Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be in range of 500 -1500mg/kg /day for reproductive toxicity, when male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
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 from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on reproductive toxicity studies on rats
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Specific details on test material used for the study:
No data available
Species:
rat
Strain:
other: 1.Charles River CD 2.Osborne-Mendel 3.Wistar
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals or test system and environmental conditions:
1.TEST ANIMALS
- Source:No data available
- Females (if applicable) nulliparous and non-pregnant: [yes/no]
- Age at study initiation: (P) x wks; (F1) x wksNo data available
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x gNo data available
- Fasting period before study:No data available
- Housing:Animals were housed individually
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum):the test diet ad libitum
- Water (e.g. ad libitum):No data available
- Acclimation period:No data available
ENVIRONMENTAL CONDITIONS
- Temperature (°C):No data available
- Humidity (%):No data available
- Air changes (per hr):No data available
- Photoperiod (hrs dark / hrs light):
IN-LIFE DATES: From: To:No data available
2.TEST ANIMALS
- Source: FDA rat breeding colony
- Age at study initiation: female: 12 - 21 wk
- Weight at study initiation:female: 220 - 270 g
- Fasting period before study: no data
- Housing: Stainless-steel hanging cages.
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): Purina Laboratory Chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25 deg C
- Humidity (%): 30 - 63%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle (8 am 8 pro).

IN-LIFE DATES: From: To:
Route of administration:
other: 1&3.oral: feed 2.oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
not specified
Details on exposure:
1.PREPARATION OF DOSING SOLUTIONS: test material mixed with food
2.test chemical was dissolved in distilled water (w/v). Fresh solutions were prepared daily and administered by gavage in a volume of 1 ml/100 g body weight, at approximately the same time each day. Controls received an equivalent volume of distilled water.
Details on mating procedure:
2.- M/F ratio per cage: 1:2
- Length of cohabitation: 1 day
- Proof of pregnancy: sperm in vaginal smear was considered day 0 of gestation
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: no data
- Further matings after two unsuccessful attempts: no data
- After successful mating each pregnant female was caged (how): no data
- Any other deviations from standard protocol: no data
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
1.Pre-Mating Exposure / Males and Females: 60 days and test material adminstered upto three generation
2.19 days
3.From F1 to F3 generation
Frequency of treatment:
Daily
Details on study schedule:
No data available
Remarks:
Doses / Concentrations:
Study 1
5, 50, 150 or 500 mg/kg bw/day
Study 2.
0, 30, 75, 150, 300, 600 or 1000mg/kg/day.
Study 3
0, 0.1, 1.0 or 3% (0, 50, 500,1500 mg/kg bw per day)
No. of animals per sex per dose:
1.Total :150
0mg/kg bw/day : 10 males and 20 females
5 mg/kg bw/day : 10 males and 20 females
50 mg/kg bw/day :10 males and 20 females
150 mg/kg bw/day :10 males and 20 females
500 mg/kg bw/day : 10 males and 20 females
2. 42-43 female/dose group
3.Total:360
0 mg/kg bw: 60 male and 60 female
50 mg/kg bw : 40male and 40 female
500 mg/kg bw : 40male and 40 female
1500 mg/kg bw : 40male and 40 female
After a nine-week test period, 24 males and 24 females from the control group, and 14 males and 14 females from each test group were used for teratogenicity studies; the remainder were used for the reproduction study.
Control animals:
yes
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
1.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
Clinical observations were recorded twice daily with at least 5 hours between observations

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: Detailed physical examinations and palpation for masses were performed weekly.



BODY WEIGHT: Yes
Time schedule for examinations: weekly for the first fourteen weeks, bi-weekly for the next 12 weeks and every 4 weeks thereafter until the end of the study.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes weekly for the first fourteen weeks, bi-weekly for the next 12 weeks and every 4 weeks thereafter until the end of the 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 ,

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

OTHER: Haematology tests, including hemoglobin, hematocrit, erythrocyte and total and differential leukocyte counts, and erythrocyte morphology, were conducted on ten randomly selected animals at months 3, 6, 12, 18 and 24 of the study.
2.The rats were weighed daily and food consumption was measured weekly. Water intake was not measured.
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
2.Each live foetus waspromptly weighed, sexed and examined for gross external malformations, and the crown-rump length was measured. Any foetus that weighed less than 70% of the average weight of the concurrent male or female controls was considered to be a runt.
Postmortem examinations (parental animals):
1.SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]

GROSS NECROPSY
Necropsies were conducted on all animals dying prior to study termination, killed in a moribund condition or killed on schedule,Tissues examined included adrenal glands, aorta, blood smear, brain, cecum, colon, duodenum, epididymus or uterus, esophagus, eyes, femur including marrow, tissue masses, gallbladder, heart, ileum, jejunum, duodenum, kidneys, liver, lungs and bronchi, mammary gland, nerves (sciatic), ovaries, lymph nodes, pancreas, parathyroids, pituitary gland, prostrate, rectum, skin, spleen, seminal vesicles, skeletal muscle, testes with ep ididymides, stomach, thymus, thyroid gland including parathyroid, trachea, urinary bladder, uterus


