/kg bodyweight

Rat

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5. TOXICITYETHANOL
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DATE: 19.11.2004

StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:

Long-Evansoral unspecified
GD 6 – 20

Result:Reliability
12.11.2004:

Offspring exerted a lower maximal suckling pressure,spent less timesuckling during test sessions and displayed an altered suckling pattern.
(4) not assignable(337)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleLong-Evansoral unspecified
throughout gestation

Result:Reliability
12.11.2004:

Litter size, litter weight and mean pup weight were lowered but there were
no gross malformations or behavioural teratogenic effects.
(4) not assignable(338)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
FemaleLong-Evansoral unspecified
Days 5-19 of gestation

Result:Reliability
12.11.2004:

Long-Evans rats administered 6 g/kg bw ethanol orally on gestation days 519 had blood ethanol concentrations of over 2.6 g/l. Fetuses had
decreased body weight, increased body water and sodium content anddecreased lipid-free solid content.
(4) not assignable(339)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses

:
:
:
:
:
:
:
:

284

liquid diets containing ethanol (35% of total calories)

throughout gestation
1 or 2 g/kg bodyweight given daily
no data specified
otherno dataas prescribed by 1.1 - 1.4

as above
6000 mg/kg achieving blood alcohol level of 260 mg/100 ml
no data specified
otherno dataas prescribed by 1.1 - 1.4

Rat
FemaleSprague-Dawleyoral unspecified
Days 1-21 of gestation
as above
36% of calorie intake achieving blood alcohol levels of 150-200 mg/100 ml.

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Control group
MethodYear
GLP
Test substance

:
:
:
:
:

no data specified
other

Result:Reliability
12.11.2004:

Sprague-Dawley rats were provided with 18, 25 and 32% protein-derived
calories and 36% ethanol-derived calories in a liquid diet on gestation days
1-21. The maternal ethanol blood levels were 1.5-2 g/l. Ethanol caused asignificant decrease in fetal body weight and brain weight
but an increase in relative brain weight, irrespective of the protein content
of the diet.
(4) not assignable(340)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

The sexually dimorphic nucleus in the preoptic area of the brain of adultmale offspring was significantly decreased in volume.
(4) not assignable(341)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

An increased incidence of resorptions and marginal effect on fetal bodyweight but no teratogenic effect were observed.
(4) not assignable(342)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability:

In contrast to studies in which gross malformations were not observed,
polydactyly and polysyndactyly were reported in the offspring of rats given
5 g/kg bw (but not in those given 6 g/kg bw) per day ethanol. Maximal
blood ethanol concentrations of 2.5-3.25 g/l were reported with the two
doses(4) not assignable

no dataas prescribed by 1.1 - 1.4

Sprague-Dawleyoral unspecified
GD 16 until postnatal day 14 or from birth until postnatal day 14
36% of total calories in a liquid dietSprague-Dawleyoral unspecified
GD 7-15
twice daily
3 day period
4g/kg

Sprague-Dawleyoral unspecified
Daily
GD 1-15 or GD 1-20
5 or 5g/kg bodyweight

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285

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

12.11.2004(343)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

ResultReliability
12.11.2004::

Treatment shortened the umbilical cord.
(4) not assignable

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Remark:ResultReliability
12.11.2004::

The most frequently reported behavioural teratogenic effect is alteration in
motor activity.
Increased motor activity of offspring was reported.
(4) not assignable(345)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Mouse
FemaleC3H
oral unspecified

Remark:Result:Reliability
12.11.2004:

The authors proposed that the effect was due to a specific action on thefertilized ovum at the time of second meiotic division, causing aneuploidy,
but the numbers of embryos available for examination in this study were
inadequate to confirm this hypothesis
Treatment of (C3H x C57Bl)F1 female mice with a single dose of 1 ml of
12.5% ethanol by gavage 2 h after a 30-min mating period produced anincrease in late (after day 11) fetal deaths. The same treatment given 1 hr
after mating did not produce this effect.
(4) not assignable(346)

Species
Sex
StrainRoute of admin.

:
:
:
:

286

Long-Evansoral unspecified

Prenatal treatment with 35% ethanol-derived calories in a liquid diet.

(344)

Long-Evansoral unspecified
GD 6-20liquid diet containing ethanol as 35% of total calories

1 x 1 ml of 12.5% alcohol; Estimate 2500 mg/kg.
otherno dataas prescribed by 1.1 - 1.4

Mouse
Male
C3H
oral unspecified

UNEP PUBLICATIONSOECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:

4 weeks before mating
Daily

Method:Result:Reliability
12.11.2004:

Male-mediated developmental effects Male C3H mice were fed ethanol (20or 30% of total calories) in a liquid diet and, after four weeks of treatment,
were mated to untreated females.
The resulting litters showed nochange in the number of implants, prenatal
mortality, fetal weight, sex ratio or soft-tissue malformations.
(4) not assignable(347)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

MonkeyResult:Reliability
12.11.2004:

An increase in pregnancy wastage (abortions and still births) was observed
but no structural malformation or facial change.
(4) not assignable(348)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

other: ferret

Result:Reliability
12.11.2004:

There was a significant increase in the number of fetuses and litters withmalformations but no effect on fetal weight or resorptions. The peak blood
ethanol concentration was 2 g/l.
(4) not assignable(349)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Method:Administration of ethanol in combination with an unspecified extract ofmarijuana containing 9-tetrahydrocanabinol.

20-30% of total calorie intake
no data specified
otherno dataas prescribed by 1.1 - 1.4

other: Cynomolgus
oral unspecified
GD 20-150
Daily
5g/kg bw

oral unspecified
GD 15-35
90 days prior to mating and during gestation or during gestation only
1.5g/kg bodyweight as a 25% solution

Long-Evansother: intragastric
GD 7-15

UNEP PUBLICATIONS

287

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Mouse

Method:Result:Reliability
12.11.2004:

Administration of ethanol in combination with an unspecified extract ofmarijuana containing 9-tetrahydrocanabinol.
Treatment produced a significant decrease in maternal weight gain and anincreased incidence of resorptions. The incidence of resorptions wasincreased with marijuana alone, but the increase was more than additive
with the combination of marijuana and ethanol.
(4) not assignable(350)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

In hooded rats given a liquid diet containing 37% ethanol-derived calories
from day 6 of gestation to time of birth (gestation day 23 for ethanolexposed rats; day 22 forcontrols), delayed and extended period of corticalneuron generation, reduced number of neurons and altered distribution ofneurons were seen.
(4) not assignable(351)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Result:Reliability
12.11.2004:

Ethanol in combination with lithium carbonate had a synergistic effect on
the induction of fetal abnormalities.
(4) not assignable(352)

Species
Sex

:
:

288

Treatment produced a significant decrease in maternal weight gain and anincreased incidence of resorptions. The incidence of resorptions wasincreased with marijuana alone, but the increase was more than additive
with the combination of marijuana and ethanol.
(4) not assignable(350)

Swiss Websters.c.
GD 1-15other: hooded

other: albino

Rat
Female

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StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:

Long-Evans

ResultReliability
12.11.2004::

An increase in congenital malformations was noted.
(4) not assignable

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Rat
male/female

Method:Result:Reliability
12.11.2004:

Preimplantation effects were studied by the examination of uterine contentsof albino rats following consumption of plum brandy (reported as 24%
ethanol) or cognac (reported as20% ethanol) for 40-45 days before mating
and during pregnancy until the rats were killed on day 5.
Development was retarded, and there was an increased number ofpathological morulae and blastocysts.
(4) not assignable(354)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat
FemaleLong-Evans

Result:Reliability
12.11.2004:

In Long-Evans rats given 4 ml/kg bw ethanol as a single oral dose between
days 6 and 15 of gestation, a variety of gross malformations was reported
in 72-100% offspring compared to 12% of controls.
(4) not assignable(355)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.

