Reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade

Endpoint summary

Description of key information

Data on the components of the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade, diethylene glycol (DEG), triethylene glycol (TEG), and tetraethylene glycol (TTEG), were used to assess its acute toxicity potential.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented publication.

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

The authors report results on the pharmacokinetics and metabolism of DEG and EG in rat, obtained with 14C-labelled DEG and non-labelled EG, using high-resolution NMR spectroscopy as a highly specific and rapid technique.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 250 - 300 g
- Housing: in stainless-steel metabolic cages
- Diet: Alma standard diet for rats and mice (H 1003, Kempten, Germany), ad libitum
- Water: ad libitum

Route of administration:

oral: gavage

Vehicle:

physiological saline

Details on oral exposure:

Diethylene glycol (DEG) was administered to rats by gavage as 50% solution in 0.9% saline (v/v), at doses of 1 (i.e., 1.12 g), 5, 10, 12.5, 15 and 17. 5 mL DEG/kg bw.

Doses:

1 (i.e., 1.12 g), 5, 10, 12.5, 15 and 17. 5 mL DEG/kg bw.

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

Observation period: 7 days

Sex:

male

Dose descriptor:

LD50

Effect level:

19 600 mg/kg bw

Clinical signs:

other: The following effects have developped consecutively: narcotic phase, diuretic phase and thirst, drop of the pH of the urine and blood, either recovery or hydrotropic degeneration of the renal tubules and anuria, accumulation of urea and uric acid in the

Interpretation of results:

GHS criteria not met

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

Not specified but between 9 Dec 1985 and 6 April 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The report does not specify about GLP/Guideline but sufficient data is available for interpretation of results

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Sprague-Dawley albino rats, weighing between 200 and 300 g were used. The rats are fasted overnight before dosing.

The animals are maintained on appropriate commercial diet and municipal water. Both are available ad libitum except during periods of fasting (rat peroral test) or manipulation.

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The maximum dosage for the peroral test is 16 ml/kg. Based on the test results, no additional dose levels were examined.

Doses:

16 ml/kg

No. of animals per sex per dose:

5 males and 5 females.

Control animals:

not specified

Details on study design:

Animal weights are recorded at 0 days (before dose), 7 days and 14 days (just prior to sacrifice). At death or sacrifice, each animal is subjected to gross pathologic evaluation.

Statistics:

LD50's and the estimated LD50 slopes are calculated by the moving average method (Thompson, 1947; Weil, 1983) and are based on a 14-day observation period.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 16 mL/kg bw

Based on:

test mat.

Mortality:

None of 5 males or 5 females died from this dosage.

Clinical signs:

other: There were no signs of toxicity observed.

Gross pathology:

There were no remarkable gross pathologic lesions observed.

Other findings:

No additional information available.

No additional information available.

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 for male and female rats receiving single peroral doses of tetraethylene glycol was greater than 16.0 ml/kg (>18000 mg/kg).

Executive summary:

The acute oral toxicity of tetraethylene glycol was examined.

The LD50 for male and female rats receiving single peroral doses of tetraethylene glycol was greater than 16.0 ml/kg, Rats gained weight with no clinical signs of toxicity. There were no remarkable gross pathological lesions observed.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

No data.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This study was conducted prior to GLP and test guidelines, but sufficient data is available for interpretation of results.

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

no guideline available

Principles of method if other than guideline:

Groups of rats were dosed with tetraethylene glycol and observed for 14 days. Body weight was recorded on days 1 and 14. Animals were observed and mortality was determined over the 14 day observation period. A gross pathologic examination was conducted.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: albino

Sex:

male

Route of administration:

oral: gavage

Vehicle:

water

Doses:

25,000, 30,000, 35,000 and 40,000 mg/kg

No. of animals per sex per dose:

2 male rats at 25,000 mg/kg, 10 male rats at 30,000 mg/kg, 10 male rats at 35,000 mg/kg and 5 male rats at 40,000 mg/kg.

Control animals:

not specified

Sex:

male

Dose descriptor:

LD50

Effect level:

33 000 mg/kg bw

Based on:

test mat.

Mortality is summarized in Table 1. Animals that died, died on day 1.

Table 1 Mortality following a single oral dose of tetraethylene glycol

Dose, mg/kg	Number dosed	Number died 14 days
25000	2	0
30000	10	4
35000	10	5
40000	5	5

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of tetraethylene glycol to male albino rats was found to be 33 grams per kilo.

Executive summary:

The acute oral toxicity of tetraethylene glycol was examined. The LD₅₀ was reported as 33,000 mg/kg.

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: basic information given

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

LD50 calculation

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

other: intubation

Vehicle:

not specified

Details on oral exposure:

Rats, weighing between 200 and 300 g, receive the test material by stomach intubation with a ball-end stainless steel needle. The sample is injected trough the needle by means of a syringe and doses are varied by adjusting the volume of the test material or its dilution.
The rats are fasted overnight before dosing. 5 males and 5 females are included on each level used for the LD50 calculation.
LD50 and the estimated LD50 slopes are calculated by the moving average method and are based on a 14-day observation period. Animals weights are recorded at 0 days (before dose), 7 days and 14 days (just prior to sacrifice). At death or sacrifice, each animal is subjected to gross pathologic evaluation.

Doses:

16 ml/kg

No. of animals per sex per dose:

5

Control animals:

not specified

Statistics:

no data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 16 mL/kg bw

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 18 000 mg/kg bw

Remarks on result:

other: No mortalities.

Mortality:

none

Clinical signs:

other: sluggishness, unsteady gait

Gross pathology:

no remarkable gross lesions

Signs of toxicity included sluggishness and an unsteady gait. Recovery occurred at 3 hours to 1 day. There were no remarkable gross lesions evident at necropsy.

