Reaction mass of carbon tetrachloride and chloroform and 1,2-dichloroethane

Endpoint summary

Description of key information

Acute toxicity, oral: Based on the harmonized classifications of the constituents and the rules of the CLP Regulation for classification of mixtures, the substance is classified as Cat.3 (H301)

Acute toxicity, inhalation: Based on the harmonized classifications of the constituents and the rules of the CLP Regulation for classification of mixtures, the substance is classified as Cat.3 (H331).

Acute toxicity, dermal: Based on the harmonized classifications of the constituents and the rules of the CLP Regulation for classification of mixtures, the substance is classified as Cat.3 (H311)

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:

experimental study

Adequacy of study:

weight of evidence

Study period:

1982

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Very few information, animals were observed for 14 days but no information on the number of animals and tested dose. Only LD50 values are reported.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

not specified

GLP compliance:

no

Test type:

other: no data

Limit test:

no

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, Mass., U.S.A.
- Age at study initiation: 6 weeks old
- Fasting period before study: 18 hours prior to dosing
- Housing: In goups of four animals in shoebox plastic cages with sawdust bedding
- Diet: Agway Lab Chow ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 70-75 degree F (about 21-24 degree C)
- Humidity: 40-60 %
- Photoperiod: 12 hours dark/ 12 hours light

Route of administration:

oral: gavage

Vehicle:

not specified

Details on oral exposure:

Solutions of dichloroethane were prepared fresh daily for acute studies and the appropriate concentrations were administered by gavage in a volume of 0.01 mL/g bw to achieve the desired dose. The chemical was administered to male and female mice by an 18-gauge stainless steel stomach tube 18 hours after fasting from food.

Doses:

no data

No. of animals per sex per dose:

no data

Control animals:

not specified

Details on study design:

Following gavage, the mice were observed for overt toxicological effects continously for 4 hours and then twice daily for 14 days. Mice dying during the experimental procedure were necropsied, and gross pathological changes were described. Mice surviving the 14-day observation period were sacrificed and gross pathology described.
Gross pathology: brain, liver, spleen, lungs, thymus,  kidneys, testes (organ weight)

Statistics:

Log probit analysis was used to determine the LD50 and the 95% confidence limits.

Preliminary study:

no data

Sex:

male

Dose descriptor:

LD50

Effect level:

489 mg/kg bw

95% CL:

424 - 552

Sex:

female

Dose descriptor:

LD50

Effect level:

413 mg/kg bw

95% CL:

337 - 499

Mortality:

The mice died over a 48-hour period. Those surviving 48 hours recovered and appeared normal at the end of the 14-day observation period.
LD50 (male) = 489 mg/kg bw (95 % CL: 424-552 mg/kg bw); LD50 (female) = 413 mg/kg bw (95 % CL = 337-499 mg/kg bw).

Clinical signs:

no data

Body weight:

no data

Gross pathology:

Target organs appeared to be liver and lungs.

Other findings:

none

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Oral administration of 1,2-dichloroethane to mice by gavage:
LD50 (male) = 489 mg/kg bw; LD50 (female) = 413  mg/kg bw

Executive summary:

In an acute oral toxicity study, groups of fasted (18 hours), 6 -week-old CD-1 mice were given a single oral dose of 1,2 -dichloroethane and observed for 14 days.

The mice died over a 48 -hour period. Those surviving 48 hours recovered and appeared normal at the end of the 14 -days observation period. Macroscopically, target organs appeared to be liver and lungs. The calculated LD50 values using log probit analysis were 489 mg/kg bw (95% C.I. 424 -552 mg/kg bw) and 413 mg/kg bw (95% C.I. 337-499 mg/kg bw) for male and female mice, respectively.

1,2 -dichloroethane is of low acute oral toxicity based on the LD50 in male and female CD-1 mice.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is in line with the relevant guideline studies (such as OECD Guideline No. 401) with acceptable restrictions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

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:

Biobreeding Labs, Ottawa, Ontario, 150-200 g, acclimated for 1 week, temperature 21 +/- 2 °C, humidity 40-70 %, caged individually and allowed free access to food and water

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

administration at a constant volume of 5 mL/kg, animals were fasted overnight before dose administration

Doses:

546, 765, 1071, 1500, 2100 mg/kg body weight

No. of animals per sex per dose:

10 males, 10 females

Control animals:

no

Details on study design:

To establish the dose levels to be used in the LD50 studies, groups of two animals were administered test compounds at 250 and 3000 mg/kg and the mortality data were recorded. Then, the main LD50 studies were carried out with doses of 546-2100 mg/kg using 10 female and 10 male rats. Clinical observations were made for 14 days after dosing and a postmortem examination was performed on animals which died during this period. All surviving animals were killed at the end of 14 days and subjected to gross pathologic examination.

