Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be 75mg /kg bw/day.When male and female rats or mice were treated with test material orally but adverse effects were observed on fertility and developmental parameters on higher dose concentration ,Thus, comparing this value with the criteria of CLP regulation test material is likely to classify as reproductive toxicant.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on reproductive toxicity studies on rats
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Species:
other: 1&2. rats 3&4. mouse
Strain:
other: 1.Wistar 2.Sprague-Dawley 3&4.CD-1
Details on species / strain selection:
No data available
Sex:
male/female
Details on test animals and environmental conditions:
2.TEST ANIMALS
- Age at study initiation: 63 days
- Weight at study initiation: 182-263 g
- Housing: Animals were housed individually under controlled conditions of temperature, humidity and lighting.
- Diet (e.g. ad libitum):Certified Rodent Chow 5002 were provided ad libitum
- Water (e.g. ad libitum):ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C):21.11±1.2 °C
- Humidity (%):57±3.8%
- Photoperiod (hrs dark / hrs light):12-h light/dark
3.Details on test animal
TEST ANIMALS
- Source: Charles River Breeding Laboratory Inc. (Kingston, N.Y.).
- Age at study initiation: (P) x wks; (F1) x wks: F0: 11 weeks
F1: 70 ± 10 days
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: F0: male- 34.5 ± 0.58 to 35.6 ± 0.76g , Female: 26.3 ± 0.66 to 27.6 ± 0.60g

F1: male- 1.65 ± 0.014 to 1.60 ± 0.023g, female: 1.53 ± 0.016 to 1.60 ± 0.014g

- Fasting period before study: No data available
- Housing: Animals were housed 2 per Polycarbonate shoebox type cages (5" x 11" x 7") with stainless steel wire bar lids, one of these animals will be punched in the left ear for identification purposes. Cages will be uniformly placed on stainless steel racks (66" x 60" x 30"). Cages will be rotated (relative placement) at least once a week. Bedded on Ab-Sorb-Dri , Approximately 100g of bedding will be used per cage.
- Diet (e.g. ad libitum): Purina certified Rodent Chow animal diet #5002, ad libitum.
- Water (e.g. ad libitum): Water, ad libitum.
- Acclimation period: F0: 1 week , F1: five weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25° C
- Humidity (%):20 to 70%
- Air changes (per hr): 10 or more air changes per hour of HEPA-filtered air.
- Photoperiod (hrs dark / hrs light): 14 hour light cycle

Route of administration:
oral: gavage
Vehicle:
other: 1.sodium carboxymethylcellulose 2.0.5 % Aqueous methyl cellulose 3.corn oil
Details on exposure:
1.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material mixed with 0.5% solution of sodium carboxymethylcellulose

DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): Test material mixed with 0.5% solution of sodium carboxymethylcellulose


- Concentration in vehicle: 0,75, 150, or 300 mg/kg- Amount of vehicle (if gavage): 5ml/kg

- Lot/batch no. (if required): No data available
- Purity: No data available
2.Test material was administered by gavage in 0.5% aqueous methyl cellulose on days 15 through 21 of gestation twice daily at a dose concentration of 0, 20, 80 or 200 mg/kg/day.

VEHICLE
0.5 % Aqueousmethylcellulose
-Justification for use and choice of vehicle (if other than water): Aqueous methyl cellulose
-Concentration in vehicle: 0, 20, 80 or 200 mg/kg bw/day
-Amount of vehicle (if gavage): Dosing volume was 10 ml/kg per dosing occasion.
-Other :One-half of the dose was given to the animals in the morning and the second half 6-8 h later.

Details on mating procedure:
1.Gravid female animals were used
2.- M/F ratio per cage: One female and one male (1:1)
- Length of cohabitation: Until confirmed pregnancy
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: The occurrence of copulation was determined by daily inspection for a copulatory plug.The day at which evidence of mating was detected was designated gestation day 0.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]: No
- After successful mating each pregnant female was caged (how):Female was placed in an individual cage.
3.- M/F ratio per cage:1: 1 ratio
- Length of cohabitation: 100 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy No data available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)] No data available
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: Litters produced during the cohabitation period (day 7 to day 107) will be humanely sacrificed. Litters produced during the period day 107 to day 127 will be allowed to remain with their mothers

Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
1.6 days ( from day 8 through day 14 of gestation )
2.6 days (Days 15 through 21 of gestation)
3&4.127 days
Frequency of treatment:
1,3&4.once daily
2.Twice daily
Details on study schedule:
1.F0:
Pregnant female rats were randomly assigned to one of three treatment groups which consisted of 25 females and received 0, 20, or 80 mg/kg/day of sodium salicylate or to one treatment group consisting of 16 females which received 200 mg/kg/day of sodium salicylate. All rats were given the respective dosage via gavage on days 15 through 21 of gestation twice daily. Beginning of day 21, the animals were observed hourly for the onset of labor. All surviving F0 females were euthanized by carbon dioxide inhalation at lactation day 1. On gestation day 25, those females which failed to deliver were euthanized. The uterus and ovaries of all dams were exposed by an abdominal incision and grossly examined to determine the number of implantation sites present.


