Dihydrogen (ethyl)[4-[[4-[ethyl(3-sulphonatobenzyl)amino]phenyl](2-sulphonatophenyl)methylene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, aluminium salt

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be 1000mg/kg /day for reproductive toxicity, when male and female rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on reproductive toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Species:

rat

Strain:

other: 1.Charles River CD 2.Long-Evans

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS
- Weight at study initiation: 601 G (Male/female) and 487 G (Female)
Age at study initiation: 38 days old at initation of F0 phase
- Source: Charles River Breeding Laboratories, Portage, MI
- Housing: housed individually in hanging wire mesh cages
- Diet (e.g. ad libitum): Purina Laboratory Rodent Chow No. 5001 (Ralston Purina Company, Inc., St Louis, MO) ad libitum
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-21°C
- Humidity (%):40-60%
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycleTEST ANIMALS

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: diet(Purina Laboratory Rodent Chow No. 5001 (Ralston Purina Company, Inc., St Louis, MO))

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly during the first 13 wk of study, and then monthly thereafter.
- Mixing appropriate amounts with (Type of food): Purina Laboratory Rodent Chow No. 5001 (Ralston
Purina Company, Inc., St Louis, MO) blended in a twin-shell blender
- Storage temperature of food: 20-21°C

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation:no data
- After successful mating each pregnant female was caged (how): housed in plastic boxes with ground corncob bedding.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Analyses of compound concentration were conducted weekly during the first 13 wk of study, and then monthly thereafter. Analyses of the basal feed for heavy metals, chlorinated hydrocarbons, and aflatoxin were conducted on all lots of feed used during the study. These analyses demonstrated that the basal feed contained acceptably low levels of contaminants, that the diets were prepared properly, and that the dietary content of the test material was stable.

Duration of treatment / exposure:

1.The maximum exposure times were 116 and 111 wk for males and females, respectively.
2.80-day ( 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations.)

Frequency of treatment:

daily

Details on study schedule:

No data available

Remarks:

Study 1
Doses / Concentrations:
0, 0.1, 1.0, 2.0 % in the diet (0,50,500,1000 mg/kg bw /day respectively)
Study 2
0, 10, 100, 300, or 1,000 mg/kg b.w./day.

No. of animals per sex per dose:

Study 1
60 males + 60 females F0; 70 males + 70 females F1
Study 2
Total:300
F0 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females
F1 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

DETAILED CLINICAL AND MORTALITY OBSERVATIONS: Yes
- Time schedule: twice daily with at least 5 hr between observations.
BODY WEIGHT: Yes
- Time schedule for examinations: weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes
- Time schedule for examinations: Weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: 3, 6, 12, 18 and 24 months of the chronic phase
HAEMATOLOGY: Yes
- Time schedule for collection of blood:
- Animals fasted: No data
- How many animals: Ten animals of each sex from each group
- Parameters checked: haemoglobin, haematocrit, erythrocyte and total and differential leucocyte counts, and erythrocyte morphology
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: 3, 6, 12, 18 and 24 months of the chronic phase and at the end of the study.
- Animals fasted: No data
- How many animals: Ten animals of each sex from each group
-Parameters checked: aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, fasting glucose, total protein and creatinine
URINALYSIS: Yes
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
-Parameters checked: gross and microscopic appearance, specific gravity, pH and for the presence of protein, glucose, ketones, bilirubin and occult blood
OTHER:
Detailed physical examinations for signs of toxicity and palpation for masses: YES,
- Time schedule for examinations: weekly.
ORGAN WEIGHT:
- How many animals: Organ weights were measured in ten rats/sex/group
-Organ examined - brain, gonads, kidneys, liver, spleen and thyroid.