Statistics:

Statistical analysis of data was carried out using one-way analysis of variance and the Student-Newman-Keuls test. LD50 values were calculated using the probit analysis (Finney 1952).

Sex:

male

Dose descriptor:

LD50

Effect level:

908 mg/kg bw

95% CL:

>= 750 - <= 1 082

Sex:

female

Dose descriptor:

LD50

Effect level:

1 117 mg/kg bw

95% CL:

>= 843 - <= 1 514

Mortality:

See also Table 1: At the highest dose level, 4 males died within 24 hours after dosing and most animals were dead after 4 days post-administration. At lower doses animals died primarily on Day 3 to Day 4 post-administration.

Clinical signs:

Common clinical signs consisted of piloerection, sedation, flaccid muscle tone, ataxia, and prostration. Chloroform elicited dacryorrhea in some animals.

Body weight:

No changes in body weight

Gross pathology:

Increased relative weights of the liver and the kidneys. For lesions see Table 2

Other findings:

A clear trend towards increased cholesterol levels was seen in all treated animals. Inhibition of lactate dehydrogenase activities were seen in all treated groups. A significant decrease in the levels of liver proteins were observed in rats fed chloroform. Microsomal hydroxylase activity of female but not male rats was activated by administration of chloroform. A reduction in lymphocytes with significant decreased was noted in female rats. Decrease in haemoglobin and haematocrit values were observed in males and females fed chloroform.

Table 1: Mortality occurring in the acute oral toxicity study:

Dose (mg/kg)	Sex	Total animals dead*	Survival time (days after oral administration)							LD50 (mg/kg)
			1	2	3	4	5	6	7
546	Male	0	--	--	--	--	--	--	--	M:908(750-1082) F:1117(843-1514)
	Female	1	--	--	1	--	--	--	--
765	Male	5	1	--	1	1	2	--	--
	Female	3	--	--	1	2	--	--	--
1071	Male	6	2	--	2	--	2	--	--
	Female	6	--	2	2	--	--	2	--
1500	Male	9	--	--	5	1	2	1	--
	Female	5	--	1	3	1	--	--	--
2100	Male	10	4	--	3	1	1	1	--
	Female	9	--	--	4	5	--	--	--

*Animals dying before the end of the 14 day observation period

Table 2: Incidence of histological changes of rats which survived single oral doses of chloroform

Doses (mg/kg)	Liver		Kidneys
	Males	Females	Males	Females
546	0/2	1/2	02	0/2
765	2/2	0/2	1/2	1/2
1071	--	0/2	--	2/2
1500	0/1	0/2	0/1	1/2
2100	--	0/1	--	0/1

Numbers denote: number of animals showing lesions/animals examined

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Due to the acute oral toxicity test in the rat chloroform is classified as harmful if swallowed.

Executive summary:

The acute oral toxicity of chloroform, solubilised in corn oil, was tested in female and male Sprague-Dawley rats (10 animals per dose group) in accordance with the OECD Guideline No. 401 with minor restrictions. Chloroform was administered at doses of 546, 765, 1071, 1500 and 2100 mg/kg body weight by oral gavage. Animals were then observed for 14 hours, clinical signs were noted and animals which were dying during the observation period or were killed after 14 days were submitted to necropsy and gross pathology. The mean LD50 values determined for male rats was 908 mg/kg body weight, that of female rats was 1117 mg/kg. Following the GHS classification system, chloroform on the basis of the present study is classified into Toxicity Category IV, receiving the hazard statement "harmful if swallowed".

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: New testing design followed (in line with OECD TG 401). No individual data, limited reporting of experimental details

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

- this survey study was conducted to evaluate a new strategy for determination of LD50 values with a lower number of animals
- nevertheless in total 11 animals were used for each dose group

GLP compliance:

no

Test type:

standard acute method

Species:

rat

Strain:

not specified

Sex:

male

Details on test animals or test system and environmental conditions:

- Acclimation period: 5 d
- no further details reported

Route of administration:

oral: gavage

Vehicle:

not specified

Details on oral exposure:

- no details reported

Doses:

1500, 2000, 2800, 3900 mg/kg bw

No. of animals per sex per dose:

in total 11

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days or longer until the surviving animals stated gaining weight again
- Frequency of observations and weighing: days 0, 7, 14 and weekly thereafter if necessary
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs daily

Statistics:

Method of Rosiello et al. (RosieUo AP, Essigmana JM, Wogan GN (1977) Rapid and accurate determination of the median lethal dose (LDSo) and its error with a smalI computer. J Toxicol Environm Health 3: 797-809), based on the method by Bliss (1938).