F1:
As soon as possible after delivery, all pups were examined for external abnormalities and the number of viable and nonviable pups was recorded for each female. Pups were sexed and weighed individually on lactation day 0. All pups were then euthanized by carbon dioxide inhalation at lactation day 1.
Remarks:
Study 1
0, 75, 150, or 300 mg/kg
Study 2
0, 20, or 80 or 200 mg/kg/day
Study 3
0, 0.1, 0.25 and 0.50 g/kg (0, 100, 250 and 500 mg/kg/day)
Study 4
0,20,50 and 100mg/kg bw/days
No. of animals per sex per dose:
1.Total:80
0 mg/kg bw/day:20
75 mg/kg bw/day:20
150 mg/kg bw/day:20
300 mg/kg bw/day:20
2.Total:86
Control:25 females
20 mg/kg/day: 25 females
80 mg/kg/day: 25 females
200 mg/kg/day: 16 females
3.Total: 200
0 mg/kg/day: 40 male, 40 female
100 mg/kg/day: 20 male, 20 female
250 mg/kg/day: 20 male, 20 female
500 mg/kg/day: 20 male, 20 female
Study 4
Total:200
0 mg/kg bw/day: 40 male and 40 female
25 mg/kg bw/day:20 male and 20 female
50 mg/kg bw/day:20 male and 20 female
100 mg/kg bw/day:20 male and 20 female



Control animals:
yes, concurrent vehicle
Details on study design:
2.- Dose selection rationale: Dose were determined in pilot study using an identical protocol which yielded an effect level of 200 mg/kg/day and a NOEL of less than 100mg/kg/day for oral exposure totest material.
Positive control:
No data available
Parental animals: Observations and examinations:
1.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: No data available
Time schedule for examinations: No data available
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

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 / No / No data: No data available


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

OTHER:
2.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: The animals were observed twice daily throughout
the study for mortality and signs of overt
toxicity.

BODY WEIGHT: Yes
Time schedule for examinations: Individual body weights were recorded on gestation days 0, 6, 15, 16, 17, 18, 19, 20 and 21
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: no
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


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

OTHER:
3.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: yes
Time schedule for examinations: All animals will be weighed once aweek from day 0 (first day oftreatment) to day127
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

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 / No / No data: No data available


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

OTHER:

Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
2.Litters’ were examined for numbers of live and stillborn pups and gross abnormalities. The pups were sexed and weighed individually on lactation day 0.
3.Number and percent of live pups per litter and Mean body weight of live offspring, Proportion of pups born alive and sex of pups born alive were observed.
4.The litters delivered during this time will be counted, sexed and weighed by sex
Postmortem examinations (parental animals):
1.Postmortem examinations (Parent Animal)
SACRIFICE : On day 20 of gestation, 15 of the animals of each group were killed; the remaining 5 were allowed to deliver
2.UTERUS
The uterus and ovaries of all dams were exposed by an abdominal incision and grossly examined to determine the number of implantation sites present.
4.Postmortem examinations (Parent Animal)
SACRIFICE : On day 20 of gestation, 15 of the animals of each group were killed; the remaining 5 were allowed to deliver

GROSS NECROPSY: No data available
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:
macroscopic examination was performed

Postmortem examinations (offspring):
Postmortem examinations (offspring)
SACRIFICE
Offspring were killed and autopsied on the 56th
day
- examinations of visceral and skeletal abnormalities
3.3.Pattern of testicular descendency in male pups were observed.
4.Postmortem examinations (offspring)
SACRIFICE:Offspring were killed and autopsied on the 56th day
- examinations of visceral and skeletal abnormalities