Interpretation of results:

GHS criteria not met

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

1974-1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: non-GLP, non-guideline

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

LD50 calculation

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 3-4 weeks
- Weight at study initiation: 90-120 g
- Fasting period before study: nonfasted
- Diet (e.g. ad libitum): ad libitum, except during period of manipulation or confinement
- Water (e.g. ad libitum): ad libitum, except during period of manipulation or confinement

Route of administration:

other: stomach intubation

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

No additional information

Doses:

16, 32 ml/kg

No. of animals per sex per dose:

5 males/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

LD50 calculated by the moving average method based on a 14-day observation period

Sex:

male

Dose descriptor:

LD50

Effect level:

22.6 mL/kg bw

Based on:

test mat.

95% CL:

ca. 16.8 - ca. 30.6

Mortality:

All animals receiving 32 ml/kg died (on days 1, 1, 1, 2, 3). No deaths occurred at 16 ml/kg.

Clinical signs:

other: Signs of toxicity in 32 ml/kg dosed animals were sluggishness, pilo-erection, and heavy breathing noted at 25 minutes. No signs of toxicity occurred in 16 ml/kg dosed animals.

Gross pathology:

In 32 ml/kg dosed animals, the following was observed at necropsy: lungs with reddening or petechiae, mottled livers, transparent stomachs, gas-filled, yellow, transparent intestines, acini of kidneys prominent, medullae slightly reddened, bladders full. In 16 mlg/kg dosed animals, reddened lungs were observed at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

Extremely low order of toxicity following acute peroral intubation in rats.

Executive summary:

The LD50 was 22.6 ml/kg for male rats exposed to DEG by stomach intubation. DEG has an extremely low order of toxicity following acute peroral intubation in rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 120 mg/kg bw

Quality of whole database:

Animal studies are available for DEG, TEG, and TTEG. However, available data in humans suggest increased sensitivity to the metabolic acidosis and nephrotoxic effects of DEG metabolites. Therefore, human data is considered for determining the acute oral dose descriptor for the registered substance. Since increased sensitivity to the metabolic acidosis and nephrotoxic effects of DEG metabolites has been noted in several intoxication incidences in humans, no key animal study was chosen for the acute oral toxicity endpoint.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

Not specified but between 9 Dec 1985 and 6 April 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The report does not specify about GLP/Guideline but sufficient data is available for interpretation of results

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Sprague-Dawley albino rats, weighing between 200 and 300 g, were used.

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

The vapor is produced by passing air (at 2.5 liters/min) through the sample and then through a 9-liter animal chamber (dynamic conditions).

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

6 h

Concentrations:

Described as substantially saturated vapor generated at 24C.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

not specified

Details on study design:

Rats were weighed prior to exposure to the test material and again on day 7 and 14. Animals were observed for clinical symptoms and mortality. At the end of the 14 day observation period, a gross pathologic examination of the rats was conducted.

Statistics:

Body weight mean and S.D. were calculated.

Sex:

male/female

Dose descriptor:

LC0

Effect level:

0.06 ppm

Based on:

test mat.

Exp. duration:

6 h

Mortality:

No mortality was observed in males or females exposed to a substantially saturated vapor concentration.

Clinical signs:

other: No clinical signs were observed in males or females exposed to a substantially saturated vapor concentration.

Body weight:

Male rats gained weight on days 7 and 14 from pre-exposure values (Table 1). Females remained essentially the same throughout the 14 day observation period.

Gross pathology:

There were no treatment-related gross pathologic observations.

Other findings:

Using a vapor pressure of 0.0000465 mm Hg at 25C corresponds to a saturated vapor concentration of 0.061 ppm.

Table 1 Body weights of Males and Females exposed to a substantially saturated vapor concentration for 6 hours.

	Day 0	Day 7	Day 14
Males	237 + 2.4	261 + 5.7	282 + 7.9
Females	256 + 7.2	257 + 8.9	256 + 8.4

Conclusions:

A single dynamic inhalation exposure to substantially saturated vapor for a 6-hour period produced no deaths or other signs of toxicity among male and female rats.

Executive summary:

Groups of 5 male and 5 female rats were exposed to a substantially saturated vapor concentration for 6 hours. There were no effects on mortality, body weight, clinical symptoms or gross pathology noted in rats.

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

Not specified but between 15 March and 17 April 1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed prior to GLP but sufficient data is available for interpretation of results.

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Wistar derived

Sex:

not specified

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

8 h

Concentrations:

Described as a substantially saturated vapor. Using a vapor pressure of 0.0000465 mm Hg at 25C corresponds to a saturated vapor concentration of 0.061 ppm (0.49 ug/L).

No. of animals per sex per dose:

6 rats

Control animals:

not specified

Sex:

not specified

Dose descriptor:

LC0

Effect level:

0.06 ppm

Based on:

test mat.

Exp. duration:

8 h

There was no mortality or clinical effects observed in rats exposed to an essentially saturated vapor for 8 hours. Rats gained weight, 65 to 97 g, during the observation period.

Conclusions:

Tetraethylene glycol had an extremely low order of acute toxicity following inhalation routes of administration. No hazard is anticipated from the infrequent inhalation of substantially saturated vapor generated at room temperature under normal handling conditions.

Executive summary:

Rats were exposed for 8 hours to an essentially saturated vapor concentration. Rats were observed for mortality, clinical symptoms, body weight gain and gross pathological changes. There were no toxicological effects noted in rats following 8 hour exposure to an essentially saturated vapor concentration of tetraethylene glycol.

Using a vapor pressure of 0.0000465 mm Hg at 25C corresponds to a saturated vapor concentration of 0.061 ppm (0.49 ug/L).