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.
GROSS EXAMINATION OF DEAD PUPS: Yes,external and internal abnormalities

Postmortem examinations (parental animals):

SACRIFICE :Yes
GROSS NECROPSY: yes
HISTOPATHOLOGY / ORGAN WEIGHTS:yes,histopathological examination of ovaries.gonads weight, histopathological examination of testes with epididymides

Postmortem examinations (offspring):

No data available

Statistics:

The one-way ANOVA was applied using the F distribution to assess significance. If significant differences among the means were indicated, Dunnett's test was used to determine which means were significantly different from control (Dunnett, 1964). If a non-parametric procedure for testing equality of means was indicated, the Kruskal-Wallis test was used (Hollander and Wolfe, 1973). If differences from the controls were indicated, a summed rank test was used to determine which groups differed from the controls.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1.Blue staining of hair , exposed skin and faeces was noted in all treatment group, other physical observations noted at low incidence in both control and treated rats included rales, red material around the eyes ,nose or anogenital region, redness around the eyes, white areas in the internal eyes corneal opacities , dilated pupils and increased distance between the pupil and cornea .Palpable masses primarily in the abdominal, thoracic and anogenital regions were present at similar incidence in control and treated rats.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, non-treatment-related

Description (incidence):

one female in the 1.0% group and one male and one female in the 1.0% group and one male in the 2.0% group died. These deaths were not related to treatment.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Group mean body weight at the end of the study were generally similar for control and treated male and female rats with the exception of the females in the 2.0% group.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Food consumption was slightly greater for males and females in the 2.0% group. These differences were statistically significant (P<0.05) for males between wk 27and 66and for female between wk 27 and 38 and between wk 79 and 90

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Ophthalmoscopic examinations at most examination intervals revealed chorioretinal degeneration and choroidal and choriorentinal hypoplasia in both control and treated rats . Conjunctivitis and keratitis were the most common finding and were seen in similar or greater numbers of control rats than of treated rats. None of these findings were compound related .

Haematological findings:

no effects observed

Description (incidence and severity):

Few of the heametological parameters showed significant difference between control and treated animals and none of the difference appeared to be compound related

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Few of the clinical chemistry parameters showed significant difference between control and treated animals and none of the difference appeared to be compound related

Urinalysis findings:

no effects observed

Description (incidence and severity):

Few of the urinalysis parameters showed significant difference between control and treated animals and none of the difference appeared to be compound related

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

The complete gross autopsies were also performed on all animals that died on test or that were killed at the end of the study. Histological evaluation revealed a variety of lesions including neoplasms among control and treated rats , These lesions were present in similar incidence in control and treated rats and appeared to be spontaneous , None of the lesions were determined to be related to the administration of test material.

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):

1.There were no compound related effects on fertility, gestation, parturition , lactation , pup survival through weaning or on number of live and stillborn pups.
2.No effects attributable to treatment were observed with respect to mating performance,pregnancy and fertility rates, gestation length.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

ophthalmological examination

haematology

clinical biochemistry

urinalysis

organ weights and organ / body weight ratios

gross pathology

reproductive performance

Remarks on result:

other: overall toxic effects on reproductive parameters

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Clinical signs:

not specified

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):

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:

no effects observed

Gross pathological findings:

not specified

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: effects: no substance-related effects on fertility, gestation, parturition, lactation, pup survival through weaning, or on the numbers of live and stillborn pups.

Remarks on result:

other: No effect observed.

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Clinical signs:

not specified

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):

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:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

There were no macroscopic or microscopic tissue abnormalities of either F3a-generation animals considered to be attributable to treatment

Dose descriptor:

NOAEL

Generation:

other: F3a

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

mortality

body weight and weight gain

organ weights and organ / body weight ratios

gross pathology

Remarks on result:

other: No developmental toxic effects were observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Mean food and compound consumption of rats that received test chemical in diet throughout their lifetime

Dietary concentration of test chemical 1(%)	Food consumption including test chemical (g/kg /day)		Compound consumption (mg/kg/day)
	Male	Female	Male	Female
0.0	51 ±18	61±16	-	-
0.0	51±19	61±17	-	-
0.1	50±18	62±18	50±18	62±18
1.0	51±18	63±17	514±179	631±173
2.0	54±19	66±17	1072±381	1319±345

 

Survival and group mean body weights and percentage difference from control values at the end of the study of rats that recived FD&C blue no.1 in the diet throughout their lifetime (in utero )

 

 

Dietary concentration of test chemical (%)	No. surviving at the end of the study	Final body weight(g)
		Mean±SD	Difference from control (IA+IB) value (%)
Male rats	18	592±88	+0.5
0.0(IA)	13	586±132	-0.5
0.0(IB)	12	595±89	-1.0
0.1	12	601±104	+2.0
1.0	18	561±65	-4.7
2.0
Female rats
0.0(IA)	28	480±78	-2.8
0.0(IB)	24	508±105	+2.8
0.1	23	487±102	-1.4
1.0	28	478±87	-3.2
2.0	10**	420±69	-15.0**

**p<0.01

Conclusions:

“No observed adverse effect level (NOAEL)” for reproductive toxicity was considered to be 2.0 % (1000 mg/kg bw /day).When Charles River CD rats were treated with test chemical orally.

