Registration Dossier

Administrative data

Description of key information

ORAL (LD50 values)
Rat: female: 15010mg/kg; male (young adult): 10600mg/kg; male (old adult); 7060mg/kg; not sex specific: old adult: 11500mg/kg; young adult: 17750mg/kg; immature animal: 6160mg/kg, ~12000mg/kg, male/female :10470mg/kg; >7692mg/kg (female).
Mouse: 8350mg/kg
Human: LD50 ~2000mg/kg
INHALATION
Rat (4hr): LC50: male: 51mg/l, female: 55mg/l
Mouse: LC50>60000ppm
DERMAL
No reliable data. Information indicates LC50>15800mg/l
INTRAPERITONEAL (LD50 values)
Rat: young animals LD50 5500-6710mg/kg. Old animals LD50: 4070-5100mg/kg
Mouse: Male 9020, 9710mg/kg. Female: 9450mg/kg.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1976-02-26 to 1976-03-19
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study according to OECD guideline 401.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: Cox CD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: not available
- Age at study initiation: not available
- Weight at study initiation: 199-215 g
- Fasting period before study: not available
- Housing: not available
- Diet (e.g. ad libitum): not available
- Water (e.g. ad libitum): not available
- Acclimation period: not available


ENVIRONMENTAL CONDITIONS
- Temperature (°C): not available
- Humidity (%): not available
- Air changes (per hr): not available
- Photoperiod (hrs dark / hrs light): not available


IN-LIFE DATES: From: 1976-02-26 To: 1976-03-19
Route of administration:
oral: unspecified
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: not applicable
- Amount of vehicle (if gavage): not applicable
- Justification for choice of vehicle: not applicable
- Lot/batch no. (if required): not applicable
- Purity: not applicable


MAXIMUM DOSE VOLUME APPLIED: 19.6 ml/kg


DOSAGE PREPARATION (if unusual): not applicable


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: not applicable
Doses:
8200, 9840, 11480 and 16070 mg/kg
No. of animals per sex per dose:
10 animals total (5 male/5 female) at 8200, 9840, 11480 and 16070 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily for observation and body weights were taken at initial and termination of the study
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs, body weight and mortalities
Statistics:
Not applicable
Preliminary study:
Not applicable
Sex:
male/female
Dose descriptor:
LD50
Effect level:
10 470 mg/kg bw
95% CL:
9 720 - 11 380
Mortality:
8200 mg/kg: 0/10
9840 mg/kg: 3 female animals/10
11480 mg/kg: 8 (5 female animals)/10
16070 mg/kg: 10 (5 female animals)/10
Clinical signs:
8200 mg/kg: moderate decrease in motor activity and respiratory rate, moderate blanching, gripping and ataxia
9840 mg/kg: moderate decrease in motor activity and respiratory rate, moderate pupillary response moderate gripping and ataxia
11480 mg/kg: extreme decrease in motor activity and respiratory rate, moderate pupillary response moderate to slight gripping and ataxia
16070 mg/kg: moderate decrease in motor activity and respiratory rate, moderate pupillary response slight blanching, abdominal gripping and ataxia
Body weight:
Gain in body weights were observed in surviving animals
Gross pathology:
No gross abnormalities were seen upon gross necropsy
Other findings:
- Organ weights: not available
- Histopathology: not available
- Potential target organs: not available
- Other observations: none

None

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
5% H2O in 95% Ethanol : OECD GHS: LD50= 10470 mg/kg (based on 95% active test material)
Executive summary:

In a guideline acute oral toxicity study, the LD50 was determined to be 10470 mg/kgbw when dosed as a 95% solution in water.

Endpoint conclusion
Dose descriptor:
LD50
10 470 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: WIGA, Sulzfeld
- Mean weight at study initiation: 185 ± 15 g
- Diet (e.g. ad libitum): Herilan MRH
- Water (e.g. ad libitum): Tap water


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 5
- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
inhalation: vapour
Type of inhalation exposure:
nose/head only
Vehicle:
other: unchanged (no vehicle)
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- The test substance was heated up to 65 °C, the vapors generated were mixed with a stream of fresh air and delived to the inhalation-system in constant amounts with the help of a dosing pump.