Sex:

male

Dose descriptor:

LD50

Effect level:

2 500 mg/kg bw

Mortality:

1500 mg/kg bw 0/11
2000 mg/kg bw 5/11
2800 mg/kg bw 6/11
3900 mg/kg bw 11/11

Clinical signs:

- not reported

Body weight:

- not reported

Gross pathology:

- not reported

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The present study (Lorke, 1983) states a LD50 (rat, oral) of 2500 mg/kg bw after single oral application via gavage.

Executive summary:

The potential of the test substance CTC (carbon tetrachloride) to induce toxicity upon exposure via the oral route was evaluated in a survey study on different substances following generally OECD TG 401. The scope of the study was to evaluate a new strategy to determine LD50 values with a lower amount of animals. Nevertheless the total amount of tested animals in each dose group allowed for CTC a classical determination of a LD50 value according to OECD TG 401. Male rats (unspecified strain) were treated at a single oral dose bw with CTC (unknown vehicle). Observation period was 14 d and further experimental details were not reported but stated to be conducted according to OECD TG 401.

The mortality incidences were 0/11 at 1500 mg/kg bw, 5/11 at 2000 mg/kg, 6/11 at 2800 mg/kg and 11/11 at 3900 mg/kg.

The LD 50(rat, oral) was therefore assessed to be 2500 mg/kg bw after single oral application via gavage.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

162 mg/kg bw

Quality of whole database:

Based on the rules of the CLP Regulation for calculation of the ATE of mixtures

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: The documentation of the study is not in accordance with current guidelines such as the method B.2 suggested by the European Commission.

Qualifier:

no guideline followed

Principles of method if other than guideline:

Male rats received whole-body exposure to chloroform vapours for 6 hours and were observed for 14 days subsequently.

GLP compliance:

no

Test type:

fixed concentration procedure

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

Rats had a body weight between 130 and 200 g.

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gas chromatograph

Duration of exposure:

6 h

No. of animals per sex per dose:

12 male rats

Control animals:

no

Details on study design:

After exposure animals were observed for 14 days. All animals were subject to necropsy and gross pathology.

Statistics:

The LD50 value and the 95 % confidence interval were determined based on the method of Bliss (1938).

Sex:

male

Dose descriptor:

LC50

Effect level:

1 849 ppm

95% CL:

1 752 - 1 955

Exp. duration:

6 h

Sex:

male

Dose descriptor:

LC50

Effect level:

9.17 mg/L air

95% CL:

8.69 - 9.7

Exp. duration:

6 h

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Chloroform is toxic when inhaled.

Executive summary:

The acute inhalation toxicity of chloroform was tested in male Sprague-Dawley rats exposed to chloroform vapours during 6 hours followed by an observation period of 14 days. The LC50(6h) value determined was 9.17 mg/L and the derived LC50(4h) value was 10.5 mg/L.

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Scientifically sound survey study, but no individual animal data, no information on necropsy data.

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:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sprague Dawley, O.F.A., Iffa-Credo
- Age at study initiation: not reported
- Weight at study end: 130 - 200 g
- Fasting period before study: not reported
- Housing: not reported
- Diet (e.g. ad libitum): not reported
- Water (e.g. ad libitum): not reported
- Acclimation period: not reported


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

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: cubic cages made of stainless steel, glass and teflon
- Exposure chamber volume: 170 L in total separated in 28 independent cages
- Method of holding animals in test chamber: whole-body in the cage
- Source and rate of air: 0 - 50 m³/h
- Method of conditioning air: dry sterile air at 10 °C is provided by a central device and is conditioned in seperate pneumatic regulation devices for each chamber to give a defined temperature, humidity and air rate
- System of generating vapour: the volatile test substance is vaporised by a thermostated injector
- Method of particle size determination: not applicable (vapour of volatile solvent)
- Treatment of exhaust air: the test substance is eliminated from the exhaust air by an undefined method
- Temperature, humidity, pressure in air chamber: 24 °C ± 1, 50 ± 1 %, - 0.294 mbar
- air changes in the exposure chamber: 60/h