HISTOPATHOLOGY / ORGAN WEIGHTS
Histological examinations were conducted on all animals from both control groups, the highest dose group (2.0 or 5.0%) from each study and also on 10 rats randomly selected from each group for an interim sacrifice at 12 months. Histology was also performed on any animal with gross lesions or masses
2.On day 20 of gestation, starting at 1 pro, the females were examined for gross abnormalities for the last time before being killed by CO, asphyxiation. Caesarean sections were performed. Corpora lutea were counted. The uterus was opened and examined in situ. The uterine positions of all implantation sites were noted and their condition (early or late resorptions, living or dead foetuses) was determined.
Postmortem examinations (offspring):
2.Approximately one-half of the foetuses were fixed in alcohol, stained with Alizarin Red S and examined under a dissecting microscope for all skeletal variations. The remaining half of the foetuses were fixed in Bouin's solution, serially sectioned by razor blade and examined under a dissecting microscope for internal variations of the soft tissues.
Statistics:
2.All data analyses were performed by the Division of Mathematics at the FDA. Data on maternal initial body weights and food consumption were analysed by straight analysis of variance (ANOVA) and two-tailed t-test, and a regression analysis. The number of dams affected was analysed by a Fisher's exact test. Data on maternal weight gain were submitted to an analysis of covariance (ANOCOVA) and a two-tailed t-test. Data on the numbers of implants, corpora lutca, total viable young and viable males and females were analysed by ANOVA followed by a one-tailed t-test. Data on implantation efficiency, early deaths, late deaths and total resorptions (early and late deaths) were transformed by using the Freeman-Tukey arc-sine transformation (Freeman and Tukey, 1950) and then analysed by ANOVA and a one-tailed t-test. Data on litters having one or more or two or more resorptions were analysed by a Fisher's exact test. Similar tests were applied to the number of runts per litter. Data on foetal body weights, crown-!o-rump lengths and foetal ossified vertebrae were analysed by nested ANOVA and a one-tailed t-test. The ANOVA included a correction for unequal sample size (Sokal and Rohlf, 1981). Data on the average number of foetuses per litter with skeletal, sternebral, combined missing plus incomplete plus bipartite sternebrae or soft-tissue variations were transformed by using the Frceman-Tukey arc-sine transformation and then analysed by ANOVA and a one-tailed t-test. Litters with foetuses with at least one, at least two, etc. skeletal, sternebral, combined missing plus incomplete plus bipartite sternebrae, or soft-tissue variations, and specific variations were analysed by Fisher's exact test.
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No treatment-related effects were reported on survival
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
2.Mean food consumption on days 0-7, 7-14, 14 -20 and 0-20 by the treated animals was similar to that of the control animals
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):
No treatment-related effects were reported on haematological finding
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were reported on clinical chemistry
Urinalysis findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were reported on urinalysis.
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
Description (incidence and severity):
1.no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats.
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
1.Histological evaluation revealed a variety of lesions, including neoplasms, present at similar incidences in control and treated animals.The authors considered the lesions to be spontaneous and not related to administration of the test material.
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
1.There were no compound related effects on fertility, gestation, pup viability or lactation indices, on reproductive organs of females, or on organ weights among parents.
2.No effect on numbers of corpora lutea and implants per female, total resorption, numbers of viable foetuses per female were observed in treated rats as compared to control. Increase in number of early deaths per litter and the number of early plus late deaths per litter were observed in in the 600 mg/kg group, but these appeared to be random occurrences. Similarly, no effect on foetal weights of males and females and crown rump lengths and number of litters with runts were observed in treated rats as compared to control.
3.No treatment-related effects on reproductive function was observed
Dose descriptor:
NOAEL
Effect level:
> 500 - <= 1 500 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
reproductive performance
other: see 'Remark'
Remarks on result:
other: No effects on reproductive performance was observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
1.no compound related effects on pup viability or lactation indices.
2.The number of litters with runts was similar in all groups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2.Mean foetal weights of males and females and crown rump lengths were similar in all groups.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
1.Food consumption was similar for control and treated animals at the lower dietary levels, but was slightly higher in the 500 mg/kg bw dose group although not statistically significant.
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):
1.No treatment-related effects were reported on Haematological finding
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
1.No treatment-related effects were reported on clinical chemistry
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
1.no compound related effects on organ weights
Gross pathological findings:
no effects observed
Description (incidence and severity):
1.Necropsies did not reveal any treatment-related gross or microscopic changes.
2.Aside from haemorrhages present in foetuses in all groups in similar numbers, there were no other external variations in any of the dosed groups. Two foetuses had multiple anomalies: one foetus from the control group had a club foot, a short tail and four digits on the hind legs, and one foetus from the 150-mg/kg group had exencephalus and hydrocephalus. Three foetuses in a single litter of a female given 75 mg/kg were oedematous.
Histopathological findings:
no effects observed
Description (incidence and severity):
1.There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from offspring.
2.No dose-related increase in sternebral variations was seen among the foetuses with reduced ossification, bipartite, missing, malaligned or fused sternebrae,
nor in the numbers of litters containing these foetuses. The incidence of specific skeletal variations was similar in all groups of foetuses, except for a significant increase in the incidence of 14th rib bud in foetuses from the 300 mg/kg group which was considered to be a random occurrence.