Executive summary:

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

Study 1

The test chemical was fed to Charles River CD rats as a dietary admixture in lifetime toxicity/carcinogenicity studies. The rat study was conducted with an in utero phase in which the compound was administered to the F0 generation rats (60/sex/group) at dietary concentrations of 0.0%, 0.0%, 0.1%, 1.0% or 2.0%.(0, 50,500,1000 mg/kg bw /day respectively). After randomly selecting the F1 animals, the lifetime phase was initiated at the same levels with 70 rats/sex/group, including two control groups. The maximum exposure times were 116 and 111week for male and females respectively. The rats were approximately 38 days old at initiation of the F0 phase .The diet were prepared weekly during the first 13 wk of study, and then monthly thereafter and stored at 20-21°C.Animals after successful mating each pregnant female was housed in plastic boxes with ground corncob bedding. All animals were observed for clinical signs twice daily with at least 5 hr between observations. Body weight observed weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter and food consumption was noted on Weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter. Ophthalmoscopic examination noted on 3, 6, 12, 18 and 24 months of the chronic phase. Organ weights were measured in ten rats/sex/group and organ examined were brain, gonads, kidneys, liver, spleen and thyroid. In urine analysis gross and microscopic appearance, specific gravity, pH and for the presence of protein, glucose, ketones, bilirubin and occult blood were noted. Haematological parameter like haemoglobin, haematocrit, erythrocyte and total and differential leucocyte counts, and erythrocyte morphology were observed.

 

Clinical signs like Blue staining of hair , exposed skin and faeces was noted in all treatment group, other physical observations noted at low incidence in both control and treated rats included rales, red material around the eyes ,nose or anogenital region, redness around the eyes, white areas in the internal eyes corneal opacities , dilated pupils and increased distance between the pupil and cornea .Palpable masses primarily in the abdominal, thoracic and anogenital regions were present at similar incidence in control and treated rats.Group mean body weight at the end of the study were generally similar for control and treated male and female rats with the exception of the females in the 2.0% group. Food consumption was slightly greater for males and females in the 2.0% group. These differences were statistically significant (P<0.05) for males between wk 27and 66and for female between wk 27 and 38 and between wk 79 and 90.No compound related gross changes, including organ weight effects were noted.Few of the heametological , clinical chemistry and urinalysis parameters showed significant difference between control and treated animals and none of the difference appeared to be compound related.Completed gross autopsies and histological evolution of the ten rats per sex from each group that were killed after 1 year revealed no compound related gross or histological changes. Complete gross autopsies were also performed on all animals that died on test or that were killed at the end of the study. Histological evaluation revealed a variety of lesions including neoplasms among control and treated rats, these lesions were present in similar incidence in control and treated rats and appeared to be spontaneous, none of the lesions were determined to be related to the administration of test material.One female in the 1.0% group and one male and one female in the 1.0% group and one male in the 2.0% group died. These deaths were not related to treatment. Ophthalmoscopic examinations at most examination intervals revealed chorioretinal degeneration and choroidal and choriorentinal hypoplasia in both control and treated rats . Conjunctivitis and keratitis were the most common finding and were seen in similar or greater numbers of control rats than of treated rats. None of these findings were compound related.There were no compound related effects on fertility, gestation, parturition, lactation, pup survival through weaning or on number of live and stillborn pups. Hence the “No observed adverse effect level (NOAEL)” for reproductive toxicity was considered to be 2.0 % (1000 mg/kg bw /day).When Charles River CD rats were treated with test chemical orally.

Study 2

Ina 3-generation reproduction study was carried out on test chemical in male and female Long-Evans rats . The test material mixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding.Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations.Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.

