Benzenamine, reaction products with aniline hydrochloride and nitrobenzene, hydrochlorides

Repeated dose toxicity: oral

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Benzenamine, reaction products with aniline hydrochloride and nitrobenzene, hydrochlorides

EC Number:

309-913-1

EC Name:

Benzenamine, reaction products with aniline hydrochloride and nitrobenzene, hydrochlorides

Cas Number:

101357-16-8

Molecular formula:

Not applicable. Multiconstituent substance.

IUPAC Name:

(2Z,7Z)-5-phenyl-2,7-bis(phenylimino)-2,7-dihydro-5λ⁵-phenazin-5-ylium (7Z)-N2,N3,5-triphenyl-7-(phenylimino)-5,7-dihydrophenazine-2,3-diamine N2,N3,5,7-tetraphenyl-5,7,12,14-tetrahydro-5,7,12,14-tetraazapentacene-2,3-diamine chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): C.I. Solvent Black 5 (analogue CAS 11099-03-9).
- Lot/batch No.: 10137087
- Physical state: Blackish brown powder
- Storage condition of test material: Room temperature.
- Stability and uniformity were property has been confirmed.

Test animals

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

Administration / exposure

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

Doses / concentrations

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.

Examinations

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.

Results and discussion

Results of examinations

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.

Effect levels

1

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)

2

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)

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

See the observed effects attached.

Applicant's summary and conclusion

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.