TEST ATMOSPHERE
- Brief description of analytical method used: gas chromatography + measurement of volume loss of vaporized solvent
- Samples taken from breathing zone: yes, every 3 minutes by automatic sampling and analysis by gas chromatography

VEHICLE
- no vehicle used

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gas chromatography analysis method of the gas chromatography not mentioned)

Duration of exposure:

6 h

Concentrations:

Not reported in detail, only visible as data point in a figure: 6000- 7000 ppm (38.4 -44.8 mg/l)

No. of animals per sex per dose:

12

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: frequently
- Frequency of weighing: once before treatment and on days 7 and 14 post treatment
- Necropsy of survivors performed: yes

Statistics:

- the LD50 calculations were done by the Bliss method

Sex:

male

Dose descriptor:

LC50

Effect level:

7 228 ppm

95% CL:

ca. 7 072 - ca. 7 378

Exp. duration:

6 h

Mortality:

- not reported

Clinical signs:

other: - hypnotic and somnolence

Body weight:

- retarded growth stated but no values presented

Gross pathology:

- analysed, but no findings, especially not in the liver the lung or the kidneys

Interpretation of results:

GHS criteria not met

Conclusions:

The present study (Bonnet, 1980) states a LC50(rat) of 7228 ppm (= 46.260 mg/L air) after single exposure to CTC for 6 h.

Executive summary:

The potential of the test substance CTC (named carbon tetrachloride in the report) to induce toxicity upon exposure via the inhalation route was evaluated in a study following, along general lines, the OECD guideline No. 403.

Male Sprague Dawley rats were treated with vapors of the test substance for 6 hours by whole body mode. The animals were observed for 14 days post exposure for mortality, body weight development and clinical signs of toxicity, but neither the clinical signs nor the body weight results or the mortality were detailed in the report.

Mortality data of these animals were analysed by Bliss' probit method, which gave an acute inhalation toxicity (LC50) value of CTC in rats of 7228 ppm air (95 % confidence level: 7072 - 7378) after an exposure for 6h.

The LC50 in rat was found to be 7228 ppm (= 46.260 mg/L air) after single exposure to CTC for 6 h.

Reference 3

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1951

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comprehensive and comparative study, basic data given, based on scientific principles apparently meeting current standards.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

not specified

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: albino rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

The albino rats were raised in this laboratory of stock originally obtained from Wistar Institute of Anatomy and Biology in 1938. They were maintained on a modified Sherman diet consisting of freshly ground whole wheat (55 %), dried whole milk (25 %), dried extracted liver (12 %), dried brewer's yeast (5 %), iodized table salt (2 %) and calcium carbonate (1 %).

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

A glass-walled chamber of about 160 L capacity was used. Two large copper tubes (closed with rubber stoppers) were soldered into the model top of the chamber so that rats could be introduced conveniently after a vapour concentration had been established. A constant air flow was maintained through the chamber, the lowest rate for any experiment being 15 L/min and the highest about 30 L/min. The desired vapour concentration was obtained by metering liquid 1,2-dichloroethane at a constant rate into the tube through which air entered the chamber, heat being applied at the point of vaporization as needed to effect complete volatilization.
The rats were introduced in groups of 5 to 12 within a period of 15 sec and were removed through the chamber door within a similar interval of time at the end of the exposure. It was shown by a continuously recording analyzer that the animals were introduced without appreciable alterations of the vapour concentration.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

All vapour concentrations with animals in the chamber were checked repeatedly from time to time by combustion analyses; the results averaged better than 90 % of the calculated theroretical concentrations of 1,2-dichloroethane.

Duration of exposure:

>= 0.1 - <= 8 h

Concentrations:

200, 300, 600, 800, 1000, 1500, 3000, 12000 and 20000 ppm (corresponding to 823, 1234, 2468, 3290, 4113, 6169, 12339, 49354 and 82256 mg/m³) at various exposure times.

No. of animals per sex per dose:

between 10 and 54 rats

Control animals:

not specified

Details on study design:

All the rats were selected on the basis of general appearance and apparent good health, males and females being used in approximately equal numbers. Animals were observed as to their behavior, body weight changes, and time of death. Survivors were observed for two to three weeks, or until it was certain that they had fully recovery of weight.
Evidence of organ pathology: Special, additional groups of animals were killed at various intervals within the time-frame of 0.2-h to 20-h exposure to determine body weight, liver and kidney weight, blood parameters (urea nitrogen concentration, plasma prothrombin clotting time, serum phosphatase activity), liver lipids, and histopathological changes (liver, kidney, adrenals).

