1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates

Endpoint summary

Description of key information

Acute oral Toxicity: 

The acute oral toxicity dose (LD50) for 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) was considered based on data available for the structurally and functionally similar read across chemicals. The LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates cannot be classified for acute oral toxicity. 

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure of humans via inhalation route is not likely taking into account due to the low vapour pressure of the substance 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3), which is reported to be 1.29E-32 mmHg. Thus, exposure to inhalable dust, mist and vapour of the chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates is highly unlikely. Therefore this study is considered for waiver. 

Acute dermal Toxicity: 

The acute dermal toxicity dose (LD50) for 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) was considered based on data available for the structurally and functionally similar read across chemicals. The LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates cannot be classified for acute dermal toxicity. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally and functionally similar read across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on two acute oral toxicity studies as- WoE-2 and WoE-3
Acute Oral toxicity test was carried out to study the effects of the test chemicals on rodents

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Specific details on test material used for the study:

- IUPAC Name: 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates
- Common Name: Basic Brown 1
- Molecular formula : C26H34N8O8
- Molecular weight : 586.597 g/mol
- Smiles notation : CC(=O)O{-}.N{+}c1ccc(N=Nc2cccc(N=Nc3ccc(N{+}.O{-}C(C)=O)cc3N{+}.O{-}C(C)=O)c2)c(N{+}.O{-}C(C)=O)c1
- Substance type : Organic
- Physical state : liquid

Species:

rat

Strain:

other: 1. Tif. RAI rats 2. not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

1. TEST ANIMALS
- Source: No data
- Age at study initiation: 6 to 7 weeks old
- Weight at study initiation: Animals were weighed 160 to 180 g
- Fasting period before study: The rats were starved during one night before starting the treatment.
- Housing: The males and females were segregated and housed in Macrolon cages (Type 3) in groups of 5.
- Diet (e.g. ad libitum): food, ad libitum.
- Water (e.g. ad libitum): water, ad libitum.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1° C
- Humidity (%): approximately 50 %
2. not specified

Route of administration:

other: 1. oral: gavage 2. oral: unspecified

Vehicle:

other: 1. CMC (carboxymethyl cellulose) 2. unchanged (no vehicle)

Details on oral exposure:

1. VEHICLE
- Concentration in vehicle: 50 and 60 %
DOSAGE PREPARATION (if unusual): Test material was suspended at 50 and 60 % with carboxymethylcellulose 2 % and administered by oral intubation.
2. not specified

Doses:

1. 6000 and 10000 mg/kg
2. 5000 mg/kg

No. of animals per sex per dose:

1. Total = 20 (sex/dose)
2. not specified

Control animals:

not specified

Details on study design:

1. - Duration of observation period following administration: 7 days
- Frequency of observations and weighing: Mortality and general symptoms was observed at 1 h, 24 h, 48 h, and 7 days.
- Necropsy of survivors performed: yes
- Other examinations performed: Animals were observed for clinical signs.
2. not specified

Statistics:

1. not specified
2. not specified

Preliminary study:

1. not specified
2. not specified

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 10 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

1. No mortality was observed at 10000 mg/kg bw.
2. No mortality was observed at 5000 mg/kg bw.

Clinical signs:

1. Within 2 hours after treatment the rats of both dosage groups showed dyspnoea, exophthalmus, curved position and ruffled fur. The animals had recovered within 4 to 6 days.
2. not specified

Body weight:

1. not specified
2. not specified

Gross pathology:

1. No substance related gross organ changes were seen.
2. not specified

Other findings:

1. not specified
2. not specified

Interpretation of results:

other: Not classified

Conclusions:

According to CLP regulation the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) cannot be classified for acute oral toxicity, as the LD50 value is >2000 mg/kg bw.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3). The studies are as mentioned below:

