Dimethyldioctylammonium chloride

Endpoint summary

Description of key information

In an unreliable acute oral toxicity study the substance was administered to mice (10 animals/dose) by oral gavage at a dose level of 375 mg/kg bw (single administration). Fifty percent of the animals died within 24 hours of treatment. Haemorrhages in stomach and intestine with pneumonia were observed during gross pathology. The oral LD50 in mice is 375 mg/kg bw.

In a reliable acute toxicity study, a structurally similar substance was administered to Sprague Dawley rats (5 animals/sex/dose) by oral gavage at a dose level of 500, 300, 200 or 100 mg/kg bw (single administration). Mortalities were observed at all dose groups except for in the 100 mg/kg bw group. The most prevalent clinical changes for all dose groups noted during the observation period included urine and fecal stains, saliva discharge and stains, dried red stains on muzzle and around eyes, eye squinting, piloerection, ataxia, body tremors, laboured and shallow respiration, slight to severe depression, viscous red blood like discharge from mouth, bloated appearance to the abdomen, and spasms in the abdominal area. The most common abnormalities found at macroscopic post-mortem examination included effects in the liver, kidneys, stomach, intestines and the spleen. The oral LD50 is 238 mg/kg bw.

In an unreliable acute inhalation toxicity study the substance was administered to mice (5 animals/sex/dose) by inhalation at a concentration of 10 mg/L air for 1 hour. No animals died. The inhalation LD50 in rats is >10 mg/L air.

There are four acute dermal toxicity studies for the substance in the rabbit. The LD50s were 259 mg/kg bw, 1.51 ml/kg bw, 247 mg/kg bw (liquid) and 191 mg/kg bw (solid). The most prevalent clinical sign of toxicity across all studies was eschar formation and the solid form of the substance was found to produce less severe toxic effects compared with the liquid form. In one study the dermal administration of the substance resulted in the animals going into physiological decline and all died within 96 hours of exposure.

In an acute dermal toxicity study a structurally similar substance was administered to New Zealand White rabbits (5 animals/sex/dose) by occlusive dressing at dose levels of 552, 1104, 3328 or 4448 mg/kg bw (single administration). A decrease in mean body weight was observed in all treatment groups in a dose response relationship. There was an increased incidence of pale kidneys in animals at the highest treatment level as compared to controls. Distended atria and/or ventricles were also noted in three animals at the highest dose levels. One animal in the 3328 mg/kg bw treatment group was noted to have a heavily pigmented gel in the large intestine. The acute dermal toxicity of the substance is 3342 mg/kg bw in the rabbit. The substance was a corrosive as evidenced by adverse signs at the site of application.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4th June 1991 - 13th August 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

On day 1 on the screening portion of the study, the test animals receiving 0.3 g/kg bw of test material inadvertently did not receive a PM observation. This devaition has not compromised any aspect of the study as reported by the study author.

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 225 - 399 g
- Fasting period before study: yes, fasted overnight prior to dosing
- Housing: groups of two in wire mesh suspension cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: four days before being used

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The test material was administered undiluted and as a 20% w/v formulation in distilled water. Based on the results from this initial phase, a dilution of 5% w/v in distilled water was selected for administration.

Doses:

0.5 g/kg bw
0.3 g/kg bw
0.2 g/kg bw
0.1 g/kg bw

No. of animals per sex per dose:

Five/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: All animals were observed closely for gross signs of systemic toxicity and mortality several times during the day of dosing, and at least twice daily thereafter for a total of 14 days. Body weights was measured for each animal on the day of dosing, on day 7 of the observation period or following the death of any animals which does not survive this period.
- Necropsy of survivors performed: A gross necropsy was performed on any animal which dies. A gross necropsy was performed on each surviving animal at the end of the study.
- Other examinations performed: clinical signs, body weight,organ weights.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

238 mg/kg bw

Based on:

test mat.