:
:
:
:
:
:

60 days
20 ml of 20% alcohol; estimated 4000 mg/kg/dayotherno dataas prescribed by 1.1 - 1.4(353)

40-45 days before mating and 5 days post fertilization
20% or 24% ethanol crudely calculated to 1100 mg/kg/day.
no data specified
otherno dataas prescribed by 1.1 - 1.4

Days 6-15 of gestation
4000 mg/kg

Rat
Sprague-Dawley

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Duration of test
Doses
Control group:::

Method:Remark:Result:Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

Remark:Result:Reliability
12.11.2004:

The most frequently reported behavioural teratogenic effect is alteration in
motor activity.
There was decreased litter weight but not litter size at birth and increased
postnatal mortality. Motor activity of neonates raised by surrogate mothers
was impaired at 16 and 20 days of age.
(4) not assignable(357)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Rat

RemarkReliability
12.11.2004::

Behavioural teratogenic effects reported.
(4) not assignable

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Mouse

Remark:Some studies in several mouse strains have shown no teratogenic effect,290

15 or 25% ethanol-derived calories

Animals were given 15 or 25% ethanol-derived calories in liquid diets 20
days before mating, throughout mating and until gestation day 19;
additional groups were pair-fed an isocaloric diet.
The effects of 15% ethanol-derived calories were attributed to ethanol,
while the effects of 25% ethanol-derived calories were attributed partly todecreased caloric intake.
There was decreased caloric intake in the group given 25% ethanolderived calories and in the pair-fed controls, and in both of these groups
there were associated decreases in fetal body weight, organ weights andDNA and protein contents compared to the pair-fed controls of the group
given 15% ethanol-derived calories.
(4) not assignable(356)

Long-Evansthroughout gestation
Daily
4 or 6g/kg bw(358) (359)other: CF-1, CD-1, C3H

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Reliability
12.11.2004:

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Remark:Reliability
12.11.2004

even at dose levels providing blood ethanol concentrations of 2 g/l or
higher. Mice given ethanol orally or in the drinking fluid had pups withminor skeletal variants or decreased fetal body weight, but there was noincrease in resorptions or malformations.
(4) not assignable(360) (361) (311)
Mouse

Studies in mice showed teratogenic effects and resorptions, typically atblood ethanol concentrations in excess of 2 g/l. The effects, such as fetal
resorptions, intrauterine growth retardation, cleft palate, alteredcraniofacial development and exencephaly, limb defects and heart defects,
varied with the strain of mice, mode of administration and stage ofgestation at which ethanol was administered.
: (4) not assignable(362) (363) (364) (365) (307) (366) (367) (368) (369) (370) (371) (372) (373)(374) (375) (376) (377) (378)Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Mouse

Result:Reliability
12.11.2004:

Combined administration of ethanol and metronidazole increased thenumber of resporptions, decreased fetal body weight and had a marginal
effect on the incidence of malformations.
(4) not assignable(379)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:
:
:
:

Mouse

Result:Reliability
02.07.2004:

Ethanol increased the incidence of cleft palate in mice administered
methylmercuric chloride and retinyl acetate.
(4) not assignable

Species
Sex
Strain:::Swiss Webster

Swiss Webster

(380)
Mouse

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Route of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group

:
:
:
:
:
:

RemarkReliability
12.11.2004::

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Miniature swine
FemaleResult:Reliability
12.11.2004:

In miniature swine given 20% ethanol in drinking-water (>3 g/kg bw/day) as
gilts (18 months old) or sows (three years old), there was a significant
decrease in mean litter size and in the birth weight of piglets and asignificant increase in the incidence of multiple malformations. There wasa slight decrease in the number of offspring per litter; there was also aslight decrease in still births. Open-field activity was significantly increased
in offspring.
(4) not assignable(383)

Species
Sex
StrainRoute of admin.
Exposure period
Frequency of treatm.
Duration of test
Doses
Control group
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

MonkeyFemaleResult:Reliability
12.11.2004:

There was an increase in abortions and stillbirths but no structural or
skeletal (facial) deformities.
(4) not assignable

Remark:Reliability:

292

Behavioural teratogenic effects reported.
(4) not assignable(381) (382)

Throughout gestation
Throughout gestation
3000 and 3600 mg/kg/day
no data specified
otherno dataas prescribed by 1.1 - 1.4

Days 20-150 of gestation
5000 mg/kg/day
no data specified
otherno dataas prescribed by 1.1 - 1.4(384)

Ethanol at high blood levels has caused significant reductions in foetal
body weight, increased resorptions and teratogenic effects in a number of
species. Some, though not all, studies in mice and rats have demonstrated
altered behavioural development following exposure to ethanol in utero.
Exposure in utero or during lactation reduced postnatal growth.
(4) not assignable

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DATE: 19.11.2004

06.07.2004(18)

Remark:Reliability
12.11.2004:

Remark:Result:Reliability
12.11.2004:

Result:Reliability
12.11.2004:

Result:Reliability
12.11.2004:

Remark:Reliability
12.11.2004:

Remark:Reliability
12.11.2004:

When 20% ethanol in water (v/v) was given as the drinking fluid for 60 days
to male Long-Evans rats, which were mated with untreated females one tothree weeks after cessation of treatment, the incidence of congenital
malformations in the offspring was increased(4) not assignable(385)

In a study to evaluate the role of zinc deficiency in the developmental
toxicity of ethanol, CBA/J mice were given a liquid diet, either fortified withzinc or deficient in zinc, and ethanol (15 or 20% of total calories).
Zinc deficiency potentiated the ethanol-induced increase in resorptions andexternal malformations and the decrease infetal weight.
(4) not assignable(386)

Among offspring of Long-Evans rats fed liquid diets containing 35%
ethanol-derived calories during gestation days 6-20, there was evidence of
behavioural deficits, which persisted until adulthood. Female offspring
showed a variety of deficits in maternal behaviour when adult, which may
have been related to prenatal hormonal alterations(4) not assignable(387)

There was an increase in the incidence of both external and internalmalformations in C57Bl/6 mice given a marginally zinc- deficient diet andethanol during gestation, in comparison with mice given a control diet andwith mice treated with ethanol alone.
(4) not assignable(388)

When ethanol is given in combination with other chemicals which tend toincrease the blood level of ethanol by reducing its metabolism, e.g. 4methylpyrazole and pyrazole, the teratogenic and fetotoxic effects are
increased.
(4) not assignable(389) (390)

Administration of ethanol with chemicals that tend to increase theacetaldehyde level, e.g. disulfiram does not increase the teratogenicity of
ethanol.
(4) not assignable
(391)

5.8.3

TOXICITY TO REPRODUCTION, OTHER STUDIES

5.9

SPECIFIC INVESTIGATIONS

Endpoint:

Neurotoxicity

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Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
VehicleExposure period
Frequency of treatm.
Doses
Control group
Observation period
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

5.8.2 Developmental Toxicity/Teratogenicity

Method:Breeding and gestation:
Proven 3 month old female breeders (~180g) mated (males ~200g) for 4
days. 12 hour dark/light cycle. Temperature 20-25C, humidity 69-70%.
SLH granules chow and water ad libitum. Vaginal sperm-positive plug
used as evidence of copulation and defined as day 1. Body weightmeasured periodically. Ethanol (12.5% in distilled water) administered byperoral intubation from day 1 onwards. Control: equicaloric sucrose.

other: learning and memory following prenatal exposureRatmale/female
WistarGavageWater
1 g/kg/day
yes, concurrent vehicle
9 weeks
poor learners
Other
1998
no dataas prescribed by 1.1 - 1.4

Cross-fostering:
Litters culled to 11 pups and cross-fostered to develop under approximately
identical conditions. Treatment continued during lactation until weaning.
Five treatment groups resulted:(Pregnant treatment/Lactation treatment)
Control/Control
Control/Ethanol(1)Ethanol/Control
Ethanol/Ethanol(1)Control/Control(2)Notes: (1) lactating dams treated with ethanol but were only feed sucrose
pre-natally. (2) Control dams used during lactation period had been fed
ethanol pre-natally.
Blood alcohol determination:
Intracardiac puncture from 6 dams on GD14 and PND14, either under lightether anasthesia 1 hour after ethanol administration. Analysis by GC.
Postnatal monitoring:
After weaning (PND23) litters segregated by gender, 10 pups per cage.
Survival monitored daily. Weights checked days 3, 10, 20, 30, 45 and at
beginning of behavioural studies (63 days).
Behavioural studies:
Two way active avoidance (Shuttle box) test used to assess
pharmacological effects on learning and memory. Conditioned stimulus:
buzzer and light. Unconditioned stimulus: electric shock (0.2mA) on grid
floor in half of box.
Training for 5 days. Memory tested over 12 days.
Positive avoidance recorded if animal avoided unconditional stimulus within
4 secs of conditioned stimulus.
All data assessed using one way ANOVA followed by group individual post-

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hoc comparisons using Scheffe test.
No effects on dam bodyweight.
Pup body weight showed significant effect following ethanol treatment but
growth recovered to control values within 9 weeks.
No visible malformations in offspring.
Blood ethanol levels in treated dams 350 +/- 57.8 mg/l
No dead pups after PND 28 in any group.
Significant increase in mortality in offspring exposed to ethanol pre-natally.
No effect with postnatal exposure. Highest mortality in group 5.
Ethanol treatment did impair learning. In worst case group 4, 60% werepoor learners compared to 33% of control group. Effects in females
disappeared by 5 months but remained in males.
(2) valid with restrictions(392)

Result:Reliability
12.11.2004:

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Neurotoxicity

Remark:Reliability
12.11.2004:

Ethanol can evoke a reduction in anxiety and a feeling ofpleasure as well as sedation, motor incoordination,
aggression, changes in other forms of social interaction andaberrations in cognition. Certain neurotransmitter systems
are more or less sensitive to ethanol with both glutamate
and GABA systems targets for ethanol's activity.
Particularly sensitive components are certain receptor-gatedion channels including GABA(A) and the N-methyl-D-aspartate
subtype of the glutamate receptor, the nicotinic cholinergic
receptor and the serotonin-3 receptor. Ethanol also elicits
an increase of dopamine in the nucleus accumbens. Some ofthese systems alter their function (adapt) during periods ofchronic drinking and such maladaptation may generate
manifestations of tolerance, physical dependence and cravingon cessation of intake. Maladaptations in systems that gatecalcium ion entry into neurons can contribute to brain
damage in some alcoholics.
See Section 1.13, entry 11.
(2) valid with restrictions