Reference 3

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented GLP study not to guideline

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

Followed the specific protocol and standard protocol amendment prepared by the Bushy Run Research Center.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague-Dawley, Inc., Indianapolis, IN
- Age at study initiation: 52-58 days
- Housing: 5/cage for prestudy quarantine and postexposure periods; individually during exposure
- Diet (e.g. ad libitum): ad libitum except during exposure
- Water (e.g. ad libitum): ad libitum except during exposure
- Acclimation period: approx. 2 weeks
- Photoperiod (hrs dark / hrs light): 12/12
- Temperature: 18-23 degrees C
- Humidity: 41-62%

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The chamber was made of glass and stainless steel. It was rectangular with a pyramidal top and bottom. The height was 1.32 meter, the length was 0.91 meter, and the width was 0.85 meter.
- Exposure chamber volume: 1300-liter
- System of generating particulates/aerosols: TEG was metered from a piston pump (Fluid Metering Inc., Oyster Bay, NY) into an atomizer (Spraying Systems Co., Wheaton, IL) fitted with a No. 1650 liquid nozzle and a No. 64 air nozzle. The atomizer was positioned in the top of the inhalation chamber turret where the liquid aerosol was diluted to the desired concentration and dispersed throughout the chamber by filtered supply air.
- Method of particle size determination: The particle size distribtuion of the TEG aerosol was obstained twice during each exposure using a Sierra 8-stage cascade impactor (Series 210, Sierra Instruments, Inc., Carmel Valley, CA). Cellulose filters (Grade 41, 47 mm, Whatman International Ltd., Maidstone, England) were sued to collect the aerosol on the stages and the filters were assayed gravimetrically. The mass median aerodynamic diameter and geometric standard deviation were determined using a probit analysis method (Finney 1952; Litchfield and Wilcoxon, 1949).
- Temperature, humidity, pressure in air chamber: 20-24 degrees C, 31-65% humidity

TEST ATMOSPHERE
- Brief description of analytical method used: The concentration of TEG in the chamber was measured approximately every 30 minutes. A known volume of air was sampled from the breathing zone of the animals through pre-weighed glass fiber filters (Type A/E, 47 mm, Gelman Sciences, Inc., Ann Arbor, MI). Following the collection of the aerosol from the chamber atmosphere, filters were weighed (without drying). The nominal concentration was determined by dividing the weight of the test material delivered by the volume of air which passed through the chamber during the exposure period.
- Samples taken from breathing zone: yes

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: The target concentration for the frist exposure was 5 mg/l based on the guidelines for "limit" testing set forth by the Toxic Substances Control Act (TSCA). The target concentrations for subsequent groups were based on the observed mortality of previous groups.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

7.5, 5, 3.9, 2.5 mg/l (target concentrations)
6.7, 5, 3.9, 2.6 (measured concentrations)
45.5, 17.1-29.3, 11.0, 7 mg/l (nominal concentration based on a specific gravity at 20/20 degrees C of 1.126 g/ml of TEG)

No. of animals per sex per dose:

5 males and 5 female/dose group
5 females only in 5 mg/l repeat group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed daily for signs of toxicity; weighed prior to exposure, on postexposure days 7 and 14
- Necropsy of survivors performed: yes

Statistics:

The mean and standard deviation of body weights, body weight changes, and exposure concentrations were calculated. No statistical comparisons were made.

Dose descriptor:

LC50

Effect level:

> 3.9 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

In the first 5 mg/l exposure, 5/5 females died on post exposure days 2 (n=3) and 3 (n=2) and 0/5 males died. No deaths were observed in the 6.7, 5 (repeat), 3.9, or 2.6 mg/l dose groups.

Clinical signs:

other: Observation of animals was obstructed by the aerosol during exposure (all groups). Clinical signs observed for all groups upon removal from the chamber included wet (oily) fur and perinasal and periocular encrustation. Additional signs observed for the 6.

Body weight:

A loss of body weight or depressed body weight gain was observed for rats of the 5.0 and 3.9 mg/L groups during the first week of the postexposure period. However, body weight gain was observed for all animals in these groups during the second week postexposure (relative to body weight gains observed during the first week postexposure). Mean body weight gain was observed for the 6.7 and 2.6 mg/L groups (both sexes) on postexposure Days 7 and 14.

Gross pathology:

Macroscopic lesions observed in female rats that died included skin covered with an oily substance (test material), periocular and perinasal redness and/or encrustation, ulceration (nares and tongue), dry shrunken eyes, and discoloration of the lungs, stomach, thymus, and pituitary. The discoloration in the stomach was observed in the glandular region and consisted of multiple red and white foci. Additionally, a black mucus was observed in the stomach in one rat. The only treatment-related macroscopic lesions observed in rats sacrificed at the end of the 2-week recovery period were brown discoloration of the kidneys (2 male rats, 5.0 mg/L group) and dark red discoloration of the caudate lobe of the liver (1 female rat, 3.9 mg/L group).

Other findings:

There were no microscopic lesions that could be attributed to exposure to TEG. The lesions encountered were incidental, inconsistent, and occurred in only a few rats. The lesions were minimal to mild, perivascular cuffings in the lungs (only one or two lobes affected) and tubular basophilia/
mineralization in the kidneys. There was no evidence of renal tubular degeneration or necrosis. Congestion was observed in the lungs of the deadfemales from the 5.0 mg/L exposure group. In some rats, fibrin emboli were present in the small pulmonary vessels and diapedesis of erythrocytes into alveolar spaces had occurred. The kidneys from three of the dead female rats were examined and all three were congested, with autolytic changes evident in two of the three. The cause of death for the female rats could not be determined from the lungs and kidneys examined.

		Deaths
Exposure Concentration of TEG (mg/L)	Sex	During Exposure	Postexposure Day 2	Postexposure Day 3	Total Incidence
6.7	M	0	0	0	0/5
6.7	F	0	0	0	0/5
5	M	0	0	0	0/5
5	F	0	3	2	5/5
5 (repeat)	F	0	0	0	0/5
3.9	M	0	0	0	0/5
3.9	F	0	0	0	0/5
2.6	M	0	0	0	0/5
2.6	F	0	0	0	0/5

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Migrated information

Conclusions:

Results of this study indicate that the 4-hr LC50 value for TEG aerosol in Sprague-Dawley rats is greater than 3.9 mg/L. The mortality observed in female rats of the 5.0 mg/L group could not be duplicated when repeated at the same or higher concentrations. One difference in these exposures was the particle size of the aerosol. A higher particle size was observed for the 5.0 and 6.7 mg/L groups in comparison to the other three groups. This factor may have resulted in a difference in deposition of the test material in the lungs.

