Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Toxicity to Reproduction

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo] naphthalen-2-amine.

Based on the data available for the read across chemicals, N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo]naphthalen-2-amine (CAS no 93964 -07 -9) does not reproductive toxicity at dose concentration > 1000 -1500mg/kg bw . Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

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 of read across substances
Justification for type of information:
Data for the target chemical is summarized based on the structurally similar read across chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on three reproductive toxicity studies via dermal and oral route on rats,
1.Reproductive toxicity study of test material was performed in mice by repeated dermal application for 18 months.
2.Reproductive toxicity of test material was assessed in Osborne-Mendel rats in 20 days study
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
other: 1. mice 2.rats
Strain:
other: 1.Swiss Webster 2.Osborne-Mendel
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
1.TEST ANIMALS
- Source: No data available
- Age at study initiation: (P) : No data available
- Weight at study initiation: (P) : 17 to 25 g
- Fasting period before study: No data available
- Housing: Animals of sex were housed five per cage
- Diet (e.g. ad libitum): Purina Laboratory Chow , ad libitum
- Water (e.g. ad libitum): Fresh water, ad libitum
- 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): No data available
Study 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. dermal 2.oral: gavage
Vehicle:
other: 1. 0.1 % solution of sodium lauryl sulfate 2.water
Details on exposure:
Study 1.
PREPARATION OF DOSING SOLUTIONS:
The test material diluted in 0.1 % solution of sodium lauryl sulfate
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): The test material dissolved in 0.1 % solution of sodium lauryl sulfate
- Concentration in vehicle: 1.0 % (1500 mg/kg bw/day)
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity:No data available
Study 2.
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
Details on mating procedure:
1. No data
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:
Study 1. 18 months
Study 2. 19 days (Gestation day 0 to day 19)
Frequency of treatment:
Study 1.Twice weekly
Study 2. Daily
Details on study schedule:
No data available
Remarks:
Study 1.1.0 % (1500 mg/kg bw/day)
Study 2.0, 30, 75, 150, 300, 600 or 1000mg/kg/day.
No. of animals per sex per dose:
1.
Total: 300
0 mg/kg/bw/day : 100 males and 100 females
1500 mg/kg bw/day: 50 males and 50 females
2.
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


Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
1.
CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations:Mean body weight was determined at 3,6, 9, 12, 15, and 18 months;

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

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

OTHER:Survival ,visible or palable growth and behavior were examined.
Study 2.
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:

OTHER:
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
2.STANDARDISATION OF LITTERS:No data available
- Performed on day 4 postpartum: [yes/no]
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 ] offspring:
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.

GROSS EXAMINATION OF DEAD PUPS:Yes
[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.]

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: No data available

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data available
Postmortem examinations (parental animals):
1.
SACRIFICE: yes after 18 months
- 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: yes
Organs were fixed in 10% formalin solution after recording any gross pathological findings


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

2.
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: On day 20 of gestation,

GROSS NECROPSY: yes ,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.
-

HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
2.
SACRIFICE: yes
- 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

HISTOPATHOLOGY / ORGAN WEIGTHS
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
Description (incidence and severity):
1. No abnormalities were observed in treated mice as compared to control.
2.No unusual behaviours were observed in the animals during the study.The external maternal findings were unremarkabke.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
1. No effect on survival of treated mice were obserevd as compared to control.
2. 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):
1. No significant effect was observed on body weight of treated mice as compared to control.
2. 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):
1.No significant effect was observed on food consumption of treated mice as compared to control.
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:
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
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
1.All grades Malignant lymphoma and Variation in nuclear morphology in Liver, Infarction, Malignant and Myeloid metaplasia, Leukocytic aggregations in Spleen, all grades Malignant lymphoma , Leukemic and round cell infiltration and Leukocytic aggregation in Kidneys , Malignant lymphoma in Lymph Nodes, Malignant lymphoma, Inflammation, pneumonitis, bronchitis, Necrotic changes in Lungs and all grades Malignant lymphoma in Thymus were observed in male and female treated miceNo significant difference in the incidence of this lesion were observed as compared to control and considered to be not treatment related.
Histopathological findings: neoplastic:
not specified
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):
2.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.
Dose descriptor:
NOAEL
Effect level:
> 1 000 - <= 1 500 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
gross pathology
histopathology: non-neoplastic
reproductive performance
Remarks on result:
other: overall no toxic effects observed
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
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Description (incidence and severity):
2. 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.
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:
no effects observed
Description (incidence and severity):
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:
not specified
Other effects:
no effects observed
Description (incidence and severity):
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.
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
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
clinical signs
mortality
body weight and weight gain
gross pathology
Remarks on result:
other: overall no developmental toxic effects observed
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
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
Conclusions:
The NOAEL for reproductive toxicity was considered to be >1000-1500 mg/kg bw/day as No effects on reproductive parameters were observed .When male and female rats were treated with N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo] naphthalen-2-amine (933964-07-9) orally.
Executive summary:

Reproductive toxicity study

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo] naphthalen-2-amine. The studies are as mentioned below:

Study1.