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals

:
:
:
:
:
:
:
:
:

Neurotoxicity

Remark:Drinking which results in a blood ethanol concentrations of 50-60mg/l toother: Literature review
Human

1999

(393)

other: literature review

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ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

12.11.2004

900mg/l (20mM) defined as 'moderate'. Such consumption selectively
affects the function of the GABA, glutamatergic, serotonergic,
dopaminergic, cholinergic, and opioid neuronal systems. Behaviouralconsequences are dose and time related and can even change on therising and falling phases of the blood ethanol curve. A number of studies
have noted a measurable diminution in neuropsychologic parameters inhabitual consumers of moderate amounts of ethanol, but others have notfound such changes. Some have even noted positive effects on congnitiveeffects in aging humans. Consumption by pregnant women can havesignificant consequences on the developing fetal nervous system.
(2) valid with restrictions
A comprehensive review document covering typical dose from drink,
pharmacokinetics, implications for nervous system, CNS effects, anxiolytic
effects, cognitive effects and developmental effects. Contains 440
references.
(394)

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
VehicleExposure period
Frequency of treatm.
Doses
Control group
Observation period
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Behavioural Effects

Method:Animals: initial body weight 180-210g, housed individually, stainless steel
cages.
Inhalation chamber: dynamic inhalation behavioural chamber ex Pradhan &Copeland. Glass chamber over a grid floor fitted with a metal plateaccomodating a lever and liquid dipper. Individual animals exposed tomultiple concentrations
Lighting: 12hr light/12hr dark
Conditions: temperature 21±1C, Humidity 55±5%Analytical: 15 minute sampling of chamber atmosphere. Samples analysedby gas chromatography.

Rat
Male
Fischer 344
inhalation

see method for details

Other
1991
no dataother TS

Behavioural schedules: Fixed ratio (FR) liquid re-inforcement and self
stimulation (SS) behaviour, with 3-4 week training period. Animals thenacclimatised to exposure chambers for 3-4 days and stability of response
times (<10%) checked before proceeding with exposure.
FR behaviour: based on rats being kept at 80% of starting weight withadditional 5% sucrose solution becoming available following pressing oflever 24 or 50x. Re-inforcement only available 5x per session. Exposure
regimes: 2 hour exposures to 140, 161, 202 and 398ppm ethanol; five hour
exposure to 140ppm; 2 hr exposure to 206ppm daily for 5 days. Control:
animal response on previous day in air. Statistical analysis: ANOVA, withsignificance at p<0.05

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SS behaviour based on electrical stimulation becoming available via
electrodes implanted in posterior hypothalamus following pressing of lever.
Exposure regime: Exposure regimes: 2 hour exposures to 129, 373, 603
and 1287ppm ethanol. Control: animal response on previous day in air.
Statistical analysis: Rate of response broken down into 20 minute periods
and consecutive periods compared with comparable control period using
ANOVA, with significance at p<0.05. For results with significant F, further
analysis using Least Signficant difference, Duncan's test and Newman
Kuels test.

Result:Blood collected from tail vein afer 2 hrs exposure in SS regimes for blood
ethanol level measurements (alcohol dehydrogenase assay).
FR behaviour: reinforcement behaviour dropped by a small but statisticallysignificantly in the 15minute exposure periods from 45mins onward and for
exposures of 202ppm and above. There was no cumulative effect(exposure for 5 hours did not produce any effects not seen in the 2hr
exposure.) In the daily repeat exposure, effects declined showing adeveloping tolerance to ethanol.
SS: there was a decline in self stimulation behaviour at exposures of600ppm and above but these were not statistically significant.

Test substance
Reliability::

Blood ethanol concentrations following 2 hr exposure: 393ug/ml after
600ppm, 545ug/ml after 1200ppm
100% anydrous ethanol from US Industrials Co.
(2) valid with restrictions
Results only presented as small graphs. No indication of total animal
numbers used.

19.11.2004(395)

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals

:
:
:
:
:
:
:
:
:

Neurotoxicity
5.8.2 Developmental Toxicity/Teratogenicity

Result:Reliability
19.11.2004:

Cognitive impairment score increased as craniofacial linear measurements
increased and craniofacial angular measurements decreased, especially inanimals exposed to ethanol on gestational days 19 or 20. Incidence andmagnitude of cognitive impairment increased with age.
(4) not assignable(396)

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
VehicleExposure period
Frequency of treatm.
Doses

:
:
:
:
:
:
:
:
:
:
:
:
:

MonkeyMacaca Fascicularis
in utero18

Neurotoxicity

Rat
FemaleSprague-Dawley
Gavage4 day(s)Daily
2, 4 and 6 g

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Control group
Observation period
Result:::

MethodYear
GLP
Test substance

:
:
:
:

Latency to move, rear and groom + more avoidance response and morecorrect discriminations in Y maze
Other
1979
no datano data

Reliability
12.11.2004:(4) not assignable

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
VehicleExposure period
Frequency of treatm.
Doses
Control group
Observation period
Result
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:
:

Neurotoxicity

Result:Reliability
19.11.2004:

Chronic exposure of rats through gestation, in particular through the last 2
weeks of gestation, had little influence on the postnatal expression of cell
type specific genes involved in brain development.
(4) not assignable(398)

Endpoint
Study descr. in chapter
Reference
Type
Species
Sex
StrainRoute of admin.
No. of animals
MethodYear
GLP
Test substance

:
:
:
:
:
:
:
:
:
:
:
:
:

Immunotoxicity

Result:High intakes of alcohol in humans may alter the production and turnoverrates of lymphocytes in the thymus or spleen, or both, with a resultant shift
in the relative concentrations of the lymphocyte subpopulations, which
include the B cells and T cells. Circulating T lymphocyte counts are
significantly reduced in alcoholics, as is the ability of these cells to undergo
blastic transformation on mitogenic stimulation. Non-specific activation of B
lymphocytes occurs in all patients who regularly drink alcohol in excess. In
vitro, alcohol alters both the development and the sensitivity of

298

(397)

other: postnatal expression of celltype specific genes.
Rat
FemaleSprague-Dawley
inhalation
16
no data
13 day(s)
Daily

Other
1993
no dataas prescribed by 1.1 - 1.4

Human

1989
other TS

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY

Reliability
12.11.2004
5.10

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

lymphocytes. Prolonged drinking affects the reticuloendothelial system of
the liver by interfering with the mobilization and activation of macrophages
and their phagocytic activity. In vitro, alcohol impairs the chemotaxis andadherence of granulocytes to capillary walls. Alcohol inhibits cell mediatedimmunitiy. It suppresses natural killer cell activity and antibody directed
cytotoxicity.
(4) not assignable(399)

EXPOSURE EXPERIENCE

Type of experience

:

Human

Remark:Reliability
29.06.2004:

Local effects: As a solvent ethanol can produce dermatitis because of its
defatting action. Whilst the majority of contact dermatitis is irritant in nature,both contact urticaria and skin sensitisation have been reported (Adams
RM, 1990; Cronin E 1980 ; Fisher AA, 1983)(4) not assignable(400) (401) (402)Type of experience

:

Human

Remark:Effects of chronic ingestion of alcohol
The study of the effects of long term ethanol ingestion has potentially been
confused by potential confounding factors such as smoking and in somecases exposure to toxic alcohol substitutes or contaminants such as
methanol.
A wide variety of conditions have been taken to be systemic effects ofethanolism and these include : Nutritional Deficiencies such as of vitamins(B1,B6 and B12)and trace elements such as zinc.
Nervous system disorders such as reduced fine motor skills, impaired
cognition, peripheral neuropathies and Wernicke- Korsakoff syndrome.
The latter is linked to thiamine deficiency which perturbs normal nerve
function.
Gastrointestinal disorders including gastritis and stomach ulceration andatrophy; gut malabsorption and motility problems, pancreatitis, fatty liver,
hepatitis, and cirrhosis which may end in hepatic failure withencephalopathy.
Endocrine Disorders. Decreased secretion of testosterone and oxytocin
and increased secretion of cortisol, aldosterone and insulin have been
associated with chronic
ethanolism.
Haematological Disorders. Bone marrow supression, thrombocytopaenia
and signs of abnormal iron metabolism may be observed.
Cardiological Disorders may include cardiomyopathy and in beer drinkers
alcoholic beri-beri. The prevalance of hypertension and cerebrovascular
accidents is higher in heavy drinkers. Arrhythmias can be induced byacute ethanol ingestion.
Fertility. Ethanolism can be associated with impotence and hypogonadism
in males and infertility in women.