Other effects:
no effects observed
Description (incidence and severity):
1.There were no compound related effects on fertility, gestation, pup viability or lactation indices, on reproductive organs of females, or on organ weights among offspring.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 500 - <= 1 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
mortality
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: Overall no developmental toxic effects were observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
1.Food consumption was similar for control and treated animals at the lower dietary levels, but was slightly higher in the 500 mg/kg bw dose group although not statistically significant.
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):
No treatment-related effects were reported on Haematological finding
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were reported on clinical chemistry
Urinalysis findings:
no effects observed
Description (incidence and severity):
No treatment-related effects were reported on urinalysis
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
no compound related effects on organ weights
Gross pathological findings:
no effects observed
Description (incidence and severity):
Necropsies did not reveal any treatment-related gross or microscopic changes.
Histopathological findings:
no effects observed
Description (incidence and severity):
There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from offspring.
Other effects:
no effects observed
Description (incidence and severity):
There were no compound related effects on fertility, gestation, pup viability or lactation indices, on reproductive organs of females, or on organ weights among offspring.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
> 500 - <= 1 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
mortality
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: overall no developmental effects were observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Conclusions:
NOAEL was considred to be in range of 500-1500mg/kg bw/day for F0, F1 and F2 generation. When male and female rats were treated with test material orally.
Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

Study 1

Three generation reproductive study of test material was performed on male and female Charles River CD rats. Animals were housed individually and fed the test diet ad libitum. The test material mixed with food in dose concentration 0, 5, 50, 150 or 500 mg/kg bw/day and 10 males and 20 females/group/generation were used .Pre-Mating Exposure / Males and Females were 60 days. Clinical observations were recorded twice daily with at least 5 hours between observations. Detailed physical examinations and palpation for masses were performed weekly. Body weights and food consumption were determined weekly for the first fourteen weeks, bi-weekly for the next 12 weeks and every 4 weeks thereafter until the end of the study. The intake of the test substance was determined from body weight, food consumption and dietary concentration. Haematology tests, including haemoglobin, haematocrit, erythrocyte and total and differential leukocyte counts, and erythrocyte morphology, were conducted on ten randomly selected animals at months 3, 6, 12, 18 and 24 of the study. Necropsies were conducted on all animals dying prior to study termination, killed in a moribund condition or killed on schedule. Histological examinations were conducted on all animals from both control groups, the highest dose group (2.0 or 5.0%) from each study and also on 10 rats randomly selected from each group for an interim sacrifice at 12 months. Histology was also performed on any animal with gross lesions or masses. Also tissues examined included adrenal glands, aorta, blood smear, brain, cecum, colon, duodenum, epididymus or uterus, esophagus, eyes, femur including marrow, tissue masses, gallbladder, heart, ileum, jejunum, duodenum, kidneys, liver, lungs and bronchi, mammary gland, nerves (sciatic), ovaries, lymph nodes, pancreas, parathyroids, pituitary gland, prostrate, rectum, skin, spleen, seminal vesicles, skeletal muscle, testes with epididymides, stomach, thymus, thyroid gland including parathyroid, trachea, urinary bladder, uterus.

 At the end of study, no treatment-related effects were reported on survival. No treatment-related changes were reported at gross necropsy. Histological evaluation revealed a variety of lesions, including neoplasms, present at similar incidences in control and treated animals. The authors considered the lesions to be spontaneous and not related to administration of the test material. There were no compound related effects on fertility, gestation, pup viability or lactation indices, on reproductive organs of females, or on organ weights among parents and offspring. There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats or from offspring. There was no toxicity to either the F1 or F2 generation. Food consumption was similar for control and treated animals at the lower dietary levels, but was slightly higher in the high-dose study, although not statistically significant. Haematological, clinical chemistry and urinalysis parameters did not differ significantly from the controls. Necropsies at one year did not reveal any treatment-related gross or microscopic changes. Hence NOAEL was considered to be 500mg/kg bw/day for F0, F1 and F2 generation. When male and female Charles River CD rats were treated with test material orally.

