2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutyramide]

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From JAN 1974 to APR 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1980

Report date:

1980

Reference 2

Reference Type:

publication

Title:

Carcinogenicity studies on different diarylide yellow pigments in mice and rats

Author:

Leuschner F

Year:

1978

Bibliographic source:

Toxicology Letters 2: 253-260

Report date:

1978

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 451 (Carcinogenicity Studies)

Deviations:

not applicable

Remarks:

The study was performed to test carcinogenicity of the test item. That's the reason why some details usually requested in a repeated dose toxicity study (e.g. data on clinical biochemistry, haematology) are missing.

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutyramide]

EC Number:

226-939-8

EC Name:

2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutyramide]

Cas Number:

5567-15-7

Molecular formula:

C36H32Cl4N6O8

IUPAC Name:

2,2'-[(3,3'-dichlorobiphenyl-4,4'-diyl)didiazene-2,1-diyl]bis[N-(4-chloro-2,5-dimethoxyphenyl)-3-oxobutanamide]

Test material form:

not specified

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: S. IVANOVAS GmbH & Co, Med. Versuchstierzucht KG (Kissleg, Germany)
- Age at study initiation: 38 (males) -42 (females) days
- Weight at study initiation: 100 - 109 g
- Housing: in groups of 2 or 3 animals in Macrolon cages (Type III)
- Diet: Altromin 1321 (Altromin, Lage), ad libitum
- Water: tap water, ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 +/- 0.5
- Humidity (%): 60 +/- 3
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

104 weeks

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

1 000 other: ppm in diet

Doses / concentrations 2

Dose / conc.:

3 000 other: ppm in diet

Doses / concentrations 3

Dose / conc.:

9 000 other: ppm in diet

No. of animals per sex per dose:

50

Control animals:

yes, plain diet

Positive control:

none

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice per day


DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: weekly


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes


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


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily


OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: once, immediately before sacrifice
- Dose groups that were examined: all animals


HAEMATOLOGY: No


CLINICAL CHEMISTRY: No

URINALYSIS: No


NEUROBEHAVIOURAL EXAMINATION: No


OTHER:
at the end of the exposure period the following investigations were performed:
- audiometry (using a simple sound test)
- inspection of denture
- organ weights of 7-8 organs (heart, liver, lungs, spleen, kidney, thymus, brain, testis)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (animals of the control and highest dose group; paraffin sections, Haematoxylin-Eosin staining):
heart, lung, liver (additionally: frozen sections with Sudan staining), kidney, spleen, adrenal, thymus, pituitary, brain, gonads, thyroid, prostate/uterus, seminal vesicle/mammary gland, stomach, duodenum, colon, salivary gland, lymph nodes, eye and optic nerve, urinary bladder, bone marrow, neoplastic lesions, bones
- histopathological investigations of animals of the lower dose groups were performed, if they died or were sacrificed in the meantime and revealed macroscopic findings

Statistics:

- variance analysis according to Peto
- Student's t-test

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 specified

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

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:

no effects observed

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

9 000 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no substance related toxicity or carcinogenicity was observed; 9000 mg/kg diet (ppm) correspond to 533.1 mg/kg bw/day and 523.0 mg/kg bw/day in male and female rats, respectively (calculated in the study report on basis of food consumption)

Target system / organ toxicity

Key result

Critical effects observed:

no

Any other information on results incl. tables

 - no effects on behaviour, appearance, faeces, feed and drinking water uptake, eyes, hearing, dentition, mortality, body weight development

- no substance induced macroscopic or histological changes

- no substance related effects on the tumour incidence

- 1000, 3000, 9000 ppm in diet correspond to 58, 174, 533 mg/kg bw/day in male and 59, 180, 523 mg/kg bw/day in female rats, respectively

 - no 3,3'-dichlorobenzidine was detected in the urine samples (limit of detection: 3 µg/10 ml urine; 0.3 ppm)

Applicant's summary and conclusion

Conclusions:

Chronic feeding of Sprague Dawley rats with up to 9000 ppm of the test item in diet did not cause any adverse effect. The NOAEL in this study was 9000 ppm in diet (corresponding to 533.1 mg/kg bw/da and 523.0 mg/kg bw/day in male and female rats, respectively).

Executive summary:

Sprague Dawley rats (50 per sex per dose) were exposed to 1000, 3000, 9000 ppm of the test item in diet (corresponding to 58, 174, 533 mg/kg bw/day and 59, 180, 523 mg/kg bw/day in male and female rats, respectively) for 104 weeks. The test item did neither induce toxicity nor tumorigenicity. The NOAEL in this study was 9000 ppm in diet.