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

Endpoint summary

Description of key information

Repeated dose: oral

NONAEL was considered to be 1000-5000 mg/kg bw for diazotized m-phenylenediamine, acetates.

Thus, comparing this value with the criteria of CLP regulation 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no 84281-74-3) is not likely to classify as repeated dose toxicant.

Repeated dose inhalation toxicity:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no 10461-98-0), which is reported as 1.28E-32 mm Hg at 25 C. 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.

Repeated dose dermal toxicity:

In accordance with coloumn 2 of Annex IX, this end point was considered for waiver since the acute toxicity by the dermal route has already been provided in section 7.2.3 (as part of the Annex VIII information requirements)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

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:

Weight of evidence approach based on structurally similar chemicals

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

WoE report is based on two repeated dose oral toxicity studies on rats.
1. 17 days repeated dose toxicity study of test chemical in F344 male and female rats were observed to evaluate its toxic potential.
2. Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening test for test chemical.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: 1. Fischer 344, 2. Crl:CD (SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

1. Details on test animal
TEST ANIMALS
- Source: Frederick Cancer Research Center (Frederick, MD)
- Age at study initiation: 56 days old
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: rats were housed five per cage throughout the study cage: Polycarbonate, solid bottom (Lab Products Inc., Garfield, NJ)
- Diet (e.g. ad libitum): NIH-07 rat, meal (Zeigler Bros., Inc., Gardners, PA), available ad libitum
- Water (e.g. ad libitum): Tap water (Birmingham Water Works)
in glass water bottles with stainless steel sippers (Edstrom Automatic Watering Systems, Waterford, WI), available ad libitum
- Acclimation period: 20 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.2-23.3 °C
- Humidity (%): 47%-61%
- Air changes (per hr): Minimum 15 change per hours
- Photoperiod (hrs dark / hrs light): 12 hours/day

2. - Source: No data
- Age at study initiation: 9 weeks old
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: No data
- Diet (e.g. ad libitum): No data
- Water (e.g. ad libitum): No data
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%):No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
IN-LIFE DATES: From: To: No data

Route of administration:

other: 1. oral : feed, 2, gavage

Vehicle:

other: 1. NIH-07 rat, meal, 2. 1 w/v% Tween 80 solution

Details on oral exposure:

1. PREPARATION OF DOSING SOLUTIONS: Not applicable

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): premix of NIH-07 rat as meal, with the appropriate amount of C.I. Pigment Red 23.
- Storage temperature of food: 2 weeks for 45 °C

VEHICLE
- Justification for use and choice of vehicle (if other than water): No data available
- Concentration in vehicle: 0, 6,000, 12,500, 25,000, 50,000 or 100,000 ppm

2. PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in 1 w/v% Tween 80 solution to give dose level of 0, 40, 200 or 1000 mg/Kg/day

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): 1 w/v% Tween 80 solution
- Concentration in vehicle: 0, 40, 200 or 1000 mg/Kg/day
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

1. Periodic analyses of the dosed-feed formulations were conducted at the study laboratory and at the analytical chemistry laboratory throughout. the studies.

2. not specified

Duration of treatment / exposure:

1. from day 15 to 17th
2. Male: 42 days / - Female: 42 - 48 days (from 14 days before mating to day 4 of lactation)

Frequency of treatment:

Daily

Remarks:

1. 0, 6,000, 12,500, 25,000, 50,000 or 100,000 ppm (0, 1200, 2500, 5000, 10000, 50000 mg/kg/day)

Remarks:

2. 0, 40, 200 or 1000 mg/Kg/day

No. of animals per sex per dose:

1. 5animals/dose
2. No data

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Observations and examinations performed and frequency:

Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice/day
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once/week and at sacrifice

BODY WEIGHT: Yes
- Time schedule for examinations: once/week and at sacrifice

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): daily/cage (calculated per animal)
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg
body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain
data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted average
s from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined.

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined.

