Benzenamine, reaction products with aniline hydrochloride and nitrobenzene, hydrochlorides

Endpoint summary

Description of key information

Key study: OECD 422. GLP study: The NOEL/NOAEL were 1000 mg/kg bw/day in rats (no effects observed at highest dose).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method according to OECD Guideline 422. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Age at study initiation: 10 weeks old

Route of administration:

oral: gavage

Vehicle:

other: Methylcellulose aqueous solution (suspended)

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg
- 0.5% w/v methylcellulose aqueous solution (suspended)

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Males: 42 days (from 14 days before mating, and through the mating period until a day before necropsy).
Females: 41-48 days (from 14 days before mating, and through the mating period until day 4 of lactation).

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0 (control), 40, 200, 1000 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

Males: 7 (control), 12 (40 mg/kg bw/day), 12 (200 mg/Kg bw/day), 7 (1000 mg/kg bw/day), 5 (control with recovery), 5 (1000 mg/kg bw/day with recovery).
Females: 12 (control), 11 (40 mg/kg bw/day), 11 (200 mg/Kg bw/day), 12 (1000 mg/kg bw/day), 5 (control with recovery), 5 (1000 mg/kg bw/day with recovery).

Control animals:

yes

Details on study design:

- Dose selection rationale: In the 14-day range finding study with 5 males and 5 females at dose levels of 0, 100, 300, 1000 mg/kg/day, there were no changes attributed to the treatment at any groups. (Inspection items: clinical signs, body weight, food consumption, necropsy, organ weight, hematology and blood chemistry).
- Post-exposure recovery period in satellite groups: 14 days.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

FOOD CONSUMPTION: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: End of the dosing and recovery period.
- How many animals: All animals
- Parameters checked: RBC, WBC, Hb, Plat., MCV, MCH, MCHC, Ret., Eosino., Baso., Mono., Lymph., Neutro., LUC, PT, APTT, Fibrin.,

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: End of the dosing and recovery period.
- How many animals: All animals
- Parameters checked: ASAT, ALTA, ALP, CPK, T. Bil., T. Prot., Albumin, T. Chol., TGL, PL, Glucosa, BUN, Creat., IP, Ca, Na, K, Cl, α1-glob., α2-glob., β-glob., γ-glob., A/G.

URINALYSIS: Yes (in males)

NEUROBEHAVIOURAL EXAMINATION: Yes
- Battery of functions tested: sensory activity / grip strength / motor activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, all animals at the end of the dosing and recovery periods.
Absolute and relative organ weight:
Males: Adrenals, testis, thymus, spleen, brain, heart, lung, liver, kidney, epididymus, sem. vesic., prostate.
Females: Adrenals, ovaries, thymus, spleen, brain, heart, lung, liver, kidneys.

HISTOPATHOLOGY: Yes, animals within control and highest dose tested (1000 mg/kg bw/day), at the end of dosing and recovery period.
Male: Femoral bone marrow, heart, kidney, liver, lung, prostate, stomach, thyroid, trachea.
Females: Adrenal, femoral bone marrow, heart, kidney, liver, lung, spleen, stomach, thymus, thyroid, trachea, uterus, adipose tissue.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
No animal died in any test article group.
No test article-related changes were observed in clinical signs.
Only test article-colored feces were observed in all groups.

BODY WEIGHT AND WEIGHT GAIN
No test related effects were observed.

FOOD CONSUMPTION.
No test related effects were observed.

HAEMATOLOGY
No test related effects were observed.

CLINICAL CHEMISTRY
No test related effects were observed.

URINALYSIS
No test related effects were observed in males.

NEUROBEHAVIOUR
No test related effects were observed.

ORGAN WEIGHTS
No test related effects were observed.

GROSS PATHOLOGY
No test related effects were observed.

HISTOPATHOLOGY: NON-NEOPLASTIC
No test related effects were observed.

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: (no effects observed at highest dose)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: (no adverse effects observed at highest dose)

Critical effects observed:

not specified

See the observed effects attached.

Conclusions:

The NOEL and NOAEL for repeated dose toxicity in rats were determined to be 1000 mg/kg bw/day since no effects were observed in the highest dose tested.

Executive summary:

A combined repeated dose toxicity study with the reproduction/developmental screening test was performed with the test item according to OECD Guideline 422 (GLP study). Based on a preliminary 14 -days range-finding study, test item was administered orally to 12 male and female rats at 0 (control), 40, 200 and 1000 mg/kg bw/day for 42 days (from 14 days before mating, and through the mating period until a day before necropsy) and for 41-48 days (from 14 days before mating, and through the mating period until day 4 of lactation) respectively. For the control and 1000 mg/kg bw/day groups, a 2 -week recovery period was set to follow the 42 -day repeated oral administration period to assess the reversibility of the toxicity. Animals were subjected to necropsy at the end of the dosing and recovery periods. No animals died in any test article group, and no test article related changes were observed in clinical signs, detailed observations, grip strength, locomotor activity, body weight, food consumption, hematology, blood chemistry, urinalysis (only perfomed in males), organ weight, necropsy, or histopathology. Based on these results, the NOEL and NOAEL for repeated dose toxicity in rats was determined to be 1000 mg/kg bw/day since no effects were observed in the highest dose tested.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study planned

Justification for type of information:

TESTING PROPOSAL ON VERTEBRATE ANIMALS:

NON-CONFIDENTIAL NAME OF SUBSTANCE: Benzenamine, reaction products with aniline hydrochloride and nitrobenzene, hydrochlorides

CONSIDERATIONS THAT THE GENERAL ADAPTATION POSSIBILITIES OF ANNEX XI OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION
[please address all points below]:
- Available GLP studies: Not available.
- Available non-GLP studies: Not available.
- Historical human data: Not available.
- (Q)SAR : Not available.
- In vitro methods: Not available.
- Weight of evidence: Not available.
- Grouping and read-across : Not available.

CONSIDERATIONS THAT THE SPECIFIC ADAPTATION POSSIBILITIES OF ANNEXES VI TO X (AND COLUMN 2 THEREOF) OF THE REACH REGULATION ARE NOT ADEQUATE TO GENERATE THE NECESSARY INFORMATION:
The specific adaptation possibilities of Column 2 of REACH Annex VIII are not adequate to generate the necessary information since:
- In a reliable short-term toxicity study (28 days) no adverse effects were observed which concluded a classification as STOT RE according to the CLP Regulation (EC) no. 1272/2008. Thus,no extrapolation towards the NOAEL-90 days would be allowed.
- There is no reliable chronic study available.
- There is no sufficient information on the cleavage products (both for systemic and effects at the site of updatake).


FURTHER INFORMATION ON TESTING PROPOSAL IN ADDITION TO INFORMATION PROVIDED IN THE MATERIALS AND METHODS SECTION:
The following study is proposed:
Test method according to OECD Guidance 408, i.e. 90 days repeated exposure by oral route, in rats.

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Species:

rat

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The key study is GLP compliant and of high quality (Klimisch score = 1).

Repeated dose toxicity: inhalation - systemic effects

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

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Key study: Experimental data.

Test method according to OECD 422. GLP study.

The NOEL and NOAEL for repeated dose toxicity in rats were determined to be 1000 mg/kg bw/day since no effects were observed in the highest dose tested.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

Only one study available.

Justification for classification or non-classification

Based on the available information, the substance is not classified for repeated dose toxicity according to CLP Regulation (EC) No. 1272/2008.