95% CL:

> 0.198 - < 0.287

Mortality:

At the top dose of 0.5 g/kg bw mortality occurred in all 10 animals.
Seven out of 10 animals died at 0.3 g/kg bw dose.
Four out of 10 animals died following dosing of 0.2 g/kg bw.
No mortality was observed at the 0.1 g/kg bw dose.

Clinical signs:

other: The most prevalent clinical changes for all dose groups noted during the observation period included urine and fecal stains, saliva discharge and stains, dried red stains on muzzle and around eyes, eye squinting, piloerection, ataxia, body tremors, labour

Gross pathology:

Gross necropsy findings in one animal which survived the observation period included an enlarged spleen, the stomach wall appeared transparent, a lobe of the liver, the stomach, and the spleen appeared to be attached together by a membrane-like structure. With the exception of the above animal, all other animals which survived the observation period exhibited no gross pathological findings. In the animals that were found dead the following common findings were observed: Lungs, spleen and liver mottled, stomach wall white in colour and distended with gas, stomach contained substance/paste, intestines were yellow and greatly distended, kidneys appeared pale and congested.

Phase 1:

Dose (g/kg bw)	Concentration	Mortality  (no. dead/no.dosed)
4.0	Undiluted	10/10
1.0	Undiluted	9/10
0.4	Undiluted	10/10
0.16	Undiluted	7/10
0.0632	20% w/v in distilled water	3/10

Phase 2:

Dose (g/kg bw)	Mortality (no. dead/no.dosed)
0.5	10/10
0.3	7/10
0.2	4/10
0.1	0/10

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The rat oral LD50 is 0.238 g/kg bw in male and female animals, with 95% confidence limites of 0.198 g/kg bw to 0.287 g/kg bw.

Executive summary:

In an acute toxicity study the substance was administered to Sprague Dawley rats (5 animals/sex/dose) by oral gavage at a dose level of 500, 300, 200 or 100 mg/kg bw (single administration). Mortalities were observed at all dose groups except for in the 100 mg/kg bw group. The most prevalent clinical changes for all dose groups noted during the observation period included urine and fecal stains, saliva discharge and stains, dried red stains on muzzle and around eyes, eye squinting, piloerection, ataxia, body tremors, laboured and shallow respiration, slight to severe depression, viscous red blood like discharge from mouth, bloated appearance to the abdomen, and spasms in the abdominal area. The most common abnormalities found at macroscopic post-mortem examination included effects in the liver, kidneys, stomach, intestines and the spleen. There were no adverse effects noted for body weight gain.The oral LD50 is 238 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

238 mg/kg bw

Quality of whole database:

Sufficient to address requirements.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

USDA Regulation 362.8, Title 7, Chapter III, Part 362, Code of Federal Regulations; Paragraph 362.116.

Deviations:

not specified

Principles of method if other than guideline:

- Principle of test: Inhaltion of substance for 1 hour.
- Short description of test conditions: Not specified.
- Parameters analysed / observed: Not specified.

GLP compliance:

not specified

Test type:

fixed concentration procedure

Limit test:

yes

Species:

rat

Strain:

not specified

Sex:

male/female

Route of administration:

inhalation

Type of inhalation exposure:

not specified

Vehicle:

not specified

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

1 h

Concentrations:

10 mg/L

No. of animals per sex per dose:

5 animals/sex/dose.

Control animals:

not specified

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 10 mg/L air

Based on:

test mat.

Exp. duration:

1 h

Remarks on result:

not determinable

Mortality:

None.

Clinical signs:

other: Not specified.

Body weight:

Not specified.

Gross pathology:

Not specified.

Dose	Animals	No.	Survivors
10 mg/L/1 hour	Male rats	5	5
10 mg/L/1 hour	Female rats	5	5

Interpretation of results:

study cannot be used for classification

Conclusions:

The LD50 is >10 mg/L air based on an acute inhalation toxicity study (1 hour exposure) in which no animals died.