Statistics:

Statistics according to the method of Litchfield and Wilcoxon was performed.

Preliminary study:

no data

Sex:

male/female

Dose descriptor:

LC50

Effect level:

7 758 mg/m³ air

95% CL:

ca. 7 485.38 - ca. 8 061.15

Exp. duration:

4 h

Remarks on result:

other: corresponding to approx. 1886 ppm

Mortality:

Mortality occurred at different exposure concentrations and exposure times. For details see the section "Remarks on results including tables and figures".

Clinical signs:

other: Please refer to the section "Remarks on results including tables and figures".

Body weight:

In special groups of animals (exposure causing 99.9, 50 or 0.01 % death), reported signs of exposure mediated toxicity were decreased body weights.

Gross pathology:

In special groups of animals (exposure causing 99.9, 50 or 0.01 % death), reported signs of exposure mediated toxicity were increased liver and kidney weights and slight parenchymatous degeneration to severe haemorrhagic necrosis (kidney, liver, adrenals), congestion (kidney, liver, adrenals, lungs) and oedema (kidney, lungs), increase in blood urea nitrogen concentration, plasma prothrombin clotting time, liver lipids, decrease in serum phospatase activity.

Other findings:

none

The 4-h LC50 corresponded to approx. 1886 ppm (approx. 7758.5 mg/m³) and was derived from the data by Standard Probit analysis (please refer to "Seuils de Toxicité aiguë 1,2-Dichloroéthane from Groupe d’Experts Toxicologues du Ministère de l’Ecologie, de l’Energie, du Développement Durable et de l’Aménagement du Territoire" dated 2008-12-03).

With 22000 ppm (approx. 90420 mg/m³), death occurred within 24 min after deep anaesthesia by depression of the central nervous system. At 12000 ppm and lower concentrations this depressant action resulted in varying degrees of "drunkenness".

In special groups of animals (exposure causing 99.9, 50 or 0.01 % death), reported signs of exposure mediated toxicity were decreased body weights, increased liver and kidney weights and slight parenchymatous degeneration to severe haemorrhagic necrosis (kidney, liver, adrenals), congestion (kidney, liver, adrenals, lungs) and oedema (kidney, lungs), increase in blood urea nitrogen concentration, plasma prothrombin clotting time, liver lipids, decrease in serum phospatase activity.

The following concentrations and exposure times were not lethal:

300 ppm (approx. 1200 mg/m³) after 7 h (20 animals)

600 ppm (approx. 2400 mg/m³) after 5 h (20 animals)

1500 ppm (approx. 6100 mg/m³) after 2 h (10 animals)

3000 ppm (approx. 12100 mg/m³) after 0.5 h (22 animals)

20000 ppm (approx. 81000 mg/m³) after 0.1 h (10 animals)

The following concentrations were void of adverse effects:

200 ppm (approx. 800 mg/m³) for 7 h

300 ppm (approx. 1200 mg/m³) for 3 h (but effects at 5.5 h)

1000 ppm (approx 4000 mg/m³) for 1.5 h (but effects at 3 h).

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The 4-h LC50 in rats derived by Standard Probit analysis was 1886 ppm (approx. 7758 mg/m³).

Executive summary:

In an acute inhalation toxicity study, groups of young adult albino rats (10-54/dose, approximately equal numbers of males and females) were whole body exposed to 1,2 -dichloroethane for 0.1 to 8 hours at concentrations of 200, 300, 600, 800, 1000, 1500, 3000, 12000 or 20000 ppm (corresponding to 823, 1234, 2468, 3290, 4113, 6169, 12339, 49354 and 82256 mg/m³). Animals then were observed for 2 to 3 weeks. At 20000 ppm, death occurred within 24 min after deep anaesthesia by depression of the central nervous system. At 12000 ppm and lower concentrations this depressant action resulted in varying degrees of "drunkenness". According to authors, deaths tended to occur at three different time intervals and in such a manner as to suggest three separate toxic actions of fatal degree:

1. At very high concentrations (e.g. 20000 ppm), deaths occurred due to depression and paralysis of CNS functions.

2. At all vapour concentrations causing death, a large proportion died rather suddenly and quietly a few hours after termination of exposure, showing marked cyanosis, reduced body temperature, stupor or coma and failing respiration. The character and sudden development of this response suggested "shock" or cardiovascular collapse.

3. All other deaths occurred delayed over a period of 2 to 7 days with progressive loss of body weight and other evidence of toxic effects, suggesting organ failure, probably due to kidney lesions.

