2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3)

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 500mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated 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 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 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 three reproductive toxicity studies via oral route on rats,1.Combined repeated dose toxicity study with reproduction and developmental screening of test material in rats.2.The reproductive effects of test material on male and female Charles River CD rats by oral (Diet) route at different dose levels was examined. 3.Multigeneration toxicity study of test material was performed on male and female wistar rats.

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated- IUPAC name: 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated- Molecular formula: C40H36N4O1 - Molecular wt. : 588.76 g/mole- smiles : Cc1ccccc1N=Nc2ccc(c(C)c2)N=Nc3c4ccc(C(C)c5ccccc5)cc4cc(C(C)c6ccccc6)c3O- Substance type: Organic- Physical state: solid

Species:

rat

Strain:

other: 1. Crj: CD(SD),2.Charles River CD3.Wistar

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.TEST ANIMALS- Source:No data available- Females (if applicable) nulliparous and non-pregnant: [yes/no]- Age at study initiation: No data available- Weight at study initiation: No 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 availableENVIRONMENTAL 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

Route of administration:

other: 1.oral: gavage 2.oral: feed 3.oral: feed

Vehicle:

other: 1.0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80

Details on exposure:

1.Details on exposurePREPARATION OF DOSING SOLUTIONS: The test material mixed with 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80VEHICLE- Justification for use and choice of vehicle (if other than water): The test material mixed with 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80- Concentration in vehicle: 0 (vehicle), 100, 300, 1000 mg/kg- Amount of vehicle (if gavage):No data available- Lot/batch no. (if required):No data available- Purity:No data available2.PREPARATION OF DOSING SOLUTIONS: test material mixed with food 3.PREPARATION OF DOSING SOLUTIONS: test material mixed with food

Details on mating procedure:

No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

1.Males, 37 daysFemales, from 14 days before mating to day 4 of lactation2.Pre-Mating Exposure / Males and Females: 60 days and test material adminstered upto three generation 3.From F1 to F3 generation

Frequency of treatment:

1. Daily

Details on study schedule:

No data available

Remarks:

Study 1.0 (vehicle), 100, 300, 1000 mg/kgStudy 2.5, 50, 150 or 500 mg/kg bw/dayStudy 30, 0.1, 1.0 or 3% (0, 50, 500,1500 mg/kg bw per day)

No. of animals per sex per dose:

Study1.Total: 960 mg/kg bw/day: 12 male, 12 female 100 mg/kg bw/day: 12 male, 12 female 100 mg/kg bw/day: 12 male, 12 female 1000 mg/kg bw/day: 12 male, 12 female Study 2.Total :1500mg/kg bw/day : 10 males and 20 females5 mg/kg bw/day : 10 males and 20 females50 mg/kg bw/day :10 males and 20 females150 mg/kg bw/day :10 males and 20 females 500 mg/kg bw/day : 10 males and 20 femalesStudy3Total:3600 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.CAGE SIDE OBSERVATIONS: Yes - Time schedule: - Cage side observations checked in table [No.?] were included. DETAILED CLINICAL OBSERVATIONS: Yes / No / No data - Time schedule: BODY WEIGHT: Yes - Time schedule for examinations: FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes - Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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): No data - Time schedule for examinations: 2.Parental animals observation and examinationsCAGE SIDE OBSERVATIONS: yes Clinical observations were recorded twice daily with at least 5 hours between observationsDETAILED CLINICAL OBSERVATIONS: Yes Time schedule: Detailed physical examinations and palpation for masses were performed weekly.BODY WEIGHT: YesTime 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.Study 3CAGE SIDE OBSERVATIONS: No data - Time schedule: - Cage side observations checked in table [No.?] were included. DETAILED CLINICAL OBSERVATIONS: Yes / No / No data - Time schedule: BODY WEIGHT: Yes - Time schedule for examinations: FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): - 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:

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

No data available

Postmortem examinations (parental animals):

1.SACRIFICE Males, day 38Females, day 5 of lactation GROSS NECROPSY: yes - Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.] 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: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.] GROSS NECROPSYNecropsies 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 WEIGHTSHistological 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 massesStudy 3Postmortem examinations (Parent Animal)SACRIFICEYes, Autopsy did of parent ratsGROSS NECROPSY:No data availableHISTOPATHOLOGY / ORGAN WEIGHTS: No data available