Executive summary:

Results of this study indicate that the 4-hr LC50 value for TEG aerosol in Sprague-Dawley rats is greater than 3.9 mg/L. The mortality observed in female rats of the 5.0 mg/L group could not be duplicated when repeated at the same or higher concentrations. One difference in these exposures was the particle size of the aerosol. A higher particle size was observed for the 5.0 and 6.7 mg/L groups in comparison to the other three groups. This factor may have resulted in a difference in deposition of the test material in the lungs.

Reference 4

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: acceptable, well-documented publication which meets basic scientific principles

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

Followed the specific protocol and standard protocol amendment prepared by the Bushy Run Research Center.

GLP compliance:

yes (incl. QA statement)

Remarks:

testing lab.

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

The rats were assigned unique identification numbers by ear tags and were housed 5 per sex in stainless steel, wire-meah cages in 2 rooms for the prestudy quarantine and postexposure periods, respectively. The rats were kept on a 12-hour photoperiod throughout the entire study. Pelletal feed and tap water were available ad libitum except during exposure.
TEG was metered from a platon pump into an atomizer fitted with liquid nozzle and a No. 64 air nozzle.
The atomizer was positioned in the top of the inhalation chamber turret where the liquid aerosol was diluted to the desired concentration and dispersed throughout the chamber by filtered supply air.
The target concentration for the exposure was 5 mg/L, based on the guidelines for limit testing set forth by the Toxic Substances Control Act (TSCA).
The concentration of TEG in the chamber was measured approximately every 30 minutes.
The nominal concentration was determined by dividing the weight of the test substance delivered by the volume of air which passed through the chamber during the exposure period.
The temperature and relative humidity in the animal housing room were recorded continuously with a 7-day recording hygrothermograph.
Animals were observed for signs of toxic effects on the day of exposure and daily following exposure.
The animals were weighed prior to exposure and on post-exposure days 7 and 14.
A complete necropsy was performed for all animals.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

other: no data

Duration of exposure:

4 h

Concentrations:

5.2 mg/l

No. of animals per sex per dose:

5

Control animals:

no

Statistics:

The mean and standard deviation of the body weights, body weight changes, and exposure concentrations were calculated. No statistical comparisons were made.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.2 mg/L air

Exp. duration:

4 h

Remarks on result:

other: No mortalities.

Mortality:

No mortality was observed.

Clinical signs:

other: Periocular wetness, blepharospasm wet (oily fur), absence of toe and tail pinch reflexes. Unkempt fur was the only sign observed during the post-exposure period.

Gross pathology:

No macroscopic lesions were observed in animals sacrificed at the end of the 2-week post-exposure period.

A mean (+/- SD) TEG concentration of 5.23 (±0.15) mg/l was obtained.

The nominal concentration was 19.3 mg/l.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The results of this study indicate that the LC50 value for TEG aerosol in rats is > 5.2 mg/l.

Reference 5

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

1985-1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Sufficient information for interpretation of results

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

Performed according to Standard Test Procedures of Bushy Run Research Center

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200-300g
- Diet (e.g. ad libitum): ad libitum except during exposure or manipulation
- Water (e.g. ad libitum): ad libitum except during exposure or manipulation

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

Sprague-Dawley albino rats, weighing between 200 and 300 g, are exposed to substantially saturated vapor for 6 hours. The vapor is produced by passing air (at 2.5 liters/min) through the sample and then through a 9~liter animal chamber (dynamic conditions). If deaths occur, exposure times are varied to determine an LT50. Five males and 5 females are Included for each exposure period.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

6 h

Concentrations:

Described as a substantially saturated vapor. Using a vapor pressure of 0.000655hPA/ 0.00049129 mm Hg at 24.7C corresponds to a saturated vapor concentration of 0.646 ppm.

No. of animals per sex per dose:

5/sex

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: day 0, 7, 14
- Necropsy of survivors performed: yes

Statistics:

No additional information.

Remarks on result:

other: Exposure to a dynamically-generated, substantially saturated vapor of TEG resulted in no deaths.

Mortality:

No deaths occurred.

Clinical signs:

other: None noted.

Body weight:

Body weight increased during post-exposure period.
Males were 257 +/- 11.4 g on day 0, 283 +/- 14.7 g on day 7, and 296 +/- 15.4 g on day 14.
Females were 211 +/- 6.8 g on day 0, 216 +/- 7.6 g on day 7, and 219 +/- 7.4 g on day 14.

Gross pathology:

No findings.

Conclusions:

Exposure to a dynamically-generated, substantially saturated vapor of TEG resulted in no deaths among 5 male or 5 female rats during or following the 6-hour test. There were no signs of toxic effect and necropsy revealed no remarkable findings.

Executive summary:

Exposure to a dynamically-generated, substantially saturated vapor of TEG resulted in no deaths among 5 male or 5 female rats during or following the 6-hour test. There were no signs of toxic effect and necropsy revealed no remarkable findings.