The 4 -h LC50 derived by Standard Probit analysis in this acute inhalation study in rats was 1886 ppm (approx. 7758 mg/m³).

1,2 -dichloroethane is of low acute oral toxicity based on the LC50 (4 hours) in male and female albino rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

3 680 mg/m³

Quality of whole database:

Based on the rules of the CLP Regulation for calculation of the ATE of mixtures

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: A scientifically sound study but no individual data, no necroscopy or histological data.

Qualifier:

no guideline followed

Principles of method if other than guideline:

0.5 and 2.0 mL of CTC were applied to guinea pigs weighting 372 ± 3 g (low dose group) and 373 ± 5 g (high dose group) to a covered skin depot; the mortality rate was determined after an observation period of 35 d

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

guinea pig

Strain:

not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: not reported
- Age at study initiation: not reported
- Weight at study initiation: 372 ± 3 g low dose group, 373 ± 5 g high dose group
- Fasting period before study: not reported
- Housing: in groups of three in Makrolon cages
- Diet (e.g. ad libitum): standard laboratory animal food (Astra Ewos, Sodcttalje, Sweden), ad libitum
- Water (e.g. ad libitum): water supplemented with ascorbic acid, ad libitum
- Acclimation period: not reported


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

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: 3.1 cm x cm
- % coverage: 0.7 %
- Type of wrap if used: A glass ring, 20 mm in inner diameter (area: 3.1 cm x cm), 4 mm in thickness, and 10 mm in height, glued on with a-cyanoncrylate glue and covered with a glued on glass cover slip (with a hole in the center to apply the substance with was finally covered with small piece glued to the cover glass).
- control animals were treated with distilled water

REMOVAL OF TEST SUBSTANCE
- Washing (if done): substance was completely absorbed
- Time after start of exposure: substance was completely absorbed


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0 and 0.5 ml pure substance



VEHICLE
- none used

Duration of exposure:

- until substance was absorbed

Doses:

- 0.5 and 2.0 ml (= 0.792 and 3.17 g, on average 2130 and 8500 mg/kg bw)

No. of animals per sex per dose:

20 per dose, sex not indicated

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 35 days

- Frequency of weighing: daily, but not reported
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: no

Statistics:

- no statictic analysis conducted

Sex:

not specified

Dose descriptor:

approximate LD50

Effect level:

ca. 2 130 - ca. 8 500 mg/kg bw

Mortality:

after 14 days: 2130 mg/kg bw 5/20
8500 mg/kg bw 12/20
after 35 days: 2130 mg/kg bw 5/20
8500 mg/kg bw 13/20

Clinical signs:

- not analyzed

Body weight:

- not reported

Gross pathology:

- not analyzed

Interpretation of results:

GHS criteria not met

Conclusions:

From the present study (Wahlberg, 1979) the LD50 (guinea pig, dermal) can be approximated to lie between 2130 and 8500 mg/kg bw. in guinea pig.

Executive summary:

The potential of the test substance CTC (named carbon tetrachloride in the study report) to induce acute dermal toxicity was evaluated in a study following no official guideline. Guinea pigs, unspecified concerning sex and strain, were treated with the test substance on the clipped skin under a special occlusive device and were observed for 35 days for mortality and body weight only, but the body weight development was not reported. No pathological or histological analysis were conducted.

Treatment with the the test substance resulted in mortalities of 25 % at 2130 mg/kg bw and 65 % at 8500 mg/kg bw after 35 d of observation with a biphasic occurence with peaks between days 2 -4 and between days 7 -14. As only two concentration were tested the LD50 could only be approximated to lie between the two tested doses. The approximated LD50 was derived to be >2000 mg/kg bw.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1948

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparative study, screening test, basic data given, based on scientific principles, results conclusive.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

not specified

GLP compliance:

no

Test type:

other: no data

Limit test:

no

Species:

rabbit

Strain:

not specified

Sex:

male

Details on test animals or test system and environmental conditions:

no details given

Type of coverage:

occlusive

Vehicle:

not specified

Details on dermal exposure:

no data

Duration of exposure:

no data

Doses:

Occluded applications : 3.16, 3.98, 4.45 and 5.0 mL/kg for 24 h (= 3972, 5000, 5594 and 6285 mg/kg bw)

No. of animals per sex per dose:

6-11 animals

Control animals:

not specified

Details on study design:

no data

Statistics:

Probit analysis

Preliminary study:

no data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

4 890 mg/kg bw

95% CL:

ca. 4 270 - ca. 5 600

Mortality:

Mortality observed within 14 days:
2/6 animals at 3972 mg/kg bw  within 5-10 days
3/11 animals at 5000 mg/kg bw within 1-5 days
8/9 animals at 5594 mg/kg bw within 1-11 days
5/6 animals at 6285 mg/kg bw within 1 day
The LD50 calculated using the method of probits was 3.89 mL/kg bw (3.40-4.46 mL/kg bw) or 4890 mg/kg bw (4270-5600 mg/kg bw).