Statistics:
2.It is indicated in the study, the data was subjected to statistical analysis.The methods used included Bartlett’s test for homogeneity of variance. Comparisons were made by one-way analysis of variance (ANOVA).Pairwise comparisons were made using the appropriate t-test (for equal and unequal variance) or pairwise comparisons were made with a Bonferroni correction to determine the significance.
3.Statistical analysis wre performed by using Pairwise comparisons for Fertility oT Pairs During Continuous Breedins, Litters per Fertile Pair and Litter Size (Live offsprins) per Fertile Pair by using Chise square Pairwise comparisons .
4.A Kruskal-Wallis analysis of variance on ranks will be employed to compare the number of litters per breeding pair among treatment groups. If this test shows significant differences (pRange Test will be utilized to make intergroup pairwise comparisons. All data reduction and analysis will utilize software from the 1979 edition of the Statistical Analysis System (Helwig and Council, 1979) and will be performed on computers at Triangle Universities Computation Center
Reproductive indices:
3.Fertility, No. Fertile/ No. Cohabited and litters per pair were observed.
Offspring viability indices:
3.Viability indices were observed.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
1.Salivation and/or piloerection were observed in this group
2.No bservations of adverse effects.
3.No distinct treatment related symptoms of toxicity were observed during routine health surveillance.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
1.The 25.7% and 100% fetal mortality was observed in animals of the 150- and 300-mg/kg dose groups as compared to controls respectively.
2.No effects on gestational body weight gain .
3.Eleven animals died, 3 in the control, 2 each in the 100mg/kg and 250mg/kg dose groups, and 4 in the 500mg/kg dose group. The cause of death varied from case to case but it was neither chemical nor dose related.
4.one parental male and female in the control group, one parental female in the 25.0 mg /kg/day group, two parental females and three parental males in the 50.0 mg /kg/day group, and three parental males in the 100.0 mg /kg/day group died prior to the conclusion of the continuous breeding phase
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
1.Maternal body weight gain was inhibited for animals of the 300-mg/kg group
2.Test material in dose level 0,100,250,500 mg/kg bw had no apparent effect on male or female body weights
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
1.Feed consumption decreased during the administation of 300 mg/kg of test material.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
1.water consumption decreased during the administation of 300 mg/kg Salicylic Acid
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
1.Decreased uterine weight was observed in animals of the 150- and 300-mg/kg dose groups as compared to controls; these groups had 25.7% and 100% fetal mortality, respectively
2.No observations of adverse effects until the onset of parturition at 0, 20 or 80 mg/kg/day.
3.No effect on Fertility Index were observed on treated rats as compared to control.
Significant decrase in litters per pair, proportion of pups born alive and sex of pups born alive when treated with 500 mg/kg/day as compared to control.
4.No effect on Fertility Index were observed on treated rats as compared to control.
Dose descriptor:
NOAEL
Effect level:
> 75 - <= 250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
gross pathology
other: Decreased uterine weight was observed in animals of the 150- and 300-mg/kg dose groups as compared to controls; these groups had 25.7% and 100% fetal mortality, respectively
Remarks on result:
other: No effects on reproductive parameters
Dose descriptor:
LOAEL
Effect level:
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
reproductive performance
Remarks on result:
other: effects observed at given dose
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
1.Litter size were significantly decreased in the 150-mg/kg dose group.
3.Significant decrase in number of live pups were observed in 500 mg/kg/day
4.no influence on the number of litters produced per pair, the number of live pups per litter, the proportion of pups born alive or sex (males/total) of pups born alive

Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
1.neonatal body weight were significantly decreased in the 150-mg/kg dose group.
2.Mean pup weight: Control: 5.84±0.62 g 20 mg/kg: 5.70±0.52 g 80 mg/kg: 5.85±0.49 g 200 mg/kg: 5.23±0.23 g
3.Significant decrease in live pup weight and adjusted live pup weight was observed in 500 mg/kg/day treated rats as compared to control.
4.Body weight weights also were unaffected by treatment of female mice
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:
effects observed, treatment-related
Description (incidence and severity):
1.The thyroid weight of male offspring from the 75-mg/kg group was significantly increased and the adrenal gland weight of male offspring from the 150-mg/kg group was significantly decreased compared to controls. The incidences of external organ, internal organ, and skeletal anomalies in offspring were 0%, 5.0%, and 0%, respectively, for the 75-mg/kg group and 13.7%, 17.2%, and 79.2%, respectively, for the 150- mg/kg group.
4.liver and pituitary weights also were unaffected by treatment of female mice
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
1.The incidences of external, internal, and skeletal anomalies in offspring autopsied at the 56th day were 1.8%, 0%, and 2.5%, respectively, for
the 75-mg/kg group and 27.8%, 12.7%, and 65.7%, respectively, for the 150-mg/kg group.
Histopathological findings:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Sperm assessment indicated no significant differences in the %motile sperm, sperm concentration or %abnormal sperm in the cauda epididymis between male mice exposed to 0.0 or 100.0 mg /kg/day
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
LOAEL
Generation:
F1
Effect level:
75 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: overall developmental 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:
No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to be 75 mg/kg/day, and the dose-level of 75 mg/kg/day was considered to be the LOAEL for embryo-foetal toxicity. When female wistar rats were treated with test material orally.
Executive summary:

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

Study 1

Reproductive toxicity study of test material was performed on female wistar rats.80 rats were divided as 20 rats /dose group.Test material mixed with0.5% solution of sodium carboxymethyl cellulosewere administered in dose concentration 0,75, 150, or 300 mg/kgfrom day 8 through day 14 by oral gavage route.The control group was given 5 ml/kg of vehicle. On day 20 of gestation, 15 of the animals of each group were killed; the remaining 5 were allowed to deliver. The offspring, which were weaned on day 21, were observed daily and weighed every 3 days. Offspring were killed and autopsied on the 56thday for examinations of visceral and skeletal abnormalities

 Maternal body weight gain was inhibited for animals of the 300-mg/kg group. Salivation and/or piloerection were observed in this group. Feed and water consumption decreased during the administration of 300 mg/kg Salicylic Acid. Three animals of this group died within a few days of the initiation of dosing. Decreased uterine weight was observed in animals of the 150- and 300-mg/kg dose groups as compared to controls; these groups had 25.7% and 100% fetal mortality, respectively. Litter size and neonatal body weight, body length, and tail length were significantly decreased in the 150-mg/kg dose group. The incidences of external, internal, and skeletal anomalies in offspring autopsied at the 56th day were 1.8%, 0%, and 2.5%, respectively, for the 75-mg/kg group and 27.8%, 12.7%, and 65.7%, respectively, for the 150-mg/kg group.The offspring from animals of 150-mg/kg Salicylic Acid group had decreased body length and tail length compared to controls. The thyroid weight of male offspring from the 75-mg/kg group was significantly increased and the adrenal gland weight of male offspring from the 150-mg/kg group was significantly decreased compared to controls. The incidences of external organ, internal organ, and skeletal anomalies in offspring were 0%, 5.0%, and 0%, respectively, for the 75-mg/kg group and 13.7%, 17.2%, and 79.2%, respectively, for the 150- mg/kg group. Hence No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to be 75 mg/kg/day, and the dose-level of 75 mg/kg/day was considered to be the LOAEL for embryo-foetal toxicity.When female wistar rats were treated with test material orally.

 

Study 2

Reproductive toxicity study of test materialwas performed on female Sprague Dawley rats. 86 rats were divided into 2 treatment group.25 female rats in treatment group 1 while 16 in treatment group 2 .Test material mixed with0.5% aqueous methyl cellulosewereadministers in dose concentration 0, 20, 80 or 200 mg/kg bw day from days 15 through 21 of gestation by oral gavage route twice daily.One-half of the dose was given to the animals in the morning and the second half 6-8 h later. Dosing volume was 10 ml/kg per dosing occasion. For mating purposes, one female and one male (from breeding stock) were housed together. The occurrence of copulation was determined by daily inspection for a copulatory plug. The day at which evidence of mating was detected was designated gestation day 0, and the female was placed in an individual cage. The animals were observed twice daily throughout the study for mortality and signs of overt toxicity. Individual body weights were recorded on gestation days 0, 6, 15, 16, 17, 18, 19, 20 and 21. Beginning on day 21, Pups were examined for external abnormalities as soon as possible following delivery. The number of viable and nonviable pups was recorded for each female. All pups were sexed and weighed individually on lactation day 0. All surviving F,, females and F, pups were euthanized by carbon dioxide inhalation at lactation day 1. On gestation day 25, those females which failed to deliver were euthanized. The uterus and ovaries of all dams were exposed by an abdominal incision and grossly examined to determine the number of implantation sites present.

 