Study 2

In a reproductive toxicity study, Osborne-Mendel male and female rats were treated with Allura Red AC in the concentration of 0, 30, 75, 150, 300, 600 and 1000 mg/kg body weight/day orally by gavage on days 0-19 of gestation. No clinical sign of toxicity, no effect on body weight and food consumption were observed in treated rats as compared to control. No effect on numbers of corpora lutea and implants per female, total resorption, numbers of viable foetuses per female were observed in treated rats as compared to control. Increase in number of early deaths per litter and the number of early plus late deaths per litter were observed in in the 600 mg/kg group, but these appeared to be random occurrences. Similarly, no effect on foetal weights of males and females and crown rump lengths and number of litters with runts were observed in treated rats as compared to control. In addition, No dose-related increase in sternebral variations, skeletal variations and soft-tissue variations were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg body weight/day when Osborne-Mendel male and female rats were treated with Allura Red AC orally by gavage on days 0-19 of gestation.

Study 3

A multigeneration reproductive toxicity study of test material was performed on male and female wistar rats. The test material mixed with feed in dose concentration 0, 0.1, 1.0 or 3% (0, 50, 500, 1500 mg/kg bw per day) and administered orally for three successive generations. Each generation having 60 animals of each sex in the control and 40 animals of each sex in test animals. No adverse effects were observed with respect to fertility, litter size and weight, general condition, male/female ratio, growth during lactation, survival or maturation. Autopsy of parent rats and pups at weaning did not reveal any treatment related changes in organ weights other than caecal enlargement in the 3% dose group. Gross and microscopic examination of the F3 generation at weaning did not reveal any abnormalities due to treatment and no adverse effects were seen in the teratology study. It was concluded that Brilliant Black PN did not exert any adverse effects on reproductive function of Wistar rats when fed at dietary levels up to 3% (1500 mg/kg bw per day) for three successive generations. Therefore, NOAEL was considered to be at 3% (1500 mg/kg bw per day) for F0 F1 and F2 generation .When male and female wistar rats were treated with test material orally.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

Study 1

Three generation reproductive study of test material was performed on male and female Charles River CD rats. Animals were housed individually and fed the test diet ad libitum. The test material mixed with food in dose concentration 0, 5, 50, 150 or 500 mg/kg bw/day and 10 males and 20 females/group/generation were used .Pre-Mating Exposure / Males and Females were 60 days. Clinical observations were recorded twice daily with at least 5 hours between observations. Detailed physical examinations and palpation for masses were performed weekly. Body weights and food consumption were determined weekly for the first fourteen weeks, bi-weekly for the next 12 weeks and every 4 weeks thereafter until the end of the study. The intake of the test substance was determined from body weight, food consumption and dietary concentration. Haematology tests, including haemoglobin, haematocrit, erythrocyte and total and differential leukocyte counts, and erythrocyte morphology, were conducted on ten randomly selected animals at months 3, 6, 12, 18 and 24 of the study. Necropsies were conducted on all animals dying prior to study termination, killed in a moribund condition or killed on schedule. Histological examinations were conducted on all animals from both control groups, the highest dose group (2.0 or 5.0%) from each study and also on 10 rats randomly selected from each group for an interim sacrifice at 12 months. Histology was also performed on any animal with gross lesions or masses. Also tissues examined included adrenal glands, aorta, blood smear, brain, cecum, colon, duodenum, epididymus or uterus, esophagus, eyes, femur including marrow, tissue masses, gallbladder, heart, ileum, jejunum, duodenum, kidneys, liver, lungs and bronchi, mammary gland, nerves (sciatic), ovaries, lymph nodes, pancreas, parathyroids, pituitary gland, prostrate, rectum, skin, spleen, seminal vesicles, skeletal muscle, testes with epididymides, stomach, thymus, thyroid gland including parathyroid, trachea, urinary bladder, uterus.

 At the end of study, no treatment-related effects were reported on survival. No treatment-related changes were reported at gross necropsy. Histological evaluation revealed a variety of lesions, including neoplasms, present at similar incidences in control and treated animals. The authors considered the lesions to be spontaneous and not related to administration of the test material. There were no compound related effects on fertility, gestation, pup viability or lactation indices, on reproductive organs of females, or on organ weights among parents and offspring. There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats or from offspring. There was no toxicity to either the F1 or F2 generation. Food consumption was similar for control and treated animals at the lower dietary levels, but was slightly higher in the high-dose study, although not statistically significant. Haematological, clinical chemistry and urinalysis parameters did not differ significantly from the controls. Necropsies at one year did not reveal any treatment-related gross or microscopic changes. Hence NOAEL was considered to be 500mg/kg bw/day for F0, F1 and F2 generation. When male and female Charles River CD rats were treated with test material orally.

Study 2

In a reproductive toxicity study, Osborne-Mendel male and female rats were treated with Allura Red AC in the concentration of 0, 30, 75, 150, 300, 600 and 1000 mg/kg body weight/day orally by gavage on days 0-19 of gestation. No clinical sign of toxicity, no effect on body weight and food consumption were observed in treated rats as compared to control. No effect on numbers of corpora lutea and implants per female, total resorption, numbers of viable foetuses per female were observed in treated rats as compared to control. Increase in number of early deaths per litter and the number of early plus late deaths per litter were observed in in the 600 mg/kg group, but these appeared to be random occurrences. Similarly, no effect on foetal weights of males and females and crown rump lengths and number of litters with runts were observed in treated rats as compared to control. In addition, No dose-related increase in sternebral variations, skeletal variations and soft-tissue variations were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg body weight/day when Osborne-Mendel male and female rats were treated with Allura Red AC orally by gavage on days 0-19 of gestation.

