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

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

Remarks:

Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data from secondary source

Data source

Reference

Reference Type:

secondary source

Title:

SIDS Initial Assessment Profile

Author:

OECD SIDS

Year:

2003

Bibliographic source:

SIAM 16, 2003

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)

Principles of method if other than guideline:

Reproductive toxicity study of test material in Rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

Not specified

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[3-oxo-N-phenylbutyramide]

EC Number:

228-787-8

EC Name:

2,2'-[(3,3'-dichloro[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[3-oxo-N-phenylbutyramide]

Cas Number:

6358-85-6

Molecular formula:

C32H26Cl2N6O4

IUPAC Name:

Pigment Yellow 12

Details on test material:

- Name of test material: Diarylanilide Yellow
- Molecular formula: C32H26Cl2N6O4
- Molecular weight: 629.5014 g/mol
- Substance type: Organic

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Age: 14 weeks
- Weight at study initiation: 503 g (males); 280 g (females)

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on exposure:

Details on oral exposure
PREPARATION OF DOSING SOLUTIONS: Test material dissolved in PEG

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): Test material dissolved in PEG
- Concentration in vehicle: 0, 50, 200 and 1000 mg/kg bw
- Amount of vehicle (if gavage):10ml/kg
- Lot/batch no. (if required):
- Purity:

Details on mating procedure:

- M/F ratio per cage:1:1
- Length of cohabitation:
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:presence of spermatozoa in a vaginal smear
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol:

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- Method: spectrophotometrically (accuracy, homogeneity and stability)
- Sampling time: week 1 and week 6

Duration of treatment / exposure:

males 4 weeks; females 6-7 weeks; animals were treated 2 weeks before mating and during mating (1:1) treatment was continued; after successful mating (presence of spermatozoa in a vaginal smear) males were treated until a total treatment time of 28 days; females were dosed during pregnancy, were allowed to litter and dosed during the lactation period until they were scheduled for necropsy (4-6 days of lactation)

Frequency of treatment:

Daily

Details on study schedule:

Not specified

Doses / concentrations

Remarks:

0, 50, 200 and 1000 mg/kg bw

No. of animals per sex per dose:

Total: 80
0 mg/kg: 10 male and 10 female
50 mg/kg: 10 male and 10 female
200 mg/kg: 10 male and 10 female
1000 mg/kg: 10 male and 10 female

Control animals:

yes, concurrent vehicle

Details on study design:

Not specified

Positive control:

Not specified

Examinations

Parental animals: Observations and examinations:

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

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:

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: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

Oestrous cyclicity (parental animals):

Not specified

Sperm parameters (parental animals):

Parameters examined in [all/P/F1/F2] male parental generations:
testis weight, epididymis weight,

Litter observations:

Examination of fetuses: as required by OECD 422

Postmortem examinations (parental animals):

- Macroscopy: accessory sex organs, epididymides, ovaries, testes and all macroscopic lesions for all animals; according to OECD 422 for 5 animals/sex/group
- Microscopy: not performed

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Statistics:

Dunnett’s test, Steel test, Fisher’s exact test

Reproductive indices:

Not specified

Offspring viability indices:

Not specified

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Clinical signs like faeces discolouration was observed in all treated females and male ; incidental animals of all dose groups showed lethargy, hunched posture, laboured respiration, salivation, chromodacryorrhoea, alopecia, scabs and piloerection. Diarrhea and erythema of the anus are attributed to the use of PEG as vehicle.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Male : : 1/10 at untreated, 50 and 200 mg/kg bw
No mortality in females was 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):

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Male : RBC increased at 50 mg/kg
Female : RBC, Hb/haematocrit increased at 50 mg/kg

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

male:decreased ALAT/ASAT and increased at 200 mg/kg
Female :ALAT/ASAT increase at 1000 mg/kg; phosphate decreased and glucose increased at 200 mg/kg; creatinine decreased at 50 mg/kg
* The increased liver enzymes in high dosed females can be attributed to a single female. Excluding this female leads to mean group values for ASAT and ALAT that are comparable with control values
Effects on blood parameters showed no relationship with the dose and were observed in one of the sexes only. Therefore they were considered to be not toxicological relevant

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

No effects on weight and no gross findings in the reproductive organs.). No additional findings on the testes were present in any of these animals. The effect on the preputial gland is a common finding in young male rats. No relationship to treatment is suspected.
- Successful mating: no treatment related effects
- % mated: 100%
- Number pregnant per dose level: 9/10 for all dose levels
- Number aborting: none
- Number of implantations: no treatment related effects
- Duration of gestation: 21-22 days

