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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian cell study: DNA damage and/or repair
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 peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Gene mutation in vivo toxicity study of the test chemical
Author:
Giri et al
Year:
1996
Bibliographic source:
Mutation Research

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
In vivo sister chromatid exchange by the oral route was performed to determine the mutagenic nature of Sodium salicylate
GLP compliance:
not specified
Type of assay:
sister chromatid exchange assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Sodium salicylate
- IUPAC name: Sodium salicylate
- Molecular formula: C7H6O3.Na
- Molecular weight: 160.1035 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
Specific details on test material used for the study:
- Name of test material: Sodium salicylate
- IUPAC name: Sodium salicylate
- Molecular formula: C7H6O3.Na
- Molecular weight: 160.1035 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Test animals

Species:
rat
Strain:
Swiss
Remarks:
albino
Details on species / strain selection:
No data
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Division of Laboratory animals, Central Drug Research Institute, Lucknow
- Age at study initiation: 10-12 week old
- Weight at study initiation: 30 g
- Assigned to test groups randomly: No data
- Fasting period before study: No data
- Housing: They were kept five per
cage with husk bedding
- Diet (e.g. ad libitum): Standard rodent
pellet diet (Gold Mohor, Lipton India Ltd., Chandigarb, India) ad libitum
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 28 ± 2°C
- Humidity (%): 60_+ 5%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): 12 h light/12 h dark

IN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: distilled water in 2% gum acacia
- Justification for choice of solvent/vehicle: The test chemical was soluble in distilled water in 2% gum acacia
- Concentration of test material in vehicle: 0 or 350 mg/Kg
- Amount of vehicle (if gavage or dermal): 0.3mL/mouse
- Type and concentration of dispersant aid (if powder): No data
- Lot/batch no. (if required): No data
- Purity: No data
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved with distilled water in 2% gum acacia (0.3 ml/mouse) at the dose of 350 mg/kg half

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data
Duration of treatment / exposure:
No data
Frequency of treatment:
Once
Post exposure period:
No data
Doses / concentrations
Remarks:
0 or 350 mg/Kg
No. of animals per sex per dose:
Total: 15 male mice
0 mg/Kg: 5 male mice
350 mg/Kg: 5 male mice
Positive control: 5 male mice
Control animals:
yes, concurrent vehicle
Positive control(s):
Mitomycin C
- Justification for choice of positive control(s): No data
- Route of administration: Oral
- Doses / concentrations: No data

Examinations

Tissues and cell types examined:
Bone marrow
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: The oral dose for the test compound was approx. 1/3 of the oral LD50 of mice reported earlier. The other two lower doses were the serial dilutions of the highest doses selected for each chemical.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Paraffin-coated (approx. 80% of the surface) BrdU tablets (50 mg each) were implanted subcutaneously in the flank of the mice under ether anaesthesia. In the single-dose oral study, SS was gavaged with distilled water in 2% gum acacia (0.3 ml/mouse) at the dose of 350 mg/kg half an hour after tablet implantation to different groups of 5 animals each. For SCE analysis, colchicine (4
mg/kg) was injected (i.p.) 22 h after Brdu-tablet implantation.

DETAILS OF SLIDE PREPARATION: Two hours later, the bone marrow was expelled with 0.075 M KCl. After hypotonic treatment (0.075 M KCl at 37°C) for 20 min, the cells were fixed 3 times with methanol/acetic acid (3 : 1). The slides were prepared, and the chromosomes were differentially stained with fluorescence-plus- Giemsa technique
All the slides were coded and 30 second division metaphase cells (40 + 2 chromosomes) per animal were scored for SCE frequencies, i.e., a total of 150 cells were scored per dose tested.

METHOD OF ANALYSIS: Randomly selected metaphase cells (100/animal) were scored for replicative indices (RI) analysis by their staining pattern as first (M 1), second (M 2) and third (M 3) division metaphases

OTHER: No data
Evaluation criteria:
The bone marrow cells were observed for sister chromatid exchanges
Statistics:
Student's t-test was used to compare the results of the treated series with the respective controls for SCE, MI and RI in the oral study conducted.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: No mutagenic potential
Additional information on results:
No data

Any other information on results incl. tables

Table: In vivo sister chromatid exchanges induced by sodium salicylate in mice after oral administration

Treatment

SCE/cell of 5 animals

SCE/cell

(mean ± SD)a

Replicative indices mean ± SD)a

Solvent control(Gum acacia)

4.4, 3.9, 5.1, 4.6, 4.7

4.54±0.43

1.83±0.10

SS (350 mg/kg)

5.6, 5.9, 4.3, 5.1, 4.7

5.12±0.64

1.82±0.07

a Mean ± SD of 5 animals (30 cells/animal). Results at each dose were compared with the control using Students, t-test (* p < 0.001).

Applicant's summary and conclusion

Conclusions:
Sodium salicylate did not induce sister chromatid exchange in the bone marrow cells of mice treated orally and hence it is not likely to classify as a gene mutant in vivo.
Executive summary:

In vivo sister chromatid exchange was performed to determine the mutagenic nature of Sodium salicylate. The study was performed using Swiss albino male mice. The test chemical was dissolved in 2% gum acacia in ditilled water at dose level of 0 or 350 mg/Kg. Paraffin-coated (approx. 80% of the surface) BrdU tablets (50 mg each) were implanted subcutaneously in the flank of the mice under ether anaesthesia. In the single-dose oral study, SS was gavaged with distilled water in 2% gum acacia (0.3 ml/mouse) at the dose of 350 mg/kg half an hour after tablet implantation to different groups of 5 animals each. For SCE analysis, colchicine (4 mg/kg) was injected (i.p.) 22 h after Brdu-tablet implantation. Two hours later, the bone marrow was expelled with 0.075 M KCl. After hypotonic treatment (0.075 M KCl at 37°C) for 20 min, the cells were fixed 3 times with methanol/acetic acid (3 : 1). The slides were prepared, and the chromosomes were differentially stained with fluorescence-plus- Giemsa technique All the slides were coded and 30 second division metaphase cells (40 + 2 chromosomes) per animal were scored for SCE frequencies, i.e., a total of 150 cells were scored per dose tested. Randomly selected metaphase cells (100/animal) were scored for replicative indices (RI) analysis by their staining pattern as first (M 1), second (M 2) and third (M 3) division metaphases. No significant increase in SCE or RI was observed for SS for all the doses tested when compared with solvent control. Sodium salicylate did not induce sister chromatid exchange in the bone marrow cells of mice treated orally and hence it is not likely to classify as a gene mutant in vivo.