TEST ATMOSPHERE
- Brief description of analytical method used: Gas-chromatography
- Samples taken from breathing zone: yes
Analytical verification of test atmosphere concentrations:
yes
Remarks:
Gas-chromatography
Duration of exposure:
4 h
Concentrations:
155.0, 115.4, 93.4, 79.1, 62.0 mg/l
No. of animals per sex per dose:
10
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical signs were observed daily, body weight were measured at the begining of the study and weekly thereafter
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Statistics:
Probitanalysis as described in DJ Finney (Probitanalysis 1971, 1 - 150).
Sex:
male/female
Dose descriptor:
LC50
Effect level:
124.7 mg/L air
Exp. duration:
4 h
Sex:
male
Dose descriptor:
LC50
Effect level:
116.9 mg/L air
Exp. duration:
4 h
Sex:
female
Dose descriptor:
LC50
Effect level:
133.8 mg/L air
Exp. duration:
4 h
Mortality:
see free text
Clinical signs:
In the top 4 dose groups: attempts to escape, reddish-watery eyes, nasal secretions, closing of eyelids, snout wiping, intermittent respiration, loss of pain reflex, narcosis, ruffled fur-coat and squatting position. The surviving animals were free from symptoms after day 1 till the end of observation period.
In the lowest dose group: attempts to escape, reddish-watery eyes, nasal secretions, closing of eyelids, snout wiping, intermittent respiration, loss of pain reflex, abdominal position and apathy. All animals were free from symptoms after day 1 till the end of observation period.
Body weight:
The body weight of males in higher dose groups was slightly decreased.
Gross pathology:
Deceased animals: acute dilation and hyperemia in heart. Lungs showed partly acute flatulence of middle grade. Additionally, lungs were partly spotted, infracted, blood-filled and edematous.
Sacrificed animals: no abnormalities

Concentration (mg/L)

Mortality

male

female

155.0

7/10

9/10

115.4

7/10

1/10

93.4

2/10

0/10

79.1

1/10

0/10

62.0

0/10

0/10

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results, the test substance do not require any classification according to EU and GHS standards.
Executive summary:

In an acute inhalation study that approximated to guideline, ethanol vapour was found to be of very low acute toxicity to male and female rats. The LC50 (4hr) was established to be around the 117 -125mg/l and the LC0 around 62mg/l. It is worthy of note that the LD50 is well above the lower explosive limit (LEL).

Endpoint conclusion
Dose descriptor:
LC50
50 000 mg/m³

Acute toxicity: via dermal route

Endpoint conclusion
Dose descriptor:
LD50
15 800 mg/kg bw

Additional information

ORAL ROUTE

In two guideline studies, the LD50 of ethanol was determined to be in the region 10000 to 10500mg/kgbw when dosed at concentrations of 80 -95% in water. In an acute oral toxicity study to guideline using the up and down procedure, the Oral LD50 in female rats of SDA Alcohol (100% ethanol) was determined to be >7692 mg/kg bw. In an acute toxicity study in female rats using a relatively large number of animals over 7 closely spaced doses, an LD50 value of 14500 -15500mg/kg was obtained. Animals were only observed for a period of 24 hours.  Another study examined the effect of age of male rats on the oral LD50. Using a relatively large number of animals and 6 -8 doses, an LD50 value of 10600mg/kg was obtained for young animals (~100 days old) whereas a significantly lower figure of 7060mg/kg was obtained for older rats (~11 -12 months old). Animals were again only observed for a period of 24 hours.  Another study, for which only basic experimental detail is reported, also examined the effect of age of rats on the oral LD50, which was obtained for three groups of animals: immature 14 day old animals, young and old adults. The study confirmed that both old and young animals are more sensitive to toxic effects than are young adults.  For those studies that only observed for 24 hours following dosing, it is unlikely that significant deaths would have occured after this point due to the known toxicokinetics of metabolism.  In these studies, those animals that died showed severe CNS effects with death due to cardiorespiratory failure that was preceded by coma. In an acute toxicity study in male and female mice, an LD50 value of 8350mg/kg was obtained. All deaths occurred within the first 24 hours of dosing. The dose response curve was relatively steep; an LD50:LD16 ratio of 1.19 was obtained. In an acute oral toxicity study , the LD50 was determined for different concentrations of ethanol in water. The LD50 was consistently found to be around 12g/kgbw and independent of the concentration dosed (range used 30 -90% ethanol in water). The value chosen for the critical parameter is the figure from a guideline study. Older rats were shown to be somewhat more sensitive but such variations are taken into account in intraspecies assessment factors.