Executive summary:

In an acute toxicity study the substance was administered to mice (5 animals/sex/dose) by inhaltion at a concentration of 10 mg/L air for 1 hour. No animals died. The inhalation LD50 in rats is >10 mg/L air.

.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

10 mg/m³ air

Quality of whole database:

Sufficient to address requirements.

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:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: As described in ''Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics'', published by the Association of Food & Drug Officials of the United States p. 54.
- Short description of test conditions: A single administration of substance onto the skin of animals and observed for 14 days.
- Parameters analysed / observed: Mortality, clinical effects and gross pathology.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

not specified

Sex:

not specified

Type of coverage:

not specified

Vehicle:

not specified

Details on dermal exposure:

The hair was removed from the back and flanks by clipping. The skins of two animals at each dose level were further prepared by making epidermal abrasions approximately 2-3 cm apart longitudinally over the treated sites. All treated animals were observed for 14 days and any surviving animals were autopsied.

Duration of exposure:

Not specified.

Doses:

1.0, 1.25, 1.5, 1.75 or 2.0 ml/kg bw.

No. of animals per sex per dose:

4 animals/dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Not specified
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

1.51 mL/kg bw

Based on:

test mat.

95% CL:

> 1.23 - <= 1.86

Mortality:

All rabbits that died did so within 8 days of dosage.

Clinical signs:

other: All treated animals stopped eating and drinking immediately after the substance was applied. Intense hyperemia and edema developed over each treated site and eschar formation was observed over each treated area.

Gross pathology:

Gross pathology in all animals showed pneumonic lungs, cloudy swelling in liver and kidneys with some petechiae, stomach, intestines, omentum and peritoneum hypermic, with the absence of perirenal fat and paucity of subcutaneous fat.

Dose (ml/kg bw)	Response
1.0	0/4
1.25	1/4
1.5	2/4
1.75	3/4
2.0	4/4

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The dermal LD50 in rabbits is 1.51 ml/kg bw. Based on the observations the substance is considered to be corrosive to the skin and toxic by the dermal route as established by the LD50.

Executive summary:

In an acute dermal toxicity study a 50% solution of the substance was administered to rabbits (4 animals/dose) by dermal administration at a dose levels of 1.0,1.25, 1.5, 1.75 or 2.0 ml/kg bw (single administration). The duration of the exposure was not specified. All animals that died did so within 8 hours of administration. Each treated animals had intense hyperemia and edema development over each treated site and eschar formation was observed over each treated area. Gross pathology in all animals showed pneumonic lungs, cloudy swelling in liver and kidneys with some petechiae, stomach, intestines, omentum and peritoneum hypermic, with the absence of perirenal fat and paucity of subcutaneous fat. The dermal LD50 in rabbits is 1.51 ml/kg bw.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3rd March 1987 - 17th March 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: to assess the short term toxicity of a test article in rabbits when administered by the dermal route.
- Short description of test conditions: The animals were acclimitised for 5 days prior to treatment. The fur from the dorsal area of the trunk of the test animals was clipped on the day of treatment. The substance was applied uniformly over the clipped area and held in contact with the skin for a 24 hour exposure period using gauze. Residual substance will be removed following the the exposure period using water (if possible).
- Parameters analysed / observed: Bodyweight, mortality, toxic effects, cage side effects (skin, fur, eyes, mucous membranes, respiratory, circulatory automic and central nervous system, somato-motor activity and behaviour patterns). Upon necropsy gross pathology examinations.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor provided the substance.

STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
- Final dilution of a dissolved solid, stock liquid or gel: The volume of liquid product administered to each animal was adjusted based on the stated 80% activity of the material to achieve the speific treatment level.

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Gota-Frisco Farms, Edon, OH (USA)
- Females (if applicable) nulliparous and non-pregnant:not specified
- Weight at study initiation: 2.704 to 2.745 kg
- Housing: Housed individually in suspended stainless steel cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle.