1. Acute oral toxicity test was conducted using test chemical in 20 Tif. RAI rats (10 males/10 females), bred under SPF conditions at the dose concentration of 6000 and 10000 mg/kg bw. The test substance was weighed into an Erlenmeyer flask on a Mettler balance. It was suspended at 50 and 60 % with carboxymethylcellulose 2 % and administered by oral intubation. Before treatment the suspension was homogeneously dispersed with an Ultra-Turrax and during treatment it was kept stable with a magnetic stirrer. Mortality and general symptoms was observed at 1 h, 24 h, 48 h, and 7 days. Animals were observed for clinical signs. No mortality was observed at any of the dose in treated rats. Within 2 hours after treatment the rats of both dosage groups showed dyspnoea, exophthalmus, curved position and ruffled fur. The animals had recovered within 4 to 6 days. They were killed and autopsied after an observation period of 7 days. No substance related gross organ changes were seen. Hence, LD50 was considered to be >10000 mg/kg bw, when Tif. RAI rats were treated with test chemical via oral gavage route.

2. Acute oral toxicity study of test chemical was conducted in rats at the concentration of 5000 mg/kg bw. No mortality was observed in treated rats at 5000 mg/kg bw. Therefore, LD50 was considered to be >5000 mg/kg bw, when rats were treated with test chemical via oral route.

Thus, based on the above summarised studies, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) and it’s structurally and functionally similar read across substance, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates cannot be classified for acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

10 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from study report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally and functionally similar read across chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on two acute dermal toxicity studies as- WoE 2.and WoE 3.
Acute dermal toxicity test was carried out to study the effects of the test chemicals on rodents

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Specific details on test material used for the study:

- IUPAC Name: 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates
- Common Name: Basic Brown 1
- Molecular formula : C26H34N8O8
- Molecular weight : 586.597 g/mol
- Smiles notation : CC(=O)O{-}.N{+}c1ccc(N=Nc2cccc(N=Nc3ccc(N{+}.O{-}C(C)=O)cc3N{+}.O{-}C(C)=O)c2)c(N{+}.O{-}C(C)=O)c1
- Substance type : Organic
- Physical state : liquid

Species:

rat

Strain:

other: 1. not specified 2. Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

1. not specified
2. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: [no]
- Age at study initiation: Young adult male and female rats aged between 6 – 9 weeks were used.
- Weight at study initiation: The weight ranges of approximately 210.2 to 245.4 grams at initiation of dosing were used.
Body weights at the start : Male Mean: 240.40 g (= 100 %); Minimum : 234.6 g (- 2.41 %); Maximum : 245.4 g (+ 2.08 %)
Female Mean: 215.98 g (= 100 %); Minimum : 210.2 g (- 2.68 %); Maximum : 220.6 g (+ 2.14 %)
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use. - Acclimation period: 5 days.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Room temperature was maintained at 20.1 to 21.9 degree centigrade.
- Humidity (%): Room humidity was maintained at 55.5% to 59.2%.
- Air changes (per hr): The animal room was independently provided with at least ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

Type of coverage:

other: 1. Dermal 2. semiocclusive

Vehicle:

other: 1. unchanged (no vehicle) 2. unchanged (no vehicle)

Details on dermal exposure:

1. not specified
2. TEST SITE
- Area of exposure: Dorsal surface and sides from scapular to pelvic area.
- % coverage: Approximately 10% of the total body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw

Duration of exposure:

1. not specified
2. 24 hours

Doses:

1. 3000 mg/kg
2. A single dose of 2000 mg of the test item per kilogram of body weight was administered to ten rats (five males and five females).

No. of animals per sex per dose:

1. not specified
2. 10 (5/sex).

Control animals:

other: 1. not specified 2. yes

Details on study design:

1. not specified
2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical Observations and General Appearance:Animals were observed for clinical signs, mortality, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.
Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.
Gross Pathology: Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

Statistics:

1. not specified
2. not specified

Preliminary study:

1. not specified
2. not specified

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

1. No mortality was observed at 3000 mg/kg bw.
2. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg: All animals survived through the study period of 14 days.
Sex : Female Group I – Animal treated at the dose level of 2000 mg/kg: All animals survived through the study period of 14 days.