Clinical signs:

No data

Body weight:

Weight loss was reported in most survivors.

Gross pathology:

Gross necropsy evaluation revealed no adverse effects.

Other findings:

none

Interpretation of results:

GHS criteria not met

Conclusions:

Acute dermal toxicity to male rabbits:
The calculated LD50 by the method of probits was 3.89 mL/kg bw (3.40-4.46 mL/kg bw) or 4890 mg/kg bw (4270-5600 mg/kg bw).

Executive summary:

In an acute dermal toxicity study, groups of male rabbits (6 -11 animals/dose level) were dermally exposed to 1,2 -dichloroethane at doses of 3972, 5000, 5594 or 6285 mg/kg bw and observed for 14 days. Mortality was observed in 2/6 at 3972 mg/kg bw within 5 -10 days, 3/11 at 5000 mg/kg bw within 1 -5 days, 8/9 at 5594 mg/kg bw within 1 -11 days and 5/6 at 6285 mg/kg bw within 1 day. Weight loss was reported in most survivors. Gross necropsy evaluation revealed no adverse effects. The calculated LD50 by the method of probits was 3.89 mL/kg bw (3.40 -4.46 mL/kg bw) or 4890 mg/kg bw (4270–5600 mg/kg bw).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

571 mg/kg bw

Quality of whole database:

Based on the rules of the CLP Regulation for calculation of the ATE of mixtures

Additional information

Some studies performed in animals on the 3 main constituents of the registered substance are reported in this dossier.

Acute toxicity, oral:

Carbon tetrachloride (CTC): Several studies performed in animals are reported in the REACH dossier of this constituent. The lowest value 2500 mg/kg bw (Lorke, 1983) in rat was chosen as the key value in the REACH dossier. In most studies, clinical signs were not reported. However, other studies show that CTC is hepatotoxic after a single oral dose. Moreover, as reported in the ATSDR Toxicological profile on carbon tetrachloride (2005), human fatalities from ingestion of carbon tetrachloride have been reported, usually following intake of a large amount of carbon tetrachloride. Fatalities occurred at lower intake in case of heavy alcohol consumption.

Carbon tetrachloride has an Harmonized classification for Acute oral toxicity: Cat.3 (H301), probably because of the cases of human poisoning following exposure to CTC.

Dichloroethane (DCE): Several studies were perfomed and reported in the REACH dossier of DCE but none of the studies fullfilled completely the requirements from internationally accepted guidelines or were documented with deficiencies. Thus, in a weight of evidence approach out of 5 studies with conclusive results the lowest LD50 of 413 mg/kg bw in mice (Munson, 1982) was chosen for classification/labelling and risk assessment in the DCE dossier.

1,2 -dichloroethane has an Harmonized classification for Acute oral toxicity: Cat.4 (H302).

This worst-case value for the LD50 was used to assess the toxicity of the registered substance based on the constituent approach.

Chloroform: The acute lethal toxicity varies depending on species, strain, sex and vehicle. In the REACH dossier of chloroform, the results from the reliable study in Sprague Dawley rats (Chu et al. 1980) which provided an oral LD50 value of 908 mg/kg body weight in male rats are used as key value for this endpoint. Acute oral exposure to chloroform causes target-organ specific local and systemic effects at lower doses with the liver and kidneys as the target organs.

Chloroform has an Harmonized classification for Acute oral toxicity: Cat.4 (H302).

Acute toxicity, inhalation:

CTC: Several studies of different quality are available in the REACH dossier for this route. In the 4 most appropriate studies, the lowest value for 6-h LC50 was 46.260 mg/m3 (Bonnet, 1980) in rat. The animals elicited signs of somnolence and at necropsy, clear hepatic injury.

Moreover, as reported in the ATSDR Toxicological profile on carbon tetrachloride (2005), inhalation exposure to high concentrations of carbon tetrachloride - most of the time concomitantly to exposure via the dermal route – was reported to be responsible for a considerable number of deaths in humans.