Type of coverage:

occlusive

Vehicle:

ethanol

Details on dermal exposure:

TEST SITE
- Area of exposure: Dorsal area of the trunk
- % coverage: 10% of the animal's total body surface.
- Type of wrap if used: A layer of 8 ply gauze dressing, a lyer of rubber dam and several wrappings of 3 inch Elastoplast tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with water.
- Time after start of exposure: 24 hours after treatment.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.06, 0.2, 0.42 or 0.83 ml/kg bw
- Concentration (if solution): 8, 160, 336 or 664 mg/kg bw
- Constant volume or concentration used: not specified

Duration of exposure:

24 hours.

Doses:

8, 160, 336 or 664 mg/kg bw

No. of animals per sex per dose:

5 animals/sex/dose

Control animals:

yes, concurrent vehicle

Remarks:

10% v/v denat. ethanol in water

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed for mortality and toxic effects for 8 hours after dosing and daily thereafter.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, gross pathology.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

259 mg/kg bw

Based on:

test mat.

95% CL:

> 82 - <= 639

Mortality:

No deaths occurred in the vehicle control animals and in the 8 mg/kg bw group (lowest dose).
In the 160 mg/kkg bw group 4 out of 10 animals had died.
In the 336 mg/kkg bw group 6 out of 10 animals had died.
In the 664 mg/kkg bw group 7 out of 10 animals had died.

Clinical signs:

other: The substance produced significant dermal irritation in the animals which was characterised by eschar formation at the treatment site. In all animals, eschar formation was evident by the day following dose administration. A dose-response related incidence

Gross pathology:

There were a number of gross pathologic findings reported throughout both the test and control groups with no apparent dose-response relationship. The findings in the dead animals could be an artefact of post-mortem changes, such as increased incidence of colour changes in kidneys and lungs. There was a single animal with white material in the lungs from a treatment group, this was atributed to pulmonary infection in this rabbit. Cardiac anomalies was noted in several animals from the treatment groups with no apparent dose-response relationship.

Treatment	Dose	No. of Deaths/No. Dosed
		Males	Females	Sexes Combined
10% Ethanol/Water	5.6 ml/kg bw	0/5	0/5	0/10
Substance	8 mg/kg bw	0/5	0/5	0/10
	160 mg/kg bw	2/5	2/5	4/10
	336 mg/kg bw	3/5	3/5	6/10
	664 mg/kg bw	4/5	3/5	7/10

Clinical Finding(s)	Incidence of Finding in Treatment Groups
	Vehicle Control	8 mg/kg bw	160 mg/kg bw	336 mg/kg bw	664 mg/kg bw
Soft stools and/or faecal stain	1/10	-	-	1/10	1/10
Few faeces	1/10	4/10	6/10	8/10	4/10
Laboured breathing	-	-	-	1/10	2/10
Prostration	-	-	-	-	1/10
Nasal discharge	-	1/10	-	-	-
Activity decreased	-	1/10	2/10	3/10	5/10
Tremors	-	-	-	-	2/10
Ataxia	-	-	-	1/10	-

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The dermal LD50 in rabbits is 259 mg/kg bw. Based on the observations the substance is considered to be corrosive to the skin and toxic by the dermal route as established by the LD50.

Executive summary:

In an acute dermal toxicity study an 80% solution of the substance was administered to rabbits (5 animals/sex/dose) by dermal administration at dose levels of 8, 160, 336 or 664 mg/kg bw (single administration) for a duration of 24 hours. A number of animals died in the top 3 dose groups and clinical toxicity was observed in the treatment animals, this included significant dermal irritation characterised by eschar formation at the treatment site. In all animals, eschar formation was evident by the day following dose administration. A dose-response related incidence of clinical effects included reduced faeces, laboured breathing and decreased activity. Prostration, nasal discharge, tremors and ataxia was observed in the top dose only. Mean body weights in the top 3 dose groups was reduced, but generally a partial or complete recovery of body weights was observed in the majority of animals that survived to termination. The dermal LD50 in rabbits is 259 mg/kg bw.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13th January 1987 - 28th January 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 434 (Acute Dermal Toxicity - Fixed Dose Procedure)