Postmortem examinations (offspring):

1.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 numbers of offspring or live offspring, the sex ratio, the live birth index, the viability index or body weights,external features, clinical signs or necropsy findings for the offspring were noted HISTOPATHOLOGY / ORGAN WEIGTHS The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.Study 3Postmortem examinations (offspring)SACRIFICEF1,F2, F3 pups were sacrificed.GROSS NECROPSYYesHISTOPATHOLOGY / ORGAN WEIGTHSYes

Statistics:

No data available

Reproductive indices:

1.mating index, the fertility, index, , the implantation index, the delivery index, gestation index, gestation length, parturition or maternal behavior were observed.

Offspring viability indices:

1.The live birth index, the viability index or body weights were observed.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1.No effect observed in treated rats

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

2.No treatment-related effects were reported on survival

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1.No effect on body weight of treated rats was observed as compared to control.

Food consumption and compound intake (if feeding study):

not specified

Description (incidence and severity):

1. No effect on food consumption of treated rats was observed as compared to control.

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 effect on haematological findings of treated rats was observed as compared to control.2.No treatment-related effects were reported on haematological finding

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

2. No treatment-related effects were reported on clinical chemistry

Urinalysis findings:

no effects observed

Description (incidence and severity):

2.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:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

1.No histopathological changes were observed in treatd rats as compared to control. 2.no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

4.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. No effects on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behavior of treated rats were observed. 2.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.3.No treatment-related effects on reproductive function was observed

Dose descriptor:

NOAEL

Effect level:

> 500 - < 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

haematology

clinical biochemistry

urinalysis

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

histopathology: neoplastic

reproductive performance

Remarks on result:

other: overall no effects on reproductive performance

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:

no effects observed

Description (incidence and severity):

1.No clinical signs of toxicity were observed in offspring as compared to cotnrol.

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

1.No significant differences in numbers of offspring or live offspring were observed as compared to cotnrol.2.no compound related effects on pup viability or lactation indices.3. no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

1.No effect on body weight of offspring were observed as compared to cotnrol.2.no effects observed on body weight

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

2.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:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

2.no compound related effects on organ weights3.no effects observed on organ weights

Gross pathological findings:

no effects observed

Description (incidence and severity):

1.No gorss pathological changes were observed in offspring as compared to cotnrol.2.Necropsies did not reveal any treatment-related gross or microscopic changes.

Histopathological findings:

no effects observed

Description (incidence and severity):

2.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):

2.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

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

organ weights and organ / body weight ratios

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

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):

2.No treatment-related effects were reported on Haematological finding

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

2.No treatment-related effects were reported on clinical chemistry

Urinalysis findings:

no effects observed

Description (incidence and severity):

2.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):

2.no compound related effects on organ weights

Gross pathological findings:

no effects observed

Description (incidence and severity):

2.Necropsies did not reveal any treatment-related gross or microscopic changes.

Histopathological findings:

no effects observed

Description (incidence and severity):

2.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

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

haematology

organ weights and organ / body weight ratios

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 between 500-1500 mg/kg bw/day as No effects on reproductive parameters were observed .When male and female rats were treated with 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3) orally.

Executive summary:

Reproductive toxicity

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3). The studies are as mentioned below:

Study 1.

In a combined repeated dose toxicity study with reproduction and developmental screening, of test material was performed according to OECD Test Guideline 422.Crj:CD(SD)IGS male and female rats were treated with test material in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.12 male and 12 female were placed in each group. Males were exposed with test chemical for 37 days while Females, from 14 days before mating to day 4 of lactation. Clinical sign, body weight, food consumption were observed. On day 38 male rats were killed while female on day 5 of lactation.

 No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test material orally by gavage.

Study 2

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 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.

 

 Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 500mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to classify as reproductive toxicant.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from secondary source

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

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3). The studies are as mentioned below:

Study 1.

In a combined repeated dose toxicity study with reproduction and developmental screening, of test material was performed according to OECD Test Guideline 422.Crj:CD(SD)IGS male and female rats were treated with test material in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.12 male and 12 female were placed in each group. Males were exposed with test chemical for 37 days while Females, from 14 days before mating to day 4 of lactation. Clinical sign, body weight, food consumption were observed. On day 38 male rats were killed while female on day 5 of lactation.