Reference 6

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study following OECD guideline 403 (1981)

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

GLP compliance:

yes

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

other: Alpk:APfSD

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Rodent Breeding Unit, Alderley park, Macclesfield, Cheshire, UK
- Age at study initiation: 8-9 wkks
- Weight at study initiation: 297.4 +/- 3.6 g (males); 214.4 +/- 3.6 g (females)
- Housing: 5/cage, sexes separately
- Diet (e.g. ad libitum): ad libitum, except during exposure
- Water (e.g. ad libitum): ad libitum, except during exposure
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 degrees C
- Humidity (%): 30-70%
- Air changes (per hr): at least 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: PERSPEX exposure chamber
- Exposure chamber volume: 27.6 liters, minimum change of 12 air changes/hour
- Method of holding animals in test chamber: restrained in polycarbonate tubes supplied by Battelle, Geneva, Switzerland that were inserted into the PERSPEX exposure chamber
- Rate of air: 26l/minute
- Method of conditioning air: dried and filtered using equipment supplied by Atlas-Copco, Sweden
- System of generating particulates/aerosols: glass concentric-jet atomiser
- Method of particle size determination: Aerodynamic particle size distribution was measured twice during the exposure period using a Marple Cascade Impactor (supplied by Schaeffer Instruments, Wantage, Oxon., UK), which aerodynamically separated airborne particles into pre-determined size ranges. The amount of aerosol, by weight, in each size range, was then used to calculate the aerodynamic particle size distribution of the aerosol. Using a microcomputer, the data were transformed using a log/probit transform and a linear regression derived from the cumulative data. Using this regression line, the mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) were calculated.
- Temperature, humidity, pressure in air chamber: temperature: 20.7-22.9 degrees C; humidity 28-32%

TEST ATMOSPHERE
- Brief description of analytical method used: The test substance was pumped to the atomiser using a peristaltic pump supplied by Watson Marlow. Clean, dry air was passed through the atomiser at a nominal flow rate of 26 l/minuted (at normal temperature and pressure) and carried the atmosphere to the exposure chamger, having an internal volume of 27.6 liters, in order to achieve a minimum of 12 air changes per hour. no diluting air was emplyed so the flow rate through the exposure chamber was the same as that employed in the generation of the test atmosphere

TEST ATMOSPHERE
- Measured particulate concentration (Mean +/- SD): 5.08 +/- 0.28 mg/l
- MMAD and GSD: at 1 hour and 12 minutes, the MMAD was 2.83 micron and the GSD was 2.05; at 2 hours and 58 minutes, the MMAD was 2.52 micron and the GSD was 2.05

CLASS METHOD
- Rationale for the selection of the starting concentration: A maximum atmospheric concentration of 5 mg/l was selected as the target exposure level as recommended by the OECD and other national and international guidelines on toxicity testing.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5 mg/l (target concentration)
5.08 +/- 0.28 mg/l (measured)

No. of animals per sex per dose:

The study consisted of 1 main group of 5 male and 5 female rats. A second group of 2 males and 2 females was used for trial exposures to the target concentration.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Frequency of weighing: on day -1, 1, 8 and prior to termination on day 15
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no deaths during the exposure or observation periods.

Clinical signs:

other: During exposure: Abnormalities generally associated with restraint (wet fur, stains around the snout and chromodacryorrhoea) were observed in most animals during exposure. All animals were salivating following 2 hours of exposure. Immediately after expos

Body weight:

All except 1 female animal had gained weight by day 8 of the study (this animal exceeded its starting weight by day 15) and animals continued to gain weight to the end of the study.

Gross pathology:

Speckled thymus was found in 1/5 males and is considered to be a common spontaneous lesion and unrelated to treatment.
Enlarged cervical lymph nodes were found in 1 male and 1 female. This may be a reactive change to changes in the nasal cavity; however, because of the low occurrence could not be ascribed to DEG exposure.

- Measured particulate concentration (Mean +/- SD): 5.08 +/- 0.28 mg/l

- MMAD and GSD: at 1 hour and 12 minutes, the MMAD was 2.83 micron and the GSD was 2.05; at 2 hours and 58 minutes, the MMAD was 2.52 micron and the GSD was 2.05

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Nose only exposure for 4 hours to a particulate concentration of 5.08 mg/l resulted in no deaths and no toxic effects. It is concluded that the median lethal concentration of diethylene glycol exceeds 5.08 mg/l.

Executive summary:

Nose only exposure for 4 hours to a particulate concentration of 5.08 mg/l resulted in no deaths and no toxic effects. It is concluded that the median lethal concentration of diethylene glycol exceeds 5.08 mg/l.

Reference 7

Endpoint:

acute toxicity: inhalation

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Sufficient information for interpretation of results.

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

Performed according to Standard Test Procedures at Bushy Run Research Center

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200-300 g
- Diet (e.g. ad libitum): ad libitum except during periods of exposure or manipulation
- Water (e.g. ad libitum): ad libitum except during periods of exposure or manipulation

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

Sprague-Dawley albino rats, weighing between 200 and 300 g, are exposed to substantially saturated vapor for 6 hours. The vapor is produced by passing air (at 2.5 liters/min) through the sample and then through a 9-liter animal chamber (dynamic conditions). If deaths occur, exposure times are varied to determine an LT50. Five males and 5 females are included for each exposure period.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

6 h

Concentrations:

Described as a substantially saturated vapor. Using a vapor pressure of 0.008 hPA/ 0.006 mm Hg at 25C corresponds to a saturated vapor concentration of 7.89 ppm.

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: day 0, 7, 14
- Necropsy of survivors performed: yes

Statistics:

No data

Remarks on result:

other: Exposure to a dynamically-generated, substantially saturated vapor of DEG resulted in no deaths.

Mortality:

No deaths were observed

Clinical signs:

other: None noted.

Body weight:

Male rats had a body weight of 257 +/- 12 on day 0, 286 +/- 15.9 on day 7 and 307 +/- 17.5 on day 14.
Female rats had a body weight of 218 +/- 9.2 on day 0, 218 +/- 7.3 on day 7 and 225 +/- 9 on day 14.

Gross pathology:

Nothing remarkable at gross pathology.

Other findings:

No additonal information.

Conclusions:

Exposure to a dynamically-generated, substantially saturated vapor of DEG resulted in no deaths among 5 male or 5 female rats during or following the 6-hour test. There were no signs of toxic effect and necropsy revealed no remarkable findings.