Version / remarks:

The top dose in this study was 1000 mg/kg bw

Deviations:

yes

Remarks:

In the 50 mg/kg bw group there were four males and six females dosed, rather than fiver per sex.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

Two products were tested in this study:
Liquid (86% active - 860 mg/ml active in 10% v/v ethanool and water)
Solid (93.5% active)

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Females (if applicable) nulliparous and non-pregnant: not specified
- Weight at study initiation: approximately 2.0 to 3.0 kg
- Housing: Suspended wire mesh bottom cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12 hour light/dark cycle

Type of coverage:

occlusive

Vehicle:

other: 10% ethanol/water

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: 10% of the body surface area
- Type of wrap if used: 8 ply gauze dressing covered with rubber dam and secured with several wrappings of 3 inch Elastoplast tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the test article will be removed with water
- Time after start of exposure: 24 hours post exposure

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0 ml/kg bw
- Concentration (if solution): 50, 100, 250, 500, 750 or 1000 mg/kg bw

Duration of exposure:

14 days.

Doses:

50, 100, 250, 500, 750 or 1000 mg/kg bw

No. of animals per sex per dose:

Five/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

A liquid and solid formulations of the substance was evaluated. The liquid was administered as provided at 860 mg/ml active in 10% v/v ethanol/water.The solid substance was dissolved in 10% v/v ethanol/water prior to dosing.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were weighed on the day of dosing (day 1) and on days 8 and 15 of the study. All animals which die on test after day 1 were also weighed. Animals were observed for mortality and toxic effects continuously for 8 hours post dose and daily thereafter until day 15. Cage side observations included evaluations of the skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nrevous systems, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, lethargy, other signs of central nervous system depression, salivation and diarrhoea.
- Necropsy of survivors performed: yes
- Other examinations performed: the contents of all body cavities were subjected to gross examinations with special attention given to the heart muscle and its auxialiary vascular system.

Statistics:

The LD50 with 95% confidence interval and the slopes of the response curve were calculated using the method of probit analysis when possible, otherwise an alternative, acceptable method was used and documented.

Key result

Sex:

female

Dose descriptor:

LD50

Remarks:

Liquid

Effect level:

183 mg/kg bw

Based on:

test mat.

95% CL:

> 56.9 - < 387.5

Key result

Sex:

male

Dose descriptor:

LD50

Remarks:

Liquid

Effect level:

336.3 mg/kg bw

Based on:

test mat.

95% CL:

> 157.6 - < 637.8

Key result

Sex:

male/female

Dose descriptor:

LD50

Remarks:

Liquid

Effect level:

247.1 mg/kg bw

Based on:

test mat.

95% CL:

> 141.7 - < 396.4

Key result

Sex:

female

Dose descriptor:

LD50

Remarks:

Solid

Effect level:

170.3 mg/kg bw

Based on:

test mat.

95% CL:

> 26.8 - < 397.1

Key result

Sex:

male

Dose descriptor:

LD50

Remarks:

Solid

Effect level:

213.6 mg/kg bw

Based on:

test mat.

95% CL:

> 71 - < 448.7

Key result

Sex:

male/female

Dose descriptor:

LD50

Remarks:

Solid

Effect level:

191.4 mg/kg bw

Based on:

test mat.

95% CL:

> 93.3 - < 321.4

Mortality:

No deaths occurred in the vehicle control animals.
Individual mortality data are presented in the tables below.

Clinical signs:

other: Dermal irritation was characterised by discolouration of the test site and subsequent eschar formation for both the solid and liquid formulations. The dermal response to the solid formulation was less severe than to the liquid version; this was particular

Gross pathology:

For both the liquid and reconstituted solid treatments, the most frequently noted findings included apparent distention of the atria and/ or ventricles and multifocal dark red foci on the thymus. Other necropsy findings which were common to the two test treatments included abdominal subcutaneous oedema, continued contractions of the heart, motttled and pale kidneys, distended large intestine, and multiple dark red foci on the lungs. In general, the occurrence of the aforementiuoned findings was similar between the liquid and the reconstituted solid treatments.