The reproductive toxicity study was considered on the bases of a repeated dose dermal toxicity study, Swiss-Webster male and female mice were treated with test material in the concentration of 1500 mg/kg bw/day in 0.1 % solution of sodium lauryl sulfate applied twice weekly on 6 cm2dorsal area of skin for 18 months. No effect was observed on survival, clinical sign and body weight of treated male and female mice as compared to control. Similarly, lesion in Mammary Gland and Subcutaneous papillary were observed gross pathologically in treated mice, but the observed effect was similar to control. In addition, All grades Malignant lymphoma and Variation in nuclear morphology in Liver, Infarction, Malignant and Myeloid metaplasia, Leukocytic aggregations in Spleen, all grades Malignant lymphoma , Leukemic and round cell infiltration and Leukocytic aggregation in Kidneys , Malignant lymphoma in Lymph Nodes, Malignant lymphoma, Inflammation, pneumonitis, bronchitis and Necrotic changes in Lungs and all grades Malignant lymphoma in Thymus were observed in male and female treated mice. No significant difference in the incidence of this lesion were observed as compared to control and considered to be not treatment related. No effects were observed on male and female reproductive organ. Therefore, NOAEL was considered to be 1500 mg/kg bw/day when Swiss-Webster male and female mice were treated with test material by dermal application.

Study 2.

The reproductive and 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 reproductive performance and development of fetus was observed, when Osborne-Mendel male and female rats were treated with test material orally in 20 days.

Based on the data available for the read across chemicals, N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo]naphthalen-2-amine (CAS no 93964 -07 -9) does not reproductive toxicity at dose concentration >1000 -1500mg/kg bw . Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
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 for the read across chemicals was reviewed to determine the reproductive toxicity of N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo] naphthalen-2-amine. The studies are as mentioned below:

Study1.

The reproductive toxicity study was considered on the bases of a repeated dose dermal toxicity study, Swiss-Webster male and female mice were treated with test material in the concentration of 1500 mg/kg bw/day in 0.1 % solution of sodium lauryl sulfate applied twice weekly on 6 cm2dorsal area of skin for 18 months. No effect was observed on survival, clinical sign and body weight of treated male and female mice as compared to control. Similarly, lesion in Mammary Gland and Subcutaneous papillary were observed gross pathologically in treated mice, but the observed effect was similar to control. In addition, All grades Malignant lymphoma and Variation in nuclear morphology in Liver, Infarction, Malignant and Myeloid metaplasia, Leukocytic aggregations in Spleen, all grades Malignant lymphoma , Leukemic and round cell infiltration and Leukocytic aggregation in Kidneys , Malignant lymphoma in Lymph Nodes, Malignant lymphoma, Inflammation, pneumonitis, bronchitis and Necrotic changes in Lungs and all grades Malignant lymphoma in Thymus were observed in male and female treated mice. No significant difference in the incidence of this lesion were observed as compared to control and considered to be not treatment related. No effects were observed on male and female reproductive organ. Therefore, NOAEL was considered to be 1500 mg/kg bw/day when Swiss-Webster male and female mice were treated with test material by dermal application.

Study 2.

The reproductive and 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 reproductive performance and development of fetus was observed, when Osborne-Mendel male and female rats were treated with test material orally in 20 days.

Based on the data available for the read across chemicals, N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo]naphthalen-2-amine (CAS no 93964 -07 -9) does not reproductive toxicity at dose concentration >1000 -1500mg/kg bw . Hence the test chemical is not likely to classify as a reproductive toxicant as per the criteria mentioned in CLP regulation.

Effects on developmental toxicity

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation N-(2-ethylhexyl)-1-[[2-methyl-4-[(4-methylphenyl)azo]phenyl]azo]naphthalen-2-amine (93964-07-9) cannot be classified as reproductive toxicant.