A comparison of the rat per oral versus intraperitoneal LD50 values suggests that first pass metabolism is significant and reduces effective systemic dose by ~50% by this route of exposure.

Human data

In a poisoning incident involving consumption of an ethanol based hand sanitiser, an estimated dose of 1.5g/kg ethanol caused marked intoxication of a 4 year old child but all effects were fully reversed within 24 hours (Engel, 2010). In another child, a dose of ~4.5g/kg caused unconciousness but the patient fully recovered with supportive treatment only. However, in a poisoning incident involving vodka consumption, an estimated dose of 2 -3g/kg ethanol proved fatal for a 5 year old child (Klys, 2008). In a case study, a 15 year of girl consumed a bottle of tequila sufficient to provide a resultant dose of 4.2 -4.8g/kg; the patient fully recovered after supportive treatment only. serum ethanol concentration of >526mg/dL proved fatal in an adult alcoholic (~3g/kg). In a poisoning case of an adult, a dose of 175g of ethanol (estimated 2.2g/kg) proved fatal. However, the patient was already intoxicated with alcohol (estimated blood ethanol concentration 188mg/l). The resulting blood ethanol concentration was at least 526mg/dL (252g ethanol for an 80kg person assuming a volume of distribution of 0.6L/kg). A retrospective study was carried out to examine ethyl alcohol poisoning related deaths in Ankara and surrounding cities between 2001 and 2011. Of the medico-legal autopsy cases performed, 39 (0.36% ) were identified as due to ethanol poisoning alone. The overwhelming majority of the cases were male (90.5%) and the mean age of the victims was 44.9 (range 10.9 to 92 years). The levels of postmortem blood alcohol levels were available for all cases and ranged from 136 to 608 mg/dL for ethanol (estimated 0.75 to 3.3g/kg) The lower end of the range may be unreliable as significant metabolism could have occured before death. (Celik, 2013). This suggests for humans the fatal dose for ethanol is variable but greater than 2g/kg along with blood ethanol concentrations in excess of 300mg/dL (which, assuming a volume of distribution of ~60% of human body weight, would equate to 1.8g/kg - remarkably consistent with the previous figure. Crudely, an LC50 of ~2g/kg in humans can be assumed.

DERMAL ROUTE

According to Annex VIII, testing by the dermal route for acute toxicity is appropriate if all of the following conditions are met: inhalation of the substance is unlikely, skin contact in production or use is likely and the physicochemical and toxicological properties suggest potential for a significant exposure.  Clearly inhalation is likely and will be the dominant route of exposure.  Whilst skin contact will occur, available data suggests that under non-occlusive conditions, evaporation is so rapid that dermal exposure is negligible.  Annex VIII indicates that testing for acute toxicity is only required by two routes of exposure. Data is available on the oral and inhalation routes. In addition, there is sufficient data available to enable reliable extrapolation from the oral to the dermal route if required.  There is data from a secondary source that cites an unreferenced result from an acute dermal toxicity study: a single dose of 15800mg/kg caused the death of 1 out of 4 rabbits. This indicates that the LD50 is greater than 15800mg/kg and is the figure used as the critical LD50.

INHALATION ROUTE

In an acute inhalation study that approximated to guideline but extended the exposure time to 6 hours, ethanol vapour was found to be of low toxicity. The LC50 (6hr) was established to be around 50 -55mg/l and the LC0 around 43 -44mg/l. In another acute inhalation study that approximated to guideline but extended the exposure time to 6 hours, the LC50 (6hr) was established to be around 50 -55mg/l and the LC0 around 43 -44mg/l. To put these results into context, it is worth noting that the lower explosive limit (LEL) is around 50mg/l.

In another study to determine non lethal as well as lethal effects, mice were exposed to ethanol for different periods of time up to 1 hour in order to determine the LC50 and the EC50 for motor performance. An LC50 could not be obtained for inhalation exposure; no deaths were reported for exposures up to 60,000ppm (114mg/l), which can be considered the saturated vapour concentration. In the motor performance test, an EC50 60 min value of 30,300ppm was established, with a slope for the dose effect curve of 12.6. This result differed only slightly from the EC50 30 min. It should be noted that all of the reported EC50 values are above the LEL reported in chapter 4.

Based on the results available, inhalation exposure poses little hazard at any feasible exposure concentration.

Justification for classification or non-classification

All LD50 and LC50 are by some margin above the thresholds for classification for acute toxicity either under directive 67/548 or the EU CLP regulations.