Necropsy findings in liquid product treated animals included distention of the gallbladder, cecum, small intestine, and stomach, occlusion of the bile duct, and fluid within the stomach.

Necropsy findings in reconstituted solid product treated animals included clear gelatinous material within the large intestine, mottled and plae lungs, multiple white foci on the lungs, and mottled thymus.

Mortality Data:

Treatment	Dose Level	No. of Deaths/No. Dosed
		Males	Females	Sexes Combined
10% Ethanol/Water (vehicle control)	2.0 ml/kg bw	0/5	0/5	0/10
Liquid	50	0/4	1/6	1/10
	100	1/5	3/5	4/10
	250	2/5	1/5	3/10
	500	2/5	3/5	5/10
	750	4/5	5/5	9/10
	1000	5/5	5/5	10/10
Solid	50	1/5	2/5	3/10
	100	1/5	2/5	3/10
	250	3/5	3/5	6/10
	500	3/5	2/5	5/10
	750	4/5	5/5	9/10
	1000	5/5	5/5	10/10

Conclusions:

The rabbit dermal LD50 of the liquid product is 336 mg/kg bw (males), 183 mg/kg bw (females) and 247 mg/kg bw (combined).
The rabbit dermal LD50 of the reconstituted solid product is 214 mg/kg bw (males), 170 mg/kg bw (females) and 191 mg/kg bw (combined).

Executive summary:

In an acute toxicity study the liquid (80%) and reconstituted solid substance (93.5%) was administered to New Zealand White rabbits (5 animals/sex/dose) by occlusive patch at dose levels of 50, 100, 250, 500, 750 or 1000 mg/kg bw (single administration). The substance was left on the test site for 24 hours and then washed off. No adverse body weight effects were observed following administration of the liquid product at 50 or 100 mg/kg bw, or following administration of the reconstituted solid product at 50 mg/kg bw. At the higher dosage groups for each test article, a dose-dependent decrease in mean body weight was noted in both the males and females. Mortality was observed in a dose-dependent manner. Signs of clinical toxicity included dermal irritation as characterised by discolouration of the test site and subsequent eschar formation for both the solid and liquid formulations. The dermal response to the solid formulation was less severe than to the liquid version. A number of adverse effects were observed during gross pathology investigations for both the liquid and reconstituted solid treatments. The most frequently noted findings included apparent distention of the atria and/ or ventricles and multifocal dark red foci on the thymus. Other necropsy findings which were common to the two test treatments included abdominal subcutaneous oedema, continued contractions of the heart, mottled and pale kidneys, distended large intestine, and multiple dark red foci on the lungs. In general, the occurrence of the aforementioned findings was similar between the liquid and the reconstituted solid treatments. Necropsy findings in liquid product treated animals included distention of the gallbladder, cecum, small intestine, and stomach, occlusion of the bile duct, and fluid within the stomach. Necropsy findings in reconstituted solid product treated animals included clear gelatinous material within the large intestine, mottled and pale lungs, multiple white foci on the lungs, and mottled thymus. The rabbit dermal LD50 of the liquid product is 336 mg/kg bw (males), 183 mg/kg bw (females) and 247 mg/kg bw (combined). The rabbit dermal LD50 of the reconstituted solid product is 214 mg/kg bw (males), 170 mg/kg bw (females) and 191 mg/kg bw (combined).

Reference 4

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

3rd March 1987 - 17th March 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

The dosages were changed to: Group Treatment Level (mg/kg bw) 1 552 2 1104 3 3328 4 4448

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

Substance was stored at room temperature at all times.

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Housing: Suspended stainless steel wire mesh bottom cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days prior to initiation of study.