No observations of adverse effects until the onset of parturition. As well, there were no apparent compound-related effects on gestational body weight gain in any of the treatment groups. One death occurred in the vehicle-treated group on gestation day 20. However, post-mortum examination revealed no abnormalities or signs of overt clinical toxicity. Both onset of labour and its duration were disrupted in the animals receiving the highest dose. Increased fetotoxicity and periparturn death also appeared to occur among foetuses from females treated with 200 mg/kg/day but this was not statistically significant,Labour times in these groups ranged from 1-14 h with a mean duration of 3.6 h for the treated group, For comparison, labor times in control animals ranged from less than 1 h to 3 h with a mean labour time of 1.5 h. There were no observed differences in mean litter size, pup size, or sex ratio, associated with test material . Since those pups which did survive delivery had no visible abnormalities or other signs of overt toxicity. Maternal perinatal death was significantly increased in animals treated with both 200 mg/kg/ day. Four animals in each of these treatment groups died or were euthanized because of extreme distress during parturition. Clinical signs of maternal physical distress during parturition included labored breathing, decreased activity, and blood staining in the abdominal, nasal, and oral areas. Notably, each of these seven animals failed to deliver some or all of their pups. Increased maternal mortality was also evidenced as a significant decrease in the gestational index (defined as the percentage of pregnant females producing live pups) for these same treatment groups. No evidence of increased maternal distress or peripatum mortality was noted in animals from either the mid-dose (80 mg/kg/day) or low-dose (20 mg/kg/day) sodium salicylate. Hence No Observed Effect Level (NOEL) was considered to be 80 mg/kg/day, and LOAEL dose-level of 200 mg/kg/day was considered to be for reproductive toxicity.When female Sprague Dawley rats were treated with test material orally.

 Study 3

 The reproductive toxicity study of test materialwas performed on male and female  CD-1 mice. Based on dose-range finding study dose levels of0,100,250,500mg/kgwere selected. The vehicle control group (40 male and 40 female) and three dose groups (20 male and 20 female).Eleven-week old male and female CD-l mice were exposed to the chemical during a7-day premating period, after which they were randomly paired (one male: one female) within each dose group. Cohabitation was continued for100days. All animals will be weighed once a week from day 0 (first day of treatment) to day127 New-born litters were evaluated and immediately sacrificed. Eleven animals died, 3 in the control, 2 each in the 100mg/kg and 250mg/kg dose groups, and 4 in the 500mg/kg dose group. The cause of death varied from case to case but it was neither chemical nor dose related. No distinct treatment related symptoms of toxicity were observed during routine health surveillance. Test material in dose level 0,100,250,500 mg/kg bw had no apparent effect on male or female body weights .The fertility index in the control and various treatment groups varied between 95 to 100% all breeding pairs except 1 in the 100mg /kg group delivered at least one litter .There was a significant decrease (p<0.05) in the mean number of litters at the 0.5 g/kg level .The average number of pups per litter, the proportion of pups born alive, and mean live pup weight values were also significantly reduced (p<0.05) in the 500mg /kg group compared to the correspondingcontrol.There was no significant effect on any of these parameters in the remaining two dose groups.HenceNo Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to be 250 mg/kg/day, and the dose-level of 500 mg/kg/day was considered to be the LOAEL for reproductive toxicity.When male and femaleCD 1 mice weretreated with test material orally.

Study 4

The reproductive toxicity study of test materialwas performed on male and female  CD-1 mice. 11 weeks of age animals were used in study. Based on dose-range finding study dose levels of 0.0, 25.0, 50.0, and 100.0 mg /kg/day were selected. Test material soluble in corn oil and administered by gavage daily during the 7-day premating, 98-day cohabitation, and 21-day segregation periods. The vehicle control group (40 male and 40 female) and three dose groups (20 male and 20 female), For the first week, animals will be housed five per cage by sex. Subsequently, the females and males from the same dose group will be randomly paired and cohabited. The adult animals will be weight weekly.

The test material had no influence on the number of litters produced per pair, the number of live pups per litter, the proportion of pups born alive or sex (males/total) of pups born alive .Female live pup weight adjusted for the total number of pups per litter, however, was significantly greater p<0.05) for the breeding pairs given 100.0 mg /kg/day than for those receiving 50.0 mg /kg/day. Further, combined (both sexes) live pup weight adjusted for the total number of pups per litter was significantly greater (p<0.05) for breeding pairs receiving 100.0 mg /kg/day than for those given 25.0 or 50.0 mg /kg/day .Since there were no other indications of toxicity during continuous breeding, these differences might be due to chance alone. It also should be noted that one parental male and female in the control group, one parental female in the 25.0 mg /kg/day group, two parental females and three parental males in the 50.0 mg /kg/day group, and three parental males in the 100.0 mg /kg/day group died prior to the conclusion of the continuous breeding phase .The random distribution of deaths across treatment groups suggested that they were not treatment-related.