Executive summary:

Exposure to a dynamically-generated, substantially saturated vapor of DEG resulted in no deaths among 5 male or 5 female rats during or following the 6-hour test. There were no signs of toxic effect and necropsy revealed no remarkable findings.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 080 mg/m³ air

Quality of whole database:

The database includes LC50 studies conducted on diethylene glycol and triethylene glycol. Studies on tetraethylene glycol involve only inhalation of substantially saturated vapor. The database is sufficiently robust to draw conclusions regarding acute inhalation toxicity of these compounds as a formulation.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1974-1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: non-GLP, non-guideline

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

LD50 calculation

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 3-5 months
- Diet (e.g. ad libitum): ad libitum, except during period of manipulation or confinement
- Water (e.g. ad libitum): ad libitum, except during period of manipulation or confinement

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Rabbits are immobilized furing the 24 hour contact period with the compound retained under impervious sheeting on the clipped intact skin of the trunk. Thereafter, excess fluid is removed to prevent ingestion. Maximum dose that can be retained is 16 ml/kg.

Duration of exposure:

24 hours

Doses:

5, 10, 20 ml/kg

No. of animals per sex per dose:

4 males/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

LD50s are calculated by the moving average method based on a 14 day observation period

Sex:

male

Dose descriptor:

LD50

Effect level:

11.2 mL/kg bw

Based on:

test mat.

95% CL:

ca. 5.28 - ca. 23.9

Mortality:

At 20 ml/kg, 3 of 4 rabbits died, all on day 5. At 10 ml/kg, 2 of 4 rabbits died on days 5 and 7. No deaths occurred at 5 ml/kg.

Clinical signs:

other: No clinical signs were observed. No skin irritation was observed.

Gross pathology:

In victims, lungs and livers were kard and kidneys were pale. In survivors, kidneys were pale and mottled.

Interpretation of results:

GHS criteria not met

Remarks:

LD50 (11.2 ml/kg) does not meet criteria for classification

Conclusions:

Slightly toxic following acute covered dermal application.

Executive summary:

The LD50 was 11.2 ml/kg (about 12500 mg/kg based on a density of 1.118 at 20 degrees C) for rabbits exposed to diethylene glycol for 24 hours under occlusive conditions.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

Not specified but between 15 March and 17 April 1972

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed prior to GLP but sufficient data is available for interpretation of results.

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

other: albino

Sex:

male

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Duration of exposure:

24 hours

Doses:

10 or 20 ml/kg

No. of animals per sex per dose:

4 male rabbits/dose level

Control animals:

not specified

Sex:

male

Dose descriptor:

LD50

Effect level:

20 mL/kg bw

Based on:

test mat.

95% CL:

5.6 - 71.5

Remarks on result:

other: CL is ml/kg

There was no mortality noted in rabbits dosed dermally with 10 ml/kg. Minimal weight change was noted in rabbits during the 14 day observation period (one rabbit lost weight (1021 grams) while the remaining rabbits gained weight (77, 406 and 521 g). Erythema and ecchymosis were noted at the application site. There were no other observations noted.

Two of 4 rabbits dosed dermally with tetraethylene glycol at 20 mL/kg died (days 5 and 9 of the study) during the 14 day observation period. Body weights of the surviving rabbits were minimally affected (one rabbit lost 95 grams while the other gained 120 g). Erythema and ecchymosis were noted at the application site. Clinical signs and symptoms observed included lethargy.

Interpretation of results:

GHS criteria not met

Remarks:

LD50 (~22600) does not meet criteria for classification of TTEG

Conclusions:

Based on the results of the study, it was concluded that Tetraethylene Glycol had an extremely low order of toxicity following covered dermal application for 24 hours. LD50 value was found to be 20.0 (5.60 to 71.5) ml/kg, undiluted.

Executive summary:

The acute dermal LD₅₀ in rabbits was examined. Based on the results of the study, it was concluded that Tetraethylene Glycol had an extremely low order of toxicity following covered dermal application for 24 hours. LD50 value was found to be 20.0 (5.60 to 71.5) ml/kg, undiluted.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: basic information given

Justification for type of information:

Read across is based on the category approach. Please refer to attached category document.

Principles of method if other than guideline:

LD50 calculation

GLP compliance:

not specified

Test type:

standard acute method

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Type of coverage:

other: intact skin of the trunk

Vehicle:

not specified

Details on dermal exposure:

Animals weighing between 2.0 and 3.0 kg are subjected to 24 hours of contact with TEG which is retained under impervious sheeting on the clilpped, intact skin of the trunk. As necessary for larger doses, gauze is wrapped around the trunk over the sample to prevent leakage. Vetrap Bandaging Tape is wrapped over the impervious sheeting and the animal is returned to its cage for the contact period. Doses are varied by adjusting the volume or weight of the test material. Solids are dosed as powders and are moistened with a sufficient amount of water or other suitable vehicle to form a paste. After the contact period, excess fluid is removed to diminish ingestion. Observations for skin reaction are made at one hour, 7 days and 14 days after the contact period. 5 males and 5 females are included on each level used for the LD50 calculation.

Duration of exposure:

24 hours

Doses:

16.0 ml/kg

No. of animals per sex per dose:

5

Control animals:

not specified

Statistics:

no data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 16 mL/kg bw

Based on:

test mat.

Remarks on result:

other: 1 female animal died.

Mortality:

males: 0/5
females: 1/5

Clinical signs:

other: There were no cutaneous effects observed on any animal during the observation period. The only abnormal signs noted were emaciation (in one female) and abdominal distention (in 2 females)

Gross pathology:

Necropsy of the animal that died revealed gas-filled intestines.

None of 5 male rabbits died from 16.0 ml/kg. One of 5 females died from this dose. There were no cutaneous effects observed on any animal during the observation period. The only abnormal signs noted were emaciation (in one female) and abdominal distention (in 2 females). One female rabbit died at 6 days. Necropsy of the animal that died revealed gas-filled intestines. The afore-mentioned signs, the one death and the necropsy findings could be attributable to a spontaneous intestinal disorder often observed in this species rather than to chemical toxicity. Gross pathologic evaluation of survivors revealed tan lungs (of one female), liquid-filled stomach and intestines (one female) and slight vascularization of the treated skin (one male).