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Foetal Alcohol Syndrome: Foetal alcohol syndrome was described byJones and Smith (1973) and named after the presumed cause of a rangeof anomalies found in the offspring of severely and chronically alcoholic
women. The minimum diagnostic criteria have been described by the
Research Society of Alcoholism (Rossett HL (1980) as being signs in eachof the following categories :1) pre-natal and/or post-natal growth retardation (weight or length below
the 10th percentile).
2) central nervous system involvement (signs of neurological abnormality
or developmental delay).
3) characteristic facial dysmorphology with at least two of the following
signsa) microcephaly (head circumference below the 3rd percentile)b) short palpebral fissuresc) poorly developed philtrum, thin upper lip, and/orflattening of themaxillary area.
Criteria for reporting infants with less extensive dysmorphology suggested
to be associated with lower levels of ethanol drinking have been outlined by
Hanson et al (1978).
Other abnormalities associated with pre-natal ethanol exposure have beencleft palate, joint and palmar crease abnormalities and septal and othercardiac defects. Dental malocclusions/malalignments and eustachian tube
defects are probably secondary to facial hypoplasia. Investigations strongly
suggest that maternal ingestion of large doses of ethanol (40 g daily) is
teratogenic and foetotoxic.
Studies of the outcome of pregnancy in women who drank heavily during
pregnancy indicate infants were small for dates and had growth retardation.
A significant decrease in birth weight has been associated with
consumption of at least one to two alcoholic drinks per day, although Millsand Graubard (1987) did not find an increased defect rate in offspring of
those drinking one to two drinks per day. It appears uncertain if stillbirth isincreased amongst heavy drinkers but there is a suggestion that the 2-4%of heaviest drinkers in some studies showed a higher risk of spontaneousabortion.
Some studies have indicated prenatal alcohol results in mental retardation
but it is unclear to what degree this might be due to pre-natal alcohol and towhat extent this may be attributable to the post-natal environment in whichchildren are raised. It is acknowledged that study of the effects of ethanol
drinking on pregnancy outcome is complicated by many potentiallyconfounding factors. (Roman E 1988)(4) not assignable(403) (404) (405) (406) (407)

Reliability
29.06.2004:

Type of experience

:

Human

Remark:Acute toxicity: Dermal exposure. Ethanol has a very low octanol: waterpartition coefficient and this is seen as contributing to the poor dermal
uptake of ethanol in intact human skin. Bowers et al (1942) demonstrated
the lack of dermal absorption in humans by occluding dressings soaked in
95% ethanol over the legs of subjects with impermeable rubber sheet.
Blood ethanol determinations made every three hours over the succeeding
12 hours showed no indication of ethanol absorption through the skin.

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On this basis the liklihood of intoxication through intact skin seems very
remote. The volatility of ethanol would suggest that inhalation exposure
would be a more relevant route of exposure.
(4) not assignable(408) (409)

Reliability
29.06.2004:

Type of experience

:

Human

Method:Remark:Ethanol liquid was vaporised by passing at a known rate through a
Rotameter into a tube heated by flame. Air was also passed through thetube at 60l/min and the ethanol laden air cooled before passing into atransparent plastic hood (33cm dia, 25cm high fitted with a cloth skirt) used
to place over the head of the volunteer. The hood was oriented so that theoutlet of the ethanol laden air was close the the mouth/nose of thevolunteer. The ethanol concentration was analysed frequently using amethod described elsewhere (Lester, unpublished.)
Acute toxicity: Inhalation: Lester and Greenberg investigated the effects ofethanol vapour in human volunteers.
At 10-20 mg/l [~5300-10,000ppm] the vapour gave transient coughing andirritation to eyes and nose lasting for 5-10 minutes. Further exposure to
this concentration gave some discomfort but could be tolerated.
At 30mg/l [~16,000ppm] ethanol vapour could be tolerated but gave
continuous lachrymation and marked coughing.
Exposure to 40mg/l [~20,300ppm] was just tolerable but volunteers
declined to be exposed for more than short periods.
Even short excursions into an atmosphere greater than 40mg/l [~20,300]
were judged impossible as this concentration was intolerable to thevolunteers.
When the ventilation rate was 30l/minute [equivalent of hardwork] anethanol concentration of 20mg/l [~10,000ppm] was just tolerable.
In more extended exposures of up to 6 hours to ethanol concentrations of
13-17mg/l [7,000-9,000 ppm],blood ethanol concentrations of 1546mg/100mls were observed. Blood ethanol concentrations were
proportional to both the concentration in air and to the respiratory rate.
After transient eye and nasal irritation had subsided, there were noreported adverse effects such as weakness, tiredness orintraocular tension.

Result:Reliability:

29.06.2004

Measured and nominal concentrations of ethanol in the air were found tobe almost identical.
Lester and Greenberg's conclusion that intolerance of the local irritation
effects would deter exposure to concentrations that would give rise tosystemic intoxication and this premis has been used as a basis for setting
workplace exposure limits.
(2) valid with restrictions
Whilst old, this study appears to be reliable.
(410)

Type of experience:

Direct observation, clinical cases

Remark:Acute toxicity: Ingestion: The widespread use and abuse of alcoholic
beverages has resulted in a large body of data about both the acute andchronic effects of ethanol ingestion. When ingested ethanol is a central
nervous system depressant. The degree of effect depends on the amount

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ingested and the susceptibility of the individual. Susceptability is dependent
upon factors such as genetic background, prior alcohol use, rate ofelevation of blood ethanol concentration, coexposure to other xenobiotics
eg pharmaceuticals, trauma and the general state of health and nutrition.
Ethanol can induce all stages of anaesthesia. The mechanism of action, aswith other anaesthetic agents appears to be interference with ion transport(sodium flux) at the cell membrane. Low concentrations of ethanol affectthe frontallobes of the brain resulting in mood changes, alterations inthinking and apparent suppression of inhibitory neurons. The latter appears
linked to the excitatory phase of intoxication.
The onset of the effects of ethanol varies, in part depending on whether theindividual is habituated to ethanol exposure.
In non-habituated individuals, reduced cognitive ability, motor coordinationand sensory perception may be seen at blood ethanol concentrations as
low as 50mg/100mls.
Attention, motor coordination and reaction time are significantly affected atblood ethanol concentrations of 135mg/100mls. Slurred speech, ataxia,
and drowsiness develop as the concentration increases. These may be
associated with a flushed face, dilated pupils, abnormal sweating andgastrointestinal disturbances.
Ethanol can give cardiac arrythmias and result in myocardial depression.
Thermoregulation can be inhibited during ethanol induced coma and theresultant hypothermia can contribute to death. Rarely some individuals
react to acute ethanol intoxication with severe hypoglycaemia which may
follow 12-16 hours after the main ingestion. This appears to be due toinhibition of hepatic gluconeogenesis and may lead to hypoglycaemic
coma. In children, particularly those aged under 5 years, ingestion ofrelatively small amounts of ethanol has resulted in hypoglycaemic
convulsions and death.
Blood concentrations of 300-500mg/100mls are generally associated withstupor or coma. Death from respiratory depression occurs at blood ethanol
levels of 500mg/100mls and above. The fatal dose has been suggested tobe 5-8g/kg body weight for adults and about 3g/kg body weight for children.
(4) not assignable(411) (412) (413)

Reliability
29.06.2004:

Type of experience:

Human - Exposure through Food

Remark:Metabolism
The metabolism of ethanol has been reviewed (IARC, 1988). Ethanol iseliminated from the body mainly by metabolism in the liver and only
minimally by urinary excretion and pulmonary exhalation. Other tissues
such as kidney, stomachand intestines oxidize ethanol to a small extent.
The hepatic metabolism of ethanol proceeds in three basic steps. First,
ethanol is oxidized within the cytosol of hepatocytes to acetaldehyde;
second, acetaldehyde is converted to acetate, mainly in the mitochondria;
and third,acetate produced in the liver is released into the blood andis
oxidized by peripheral tissues to carbon dioxide, fatty acids and water.
The main pathway for ethanol metabolism proceeds via alcohol
dehydrogenase. However other pathways for ethanol oxidation have been
described including a microsomal ethanol-oxidizing system located in theendoplasmic reticulum and a catalase system located in the peroxisomes.
The rate of hepatic metabolism of ethanol is concentration independent
except at very low or very high concentrations. Blood ethanol in humans
decreases more rapidly at concentrations over 300 mg/dl than at

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concentrations below this level, possibly due to oxidation by the
microsomal ethanol oxidizing system. The maximum rate of metabolism is
100 - 125 mg/kg body weight/hour, although tolerant individuals may havehigher metabolic rates (up to 175 mg/kg/hour) due to enzyme induction.
Adults metabolize 7 - 10 g ethanol/hour reducing blood ethanol
concentrations at a rate of 15 - 20 mg/100 ml/hour. Ethanol is metabolized
more rapidly in chronic alcohol abusers (up to 40 mg/100 ml/hour)and inchildren (up to 28 mg/100 ml/hour) (Ellenhorn & Barceloux, 1988).
(4) not assignable(414)

Reliability
01.10.2003:

Type of experience

:

Human

Method:Remark:Result:Reliability
16.01.2004:

Skin absorption: In order to assess the potential systemic dose, threeparameters were measured: the evaporation of [14C]ethanol from the skinsurface, the in vitro penetration of [14C]ethanol through excised pig skinand the ethanol concentration in the blood of human volunteers following
simulated use of an alcohol based deodorant spray. 20ul labelled ethanol
was dosed on to each substrate to give a dose of 2.5ul/cm2. Substrates
and residual ethanol were taken at intervals of 10s (up to 90s) and placedin scintillation vials containing 5ml StarcScint and counted on a Beckman
LS6000 liquid scintillation counter. In the case of pig skin, the residual skinradioactivity was also assessed (counted for 1 hr). The skin was thenremoved, solubilised using 5ml Soluene 350 and counted again (after
addition of 10ml HionicFluor) to measure activity within the skin. The
original vials now without skin, were also recounted. Statistical analysis
was performed using JMP v3.1 software.
These studies provide evidence that a systemic dose of ethanol is likely to
be very low after the use of formulations delivering ethanol to the skin.
The rate of evaporation from Benchkote and whole pig skin was similar(t1/2 = 13.6s and 11.7s respectively) whilst that from glass was longer (t1/2= 24.8s). Ethanol penetration through pig skin in vitro was greater in
occluded cells than in non-occluded cells (2.19mg/cm2 and 0.10mg/cm2 in
24 hours respectively). At the maximum flux seen in this experiment underocclusion, the amount of ethanol penetrating from a 1m2 area of skin would
give a blood alcohol level of about 4mg% in a 70kg man. In the human use
study, none of the blood samples taken from sixteen human volunteers
exhibited a detectable level of alcohol.
(2) valid with restrictions(415)

Type of experience:

Human - Medical Data

Remark:Reliability
29.06.2004:

Ethanol is excreted in the unmetabolized form in urine, exhaled air andsweat. Its metabolic products are also excreted by exhalation and in theurine. The major route of excretion of ethanol is in the urine.
Excretion: The kidneys and lungs excrete only 5 - 10% of an absorbed
dose of ethanol unchanged (Ellenhorn & Barceloux, 1988).
(4) not assignable(416) (414)Type of experience:

Human – Epidemiology

Method:Epidemiological data published between 1966 and 2000 were subjected tometa-regression analysis in which models were fitted with linear and nonlinear effects of alcohol intake.

Result:235 Studies including 117,000 cases were considered and selectedcovariates were tobacco smoking and gender.
Strong trends in risk were observed for cancers of the oral cavity, pharynx,
oesophagus and larynx. Alcohol-related effects were less strong for

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cancers of the stomach, colonand rectum, liver, breast and ovary. For all these increased risks were forfound for ethanol intake of 25 g/day. Allowance for tobacco smoke
modified association with laryngeal, lung and bladder cancers but not oral,
oesophageal or colorectal cancers.
There was no threshold effect for most alcohol-related neoplasms and theinference was limited by absence of distinction between lifelong abstainers
and former drinkers.
(4) not assignable(417)

Reliability
29.06.2004:

Type of experience

:

Human

Remark:Reliability
29.06.2004:

Distribution
Once absorbed, alcohols are contained in the water compartment of the
body. They are not stored or accumulated to any degree, so that the bodyburden at any point in time is a result of recent absorption, usually within
the previous 12 hours (Conibear, 1988).
Since it is both water and lipid soluble, ethanol easily penetrates the bloodbrain barrier and placenta (Ellenhorn & Barceloux, 1988).
(4) not assignable(418) (419)Type of experience

:

Human

Remark:Reliability
01.10.2003:

Absorption
The most important routes of exposure in terms of occupational exposure
to ethanol are the pulmonary and dermal routes. Ethanol is not wellabsorbed through intact skin, but is well absorbed by inhalation. Significant
intoxication from inhalation is unlikely to occur because ethanol becomes
irritating to the eyes and mucus membranes before concentrations that
could result in CNS depression are reached. Persistant contact withalcohols can result in the removal of the skin's protective fatty barrier which
results in increased absorption (Conibear, 1988).
In man, the small intestine absorbs about 80% of an oral ethanol dose, withthe stomach absorbing the remainder. Since ethanol is poorly absorbed
from the stomach, factors that delay gastric emptying decrease absorption.
In healthy adults, 80 - 90% of absorption occurs within 30 to 60 minutes,
but food may delay complete absorption for 4 to 6 hours (Ellenhorn &Barceloux, 1988).
(4) not assignable(420) (419)Type of experience:

Human – Epidemiology

Remark:Contact sensitivity has been reported in 8 women:

Reliability
22.08.2003:

1. Eczema after washing with spirit (Haxthausen, 1944)
2. Eczema after splashing with spirit (Martin-Scott, 1960)
3. Bullous eczema after splashing with alcohol (Fregert et al. 1969).
4. Eczema (Drevets and Seebohm, 1961)
5. Burning and discomfort of the mouth (Fregert et al. 1969).
6. Reaction to allergens diluted in ethanol (Fregert et al. 1963;1969).
7. Post-operative eruption (Ketel and Tan-Lim, 1975).
(4) not assignable

Type of experience:

Direct observation, clinical cases

Remark:Contact dermatitis as a respose to topically applied alcohols is briefly

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reviewed. Dermatitis may take the form of eczematous eruption orerythematous flush or contact urticaria. This may follow dermal contact or,
in sensitized people, following ingestion of alcoholic beverage. Methods oftesting are discussed.
(4) not assignable
Critical study for SIDS endpoint(402)

Reliability
Flag
12.11.2004::

Type of experience:

Direct observation, clinical cases

Remark:Reliability
12.11.2004:

A case history of contact dermatitis in a 62 yr-old housewife is presented.
She gave positive eczematous reactions to topical formulations in ethanol
and to ethanol alone.
(4) not assignable(422)

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Personal exposure to solvents including ethanol was studied among
hairdressers in 28 salons in 2 regions of Netherlands in 2 seasons andfound to be 200 times below the occupational exposure limit. The average
exposures differed by an factor of 30 between salons. There were alsoseasonal and metereological differences.
(2) valid with restrictions(423)

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Drager badge readings from three employees at a liquid inks blending
factory gave working day exposure levels of 18, 22 and 51 mg/m^3.
(2) valid with restrictions

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Solvent vapour concentrations in the printing area at BP Chemicals Ltd.,
Darton Factory 4 taken on mid-summer dates in 1997, 1998 and 1999
showed concentrations (mg/m^3) of ethanol well below the UK 8-hr TWA
reference period OES of 1920 mg/m^3.
(2) valid with restrictions(30)

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Organic vapour exposures measured in a BP Group Occupational Hygiene
Survey of a flexible packaging area of BP Chemicals Ltd., Darton in 1992
showed exposure values in the range 36 to 337 mg/m^3 for printers and 80
to 250 mg/m^3 during various cleaning activities. These values are well
below the 8-hr TWA OES of 1920 mg/m^2.
(2) valid with restrictions(425)

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Concentrations of ethanol in air at the mixing location in 6 Norwegian
hairdressing salons ranged from 4 to 36 mg/m^3. The exposure level was
significantly lower in salons with local exhaust ventilation than in salons
without ventilation.
(2) valid with restrictions
(426)

(424)

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ID: 64-17-5
DATE: 19.11.2004

Type of experience:

Direct observation, other

Remark:Reliability
12.11.2004:

Both stationary and personal monitoring of ethanol exposure conducted in
6 Norwegain car painting facilities showed very low exposures to ethanol(0.1 to 8.1 ppm versus the Norwegian OES of 500 ppm) and no exposurerelated symptoms.
(2) valid with restrictions(427)

Type of experience:

Human – Epidemiology

Remark:Reliability
12.11.2004:

In a study of the respiratory hazards associated with exposure toformaldehyde and solvents in acid-curing paints a mean exposure to 17
mg/m^3 of ethanol was recorded. This value is far below the Swedish
National Board of Occupational Health and Safety Threshold Value of 1400
mg/m^3.
(2) valid with restrictions(428)

Type of experience:

Direct observation, other

Method:Result:Sampling method : Active sampling using SKC manufactured commercial
sorbing Silica Gel tubes. Sampling rate 0,1l.min-1.
Measurement of ethanol: Gas chromatography and FID detector was usedas the measurement technique.
Long-term stationary and personal measurement were applied. Time ofsampling was>70% of the shift time, the measurement represent the whole
shift exposure
Type of company: ELECTRO-TECHNICAL COMPANY
Profession: Operator (assembling department)
Activity: soldering, electric test of technological unit
Chemical identity: Aktivátor 077 (Ethanol 97%, N,N-dimetyl-p-toluidín 3%)
PPE worn? Yes (gloves only)
Method: Personal sampling
Results 1: TWA 3.7, LOD 0.1, LOQ 0.3 (mg/m3)
Results 2: TWA 11.1, LOD 0.1, LOQ 0.3 (mg/m3)
Profession: Operator (repair workplace of technological unit)
Activity: mechanical repair-analyses of technological unit
Chemical identity: Denatured alcohol (Ethanol 100%)
PPE worn? Yes (gloves only)
Method: Personal sampling
Results : TWA 2.3, LOD 0.1, LOQ 0.3 (mg/m3)
Type of company: WOOD PROCESSING
Profession: PainterActivity: Spraying of wood stain
Chemical identity: Micro Ton SP NK 173-14991 (no further data available)
PPE worn? No
Method: Personal sampling
Results 1: Range 165.02 to 311.34, TWA 200.27, LOD 3.35 to 7.88, LOQ
11.16 to 26.28 (mg/m3)
Profession: PainterActivity: Using spray gun in spray booth with air evacuation
Chemical identity: Ethanol, Toluene ,Xylene, Butylacetate,
Ethylmethylketone, 4-methyl-2-pentanone.
PPE worn? not applicable