Study 3

A multigeneration reproductive toxicity study of test material was performed on male and female wistar rats. The test material mixed with feed in dose concentration 0, 0.1, 1.0 or 3% (0, 50, 500, 1500 mg/kg bw per day) and administered orally for three successive generations. Each generation having 60 animals of each sex in the control and 40 animals of each sex in test animals. No adverse effects were observed with respect to fertility, litter size and weight, general condition, male/female ratio, growth during lactation, survival or maturation. Autopsy of parent rats and pups at weaning did not reveal any treatment related changes in organ weights other than caecal enlargement in the 3% dose group. Gross and microscopic examination of the F3 generation at weaning did not reveal any abnormalities due to treatment and no adverse effects were seen in the teratology study. It was concluded that Brilliant Black PN did not exert any adverse effects on reproductive function of Wistar rats when fed at dietary levels up to 3% (1500 mg/kg bw per day) for three successive generations. Therefore, NOAEL was considered to be at 3% (1500 mg/kg bw per day) for F0 F1 and F2 generation .When male and female wistar rats were treated with test material orally.

Based on the data available from different studies, NOAEL for test material was considered to be in range of 500 -1500mg/kg /day for reproductive toxicity, when male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAELfor test chemical was considered to be in range of 1000 -2500mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria ofCLP regulation test chemical is not likelyto classify as reproductive and developmental toxicant.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
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 from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on developmental toxicity studies on rats
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
No data available
Species:
rat
Strain:
other: 1.Osborne-Mendel 2.Wistar
Details on test animals or test system and environmental conditions:
1.TEST ANIMALS
- Source: FDA rat breeding colony
- Age at study initiation: female: 12 - 21 wk
- Weight at study initiation: female: 220 - 270 g
- Fasting period before study: no data
- Housing: Stainless-steel hanging cages.
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): Purina Laboratory Chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25 °C
- Humidity (%): 30 - 63%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle (8 am 8 pro).

IN-LIFE DATES: From: To:
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
test material , Certified Batch No. AA-4181 was dissolved in distilled water (w/v). Fresh solutions were prepared daily

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

VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 30, 75, 150, 300, 600 or 1000mg/kg/day.
- Amount of vehicle (if gavage): 1ml/100g body weight
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- M/F ratio per cage: 1:2
- Length of cohabitation: 1 day
- Proof of pregnancy: sperm in vaginal smear was considered day 0 of gestation
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: no data
- Further matings after two unsuccessful attempts: no data
- After successful mating each pregnant female was caged (how): no data
- Any other deviations from standard protocol: no data
Duration of treatment / exposure:
19 days (Gestation day 0 to day 19)
Frequency of treatment:
Daily
Duration of test:
19 days
Remarks:
Study 1
0, 30, 75, 150, 300, 600 or 1000mg/kg/day.
Study 2
0, 250, 500 or 2500 mg/kg
No. of animals per sex per dose:
1.Total: 294-301
0 mg/kg/bw/day : 42-43 females
30mg/kg/bw/day : 42-43 females
75mg/kg/bw/day : 42-43 females
150 mg/kg/bw/day : 42-43 females
300mg/kg/bw/day : 42-43 females
600mg/kg/bw/day : 42-43 females
1000 mg/kg/bw/day : 42-43 females
2.Total:120
0 mg/kg bw: 30 female
250 mg/kg bw : 30 female
500 mg/kg bw : 30 female
2500 mg/kg bw : 30 female



Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS:No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:The rats were weighed daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes ,food consumption was measured weekly.
- 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