 No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test material orally by gavage.

Study 2

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 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.

 

 Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 500mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to classify as reproductive toxicant.

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Data available for the read across chemicals was reviewed to determine the Developmental toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3)

Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 500mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to 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 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 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 two developmental toxicity studies via oral route on rats,1.Combined repeated dose toxicity study with reproduction and developmental screening of test material in rats2.Developmental study perfored on SPF-derived Wistar rats when treated with test material 3.The teratogenic toxicity study of test material was performed on rat

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated- IUPAC name: 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl) azo] phenyl] azo]-, ar-styrenated- Molecular formula: C40H36N4O1 - Molecular wt. : 588.76 g/mole- smiles : Cc1ccccc1N=Nc2ccc(c(C)c2)N=Nc3c4ccc(C(C)c5ccccc5)cc4cc(C(C)c6ccccc6)c3O- Substance type: Organic- Physical state: solid

Species:

rat

Strain:

other: 1.Crj:CD(SD)IGS 2.Wistar 3.Osborne-Mendel

Details on test animals or test system and environmental conditions:

3.EST 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 dataENVIRONMENTAL 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:

other: 1.0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 2. 2.water

Details on exposure:

1.Details on exposurePREPARATION OF DOSING SOLUTIONS: The test material mixed with 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80DIET 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 availableVEHICLE- Justification for use and choice of vehicle (if other than water): The test material mixed with 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80- Concentration in vehicle: 0 (vehicle), 100, 300, 1000 mg/kg- Amount of vehicle (if gavage):No data available- Lot/batch no. (if required):No data available- Purity:No data available3.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:

3.- 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:

1.Males, 37 daysFemales, from 14 days before mating to day 4 of lactation2.19 days (from day 0-19 of pregnancy)3.19 days (Gestation day 0 to day 19)

Frequency of treatment:

Daily

Duration of test:

1.Approx 60 days 2.Till F1 generation3.19 days

Remarks:

Study1.0 (vehicle), 100, 300, 1000 mg/kgStudy2.0, 250, 500 or 2500 mg/kgStudy 3.0, 30, 75, 150, 300, 600 or 1000mg/kg/day.

No. of animals per sex per dose:

Study1.Total: 960 mg/kg bw/day: 12 male, 12 female 100 mg/kg bw/day: 12 male, 12 female 100 mg/kg bw/day: 12 male, 12 female 1000 mg/kg bw/day: 12 male, 12 female Study2.Total:1200 mg/kg bw: 30 female 250 mg/kg bw : 30 female500 mg/kg bw : 30 female 2500 mg/kg bw : 30 femaleStudy 3.Total: 294-3010 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 females300mg/kg/bw/day : 42-43 females600mg/kg/bw/day : 42-43 females1000 mg/kg/bw/day : 42-43 females

Control animals:

yes

Details on study design:

No data available

Maternal 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: 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: POST-MORTEM EXAMINATIONS: yes - Sacrifice on gestation day:day 5 of lactation - Organs examined: numbers of corpora lutea or implantations 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: No data - Time schedule for examinations: FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data - 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): No data - Time schedule for examinations: POST-MORTEM EXAMINATIONS: No data - Sacrifice on gestation day - Organs examined: 3.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:

1.The ovaries and uterine content was examined after termination: Yes Examinations included: - Gravid uterus weight: No data - Number of corpora lutea: Yes - Number of implantations: Yes - Number of early resorptions: No data - Number of late resorptions: No data2.The number of corpora lutea in each ovary was recorded and the foetuses examined.3.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:

1.- External examinations: Yes: - Soft tissue examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data - Skeletal examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data - Head examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data2.Live foetuses, embryonic and foetal resorptions and dead foetuses were counted and the number and position of implantation sites were recorded.3.- 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:

3.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:

1.reproductive parameters such as the mating index, the fertility index, the implantation index, the delivery index, the gestation index,the live birth index, the viability index were observed

Historical control data:

No data available

Clinical signs:

no effects observed

Description (incidence and severity):

2.No abnormalities in condition or behaviour of the dams were observed 3.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):

3.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):

3.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):

not specified

Description (incidence and severity):

3.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

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:

effects observed, non-treatment-related

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Number of abortions:

no effects observed

Description (incidence and severity):

3.No litters were totally resorbed

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

3.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):

3.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):

3.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:

not specified

Description (incidence and severity):

3.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):

3.The pregnancy rate ranged from 85.71 to 95.35% with no evidence of dose correlation.
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:

not specified

Other effects:

not specified

Details on maternal toxic effects:

2.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.