 

Due to the lack of an effect of test material on fertility and reproductive performance in the parental mice (F0 generation), it was decided to assess fertility and reproductive performance in the F1 offspring. The litters of the 0.0 and 100.0 mg /kg/day. groups were weaned at 21 days of age, and one or two females and male pups from each surviving litter were randomly selected .Each weanling was housed individually and maintained on the same treatment as their parents. At 90±10 days of age, a male and female from different litters within treatment groups were cohabited from 1 to 7 days, depending upon when a copulatory plug was detected. The pairs were separated and the females were allowed to deliver their litters. Continuous exposure of CD-1 mice to test material in utero, via the mother's milk and by gavage (100.0 mg /kg/day) from weaning at 21 days of age to sacrifice at 127±8 days of age was found to have no statistically significant effects on mating behavior , fertility rate or reproductive performance

The 0.0 and 100.0 mg/kg/day F1 parental mice were necropsied .Sperm assessment indicated no significant differences in the %motile sperm, sperm concentration or %abnormal sperm in the cauda epididymis between male mice exposed to 0.0 or 100.0mg /kg/day .In addition, there were no significant differences in body weights or organ weights between the male mice in these two groups .Body weight and liver and pituitary weights also were unaffected by treatment of female mice .In contrast, brain weight was significantly increased (p<0.05)and reproductive tract weight significantly decreased (p<0.05) in female mice given 100.0 mg /kg/day versus control female mice .The reproductive tract weights of the two females in the control group that did not deliver pups were unexpectedly heavier on average than those from the 17 control females that did deliver pups. This was in contrast to the findings for the females treated with100.0 mg/kg/day. That is, the six females in this group that did not give birth had lighter reproductive tracts on average than did their eleven post gravid cohorts .This difference,along with the fact that the control group had a greater proportion of females with postgravid reproductive tracts than did the treated group (17/19 versus 11/17, respectively), might well explain the significantly heavier reproductive tracts in the control group as compared to those in the treated group. Thus, since the difference between the two groups appeared to be due to biological variation, it was not considered to be of toxicological significance. Finally, no-treatment-related gross or histopathologic lesions were noted for the pituitary, testis, epididymis, prostate or seminal vesicles in male mice, or for the pituitary, ovary, oviduct, uterus or vagina in female mice. Hence No Observed Adverse Effect Level (NOAEL) for reproductive toxicity for F0 and F1 generation was considered to be 100 mg/kg/day.When male and female  CD-l mice were treated with test material orally.

 

 Based on the data available from different studies ,test chemical did not showed reproductive toxicity at dose concentration 75 mg/kg bw/day.When rats or mice were treated with test material orally. but adverse effects were observed on fertility and developmental parameters on higher dose concentration ,Thus, comparing this value with the criteria of CLP regulation test material is likely to classify as reproductive toxicant.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
75 mg/kg bw/day
Study duration:
subacute
Species:
other: rats or mice
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

Reproductive toxicity study of test materialwas performed on female wistar rats.80 rats were divided as 20 rats /dose group.Test material mixed with0.5% solution of sodium carboxymethyl cellulosewere administered in dose concentration 0,75, 150, or 300 mg/kgfrom day 8 through day 14 by oral gavage route.The control group was given 5 ml/kg of vehicle. On day 20 of gestation, 15 of the animals of each group were killed; the remaining 5 were allowed to deliver. The offspring, which were weaned on day 21, were observed daily and weighed every 3 days. Offspring were killed and autopsied on the 56thday for examinations of visceral and skeletal abnormalities

 Maternal body weight gain was inhibited for animals of the 300-mg/kg group. Salivation and/or piloerection were observed in this group. Feed and water consumption decreased during the administration of 300 mg/kg Salicylic Acid. Three animals of this group died within a few days of the initiation of dosing. Decreased uterine weight was observed in animals of the 150- and 300-mg/kg dose groups as compared to controls; these groups had 25.7% and 100% fetal mortality, respectively. Litter size and neonatal body weight, body length, and tail length were significantly decreased in the 150-mg/kg dose group. The incidences of external, internal, and skeletal anomalies in offspring autopsied at the 56th day were 1.8%, 0%, and 2.5%, respectively, for the 75-mg/kg group and 27.8%, 12.7%, and 65.7%, respectively, for the 150-mg/kg group.The offspring from animals of 150-mg/kg group had decreased body length and tail length compared to controls. The thyroid weight of male offspring from the 75-mg/kg group was significantly increased and the adrenal gland weight of male offspring from the 150-mg/kg group was significantly decreased compared to controls. The incidences of external organ, internal organ, and skeletal anomalies in offspring were 0%, 5.0%, and 0%, respectively, for the 75-mg/kg group and 13.7%, 17.2%, and 79.2%, respectively, for the 150- mg/kg group. Hence No Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to be 75 mg/kg/day, and the dose-level of 75 mg/kg/day was considered to be the LOAEL for embryo-foetal toxicity.When female wistar rats were treated with test material orally.