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle.

Type of coverage:

occlusive

Vehicle:

other: 10% v/v solution of ethanol/water.

Details on dermal exposure:

On the day prior to dosing, the fur was clipped from the dorsal area of the trunk of each rabbit. The exposed area on the trunk of each animal measured approximately 12x20 cm and constitutred 10% of the animal's total body surface. On the following day, the appropriate material was applied uniformly over the exposed skin, or as much of the area as possible. An occlusive binder was secured around the trunk of each animal immediately after treatment.

Duration of exposure:

24 hours.

Doses:

552, 1104, 3328 or 4448 mg/kg bw.

No. of animals per sex per dose:

Five/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed for mortality and toxic effects for 8 hours after dosing and daily thereafter until day 15. Particular attention was given to observations of tremours, convulsions, seizures, lethargy and other signs of cardiac and/or central nervous system toxicity. Individual body weights were determined on days 1, 8 and 15 or at death.
- Necropsy of survivors performed: yes. Animals which died during the study were also subjected to a gross necropsy examination at the time of death or when found dead. During necropsy examinations, particular attention was directed to abnormalities of the heart and its auxialiary vascular system.

Statistics:

LD50 was calculated using the Probit Analysis method.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

3 861 mg/kg bw

Based on:

test mat.

95% CL:

> 0 - < 4 292

Mortality:

Individual mortality data is presented below in the table.
No deaths occurred among the vehicle control animals.

Clinical signs:

other: No clinical effects related to the treatment with the vehicle were noted. The substance was irritating to the skin in all animals at all doses, these effects were characterised by eschar formation at the treatment sites within 24 hours after administratio

Gross pathology:

Gross necropsy findings were minimal. There was an increased incidence of pale kidneys in animals at the highest treatment level as compared to controls.Distended atria and/or ventricles were also noted in three animals at the highest dose levels.One animal in the 3328 mg/kg bw treatment group was noted to have a heavily pigmented gel in the large intestine.

Mortality data:

Treatment	Dose level	No. of Deaths/No. Dosed
		Males	Females	Sexes Combined
10% Ethanol/Water	5.56 ml/kg bw	0/5	0/5	0/10
Substance (80% active)	552 mg/kg bw	0/5	0/5	0/10
	1104 mg/kg bw	0/5	0/5	0/10
	3328 mg/kg bw	3/5	2/5	5/10
	4448 mg/kg bw	5/5	3/5	8/10

Conclusions:

The acute dermal toxicity of the substance is 3342 mg/kg bw in the rabbit. The substance was a corrosive as evidenced by adverse signs at the site of application.

Executive summary:

In an acute dermal toxicity study an 80% solution of the substance was administered to New Zealand White rabbits (5 animals/sex/dose) by occlusive dressing at a dose levels of 552, 1104, 3328 or 4448 mg/kg bw (single administration). The exposure was for 24 hours after which the animals were observed for 14 days and observed for mortlaity, signs of clinical toxicity, changes to body weight gain and gross pathology was conduted at necropsy. There was dose-dependent mortalities and signs of clinical toxicity. A decrease in mean body weight was observed in all treatment groups in a dose response relationship. Gross necropsy findings were minimal. There was an increased incidence of pale kidneys in animals at the highest treatment level as compared to controls.Distended atria and/or ventricles were also noted in three animals at the highest dose levels.One animal in the 3328 mg/kg bw treatment group was noted to have a heavily pigmented gel in the large intestine. The acute dermal toxicity of the substance is 3342 mg/kg bw in the rabbit. The substance was a corrosive as evidenced by adverse signs at the site of application.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

191 mg/kg bw

Quality of whole database:

Sufficient to address requirements.

Justification for classification or non-classification

On the basis of the results of a reliable acute oral toxicity study on a structurally related substance, the substance is classified as Acute tox 3 (H301). On the basis of the lowest acute dermal LD50 value, the substance is classified as Acute tox 2 ( H310).