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ID: 64-17-5
DATE: 19.11.2004
Method: Area monitoring of source of emissions
Results 1: Range 69 to 158.08, TWA 110.22, LOD 2.44, LOQ 8.14 (mg/m3)
Type of company: MACHINERY
Profession: PainterActivity: spraying of paint using spray gun in spray booth with air
evacuation
Chemical identity: Sigma Wash (paint) (Ethanol 12,10%, Toluene 14,45%,
1-butanol 2,89%, xylene 0,16%, fenol 0,25% iso-butanol 0,66%, butanone
2,51%, 4-metylpentan-2-one 7,64%, propan-2-ol 22,35%.) Diluent C6000(Ethanol 5,11%, Ethylacetate 3-7%, Butylacetate 3-7%, Methylacetate 1015%, Toluene 50-70%.)
PPE worn? Yes (gloves only)
Method: Personal sampling
Result 1: Not detected, LOD 3.7 to 4.5, LOQ 7.0 to 9.0 (mg/m3)
Result 2: Not detected, LOD 3.7 to 4.4, LOQ 7.5 to 8.7 (mg/m3)
Result 3: Not detected, LOD 3.7 to 4.5, LOQ 7.3 to 9.0 (mg/m3)
Result 1: Not detected, LOD 4.1 to 5.4, LOQ 8.3 to 10.9 (mg/m3)
Profession: Technologist
Activity: preparation of paints (mixing)
Chemical identity: not specified
PPE worn? Yes (gloves only)
Method: Personal sampling
Result 1: Not detected, LOD 3.9 to 4.3, LOQ 7.8 to 8.5 (mg/m3)
Type of company: BUS TRANSPORT
Profession: PainterActivity: spraying of paint using spray gun in spray booth without air
evacuation
Chemical identity: Hardener 590 (hardener adding into paint) (Ethanol 1025%, Toluene 10-25%, Methanol pod 2,5%, Propanol 25-50%, Butanol 2550%, 4-metylpentan-2-one 10-25%.)
PPE worn? Yes (inhalation only - Respirator 3M model)
Method: Personal sampling
Result 1: Range 19.5 to 579, TWA 190.1, LOD 3.0 to 7.5, LOQ 10.02 to
24.92 (mg/m3)
Result 2: Range 0 to 249.6, TWA 13.0, LOD 10.0 to 19.2, LOQ 33.39 to
64.25 (mg/m3)
Profession: upholsterer
Activity: swabbing adhesive
Chemical identity: Adhesive Chemoprén Univerzál (Ethanol, Xylene,
Butylacetate, ethylacetate, Butanol)
PPE worn? Yes (inhalation only - Respirator 3M model)
Method: Personal sampling
Result: not detected, LOD 4.3 to 5.1, LOQ 8.6 to 10.2 (mg/m3)
Type of company: PHARMACEUTICAL BIOTECHNICAL
Profession: Pharmaceutical production chemist
Activity: operator of centrifuge during fractionation with ethanol
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result 1: Range 363.8 to 2539.7, TWA 963.4, LOD/LOQ not available(mg/m3)Result 2: Range 450.8 to 2383.3, TWA 1435, LOD/LOQ not available

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ID: 64-17-5
DATE: 19.11.2004(mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of equipment during precipitation with ethanol
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result 1: Range 128.9 to 2237.5, TWA 980.0, LOD/LOQ not available(mg/m3)Result 2: Range 146.3 to 4692.1, TWA 1783.2, LOD/LOQ not available(mg/m3)
Profession: Maintenance engineer
Activity: maintenanceChemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 114.4 to 262.5, TWA 204.9, LOD/LOQ not available(mg/m3)Background measurement
Method: Stationary sampling
Result 1: Range 34.0 to 54.2, TWA 46.2, LOD/LOQ not available (mg/m3)
Result 2: Range 72.0 to 133.5, TWA 99.5, LOD/LOQ not available (mg/m3)
Profession: Chemist operator
Activity: control tasks in regeneration of ethanol
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 11.3 to 293.7, TWA 63.2, LOD/LOQ not available (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator, control of drying equipment, homogenisation
Chemical identity: Ethanol 96%
PPE worn? Yes (gloves only)
Method: Personal sampling
Result: Range 175.5 to 405.6, TWA 322.7, LOD 0.72 to 1.29, LOQ 2.398 to
4.292 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of centrifugal, separation of used spirits, washing withethanolChemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 499.6 to 564.5, TWA 530.1, LOD 0.86 to 1.1, LOQ 2.869 to
3.675 (mg/m3)
Background measurement
Method: Stationary sampling
Result 1: TWA 89.0, LOD 0.89, LOQ 2.953 (mg/m3)
Result 2: Range 43.9 to 51.6, TWA 46.9, LOD 1.77 to 2.81, LOQ 5.895 to
6.699 (mg/m3)
Profession: Chemist operator
Activity: washing of used spirits, pumping of ethanol
Chemical identity: Ethanol 96%
PPE worn? Yes (gloves only)
Method: Personal sampling
Result 1: Range 2504.6 to 4990.2, TWA 3574.6, LOD 1.73 to 2.29, LOQ

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5.755 to 7.613 (mg/m3)
Result 2: Range 51.9 to 138.0, TWA 95.9, LOD 2.2 to 2.3, LOQ 7.4 to 7.7(mg/m3)
Profession: Chemist operator
Activity: pumping of ethanol into colons, distillation of ethanol
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result 1: Range 90.1 to 127.7, TWA 109.1, LOD 2.2 to 2.3, LOQ 7.4 to 7.6(mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of fermentation (control, pumping, sampling of ethanol
and mixing)
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result 1: Range 58.4 to 6590.9, TWA 3197.7, LOD/LOQ not available(mg/m3)Result 2: Range 18.8 to 5097.6, TWA 2979.2, LOD 16.7 to 41.8, LOQ 51.9
to 125.5 (mg/m3)
Background measurement
Chemical identity: Ethanol 96%
Method: Stationary sampling
Result: Range 33.0 to 72.8, TWA 53.5, LOD 26.2 to 33.7, LOQ 78.7 to101.2 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of fractional (adding of ethanol, sampling)
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 282.6 to 1048.1, TWA 495.1, LOD 22.5 to 39.4, LOQ 67.5 to118.1 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of fractional (adding of ethanol, sampling) between
fractional tank
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 282.6 to 1048.1, TWA 495.1, LOD 22.5 to 39.4, LOQ 67.5 to118.1 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of fractional (adding of ethanol, sampling) between
fractional tank
Chemical identity: Ethanol 96%
PPE worn? No
Method: Stationary sampling
Result 2: Range 47.7to 1321.6, TWA 437.4, LOD 29.5 to 52.5, LOQ 88.6 to
157.6 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: operator of centrifuge, washing of tubes with ethanol
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 792.8 to 2636.8, TWA 1369.6, LOD 29.38 to 88.1, LOQ 75.4

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to 264.2 (mg/m3)
Background measurement
Chemical identity: Ethanol 96%
Method: Stationary sampling
Result: Range 711.4 to 1634.5, TWA 1003.0, LOD 21.2 to 88.1, LOQ 75.4
to 264.2 (mg/m3)
Profession: Chemist operator
Activity: sampling of ethanol in the processing hall (temperature control)
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 26.4 to 831.8, TWA 209.6, LOD 11.4 to 18.1, LOQ 34.3 to54.2 (mg/m3)
Profession: Foreman
Activity: supervision of manufacturing operation
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 255.5 to 2419.8, TWA 1286.7, LOD 17.8 to 24.6, LOQ 53.3
to 73.8 (mg/m3)
Profession: Maintenance engineer
Activity: repairs
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 2822.2 to 4686.4, TWA 4063.2, LOD 16.4 to 24.6, LOQ 49.2
to 81.4 (mg/m3)
Background measurement
Chemical identity: Ethanol 96%
Method: Stationary sampling
Result: Range 77.1 to 228.8, TWA 170.5, LOD 16.4 to 34.4, LOQ 52.4 to103.1 (mg/m3)
Profession: Pharmaceutical production chemist
Activity: adding of material, opening of the processing tank containing
ethanolChemical identity: Ethanol 96%
PPE worn? Yes (inhalation - face shield respirator)
Method: Personal sampling
Result: Range 11.9 to 102.5, TWA 30.6, LOD 1.6 to 6.0, LOQ 3.1 to 11.9
(mg/m3)
Profession: Foreman
Activity: supervision of manufacturing operation
Chemical identity: Ethanol 96%
PPE worn? No
Method: Personal sampling
Result: Range 9.3 to 344.6, TWA 78.6, LOD 1.6 to 6.0, LOQ 3.2 to 11.9
(mg/m3)12.11.2004

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Background measurement from drying equipment
Chemical identity: Ethanol 96%
Method: Stationary sampling
Result: Range 3077.6 to 3223.8, TWA 3151.1, LOD 15.2 to 15.4, LOQ 30.4
to 30.7 (mg/m3)(429)

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5. TOXICITY

5.11

ETHANOL
ID: 64-17-5
DATE: 19.11.2004

ADDITIONAL REMARKS

Type

:

Metabolism

Method:Result:

The PBPK model used for this work was modified from that described by
Pastino et al. (1997). The parameters describing Michaelis-Menten
metabolism of ethanol in the liver (VMAX and KM) were varied using thesimulation and optimization software ACSLopt to fit the data. For eachexposure scenario the simulation was run for male workers in "sitting
awake" and "light exercise" activity levels as designated by ICRP. Mixedvenous blood concentration (mM) and area under the curve for the venous
blood concentration (mM-hr) are reported.
A reasonably good overall description of the data was obtained). An exact
fit would not be expected due to, for example, (a) lack of subject-specificinformation on body composition (e.g., fat volume, capacity formetabolizing ethanol) and breathing rates throughout the exposure and, (b)lack of data needed for a more realistic, multi-enzyme specification of
ethanol metabolism (e.g., MEOSand ADH). The model-predicted timecourses of the mixed venous blood concentrations for the 12 exposure
scenarios.
The model predictions are that for men exposed to ethanol, at 0.942 and1.88 mg/L for 8 hr and for the lower breathing rate in men exposed to 9.42
mg/L the liver is able to metabolize ethanol at the rate it enters the body.
However,for the higher breathing rate in men exposed to 9.42 mg/L and formen exposed to 37.6 or 63.6 mg/L the rate of ethanol delivery via breathing
exceeds metabolic capacity and ethanol blood levels consequently rise forthe duration of the exposure. Men exposed to 20 mg/L ethanol for 4 hr
also showed a continued accumulation during exposure at the higher
breathing rate but little or no accumulation at the lower breathing rate.
(4) not assignable(430)

Reliability
12.11.2004

:

Type

:

Chemobiokinetics general studies

Method:Result:A comparative reprotoxic risk assessment was derived for ethanol
exposure due to endogenous generation, alcoholic beverage consumption
and food consumption versus a number of occupational and domestic
exposure scenarios.
Risk assessment
Reprotoxic effects of ethanol are generally thought to have a no observed
effect level below. Although these are not known and are difficult todetermine precisely for each individual end point, it can reasonably be
assumed that the threshold is related to blood alcohol concentration (BAC),
that the critical effect is developmental toxicity and that the recognised safe
level of drinking for pregnant women can be used as a benchmark.
Blood alcohol levels from different activities:
Drink driving limit in Europe
500-850mg/l
No effect level for pregnant women
200mg/lSafe drinking advice from UK Healthuthorities (one drink)
80mg/l100g vanilla ice cream
7.0mg/lMouthwash use
3.5-15mg/l
Pharmaceutical use
3.5-15mg/l
Working at 1000ppm
2.9mg/lUNEP PUBLICATIONS

311

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004
Endogenous blood alcohol level0.035-0.68mg/l
Dermal exposure at work (poor practice) 0.5mg/l
Spray window cleaner
0.2mg/lConsumer toiletry use
0.12mg/l
Consumer hairspray use
0.09mg/l
From this table it can be seen that the level of risk to pregnant women fromeither working with ethanol occupationally or using ethanol containing
consumer products is negligible. Even assuming the rather absurd case ofa female worker maximally exposed to ethanol who, on leaving work,
immediately takes a dose of medicine, sets her hair and cleans thewindows followed by mouthwash would reach a BAC less than one quarterof the level that would be seen through consumption of a single alcoholic
drink. A single drink is recognised as a dose which poses no adverse
health risks by the authorities. This latter dose cannot be approachedfrom any conceivable use or misuse of ethanol containing products or any
imaginable occupational scenario. The reprotoxic hazard of alcoholic
beverages does not exist during the 'normal handling and use' of ethanol
and ethanol containing products. Indeed, developing a craving for vanilla
ice cream would pose a greater risk for a pregnant woman than working
occupationally with ethanol.
(4) not assignable(431)

Reliability
01.10.2003

:

Type

:

Distribution

Remark:Reliability
01.10.2003:

The diffusion of ethanol through cell boundaries is slow and is affected byblood flow. Ethanol in the blood passes almost immediately into brain
tissue, while its distibution to resting muscle is particularly slow. After oral
dosing, ethanol disappeared from the blood of dogs linearly in the post
absorption phase, irrespective of the concentration of ethanol in the body;zero-order kinetics were also found in dogs, cats, rabbits, pigeons andchickens after i.v. administration.
(4) not assignable
(18)

Type

:

Metabolism

Remark:Reliability
29.06.2004:

The metabolism of ethanol has been reviewed (IARC, 1988). Ethanol iseliminated from the body mainly by metabolism in the liver and only
minimally by urinary excretion and pulmonary exhalation. Other tissues
such as kidney, stomach and intestines oxidize ethanol to a small extent.
The hepatic metabolism of ethanol proceeds in three basic steps. First,
ethanol is oxidized within the cytosol of hepatocytes to acetaldehyde;
second, acetaldehyde is converted to acetate, mainly in the mitochondria;
and third,acetate produced in the liver is released into the blood andis
oxidized by peripheral tissues to carbon dioxide, fatty acids and water.
The main pathway for ethanol metabolism proceeds via alcohol
dehydrogenase. However other pathways for ethanol oxidation have been
described includinga microsomal ethanol-oxidizing system located in theendoplasmic reticulum and a catalase system located in the peroxisomes.
(4) not assignable
(18)

Type

:

Metabolism

Remark:The activities of alcohol dehydrogenase (ADH), catalase, microsomal
ethanol-oxidizing system (MEOS) and aldehyde dehydrogenase (ALDH)
were measured in gastric, small intestinal, colonic and rectal mucosal
samples of rats fed on a liquid alcohol diet for 1 month.

312

UNEP PUBLICATIONSOECD SIDS
5. TOXICITY

Conclusion

ETHANOL
ID: 64-17-5
DATE: 19.11.2004:

In the rectum and large intestine of control animals, the activities of ADH,
MEOS and catalase were maximal, whereas the activity of ALDH wasminimal. After chronic alcohol intoxication, MEOS activity increased
significantly in the stomach. An activation of catalase and MEOS and adecrease of the low-K(M) ALDH activity were observed in the rectum. In
rats consuming the alcohol diet, hypertrophy of crypts and an increased
number of mitoses were noticed in colonic and rectal mucosa. Acutealcohol intoxication (2 g/kg, intragastrically) produced significantly higher
acetaldehyde concentrations in the contents of the large intestine andrectum of rats receiving alcohol chronically compared tocontrols.
After chronic alcohol intoxication, the large intestine regions show a greater
imbalance between the activities of acetaldehyde-producing andacetaldehyde-oxidizing enzymes, which results in accumulation of
acetaldehyde.
This mechanism can account for the local toxicity of ethanol after its
chronic consumption, and relates the development of mucosal damage andcompensatory hyper-regenerative processes, and possibly carcinogenesis,in the colonic and rectal mucosae of alcoholics to the effects ofacetaldehyde.
(4) not assignable(432)

Reliability
01.10.2003

:

Type

:

Toxicokinetics

Remark:

The teratogenic effects of ethanol are reviewed in terms of the differences
between direct and indirect effects of ethanol on the developing foetus.
Direct effects of ethanol are caused by ethanol interacting with the foetal
cell whereas indirect effects of ethanol teratogenicity are defined as any
perturbation of the developing fetus resulting from ethanol exposure, but
not caused by ethanol's interacting with the fetal cell.
Indirect effects of ethanol teratogenicity include: ethanol-induced maternal
undernutrition, ethanol-induced placental dysfunction and acetaldehyde
teratogenicity.
(4) not assignable(433)

Reliability
01.10.2003

:

Type

:

other: Percutaneous absorption

Method:Result:Conclusion:

Reliability
01.10.2003:

The absorption of 14C-ethanol was determined in fresh, metabolically
active human breast skin in vitro over 6 hr. Detection of CYP 2E1 was byimmunoblotting.
Absorption over 6 hr was low (0.5 to 2.2%) with a 4-fold variation between
the 3 individuals. Absorption through occluded skin was higher (5.5 to
13.4%).
Absorption of ethanol through occluded human skin over 6 hr was poor.
Occlusion enhanced absorption and increased skin residue.
(4) not assignable
(434)

Type

:

other: Reviews (II)Method:Remark:Pregnant ICR mice were treated with 100 mg/kg pyrazole, an alcohol
dehydrogenase inhibitor, prior to i.p. ethanol injection. In anotherexperiment pregnant mice were housed in an ethanol-vapour box for 3 or 6
days in order to examine the effects of prolonged low level exposure toalcohol. The maternal blood alcohol concentration was maintained at
approximately 0.03 mg/mL during inhalation.
These results suggest that ethanol rather than its

UNEP PUBLICATIONS

313

OECD SIDS
5. TOXICITYETHANOL
ID: 64-17-5
DATE: 19.11.2004

Result:Reliability
29.06.2004:

314

metabolites is mainly responsible for the embryotoxicity.
I.p. treatment with 2 or 4 g/kg ethanol on day 7 ofgestation increased the prenatal mortality rate andproducedexternal and skeletal malformations in theoffspring, and the embryotoxic effects were potentiated bypyrazole pretreatment. The inhalation treatment with ethanolincreased the prenatal mortality rate, althoughteratogenicity was not shown.
(4) not assignable(435)