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Water intake was not measured.
- Time schedule for examinations:
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:
Fetal examinations:
- External examinations: Yes: [all per litter ]
- Soft tissue examinations: Yes:The remaining half of the foetuses were fixed in Bouin's solution, serially sectioned by razor blade and examined under a dissecting microscope for internal variations of the soft tissues.
- Skeletal examinations: Yes:Approximately one-half of the foetuses were fixed in alcohol, stained with Alizarin Red S and examined under a dissecting microscope for all skeletal variations
- Head examinations: No data
Statistics:
All data analyses were performed by the Division of Mathematics at the FDA. Data on maternal initial body weights and food consumption were analysed by straight analysis of variance (ANOVA) and two-tailed t-test, and a regression analysis. The number of dams affected was analysed by a Fisher's exact test. Data on maternal weight gain were submitted to an analysis of covariance (ANOCOVA) and a two-tailed t-test. Data on the numbers of implants, corpora lutca, total viable young and viable males and females were analysed by ANOVA followed by a one-tailed t-test. Data on implantation efficiency, early deaths, late deaths and total resorptions (early and late deaths) were transformed by using the Freeman-Tukey arc-sine transformation (Freeman and Tukey, 1950) and then analysed by ANOVA and a one-tailed t-test. Data on litters having one or more or two or more resorptions were analysed by a Fisher's exact test. Similar tests were applied to the number of runts per litter. Data on foetal body weights, crown-!o-rump lengths and foetal ossified vertebrae were analysed by nested ANOVA and a one-tailed t-test. The ANOVA included a correction for unequal sample size (Sokal and Rohlf, 1981). Data on the average number of foetuses per litter with skeletal, sternebral, combined missing plus incomplete plus bipartite sternebrae or soft-tissue variations were transformed by using the Frceman-Tukey arc-sine transformation and then analysed by ANOVA and a one-tailed t-test. Litters with foetuses with at least one, at least two, etc. skeletal, sternebral, combined missing plus incomplete plus bipartite sternebrae, or soft-tissue variations, and specific variations were analysed by Fisher's exact test.
Indices:
No data available
Historical control data:
No data available
Clinical signs:
no effects observed
Description (incidence and severity):
1.No unusual behaviours were observed in the animals during the study.The external maternal findings were unremarkabke.
2.No abnormalities in condition or behaviour of the dams were observed
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One female inthe group given 300mg/kg died at day 12 of gestation as a result of gavage difficulties unrelated to dosage
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Initial body weight at day 0 and maternal body-weight gain during gestation were similar in all groups
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Mean food consumption on days 0-7, 7-14, 14 -20 and 0-20 by the treated animals was similar to that of the control animals
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Number of abortions:
no effects observed
Description (incidence and severity):
No litters were totally resorbed
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
The mean numbers of corpora lutea and implants per female were similar in all groups
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
The percentage of females with at least one resorption was similar in all groups. The percentage of females given 600mg/kg that had at least two resorptions was significantly greater than the percentage in the control females, but there was no dose related effect in the percentage of females with at least two resorptions.
Early or late resorptions:
no effects observed
Description (incidence and severity):
The number of early deaths per litter and the number of early plus late deaths per litter were greatest in the 600 mg/kg group, but these appeared to be random occurrences. The percentage of females with at least one resorption was similar in all groups. The percentage of females given 600mg/kg that had at least two resorptions was significantly greater than the percentage in the control females, but there was no dose related effect in the percentage of females with at least two resorptions.
Dead fetuses:
no effects observed
Description (incidence and severity):
The mean number of viable foetuses per female was similar in all groups. The number of viable male foetuses was increased over the control value in the groups given 30 and 1000mg/kg, but because of the lack of relation to dosage, these increases were considered to be random. The number of viable female foetuses was not affected in any group.
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
The pregnancy rate ranged from 85.71 to 95.35% with no evidence of dose correlation.
Other effects:
not specified
Details on maternal toxic effects:
No abnormalities in condition or behaviour of the dams were observed in either study
Dose descriptor:
NOAEL
Effect level:
> 1 000 - <= 2 500 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
changes in number of pregnant
clinical signs
dead fetuses
early or late resorptions
food consumption and compound intake
mortality
number of abortions
pre and post implantation loss
total litter losses by resorption
Remarks on result:
other: No maternal toxic effects on was observed
Abnormalities:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Mean foetal weights of males and females and crown rump lengths were similar in all groups. The number of litters with runts was similar in all groups.
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
1. Aside from haemorrhages present in foetuses in all groups in similar numbers, there were no other external variations in any of the dosed groups. Two foetuses had multiple anomalies: one foetus from the control group had a club foot, a short tail and four digits on the hind legs, and one foetus from the 150-mg/kg group had exencephalus and hydrocephalus. Three foetuses in a single litter of a female given 75 mg/kg were oedematous
2.One foetus in the 2500mg/kg bw dose group showed a complexity of malformations but this was considered to be a fortuitous finding
Skeletal malformations:
no effects observed
Description (incidence and severity):
No dose-related increase in sternebral variations was seen among the foetuses with reduced ossification, bipartite, missing, malaligned or fused sternebrae,nor in the numbers of litters containing these foetuses. The incidence of specific skeletal variations was similar in all groups of foetuses, except for a significant increase in the incidence of 14th rib bud in foetuses from the 300 mg/kg group which was considered to be a random occurrence.
Visceral malformations:
no effects observed
Description (incidence and severity):
The numbers of soft-tissue variations in foetuses and their incidences per litter were similar in all groups except for a significant increase in the 150-mg/kg group in the number of litters with at least one variation; this increase was considered a random occurrence
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
At autopsy, no signs of embryo-toxicity or teratogenicity were observed
Dose descriptor:
NOAEL
Effect level:
> 1 000 - <= 2 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
external malformations
skeletal malformations
visceral malformations
Remarks on result:
other: No developmental toxic effects was observed
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
In developmental toxicity study, The NOAEL was considered to be in range of 1000 -1500 mg/kg bw as no effects on development of fetus was observed , when male and female rats were treated with test material orally in 20 days
Executive summary:

Data available from different studies for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

The developmental toxicity study of test material was performed on male and female Osborne-Mendel rats. The test material was dissolved in distilled water (w/v). Fresh solutions were prepared daily. The dose concentration of 0, 30, 75, 150, 300, 600 or 1000mg/kg/day was administered by oral gavage route in volume 1ml/100g body weight .On mating days , two females were randomly mated with one male at approximately 4.30pm. The following morning, a vaginal smear was obtained from each female to determine whether copulation had taken place. Sperm positive dams were considered to be at day 0 of gestation. 42-43 female/dose groups were used. All the animals were observed for signs of toxicity. The rats were weighed daily and food consumption was measured weekly. Water intake was not measured. On day 20 of gestation, the females were examined for gross abnormalities for the last time before being killed by CO, asphyxiation. Caesarean sections were performed. Corpora lutea were counted. The uterus was opened and examined in situ. The uterine positions of all implantation sites were noted and their condition (early or late resorptions, living or dead foetuses) was determined. Each live foetus was promptly weighed, sexed and examined for gross external malformations, and the crown-rump length was measured. Any foetus that weighed less than 70% of the average weight of the concurrent male or female controls was considered to be a runt. Approximately one-half of the foetuses were fixed in alcohol, stained with Alizarin Red S and examined under a dissecting microscope for all skeletal variations. The remaining half of the foetuses were fixed in Bouin's solution, serially sectioned by razor blade and examined under a dissecting microscope for internal variations of the soft tissues.

No unusual behaviours were observed in the animals during the study. The external maternal findings were unremarkable. One female in the group given 300mg/kg died at day 12 of gestation as a result of gavage difficulties unrelated to dosage. Initial body weight at day 0 and maternal body-weight gain during gestation were similar in all groups. Mean food consumption on days 0-7, 7-14, 14 -20 and 0-20 by the treated animals was similar to that of the control animals. The pregnancy rate ranged from 85.71 to 95.35% with no evidence of dose correlation.

The mean numbers of corpora lutea and implants per female were similar in all groups. No litters were totally resorbed. The mean number of viable foetuses per female was similar in all groups. The number of viable male foetuses was increased over the control value in the groups given 30 and 1000mg/kg, but because of the lack of relation to dosage, these increases were considered to be random. The number of viable female foetuses was not affected in any group. The number of early deaths per litter and the number of early plus late deaths per litter were greatest in the 600 mg/kg group, but these appeared to be random occurrences. The percentage of females with at least one resorption was similar in all groups. The percentage of females given 600mg/kg that had at least two resorptions was significantly greater than the percentage in the control females, but there was no dose related effect in the percentage of females with at least two resorptions.Mean foetal weights of males and females and crown rump lengths were similar in all groups. The number of litters with runts was similar in all groups.Aside from haemorrhages present in foetuses in all groups in similar numbers, there were no other external variations in any of the dosed groups. Two foetuses had multiple anomalies: one foetus from the control group had a club foot, a short tail and four digits on the hind legs, and one foetus from the 150-mg/kg group had exencephalus and hydrocephalus. Three foetuses in a single litter of a female given 75 mg/kg were oedematous. No dose-related increase in sternebral variations was seen among the foetuses with reduced ossification, bipartite, missing, malaligned or fused sternebrae, nor in the numbers of litters containing these foetuses. The incidence of specific skeletal variations was similar in all groups of foetuses, except for a significant increase in the incidence of 14th rib bud in foetuses from the 300 mg/kg group which was considered to be a random occurrence. Hence the NOAEL was considered to be 1000mg/kg bw as no effects on development of fetus was observed, when Osborne-Mendel male and female rats were treated with test material orally in 20 days.

Study 2

A teratogenicity study of test material was performed on SPF-derived Wistar female rats. Study were conducted in two stages as the preliminary study four groups of 15 pregnant rats, the test material in dose concentration 0, 250, 500 or 2500 mg/kg bw administered orally by gavage from day 0-19 of pregnancy. In second study, 30 pregnant rats were used with same protocol. On day 21, the animals were killed and ovaries and uterus removed. The number of corpora lutea in each ovary was recorded and the foetuses examined. Live foetuses, embryonic and foetal resorptions and dead foetuses were counted and the number and position of implantation sites were recorded, In the second study, half the foetuses in the control and top dose groups were examined for skeletal malformations and half for visceral defects. No abnormalities in condition or behaviour of the dams were observed in either study. At autopsy, no signs of embryo-toxicity or teratogenicity were observed. One foetus in the 2500mg/kg bw dose group showed a complexity of malformations but this was considered to be a fortuitous finding. Hence NOAEL was considered to be at 2500 mg/kg bw as no signs of embryo-toxicity or teratogenicity were observed. When female wistar rats were treated with test material orally.