Basis for effect level:

body weight and weight gain

clinical signs

gross pathology

mortality

number of abortions

organ weights and organ / body weight ratios

pre and post implantation loss

total litter losses by resorption

Remarks on result:

other: overall no developmental toxic effects observed

Abnormalities:

not specified

Localisation:

not specified

Fetal body weight changes:

no effects observed

Description (incidence and severity):

3.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.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified

Reduction in number of live offspring:

no effects observed

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):

2.One foetus in the 2500mg/kg bw dose group showed a complexity of malformations but this was considered to be a fortuitous finding.3. 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

Skeletal malformations:

not specified

Description (incidence and severity):

3.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:

not specified

Description (incidence and severity):

3.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

Dose descriptor:

NOAEL

Effect level:

> 1 000 - <= 2 500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

fetal/pup body weight changes

external malformations

Remarks on result:

other: No effects on external malformation

Abnormalities:

not specified

Localisation:

other: not specified

Developmental effects observed:

not specified

Treatment related:

not specified

Relation to maternal toxicity:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

The NOAEL for reproductive toxicity was considered to be betwwen 1000-2500 mg/kg bw/day as No effects on reproductive parameters were observed .When male and female rats were treated with 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3) orally.

Executive summary:

Developmental toxicity study

Data available for the read across chemicals was reviewed to determine the Developmental toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3). The studies are as mentioned below:

Study 1.

In a combined repeated dose toxicity study with reproduction and developmental screening, of test material was performed according to OECD Test Guideline 422.Crj:CD(SD)IGS male and female rats were treated with test material in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.12 male and 12 female were placed in each group. Males were exposed with test chemical for 37 days while Females, from 14 days before mating to day 4 of lactation. Clinical sign, body weight, food consumption were observed. On day 38 male rats were killed while female on day 5 of lactation.

 No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test material orally by gavage.

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.

Study 3.

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.

 

  Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 1000mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to classify as developmental toxicant.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from secondary source

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 for the read across chemicals was reviewed to determine the Developmental toxicity of 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated (85203-90-3). The studies are as mentioned below:

Study 1.

In a combined repeated dose toxicity study with reproduction and developmental screening, of test material was performed according to OECD Test Guideline 422.Crj:CD(SD)IGS male and female rats were treated with test material in the concentration of 0, 100, 300, 1000 mg/kg bw/day in 0.5 % Sodium carboxymethylcellulose mixed with 0.1 % Tween 80 orally by gavage.12 male and 12 female were placed in each group. Males were exposed with test chemical for 37 days while Females, from 14 days before mating to day 4 of lactation. Clinical sign, body weight, food consumption were observed. On day 38 male rats were killed while female on day 5 of lactation.

 No effect on clinical sign, body weight, food consumption, haematology and clinical chemistry of treated rats as compared to control. Similarly, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations, implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour were observed in treated rats. Increase in liver weight was observed in male and female rats. No gross pathological and histopathological changes were observed in treated rats. In addition, no significant effect on mating index, fertility index, numbers of corpora lutea or implantations implantation index, delivery index, gestation index, gestation length, parturition or maternal behaviour and numbers of offspring or live offspring, sex ratio, live birth index, viability index or body weights of offspring were observed. No gross pathological changes were observed in offspring as compared to control. Therefore, NOAEL was considered to be 1000 mg/kg bw/day when Crj:CD(SD)IGS male and female rats were treated with test material orally by gavage.

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.

Study 3.

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.

 

  Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 1000mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to classify as developmental toxicant.

 

 

Justification for classification or non-classification

 Thus, Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 500mg/kg bw . Thus, comparing this value with the criteria of CLP regulation 2-Naphthalenol, 1-[[2-methyl-4-[(2-methylphenyl)azo]phenyl]azo]-, ar-styrenated is not likely to classify for reproductive and developmental toxicity.