UNEP PUBLICATIONSOECD SIDS
6. ANALYTICAL METHODS FOR DETECTION AND IDENTIFICATION

6.1

ANALYTICAL METHODS

6.2

DETECTION AND IDENTIFICATIONUNEP PUBLICATIONSETHANOL
ID: 64-17-5
DATE: 19.11.2004315

OECD SIDS
7. EFFECTS AGAINST TARGET ORGANISMS AND INTENDED USES

7.1

FUNCTION

7.2

EFFECTS ON ORGANISMS TO BE CONTROLLED

7.3

ORGANISMS TO BE PROTECTED

7.4

USER

7.5

RESISTANCE

316

UNEP PUBLICATIONSETHANOL
ID: 64-17-5
DATE: 19.11.2004

OECD SIDSETHANOL
8. MEASURES NECESSARY TO PROTECT MAN, ANIMALS, ENVIRONMENT ID: 64-17-5
DATE: 19.11.2004
8.1

METHODS HANDLING AND STORING

Safe handling
Fire/exp. protection::

Storage requirement
Common storage
Container
Unsuitable container
Add. information
Transport code

:
:
:
:
:
:

Do not smoke eat or drink in areas of use and storage.
Keep away from heat, sources of ignition and incompatible substances.
Earth (ground) lines and equipment used during transfer to reduce
possibility of static spark initiated fire or explosion.
Store in tightly closed containers in cool, dry, isolated, well-ventillated area.
Aluminium
UN 1170

12.11.2004
8.2

(1)

FIRE GUIDANCE

Hazards
Protective equipment::

Extinguishing medium
Unsuit. exting. medium
Add. information:::

Fire class
Products arising::

Flammable liquid and vapour
Approved positive pressure self-contained breathing apparatus with fullface mask and full protective clothing.
Dry chemical, alcohol foam, all-purpose AFFF or carbon dioxide.
Water for large volumes of ethanol.
Use water to cool fire-exposed containers and to disperse vapour. Avoid
vapour flash-back.
Oxides of carbon

12.11.2004
8.3

(1)

EMERGENCY MEASURES

Type

:

accidental spillage

Remark:Keep unecessary people away; isolate hazard area and deny entry.
Stay up wind and keep out of low areas where vapour may accumulate andignite.
Shut off all sources of ignition.
Stop leak if this can be achieved without risk.
For small spills take up with and or non-combustible
absorbant.
For large spills, Dike for later disposal.
(1)

Type

:

injury to persons (skin)

Remark:Avoid skin contact by using appropriate chemical protective gloves, faceshield, apron and armcovers. Use closed-system transfers wherever
possible.

11.01.2002

29.06.2004

(1)

Type

:

injury to persons (eye)Remark:Prevent eye contact by wearing chemical tight goggles.
Provide an eyewash station in the immediate vicinity of use.

29.06.2004(1)

UNEP PUBLICATIONS

317

OECD SIDSETHANOL
8. MEASURES NECESSARY TO PROTECT MAN, ANIMALS, ENVIRONMENT ID: 64-17-5
DATE: 19.11.2004
Type:

injury to persons (oral)

Remark:Accidental ingestion at a level high enough to be dangerous to health isunlikely. Be aware of 'recreational ingestion' and workplace alcoholism.

29.06.2004

(1)

Type

:

injury to persons (inhalation)

Remark:In circumstances where physical controls cannot be effectively applied use
a respirator approved for organic vapours.
(1)

Type

:

other: diagnosis of intoxication

Remark

:Toxicity from ethanol, methanol, ethylene glycol, and isopropyl alcohol
varies widely, and appropriate use of the available laboratory tests can aid
in timely and specific treatment. Available testing includes direct
measurements of serum levels of these alcohols; however, these levels
often are not available rapidly enough for clinical decision making. This
article discusses the indications and methods for both direct and indirect
testing for ethanol, methanol, ethylene glycol, and isopropanol toxicity. Also
discussed are the costs, availability, and turn-around times for these tests.
(436)

11.01.2002

29.06.2004
8.4

POSSIB. OF RENDERING SUBST. HARMLESS

DomainProcess
Type of destruction:::

Industry/skilled trades
Recycling
Incineration

12.11.2004
DomainProcess
Type of destruction

(1)
:
:
:

Public at large
Destruction
Incineration

12.11.2004
8.5

(1)

WASTE MANAGEMENT

Memo

:

Possibility of recovery/recycling

11.01.2002
Memo

(1)
:

Possibility of destruction: water purification

11.01.2002
Memo

(1)
:

Possibility of destruction: incineration

12.11.2004
8.6

318

(1)

SIDE-EFFECTS DETECTION

UNEP PUBLICATIONSOECD SIDSETHANOL
8. MEASURES NECESSARY TO PROTECT MAN, ANIMALS, ENVIRONMENT ID: 64-17-5
DATE: 19.11.2004
8.7

SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER

8.8

REACTIVITY TOWARDS CONTAINER MATERIAL

Memo

:

Aluminium at higher temperatures.

12.11.2004(1)

UNEP PUBLICATIONS

319

OECD SIDS
9. REFERENCESETHANOL
ID: 64-17-5
DATE: 19.11.2004(1)

CEFIC Ethyl Alcohol Group (2003)(2)

Ministry of Environment, Prague (2004)(3)

UK Health and Safety Executive, EH40 (2002)(4)

Deutche Forschungsgemeinschaft (2003)(5)

HSDB (2003) Hazardous Substances Databank. US National Library for Medicine.

(6)

Centre for Chemical Substances and Preparations, Bratislava(7)

http://www.epa.gov/ttn/atw/orig189.html(8)IARC Monographs on Evaluation of Carcinogenic Risks to Humans Vol.44 (1988)(9)

Sprung, R., Bonte, W., Rudell, E., Domke, M., Frauenrath, C. (1981). On the problem ofendogeneous alcohol. Endogenous ethanol: Further investigations. Alcohol Drugs Behav.
1981;18(2):65-70.

(10)Lerici, C.R., Nicoli, M.C. (1996). Chemical and physico-chemical properties affecting thequality and stability of bakery products. Adv. Food Sci 18(5/6):229-233(11)Greubet, S. (1997). Versteckter Alkohol in Lebensmittel? Medizische Monatsschrift fur
Pharmazeuten 1997;20(8):218-219.

(12)

Anonymous (1986). Hearings Before the SubCommittee on Select Revenue Measures of
the Committee on Ways and Means, House of Representatives, Ninety-ninth Congress,
Second Session, May 12 and 19, 1986.

(13)

Association of Industry of Juices and Nectars from Fruits and Vegetables of the EU (1996)
Code of Practice for Evaluation of Fruit and Vegetable Juices.

(14)

SPIN database http://www.spin2000.net/spin.html(15)

Verschueren, K. (1996) Handbook of environmental data on organic chemicals. 3rd Edition.
Van Nostrand Reinhold Company, New York.

(16)

http://www.epa.gov/medecotx/searches/aquatic. U.S. EPA online ECETOX database.

(17)

http://www.epa.gov/medecotx/searches/terrestrial.

(18)

IARC Monographs. (1988) Volume 44. Alcohol Drinking. Lyon, France.

(19)

Irvine, L. (2003) Relevance of the Developmental Toxicity of Ethanol in the Occupational
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Phillips P. J., Jenkinson, P. (2001) Is ethanol genotoxic? A review of the published data.
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Ophaswongse, S., Maibach, H.J. (1994). Alcohol dermatitis and contact dermatitis
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(22)

Sensible Drinking. The report of an Inter-Departmental Working Group. December 1995.
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(23)

Macdonald, I. (Ed). (1999). Health issues related to alcohol consumption. 2nd ed.
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(25)

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(26)

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(27)

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(30)

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(31)

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(32)

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Howard, P. (1990). Handbook of Environmental Fate and Exposure Data for Organic
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(37)

Kavanaugh, M.C; , Stocking, A. (1999). Fate and Transport of Ethanolin the Environment.
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(38)

Maher PJ, Smith BD (1979) A new total pressure vapor-liquid equilibrium apparatus. The
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(40)

Kavanaugh, M.C; , Stocking, A. (1999). Fate and Transport of Ethanol in the Environment.
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Campbell, I.M. et al. (1976) Rate constants for reactions ofhydroxyl radicals with alcohol
vapours at 292 K. Chem. Phys.Letters 38, 362 - 364.

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Carter, PL (1994) Development of ozone reactivity scales for volatile organic compounts. J
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Carter, W.P.L., The SAPRC-99 Chemical Mechanism and Updated VOC Reactivity Scales(2000). URL http://www.cert.ucr.edu/~carter/reactdat.htm(53)

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DATE: 19.11.2004(82)

In a similar study, a 96-hour LC50 of 10000 - 11500 mg/l wasreported in the bleak(Alburnus alburnus) in a static test at 10 °C. Linden, E. et al. (1979) The acute toxicity of
78 chemicals and pesticide formulations against two brackish water organisms, the bleak(Alburnus alburnus) and the harpacticoid Nitrocra spinipes. Chemosphere 11/12, 843 851.

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(84)

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(85)

O'Connor, C.S., Crawshaw, L.I., Bedichek, R.C., Crabbe, J.C.The effect of ethanol on
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