Interpretation of results:

GHS criteria not met

Remarks:

LD50 (18000 mg/kg) does not meet criteria for classification

Conclusions:

The LD50 for rabbits exposed to TEG dermally for 24 hours under occlusive conditions is >16 ml/kg (~ >18000 mg/kg).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

12 500 mg/kg bw

Quality of whole database:

The database includes LD50 studies conducted on all the ethylene glycol components and is sufficiently robust to draw conclusions regarding acute dermal toxicity of these compounds as a formulation.

Additional information

Acute Oral Toxicity

Acute oral toxicity of DEG, TEG, and TTEG has been assessed in rat studies. However, clinical data following DEG intoxication is also available and is the basis for the harmonized acute oral H302 classification based on the human findings for DEG. Overall, humans demonstrate lower threshold for acute toxicity than the laboratory animals do.

In a study by Carnegie-Mellon University (1979), administration of 16 and 32 ml/kg DEG to groups of five male Wistar rats/dose by stomach intubation resulted in the deaths of all 32 ml/kg exposed rats and no 16 ml/kg exposed rats. The LD50 was >17900 mg/kg. In groups of ten male Sprague Dawley rats/dose exposed to 1, 5, 10, 12.5, 16, and 17.6 ml/kg DEG, death of 5 rats at 17.6 ml/kg resulted in a LD50 of 19600 mg/kg (Lenk et al., 1989). Hydrotropic degeneration of the renal tubules was observed in some animals at high doses. Laug et al., (1939) reported an observed LD50 value of 1120 mg/kg in humans, based on a mass poisoning incident with Sulfanilamid (10%) diluted in 72% DEG.

In an acute oral toxicity study with TEG by Bushy Run Research Center (1990), five male and five female Sprague-Dawley rats were exposed to 16 ml/kg TEG by intubation. No mortality was observed; the LD50 was >18000 mg/kg.

In a study by Bushy Run Research Center (1986), no deaths were observed in five male and five female Sprague-Dawley rats exposed to 16 ml/kg TTEG by oral gavage; the LD50 was >18000 mg/kg. In another study of male rats exposed to TTEG, deaths occurred in 6/10, 5/6 and 3/4 at doses of 37500, 40000, and 50000 mg/kg, respectively (Mellon Institute for Union Carbide Corporation, 1940). The LD50 was reported as 30000 mg/kg. In a study by the Mellon Institute for Union Carbide Corporation (1938), an LD50 of 33000 mg/kg was found in male albino rats. An acute oral LD50 of > 3980 mg/kg for Polyol PS was found in a study by The Dow Chemical Company (1961), in which no deaths were observed but very slight liver and kidney injury was noted at the high dose of 3980 mg/kg. In a study by the Mellon Institute for Union Carbide Corporation (1972), an LD50 of 30.8 ml/kg (~34000 mg/kg) was found for male Wistar derived rats. A 1976 study by BASF determined the LD50 of TTEG to be >11240 mg/kg for male and female rats; no mortalities were observed.

While animal data do not suggest an acute oral toxicity hazard from DEG, TEG, or TTEG, there have been multiple accounts of poisoning from DEG exposure, demonstrating that humans are more sensitive to acute oral DEG exposure than rats. In a case-control study with laboratory toxic evaluations by O’Brien (1989), the outcome of a poisoning incident in Haitian children is described. 24% DEG from contaminated glycerin in acetaminophen syrup was associated with the deaths of 88 of 109 patients who presented with renal failure, hepatitis, pancreatitis, central nervous system impairment, and/or coma. The mean estimated toxic dose of DEG was 1.34 ml/kg (0.22-4.42 ml/kg, range). Ferrari and Giannuzzi (2005) describe an intoxication incident in Argentina where 15 victims used propolis syrup as an upper respiratory infection medicinal agent. Three victims survived 3 days, three between 4 and 5 days, and nine survived between 6 and 21 days. DEG was found in the viscera and blood of 3 victims. Syrup was found to contain 65% DEG and 32% propylene glycol. The lethal dose was estimated as 0.014 to 0.170 mg DEG/kg bw. A review article by Schep et al. (2009) notes that most documented cases of DEG poisoning have occurred where DEG was substituted in pharmaceutical preparations. The clinical effects of DEG poisoning are separated into three stages: the first phase consists of gastrointestinal symptoms with evidence of inebriation and developing metabolic acidosis. If poisoning is pronounced, patients can progress to a second phase with more severe metabolic acidosis and evidence of emerging renal injury which, in the absence of appropriate supportive care, can lead to death. If patients are stabilized, they may then enter the final phase with various delayed neuropathies and other neurological effects, sometimes fatal. Initial treatment consists of appropriate airway management and attention to acid-base abnormalities. Schep et al. (2009) further state that prompt use of fomepizole or ethanol is important in preventing the formation of the toxic metabolite 2-hydroxyethoxyacetic acid (HEAA) and that hemodialysis and assisted ventilation can also be necessary.

Though animal studies do not suggest acute oral toxicity for DEG, TEG, or TTEG, evidence of DEG toxicity in humans suggests that the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade will be acutely toxic to humans via the oral route, as well.

Justification for selection of acute toxicity – oral endpoint
Of the di-tri- and tetraethylene glycols (DEG, TEG, and TTEG, respectively) comprising this mixture, DEG has harmonized classification of Acute Toxicity 4, H302 based on the human findings. Therefore, the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade is likewise classified as acutely toxic category 4, H302 via the oral route. 