Clinical signs:

1. not specified
2. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.

Body weight:

1. not specified
2. Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 8.88% and 18.87% respectively.
Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.29% and 9.60% respectively.

Gross pathology:

1. not specified
2. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.

Other findings:

1. not specified
2. - Other observations: Evaluation of Dermal Reaction
Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Interpretation of results:

other: Not classified

Conclusions:

According to CLP regulation the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) cannot be classified for acute dermal toxicity, as the LD50 value is >2000 mg/kg bw

Executive summary:

Data available for the structurally similar read across chemicals has been reviewed to determine the acute dermal toxicity of the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3). The studies are as mentioned below:

1. Acute Dermal toxicity study of test chemical was conducted in rats at the concentration of 3000 mg/kg bw. No mortality was observed in treated rats at 3000 mg/kg bw. Therefore, LD50 was considered to be >3000 mg/kg bw, when rats were treated with test chemical by dermal application to the skin.

2. The acute dermal toxicity study was designed and conducted for the test chemical in Sprague Dawley rats. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Thus, based on the above summarised studies, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) cannot be classified for acute dermal toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

3 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from secondary source.

Additional information

Acute oral Toxicity: 

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3). The studies are as mentioned below:

1. Acute oral toxicity test was conducted using test chemical in 20 Tif. RAI rats (10 males/10 females), bred under SPF conditions at the dose concentration of 6000 and 10000 mg/kg bw. The test substance was weighed into an Erlenmeyer flask on a Mettler balance. It was suspended at 50 and 60 % with carboxymethylcellulose 2 % and administered by oral intubation. Before treatment the suspension was homogeneously dispersed with an Ultra-Turrax and during treatment it was kept stable with a magnetic stirrer. Mortality and general symptoms was observed at 1 h, 24 h, 48 h, and 7 days. Animals were observed for clinical signs. No mortality was observed at any of the dose in treated rats. Within 2 hours after treatment the rats of both dosage groups showed dyspnoea, exophthalmus, curved position and ruffled fur. The animals had recovered within 4 to 6 days. They were killed and autopsied after an observation period of 7 days. No substance related gross organ changes were seen. Hence, LD50 was considered to be >10000 mg/kg bw, when Tif. RAI rats were treated with test chemical via oral gavage route.

2. Acute oral toxicity study of test chemical was conducted in rats at the concentration of 5000 mg/kg bw. No mortality was observed in treated rats at 5000 mg/kg bw. Therefore, LD50 was considered to be >5000 mg/kg bw, when rats were treated with test chemical via oral route.

Thus, based on the above summarised studies, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) and it’s structurally and functionally similar read across substance, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure of humans via inhalation route is not likely taking into account due to the low vapour pressure of the substance 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3), which is reported to be 1.29E-32 mmHg. Thus, exposure to inhalable dust, mist and vapour of the chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates is highly unlikely. Therefore this study is considered for waiver. 

Acute dermal Toxicity: 

Data available for the structurally similar read across chemicals has been reviewed to determine the acute dermal toxicity of the test chemical 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3). The studies are as mentioned below:

1. Acute Dermal toxicity study of test chemical was conducted in rats at the concentration of 3000 mg/kg bw. No mortality was observed in treated rats at 3000 mg/kg bw. Therefore, LD50 was considered to be >3000 mg/kg bw, when rats were treated with test chemical by dermal application to the skin.

2. The acute dermal toxicity study was designed and conducted for the test chemical in Sprague Dawley rats. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Thus, based on the above summarised studies, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no.: 84281-74-3) and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw for acute oral and dermal toxicity. Thus, comparing these values with the criteria of CLP regulation, 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates cannot be classified for acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.