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: No data sensory activity / grip strength / motor activity / other: No data

2. CAGE SIDE OBSERVATIONS: No data
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/ kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain
data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted av
erages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: Yes
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: FOB test was performed

Sacrifice and pathology:

1. GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

2. GROSS PATHOLOGY: Yes, organ weight was measured
HISTOPATHOLOGY: Yes

Statistics:

1. Mean ±Standard deviation was observed.

Clinical signs:

no effects observed

Description (incidence and severity):

1. No significant effect were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Mortality:

no mortality observed

Description (incidence):

1.No significant mortality were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

1. Differences in mean body weight change and weight gain of exposed group than control were not significant.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

1. No significant effect were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Description (incidence and severity):

Decreased erythrocyte count observed in all male dose groups and the two highest female dose groups, indicating a mild anemia

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

1. No significant effect were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Gross pathological findings:

no effects observed

Description (incidence and severity):

1. No significant effect were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

1. No significant effect were observed at all dose level 0, 1200, 2500, 5000, 10000, 50000 mg/kg/day of treated group compare to control.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Dose descriptor:

NOAEL

Effect level:

1 000 - 5 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

urinalysis

Remarks on result:

other: No effect observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

NONAEL was considered to be 1000-5000 mg/kg bw for diazotized m-phenylenediamine, acetates.

Executive summary:

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no. 84281-74-3). The studies are as mentioned below:

Study 1:

Repeated dose oral toxicity study for the test compound was studied F344 male and female rats by administrating test compound orally at doses 0, 600, 1200, 2500, 5000, 10000 mg/kg/day. There were no clinical and mortality was observed. No biologically significant differences recorded in organ weights among exposed and control rats. Decreased erythrocyte count observed in the two highest female and male dose groups, indicating a mild anemia. Therefore, NOAEL value for test substance is considered to be 5000 mg/kg in F344 female rats.

Study 2:

Sub-chronic repeated dose oral toxicity test for test chemical was studied in male and female Crl:CD (SD) rats. The males were treated for 42 days and the females were treated from 42 - 48 days (from 14 days before mating to day 4 of lactation). The test chemical was mixed 1 w/v% Tween 80 solution to give dose level of 0, 40, 200 or 1000 mg/Kg/day. During the study period, the animals were observed for clinical signs, body weight and food consumption changes, hematology, clinical chemistry and urinalaysis was performed and the animals were subjected to gross pathology and histopathology. No effects were observed during the study period at the highest dose level of 1000 mg/Kg/day. Based on the observations made, No observed adverse effect level for test chemical is considered to be 1000 mg/Kg/day.

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 1000-5000 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) is not likely to classify as repeated dose toxicant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

5 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimisch 2 and from publication

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: inhalation, other

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: dermal, other

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose: oral

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no. 84281-74-3). The studies are as mentioned below:

Study 1:

Repeated dose oral toxicity study for the test compound was studied F344 male and female rats by administrating test compound orally at doses 0, 600, 1200, 2500, 5000, 10000 mg/kg/day. There were no clinical and mortality was observed. No biologically significant differences recorded in organ weights among exposed and control rats. Decreased erythrocyte count observed in the two highest female and male dose groups, indicating a mild anemia. Therefore, NOAEL value for test substance is considered to be 5000 mg/kg in F344 female rats.

Study 2:

Sub-chronic repeated dose oral toxicity test for test chemical was studied in male and female Crl:CD (SD) rats. The males were treated for 42 days and the females were treated from 42 - 48 days (from 14 days before mating to day 4 of lactation). The test chemical was mixed 1 w/v% Tween 80 solution to give dose level of 0, 40, 200 or 1000 mg/Kg/day. During the study period, the animals were observed for clinical signs, body weight and food consumption changes, hematology, clinical chemistry and urinalaysis was performed and the animals were subjected to gross pathology and histopathology. No effects were observed during the study period at the highest dose level of 1000 mg/Kg/day. Based on the observations made, No observed adverse effect level for test chemical is considered to be 1000 mg/Kg/day.

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 1000-5000 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) is not likely to classify as repeated dose toxicant.

Repeated dose inhalation toxicity:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. Taking into account the low vapour pressure of the substance 1,3-Benzenediamine, coupled with diazotized m-phenylenediamine, acetates (CAS no 10461-98-0), which is reported as 1.28E-32 mm Hg at 25 C. 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.

Repeated dose dermal toxicity:

In accordance with coloumn 2 of Annex IX, this end point was considered for waiver since the acute toxicity by the dermal route has already been provided in section 7.2.3 (as part of the Annex VIII information requirements)

Justification for classification or non-classification

Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 1000-5000 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) is not likely to classify as repeated dose oral, dermal and inhalation toxicant.