Acute Inhalation Toxicity

Acute toxicity via the inhalation route was determined for DEG, TEG, and TTEG. In a key study by Central Toxicology Laboratory (2000), five male and five female Alpk:APfSD rats were exposed to 5.08 mg/L DEG for 4 hours via nose only aerosol inhalation. No deaths were observed. Similarly, a supporting study by Bushy Run Research Center (1986) showed no deaths in Sprague Dawley rats exposed, whole body, to substantially saturated DEG vapor for 6 hours and MAK documentation (1995) defines the LC50 for DEG to be above 4.6 mg/L in rats.

In a key study by Bushy Run Research Center (1991), five male and five female Sprague-Dawley rats received whole body exposure to 5.2 mg/L TEG for 4 hours. No mortality occurred. In a supporting study by Bushy Run Research Center (1990), five Sprague-Dawley rats/sex/dose were exposed to whole body TEG at concentrations of 2.6, 3.9, 5, and 6.7 mg/L for 4 hours. 100% mortality occurred in females exposed to 5 mg/L. No mortality occurred in a repeat exposure of five female rats to 5 mg/L or at the higher concentration of 6.7 mg/L. The 4 hr LC50 was considered greater than 3.9 mg/L. An additional supporting study by Bushy Run Research Center (1986) showed that no mortality occurred in male and female Sprague-Dawley rats exposed, whole body, to substantially saturated TEG vapor for 6 hours.

A supporting study by Bushy Run Research Center (1986) showed no mortality in male and female Sprague Dawley rats exposed for 6 hours to substantially saturated TTEG vapor. Similarly, Carnegie-Mellon University (1972) showed no mortality in Wistar rats exposed to substantially saturated TTEG vapor for 8 hours. A supporting study by The Dow Chemical Company (1967) showed that acute inhalation exposure of rats for 2 hours to saturated TTEG vapors generated at 200ºC produced total mortality. However, no fatalities were observed when the duration of exposure was one hour. L(Ct)50 was estimated to be 671 mg.min./L. Although the authors described the exposure as essentially a saturated vapor atmosphere, the nominal concentration of 6.1 mg/L is substantially higher than that based on vapor pressure. Using a vapor pressure of 0.0000465 mm Hg at 25^oC corresponds to a saturated vapor concentration of 0.061 ppm (0.49 ug/L). Thus either the difference is mist (aerosol) or test material settled out of the airstream. Dow (1961) also described the results of a 7 hour inhalation of TTEG substantially saturated vapor generated at 110ºC. In this study, no mortality was observed, but some lung congestion and hemorrhage was observed after 1 to 3 days and slight liver and kidney injury were noted. The results of this supporting study suggest some slight upper respiratory and lung irritation from prolonged inhalation of test material heated to 110ºC. 

Based on these data, low inhalation toxicity is expected for the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade.

Justification for selection of acute toxicity – inhalation endpoint

Acute inhalation studies provide LC50s of >5.08 mg/L and >5.2 mg/L for DEG and TEG, respectively, both above the limit dose for acute inhalation toxicity studies. The lower LC50 value was applied to the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade.

Acute Dermal Toxicity

Acute dermal toxicity has been determined for diethylene glycol (DEG), triethylene glycol (TEG), and tetraethylene glycol (TTEG).

In the key acute dermal toxicity study by Carnegie-Mellon University (1979), 5 male New Zealand White rabbits were exposed to 5, 10 or 20 ml/kg DEG under occluded conditions. A LD50 of 11.2 ml/kg (~12500 mg/kg) was observed. MAK documentation (1995) provides a similar LD50 of 13300 mg/kg bw for rabbits exposed to DEG under occluded conditions.

In a supporting acute dermal toxicity study by Bushy Run Research Center (1990), five New Zealand White rabbits/sex were dermally exposed to 16 ml/kg TEG under occlusion for 24 hours. The only death occurred in one female rabbit. The LD50 was >16 ml/kg (~18000 mg/kg).

In an acute dermal toxicity study with TTEG by Carnegie-Mellon University (1972), 24 hour occluded dermal exposure to the test material in male rabbits resulted in death of 2 rabbits at the high dose (20 ml/kg) and no deaths at the low dose (10 ml/kg). The LD50 for TTEG in this supporting study was 20 ml/kg (~22600 mg/kg). An additional supporting study by Bushy Run Research Center (1986), found that 24 hour exposure of male and female New Zealand White rabbits to 16 ml/kg TTEG under occluded conditions resulted in no mortality.

Based on these data, low dermal toxicity is expected for the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade.

Justification for selection of acute toxicity – dermal endpoint
All the individual di-tri- and tetra ethylene glycols (DEG, TEG, and TTEG, respectively) comprising this mixture, have dermal LD50 values many times the current limit dose of guideline studies. Therefore, no acute dermal classification is proposed for the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade.

Justification for classification or non-classification

Acute toxicity: via the oral route

DEG, TEG, and TTEG, components of the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade, demonstrate low acute toxicity after oral exposure in rats Based on known acute toxicity to humans, DEG is classified as Acute Tox 4, H302 according to Annex VI under Regulation (EC) No 1272/2008. Therefore, the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade is likewise classified as Acute Tox 4, H302 according to Annex VI of (EC) No 1272/2008.

Acute toxicity: via the inhalation route

No deaths were observed upon inhalation of substantially saturated vapor of individual DEG, TEG, or TTEG mixture components in acute inhalation toxicity studies. Moreover, no deaths were observed after 4 hr nose only inhalation of 5.08 mg/L DEG by male and female Alpk:APfSD rats or after 4 hr whole body exposure to 5.2 mg/L TEG by male and female Sprague-Dawley rats. Therefore, the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade does not require classification under GHS.

Acute toxicity: via the dermal route

The individual DEG, TEG, and TTEG mixture components demonstrate low acute dermal toxicity. Their observed LD50 values were many times the current limit dose of guideline studies Therefore, the reaction mass of 3,6,9-trioxaundecane-1,11-diol and 2,2'-oxydiethanol and 2,2'-(ethylenedioxy)diethanol and 3,6,9,12-tetraoxatetrade is not expected to be acutely toxic via the dermal route and should not be classified according to GHS.