 

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from authoritative database
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Developmental toxicity study

Data available from different studies for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

The developmental toxicity study of test material was performed on male and female Osborne-Mendel rats. The test material was dissolved in distilled water (w/v). Fresh solutions were prepared daily. The dose concentration of 0, 30, 75, 150, 300, 600 or 1000mg/kg/day was administered by oral gavage route in volume 1ml/100g body weight .On mating days , two females were randomly mated with one male at approximately 4.30pm. The following morning, a vaginal smear was obtained from each female to determine whether copulation had taken place. Sperm positive dams were considered to be at day 0 of gestation. 42-43 female/dose groups were used. All the animals were observed for signs of toxicity. The rats were weighed daily and food consumption was measured weekly. Water intake was not measured. On day 20 of gestation, the females were examined for gross abnormalities for the last time before being killed by CO, asphyxiation. Caesarean sections were performed. Corpora lutea were counted. The uterus was opened and examined in situ. The uterine positions of all implantation sites were noted and their condition (early or late resorptions, living or dead foetuses) was determined. Each live foetus was promptly weighed, sexed and examined for gross external malformations, and the crown-rump length was measured. Any foetus that weighed less than 70% of the average weight of the concurrent male or female controls was considered to be a runt. Approximately one-half of the foetuses were fixed in alcohol, stained with Alizarin Red S and examined under a dissecting microscope for all skeletal variations. The remaining half of the foetuses were fixed in Bouin's solution, serially sectioned by razor blade and examined under a dissecting microscope for internal variations of the soft tissues.

No unusual behaviours were observed in the animals during the study. The external maternal findings were unremarkable. One female in the group given 300mg/kg died at day 12 of gestation as a result of gavage difficulties unrelated to dosage. Initial body weight at day 0 and maternal body-weight gain during gestation were similar in all groups. Mean food consumption on days 0-7, 7-14, 14 -20 and 0-20 by the treated animals was similar to that of the control animals. The pregnancy rate ranged from 85.71 to 95.35% with no evidence of dose correlation.

The mean numbers of corpora lutea and implants per female were similar in all groups. No litters were totally resorbed. The mean number of viable foetuses per female was similar in all groups. The number of viable male foetuses was increased over the control value in the groups given 30 and 1000mg/kg, but because of the lack of relation to dosage, these increases were considered to be random. The number of viable female foetuses was not affected in any group. The number of early deaths per litter and the number of early plus late deaths per litter were greatest in the 600 mg/kg group, but these appeared to be random occurrences. The percentage of females with at least one resorption was similar in all groups. The percentage of females given 600mg/kg that had at least two resorptions was significantly greater than the percentage in the control females, but there was no dose related effect in the percentage of females with at least two resorptions.Mean foetal weights of males and females and crown rump lengths were similar in all groups. The number of litters with runts was similar in all groups.Aside from haemorrhages present in foetuses in all groups in similar numbers, there were no other external variations in any of the dosed groups. Two foetuses had multiple anomalies: one foetus from the control group had a club foot, a short tail and four digits on the hind legs, and one foetus from the 150-mg/kg group had exencephalus and hydrocephalus. Three foetuses in a single litter of a female given 75 mg/kg were oedematous. No dose-related increase in sternebral variations was seen among the foetuses with reduced ossification, bipartite, missing, malaligned or fused sternebrae, nor in the numbers of litters containing these foetuses. The incidence of specific skeletal variations was similar in all groups of foetuses, except for a significant increase in the incidence of 14th rib bud in foetuses from the 300 mg/kg group which was considered to be a random occurrence. Hence the NOAEL was considered to be 1000mg/kg bw as no effects on development of fetus was observed, when Osborne-Mendel male and female rats were treated with test material orally in 20 days.

Study 2

A teratogenicity study of test material was performed on SPF-derived Wistar female rats. Study were conducted in two stages as the preliminary study four groups of 15 pregnant rats, the test material in dose concentration 0, 250, 500 or 2500 mg/kg bw administered orally by gavage from day 0-19 of pregnancy. In second study, 30 pregnant rats were used with same protocol. On day 21, the animals were killed and ovaries and uterus removed. The number of corpora lutea in each ovary was recorded and the foetuses examined. Live foetuses, embryonic and foetal resorptions and dead foetuses were counted and the number and position of implantation sites were recorded, In the second study, half the foetuses in the control and top dose groups were examined for skeletal malformations and half for visceral defects. No abnormalities in condition or behaviour of the dams were observed in either study. At autopsy, no signs of embryo-toxicity or teratogenicity were observed. One foetus in the 2500mg/kg bw dose group showed a complexity of malformations but this was considered to be a fortuitous finding. Hence NOAEL was considered to be at 2500 mg/kg bw as no signs of embryo-toxicity or teratogenicity were observed. When female wistar rats were treated with test material orally.

 

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAELfor test chemical was considered to be in range of 1000 -2500mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria ofCLP regulation testchemical isnot likelyto classify as reproductive and developmental toxicant.

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Additional information