Carbon tetrachloride has an Harmonized classification for Acute inhalation toxicity: Cat.3 (H331), probably because of the cases of human poisoning following exposure to CTC.

DCE: After acute inhalation exposure to 1,2-dichloroethane, LC50-values obtained in rats ranged from 4100 mg/m³/7.2 h to 49400 mg/m³/0.5 h (Spencer et al., 1951). In compliance with these results, another 6-h LC50 was determined at about 1650 ppm (= 6670 mg/m³) in a study reported in the DCE REACH dossier. A 4 -h LC50 in rats of 7758 mg/m³ (= 1886 ppm) was derived from a concentration-response graph (Spencer et al., 1951) and used as the key value.

1,2 -dichloroethane has an Harmonized classification for Respiratory irritation: STOT-SE 3 (H335).

Chloroform: Depression of the central nervous system is the dominant symptom of acute inhalation toxicity of chloroform. The available rodent studies on the acute inhalation toxicity of chloroform are not reliable. However, in sum they give evidence that chloroform exhibits low acute inhalation toxicity. The study performed in male Sprague Dawley rats (Bonnet et al. 1980) resulted in an acute inhalation LC50 value of 9.2 mg/L after 6 hours of exposure. For classification, the relevant duration of exposure is 4 hours. Applying Haber’s law a corrected LC50 value for acute inhalation toxicity of 10.5 mg/L is calculated for chloroform. Acute inhalation exposure to chloroform causes adverse systemic effects also at lower doses in some studies available in the Chlorform dossier.

Chloroform has an Harmonized classification for Acute inhalation toxicity: Cat.3 (H331).

Acute toxicity, dermal:

CTC: Several studies are available to estimate the acute LD50 in its REACH dossier. The lowest value for the acute dermal LD50 was 2130 mg/kg bw in guinea pigs (Wahlberg 1979). No information is available on rats.

In the past, exposure via the dermal route (concimitantly to inhalation exposure) was reported to be responsible for a considerable number of deaths in humans.

Carbon tetrachloride has an Harmonized classification for Acute dermal toxicity: Cat.3 (H311), probably because of the cases of human poisoning following exposure to CTC.

DCE: The mean LD50-value for acute dermal toxicity of 1,2-dichloroethane after application under occluded conditions to rabbits was 4890 mg/kg bw with a 95 % confidence interval of 4270–5600 mg/kg bw (Mellon Inst. Industr. Research, 1948).

Chloroform: No key study was found. Not required based on the Annex VIII of the REACH regulation (EC) No 1907/2006 as inhalation and oral routes were assessed.

Based on the overall animal and human data, the 3 main constituents have both harmonized and self classifications. Following the rules of the CLP for classification of mixtures, the registered substance is therefore considered to be toxic (cat. 3) by inhalation, oral and dermal routes.

For additional information on these constituents, their dossier are available on the ECHA website.

Justification for classification or non-classification

Harmonized classification

The registered substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 (CLP).

Self-classification

Acute toxicity, oral:

Based on the overall data on its 3 main constituents, the registered substance is classified as Cat.3 (H301: Toxic if swallowed) according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Acute toxicity, inhalation:

Based on the overall data on its 3 main constituents, the registered substance is classified as Cat.3 (H331: Toxic if inhaled) according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Acute toxicity, dermal:

Based on the overall data on its 3 main constituents, the registered substance is classified as Cat.3 (H311: Toxic in contact with skin) according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Specific target organ toxicity: single exposure, oral:

Based on the overall data on its 3 main constituents, the registered substance is not classified regarding specific target organ toxicity after single exposure via oral route according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

Specific target organ toxicity: single exposure, inhalation:

Based on the overall data on its 3 main constituents, the registered substance can be classified as STOT-SE Cat.3 (H335: may cause respiratory irritation) regarding specific target organ toxicity after single exposure via inhalation but this classification is deemed unnecessary because more severe organ effects including in the respiratory system are observed in the Acute Toxicity study by inhalation, and is considered covered by the ATI Cat.3 (H331).

Based on the well recognised narcotic effects of chloroform, its self-classification as STOT-SE Cat.3 (H336) in the REACH dossier and its concentration in the registered substance, the multi-constituent substance should be classified as STOT (H336: may cause drowsiness or dizziness).

Specific target organ toxicity: single exposure, dermal:

Based on the overall data on its 3 main constituents, the registered substance is not classified regarding specific target organ toxicity after single exposure via dermal route according to the Annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.