 

Study 2

Reproductive toxicity study of test materialwas performed on female Sprague Dawley rats. 86 rats were divided into 2 treatment group.25 female rats in treatment group 1 while 16 in treatment group 2 .Test material mixed with0.5% aqueous methyl cellulose were administers in dose concentration 0, 20, 80 or 200 mg/kg bw day from days 15 through 21 of gestation by oral gavage route twice daily.One-half of the dose was given to the animals in the morning and the second half 6-8 h later. Dosing volume was 10 ml/kg per dosing occasion. For mating purposes, one female and one male (from breeding stock) were housed together. The occurrence of copulation was determined by daily inspection for a copulatory plug. The day at which evidence of mating was detected was designated gestation day 0, and the female was placed in an individual cage. The animals were observed twice daily throughout the study for mortality and signs of overt toxicity. Individual body weights were recorded on gestation days 0, 6, 15, 16, 17, 18, 19, 20 and 21. Beginning on day 21, Pups were examined for external abnormalities as soon as possible following delivery. The number of viable and nonviable pups was recorded for each female. All pups were sexed and weighed individually on lactation day 0. All surviving F,, females and F, pups were euthanized by carbon dioxide inhalation at lactation day 1. On gestation day 25, those females which failed to deliver were euthanized. The uterus and ovaries of all dams were exposed by an abdominal incision and grossly examined to determine the number of implantation sites present.

 

No observations of adverse effects until the onset of parturition. As well, there were no apparent compound-related effects on gestational body weight gain in any of the treatment groups. One death occurred in the vehicle-treated group on gestation day 20. However, post-mortum examination revealed no abnormalities or signs of overt clinical toxicity. Both onset of labour and its duration were disrupted in the animals receiving the highest dose. Increased fetotoxicity and periparturn death also appeared to occur among foetuses from females treated with 200 mg/kg/day but this was not statistically significant,Labour times in these groups ranged from 1-14 h with a mean duration of 3.6 h for the treated group, For comparison, labor times in control animals ranged from less than 1 h to 3 h with a mean labour time of 1.5 h. There were no observed differences in mean litter size, pup size, or sex ratio, associated with test material . Since those pups which did survive delivery had no visible abnormalities or other signs of overt toxicity. Maternal perinatal death was significantly increased in animals treated with both 200 mg/kg/ day. Four animals in each of these treatment groups died or were euthanized because of extreme distress during parturition. Clinical signs of maternal physical distress during parturition included labored breathing, decreased activity, and blood staining in the abdominal, nasal, and oral areas. Notably, each of these seven animals failed to deliver some or all of their pups. Increased maternal mortality was also evidenced as a significant decrease in the gestational index (defined as the percentage of pregnant females producing live pups) for these same treatment groups. No evidence of increased maternal distress or peripatum mortality was noted in animals from either the mid-dose (80 mg/kg/day) or low-dose (20 mg/kg/day) sodium salicylate. Hence No Observed Effect Level (NOEL) was considered to be 80 mg/kg/day, and LOAEL dose-level of 200 mg/kg/day was considered to be for reproductive toxicity.When female Sprague Dawley rats were treated with test material orally.

 Study 3

 The reproductive toxicity study of test materialwas performed on male and female  CD-1 mice. Based on dose-range finding study dose levels of0,100,250,500mg/kgwere selected. The vehicle control group (40 male and 40 female) and three dose groups (20 male and 20 female).Eleven-week old male and female CD-l mice were exposed to the chemical during a7-day premating period, after which they were randomly paired (one male: one female) within each dose group. Cohabitation was continued for100days. All animals will be weighed once a week from day 0 (first day of treatment) to day127 New-born litters were evaluated and immediately sacrificed. Eleven animals died, 3 in the control, 2 each in the 100mg/kg and 250mg/kg dose groups, and 4 in the 500mg/kg dose group. The cause of death varied from case to case but it was neither chemical nor dose related. No distinct treatment related symptoms of toxicity were observed during routine health surveillance. Test material in dose level 0,100,250,500 mg/kg bw had no apparent effect on male or female body weights .The fertility index in the control and various treatment groups varied between 95 to 100% all breeding pairs except 1 in the 100mg /kg group delivered at least one litter .There was a significant decrease (p<0.05) in the mean number of litters at the 0.5 g/kg level .The average number of pups per litter, the proportion of pups born alive, and mean live pup weight values were also significantly reduced (p<0.05) in the 500mg /kg group compared to the correspondingcontrol.There was no significant effect on any of these parameters in the remaining two dose groups.HenceNo Observed Adverse Effect Level (NOAEL) for reproductive toxicity was considered to be 250 mg/kg/day, and the dose-level of 500 mg/kg/day was considered to be the LOAEL for reproductive toxicity.When male and femaleCD 1 mice weretreated with test materialorally.