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

haematology

clinical biochemistry

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

reproductive performance

Remarks on result:

other: overall no toxic effetcs was observed

Target system / organ toxicity (P0)

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

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Viability: Number of litters: 9 at all doses
- Number of litters: 9 at all doses
- Number of dead pups/no litters: at 0, 50, 200 and 1000 mg/kg bw 3/2, 3/2, 1/1 and 0/0, respectively
- Mean live pups/litter: 0, 50, 200 and 1000 mg/kg bw 13, 12, 15 and 14, respectively
- Postnatal loss/no litters: 0, 50, 200 and 1000 mg/kg bw 5/2, 17/4, 9/3 and 6/5, respectively

*. At 50 mg/kg bw 13 pups were lost in one litter

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

- Macroscopy pups: at 200 mg/kg one pup with malformations was found (a.o. open back/skull); yellow skin in 2 of 10 litters at 1000 mg/kg.The discolourations observed in parents and pups are considered to be related to the staining effect of test substance. From their location it can be concluded that the test substance did not become available systemically, which is in line with other publications on Diarylide Yellow Pigments

Histopathological findings:

not specified

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

mortality

body weight and weight gain

gross pathology

Remarks on result:

other: overall no effects on developmental parameters

Target system / organ toxicity (F1)

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

Overall reproductive toxicity

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 1000 mg/kg bw for P and F1 generation when wistar rats were treated with test material orally by gavage according to OECD 422.

Executive summary:

In a reproductive toxicity study, male andfemale wistar rats were treated with test material in the concentration of 0, 50, 200 and 1000 mg/kg bw orally by gavage formales 4 weeks; females 6-7 weeks; animals were treated 2 weeks before mating and during mating (1:1) treatment was continued; after successful mating (presence of spermatozoa in a vaginal smear) males were treated until a total treatment time of 28 days; females were dosed during pregnancy, were allowed to litter and dosed during the lactation period until they were scheduled for necropsy (4-6 days of lactation). The test material dissolved in PEG and analyzed by spectrophotometrically (accuracy, homogeneity and stability).10 animals/sex /dose group were used. All the animals were observed for Clinical signs, Body weight gain, Food consumption and Examination of uterine content and fetuses were carried out. Macroscopic examination of accessory sex organs, epididymides, ovaries, testes and all macroscopic lesions for all animals; according to OECD 422 for 5 animals/sex/group were done were as Microscopy not performed.

No mortality in females was observed while in male 1/10 at untreated, 50 and 200 mg/kg bw

Were observed. Clinical signs like faeces discolouration was observed in all treated females and male ; incidental animals of all dose groups showed lethargy, hunched posture, laboured respiration, salivation, chromodacryorrhoea, alopecia, scabs and piloerection. Diarrhea and erythema of the anus are attributed to the use of PEG as vehicle. No treatment related effects onBody weight, food consumption, organweights andhistopathology. Gross pathology in female showed 1/10 greenish contents of the caecum at 1000 mg/kg

 

 In male hematological examination showed RBC increased at 50 mg/kg while decreased ALAT/ASAT and increased at 200 mg/kg.In female hematological examination showed RBC, Hb/hematocrit increased at 50 mg/kg while ALAT/ASAT increase at 1000 mg/kg; phosphate decreased and glucose increased at 200 mg/kg; creatinine decreased at 50 mg/kg. Effects on blood parameters showed no relationship with the dose and were observed in one of the sexes only. Therefore they were considered to be not toxicological relevant,

The increased liver enzymes in high dosed females can be attributed to a single female. Excluding this female leads to mean group values for ASAT and ALAT that are comparable with control values. The inflammation of the preputial glands seen in 4 of 5 high dosed males was minimal (grade 1-2). No additional findings on the testes were present in any of these animals. The effect on the preputial gland is a common finding in young male rats. No relationship to treatment is suspected

No effects on weight and no gross findings in the reproductive organs. Successful mating as no treatment related effects was observed. Number pregnant per dose level: 9/10 for all dose levels. No abortion was observed. No treatment related effects on Number of implantations and duration of gestation. No effects onViability,Number of dead pups/no litters and Mean live pups/litter was observed. Body weight/sex pups were unchanged.
In macroscopic examination of pups at 200 mg/kg one pup with malformations was found (a.o. open back/skull); yellow skin in 2 of 10 litters at 1000 mg/kg was observed. The discolourations observed in parents and pups are considered to be related to the staining effect of test substance. From their location it can be concluded that the test substance did not become available systemically, which is in line with other publications on Diarylide Yellow Pigments. HenceNOAEL was considered to be 1000 mg/kg bw for P and F1 generation whenwistar rats were treated with test material orally by gavage according to OECD guideline 422.