Registration Dossier

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

Administrative data

Endpoint:
developmental toxicity
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:
Developmental toxicity study was performed on Sprague Dawley rats to evaluate and assess toxicological potential of the test chemical.
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
- Name of test material (as cited in study report):sodium salicylate
- Molecular formula :C7H6O3.Na
- Molecular weight : 160.1035 g/mol
- Substance type: organic
- Physical state: solid
- Smilies notation: c1(c(cccc1)O)C(=O)[O-].[Na+]
- InChI: 1S/C7H6O3.Na/c8-6-4-2-1-3-5(6)7(9)10;/h1-4,8H,(H,9,10);/q;+1/p-1
Specific details on test material used for the study:
- Molecular formula :C7H6O3.Na
- Molecular weight : 160.1035 g/mol
- Substance type: organic
- Physical state: solid
- Smilies notation: c1(c(cccc1)O)C(=O)[O-].[Na+]
- Inchi: 1S/C7H6O3.Na/c8-6-4-2-1-3-5(6)7(9)10;/h1-4,8H,(H,9,10);/q;+1/p-1

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Laboratories
(Portage, MI).
- Age at study initiation: 63 days
- Weight at study initiation: 182-263g
- Fasting period before study: No data available

- Housing: The animals were housed individually
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): Food (Certified Rodent
Chow 5002, Purina Mills, Inc., St. Louis, MO) ad libitum
.
- Water (e.g. ad libitum): water were provided ad libitum
- Acclimation period: No data available


ENVIRONMENTAL CONDITIONS
- Temperature (°C):21.11±1.2 °C
- Humidity (%):57±3.8%
- Air changes (per hr): No data available

- Photoperiod (hrs dark / hrs light): No data available


IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% aqueous methyl cellulose
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test chemical was mixed with 0.5% aqueous methyl cellulose to form dosing solution
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 chemical was mixed with 0.5% aqueous methyl cellulose
- Concentration in vehicle: 0, 20, 80 or 200 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg

- Lot/batch no. (if required): No data available
- Purity: No data available
Other :
One-half of the dose was given to the animals in the morning and the second half 6-8 h later
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Details on mating procedure:
- Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]
- If cohoused: No Data Available
- M/F ratio per cage: 1:1
- Length of cohabitation: No Data Available
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- Verification of same strain and source of both sexes: [yes / no (explain)]
- 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,
- Any other deviations from standard protocol: No Data Available
Duration of treatment / exposure:
7 days ( days 15 through 21 of gestation. )
Frequency of treatment:
twice daily
Duration of test:
25 days
Doses / concentrations
Remarks:
0, 20, 80 or 200 mg/kg bw/day
No. of animals per sex per dose:
Total: 86 animals

Treatment group 1
0 mg/kg bw/day: 25 animals
20 mg/kg bw/day: 25 animals
80 mg/kg bw/day:25 animals

Treatment group 2
200 mg/kg bw/day: 16 animals
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 100 mg/kg/day for oral exposure to the test chemical.

- Rationale for animal assignment (if not random):

- Other:

Examinations

Maternal 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:

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of early resorptions: No data
- Number of late resorptions: No data
- Other: No Data
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: Yes
Statistics:
All measured parameters were subjected to Bartlett’s test for hlomogeneity of variance. Then all treatment groups and the vehicle-treated control
group were compared by one-way analysis of variance (ANOVA). Pairwise comparisons were made using the appropriate t-test (for equal and unequal variance) as described by Steel and Torrie using Dunnett’s multiple comparison tables, or pairwise comparisons were made with a Bonferroni correction to determine the significance.
Indices:
No data available
Historical control data:
No data available

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No observations of adverse effects until the onset of 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.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One death occurred in the vehicle-treated group on gestation day 20.

Maternal perinatal death was significantly increased in animals treated with 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.

No evidence of increased 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.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
There were no apparent compound-related effects on gestational body weight gain in any of the treatment groups
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
Post-mortem examination revealed no abnormalities in the animals.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Post-mortem examination revealed no abnormalities in the animals.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Maternal developmental toxicity

Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): effects observed, treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): Both onset of labour and its duration were disrupted in the animals receiving the highest dose 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 hour to 3 hour with a mean labour time of 1.5 hours.
Changes in number of pregnant:
not specified
Other effects:
not specified

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
effects on pregnancy duration
Remarks on result:
other: No toxic effects observed
Dose descriptor:
LOAEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
effects on pregnancy duration
Remarks on result:
other: effcts observed

Maternal abnormalities

Abnormalities:
not specified
Localisation:
not specified

Results (fetuses)

Fetal body weight changes:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
effects observed, non-treatment-related
Description (incidence and severity):
Increased fetotoxicity and periparturn 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.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
There were no observed differences in sex ratio, associated with the test chemical.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
There were no observed differences in mean litter size, pup size associated with the test chemical.
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
The pups which survived at the delivery had no visible abnormalities or other signs of overt toxicity
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
not specified

Effect levels (fetuses)

open allclose all
Dose descriptor:
NOAEL
Effect level:
80 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
changes in sex ratio
changes in litter size and weights
external malformations
Remarks on result:
other: No toxic effects observed
Dose descriptor:
LOAEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
reduction in number of live offspring
changes in sex ratio
changes in litter size and weights
external malformations
Remarks on result:
other: effects observed in highest dose tested

Fetal abnormalities

Abnormalities:
not specified
Localisation:
other: not specified

Overall developmental toxicity

Developmental effects observed:
not specified
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

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 and developmental toxicity. When female Sprague Dawley rats were treated with the test chemical orally.
Executive summary:

The experimental study conducted on the test chemical to evaluate 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 chemical mixed with 0.5% aqueous methyl cellulose were 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 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 animals were 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 periparturn 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. Labour durations in these groups ranged from 1-14 h with a mean duration of 3.6 h for the SA-treated. For comparison, labor duration 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 prenatal 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 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 and developmental toxicity, when  female Sprague Dawley rats were treated with the test chemical orally.