Study 4

The reproductive toxicity study of test materialwas performed on male and female  CD-1 mice. 11 weeks of age animals were used in study. Based on dose-range finding study dose levels of 0.0, 25.0, 50.0, and 100.0 mg /kg/day were selected. Test material soluble in corn oil and administered by gavage daily during the 7-day premating, 98-day cohabitation, and 21-day segregation periods. The vehicle control group (40 male and 40 female) and three dose groups (20 male and 20 female), For the first week, animals will be housed five per cage by sex. Subsequently, the females and males from the same dose group will be randomly paired and cohabited. The adult animals will be weight weekly.

The test material had no influence on the number of litters produced per pair, the number of live pups per litter, the proportion of pups born alive or sex (males/total) of pups born alive .Female live pup weight adjusted for the total number of pups per litter, however, was significantly greater p<0.05) for the breeding pairs given 100.0 mg /kg/day than for those receiving 50.0 mg /kg/day. Further, combined (both sexes) live pup weight adjusted for the total number of pups per litter was significantly greater (p<0.05) for breeding pairs receiving 100.0 mg /kg/day than for those given 25.0 or 50.0 mg /kg/day .Since there were no other indications of toxicity during continuous breeding, these differences might be due to chance alone. It also should be noted that one parental male and female in the control group, one parental female in the 25.0 mg /kg/day group, two parental females and three parental males in the 50.0 mg /kg/day group, and three parental males in the 100.0 mg /kg/day group died prior to the conclusion of the continuous breeding phase .The random distribution of deaths across treatment groups suggested that they were not treatment-related.

 

Due to the lack of an effect of test material on fertility and reproductive performance in the parental mice (F0 generation), it was decided to assess fertility and reproductive performance in the F1 offspring. The litters of the 0.0 and 100.0 mg /kg/day. groups were weaned at 21 days of age, and one or two females and male pups from each surviving litter were randomly selected .Each weanling was housed individually and maintained on the same treatment as their parents. At 90±10 days of age, a male and female from different litters within treatment groups were cohabited from 1 to 7 days, depending upon when a copulatory plug was detected. The pairs were separated and the females were allowed to deliver their litters. Continuous exposure of CD-1 mice to test material in utero, via the mother's milk and by gavage (100.0 mg /kg/day) from weaning at 21 days of age to sacrifice at 127±8 days of age was found to have no statistically significant effects on mating behavior , fertility rate or reproductive performance

The 0.0 and 100.0 mg/kg/day F1 parental mice were necropsied .Sperm assessment indicated no significant differences in the %motile sperm, sperm concentration or %abnormal sperm in the cauda epididymis between male mice exposed to 0.0 or 100.0mg /kg/day .In addition, there were no significant differences in body weights or organ weights between the male mice in these two groups .Body weight and liver and pituitary weights also were unaffected by treatment of female mice .In contrast, brain weight was significantly increased (p<0.05)and reproductive tract weight significantly decreased (p<0.05) in female mice given 100.0 mg /kg/day versus control female mice .The reproductive tract weights of the two females in the control group that did not deliver pups were unexpectedly heavier on average than those from the 17 control females that did deliver pups. This was in contrast to the findings for the females treated with100.0 mg/kg/day. That is, the six females in this group that did not give birth had lighter reproductive tracts on average than did their eleven post gravid cohorts .This difference,along with the fact that the control group had a greater proportion of females with postgravid reproductive tracts than did the treated group (17/19 versus 11/17, respectively), might well explain the significantly heavier reproductive tracts in the control group as compared to those in the treated group. Thus, since the difference between the two groups appeared to be due to biological variation, it was not considered to be of toxicological significance. Finally, no-treatment-related gross or histopathologic lesions were noted for the pituitary, testis, epididymis, prostate or seminal vesicles in male mice, or for the pituitary, ovary, oviduct, uterus or vagina in female mice. Hence No Observed Adverse Effect Level (NOAEL) for reproductive toxicity for F0 and F1 generation was considered to be 100 mg/kg/day.When male and female  CD-l mice were treated with test material orally.

 

 Based on the data available from different studies ,test chemical did not showedreproductive toxicityat dose concentration 75 mg/kg bw/day.When rats or mice were treated with test material orally. but adverse effects were observed on fertility and developmental parameters on higher dose concentration ,Thus, comparing this value with the criteria of CLP regulation test material is likely to classify as reproductive toxicant.

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 likely to classify as reproductive toxicant.