 

Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment. Hence ,Therefore the “No observed adverse effect level (NOAEL)” for reproductive toxicity was considered to be 1000 mg/kg bw /day.Asthere were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings were observed in rat at dose concentration 1000 mg/kg bw /day given orally.

 

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

Study 1

The test chemical was fed to Charles River CD rats as a dietary admixture in lifetime toxicity/carcinogenicity studies. The rat study was conducted with an in utero phase in which the compound was administered to the F0 generation rats (60/sex/group) at dietary concentrations of 0.0%, 0.0%, 0.1%, 1.0% or 2.0%.(0, 50,500,1000 mg/kg bw /day respectively). After randomly selecting the F1 animals, the lifetime phase was initiated at the same levels with 70 rats/sex/group, including two control groups. The maximum exposure times were 116 and 111week for male and females respectively. The rats were approximately 38 days old at initiation of the F0 phase .The diet were prepared weekly during the first 13 wk of study, and then monthly thereafter and stored at 20-21°C.Animals after successful mating each pregnant female was housed in plastic boxes with ground corncob bedding. All animals were observed for clinical signs twice daily with at least 5 hr between observations. Body weight observed weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter and food consumption was noted on Weekly for the first 14 wk, biweekly for the next 12 wk, and then every 4 wk thereafter. Ophthalmoscopic examination noted on 3, 6, 12, 18 and 24 months of the chronic phase. Organ weights were measured in ten rats/sex/group and organ examined were brain, gonads, kidneys, liver, spleen and thyroid. In urine analysis gross and microscopic appearance, specific gravity, pH and for the presence of protein, glucose, ketones, bilirubin and occult blood were noted. Haematological parameter like haemoglobin, haematocrit, erythrocyte and total and differential leucocyte counts, and erythrocyte morphology were observed.

 

Clinical signs like Blue staining of hair , exposed skin and faeces was noted in all treatment group, other physical observations noted at low incidence in both control and treated rats included rales, red material around the eyes ,nose or anogenital region, redness around the eyes, white areas in the internal eyes corneal opacities , dilated pupils and increased distance between the pupil and cornea .Palpable masses primarily in the abdominal, thoracic and anogenital regions were present at similar incidence in control and treated rats.Group mean body weight at the end of the study were generally similar for control and treated male and female rats with the exception of the females in the 2.0% group. Food consumption was slightly greater for males and females in the 2.0% group. These differences were statistically significant (P<0.05) for males between wk 27and 66and for female between wk 27 and 38 and between wk 79 and 90.No compound related gross changes, including organ weight effects were noted.Few of the heametological , clinical chemistry and urinalysis parameters showed significant difference between control and treated animals and none of the difference appeared to be compound related.Completed gross autopsies and histological evolution of the ten rats per sex from each group that were killed after 1 year revealed no compound related gross or histological changes. Complete gross autopsies were also performed on all animals that died on test or that were killed at the end of the study. Histological evaluation revealed a variety of lesions including neoplasms among control and treated rats, these lesions were present in similar incidence in control and treated rats and appeared to be spontaneous, none of the lesions were determined to be related to the administration of test material.One female in the 1.0% group and one male and one female in the 1.0% group and one male in the 2.0% group died. These deaths were not related to treatment. Ophthalmoscopic examinations at most examination intervals revealed chorioretinal degeneration and choroidal and choriorentinal hypoplasia in both control and treated rats . Conjunctivitis and keratitis were the most common finding and were seen in similar or greater numbers of control rats than of treated rats. None of these findings were compound related.There were no compound related effects on fertility, gestation, parturition, lactation, pup survival through weaning or on number of live and stillborn pups. Hence the “No observed adverse effect level (NOAEL)” for reproductive toxicity was considered to be 2.0 % (1000 mg/kg bw /day).When Charles River CD rats were treated with test chemical orally.

Study 2

Ina 3-generation reproduction study was carried out on test chemical in male and female Long-Evans rats . The test material mixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding.Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations.Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.

 

Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment. Hence ,Therefore the “No observed adverse effect level (NOAEL)” for reproductive toxicity was considered to be 1000 mg/kg bw /day.Asthere were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings were observed in rat at dose concentration 1000 mg/kg bw /day given orally.

 

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 test material is not likely to classify as reproductive toxicant.