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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
toxicity to reproduction
Remarks:
other: one-generation study
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from a peer reviewed journal.

Data source

Reference
Reference Type:
publication
Title:
Maternal reproductive effects of oral salicylic acid in Sprague-Dawley rats
Author:
D.P. Davis, G.P. Daston, M.R. Odio, R.G. York, A.L. Krausa
Year:
1996
Bibliographic source:
Toxicology Letters 84 (1996) 135-141

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Maternal reproductive effects of the test chemical in Sprague-Dawley Rats by oral route were assessed in a one generation of rats in an overall estimation period of 15 through 21 days of gestation.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Crystal powder
Details on test material:
- Name of test material (as cited in study report): Sodium salicylate
- Molecular formula : C7H5NaO3
- Molecular weight : 160.10467
- Substance type: Organic
- Physical state: Solid
Specific details on test material used for the study:
- Name of test material (as cited in study report): Sodium salicylate
- Molecular formula : C7H5NaO3
- Molecular weight : 160.10467
- Substance type: Organic
- Physical state: Solid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
No data available
Sex:
female
Details on test animals and environmental conditions:
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

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5 % Aqueous methyl cellulose
Details on exposure:
Test chemical 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 % Aqueous methyl cellulose
- 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:
- 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.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
6 days (Days 15 through 21 of gestation)
Frequency of treatment:
Twice daily
Details on study schedule:
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 the test chemical or to one treatment group consisting of 16 females which received 200 mg/kg/day of the test chemical. 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.
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 20, or 80 or 200 mg/kg/day
Basis:

No. of animals per sex per dose:
Total:86
Control:25 females
20 mg/kg/day: 25 females
80 mg/kg/day: 25 females
200 mg/kg/day: 16 females
Control animals:
yes, concurrent vehicle
Details on study design:
- 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 to the test chemical.

- Rationale for animal assignment (if not random): No Data Available

- Other:
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
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: No Data Available
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
Litters’ were examined for numbers of live and stillborn pups and gross abnormalities. The pups were sexed and weighed individually on lactation day 0.
Postmortem examinations (parental animals):
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.
Postmortem examinations (offspring):
No data available
Statistics:
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.
Reproductive indices:
No data available
Offspring viability indices:
No data available

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
No observations of adverse effects.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
When pregnant female rats were treated with 200 mg/kg/day of the test chemical, the maternal perinatal death significantly increased while the gestational index decreased. At 20 or 80 mg/kg/day, no evidence of increased maternal distress or peripatum mortality was noted.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effects on gestational body weight gain .
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not specified
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No observations of adverse effects until the onset of parturition at 0, 20 or 80 mg/kg/day.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: No effect on reproductive parameters
Dose descriptor:
LOAEL
Effect level:
200 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: When pregnant female rats were treated with 200 mg/kg/day, the maternal perinatal death significantly increased while the gestational index decreased.

Target system / organ toxicity (P0)

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

Results: P1 (second parental generation)

General toxicity (P1)

Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Mean pup weight was found to be reduced.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Details on results (F1)

Litter Observation: There were no observed differences in mean litter size, pup size, or sex ratio, associated with the test chemical treatment. Increased fetotoxicity and periparturition death also appeared to occur among fetuses from females treated with 200 mg/kg/day of the test chemical but this was not statistically significant.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
viability
clinical signs
body weight and weight gain
Remarks on result:
other: overall developmental effects

Target system / organ toxicity (F1)

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

Overall reproductive toxicity

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

Any other information on results incl. tables

I] Fetal observations in offspring of Sprague-Dawley rats treated orally with the test chemical on days 15-21 of gestation.

 

Control

Test Chemical

 

 0 mg/kg/day

20 mg/kg/day

80 mg/kg/day

200 mg/kg/day

Litters with livebom pups

20

20

18

6

Pups delivered (total)

255

270

230

62

Mean pups/litter

12.75 ± 3.82

13.50 ± 2.80

12.78 ± 3.08

10.33 ±4.46

Stillborn pups

8

7

1

6

Live pups/litter

 

 

 

 

Lactation day 0

12.35 ±3 .82

13.15 ±2.66

12.72 ±3.04

9.33 ± 4.76

Lactation day 1

12.30 ±3.88

13.15 ± 2.66

12.72 ±3.04

8.67 ±5.32

Mean pup weight (g)

5.84 ±0.62

5.70 ± 0.52

5.85 ± 0.49

5.23 ± 0.23

Pups dying, missing, and/or

cannibalized

 

 

 

 

Lactation day

0

0

0

2

Lactation day

1

0

0

2

Sex ratio (male pups:total pups)

55.1

47.5

50.2

51.8

 

  II] Maternal and reproductive observations in Sprague-Dawley rats treated orally with the test chemicals on days 15-21 of gestation.

 

Control

Test chemical

 

 0 mg/kg/day

20 mg/kg/day

80 mg/kg/day

200 mg/kg/day

Number of females mated

25

25

25

I6

Number of females pregnant

21

20

I8

10

Mean gestational weight change (g)”

133.6 ± 20.3

136.9 ±1 9.8

135.1 ±18.8

131.7 ±9.7

Females with liveborn pups

20

20

I8

6

Gestation Index

95.2

100

100

60

Females surviving delivery

20

20

I8

6*

 

 

Applicant's summary and conclusion

Conclusions:
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 the test chemical orally.
Executive summary:

The experimental study conducted for assessment and evaluation of Reproductive and developmental toxicity study of the test chemical was 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 with 0.5% aqueous methyl cellulose as the vehicle and was administered 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 maintained at 10 ml/kg per dosing. 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, after which the females 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 11 pups were sexed and weighed individually on lactation day 0. All surviving F0 females and F1 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-mortem 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 periparturition death also appeared to occur among foetuses from females treated with 200 mg/kg/day of the test chemical but this was not statistically significant of the test chemical (200 mg/kg) ,Labour times in these groups ranged from 1-14 h with a mean duration of 3.6 h for the SA-treated 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 the test chemical. 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 of the test chemical. 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) of the test chemical. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 80 mg/kg/day, and LOAEL dose-level of 200 mg/kg/day was considered to be for reproductive and developmental toxicity. When  female Sprague Dawley rats were treated with the test chemical orally.