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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study published in a peer-reviewed journal, according to scientific standards. Experimental details well documented, no explicit reference to guideline.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Comparative genotoxicity of six salicylic acid derivatives in bone marrow cells of mice
Author:
Giri AK, Adhikari N & Khan KA
Year:
1996
Bibliographic source:
Mutat. Res., 1996, 370, 1-9

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
Deviations:
yes
Remarks:
Housing temperature (28+-2°C) too high. Only male mice tested (5 per dose) - no justification given.
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.5385 (In Vivo Mammalian Cytogenetics Tests: Bone Marrow Chromosomal Analysis)
Deviations:
yes
Remarks:
Only male mice tested (4 per dose i.p., 5 for oral gavage----) - no justification given.
GLP compliance:
not specified
Type of assay:
chromosome aberration assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Substance type: pure active substance
- Physical state: solid
- Supplier: Sigma Chemical Co, St. Louis MO, USA
- Analytical purity: no data

Test animals

Species:
mouse
Strain:
Swiss
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Division of Laboratory animals, Central Drug Res. Institute, Lucknow, India
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 30 g
- Assigned to test groups randomly: no data
- Fasting period before study: no
- Housing: five per cage, husk bedding
- Diet: Standard rodent pellet diet, Gold Mohor, Lipton India Ltd, Chandigarh, India (ad libitum)
- Water: ad libitum
- Acclimation period: no data

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

IN-LIFE DATES: no data

Administration / exposure

Route of administration:
other: intraperitoneal (3 dose levels); or oral (gavage) 1 dose level
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO (for i.p. application), suspension in 2% gum acacia in distilled water (for gavage application)
- Justification for choice of solvent/vehicle: solubility of the test substance
- Concentration of test material in vehicle: dose-dependent: approx. 20, 40, and 80 mg/mL (i.p.), 35 mg/mL (gavage)
- Amount of vehicle (if gavage or dermal): DMSO 75 microliter/mouse for i.p., 2% gum acacia in water 0.3 mL per mouse for gavage
- Type and concentration of dispersant aid (if powder): 2% gum acacia
- Lot/batch no. (if required): no data
- Purity: no data
Duration of treatment / exposure:
i.p. administration: 24 hours, gavage administration: 24 hours
colchicine: 2 hours before sacrifice
Frequency of treatment:
single treatment
Post exposure period:
none (time between administration and sacrifice: i.p. 24 hours, gavage 24 hours)0
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
i.p. 50, 100, 200 mg/kg body weight
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
oral (gavage) 350 mg/kg body weight
Basis:
nominal conc.
No. of animals per sex per dose:
4 males per dose, i.p. administration
5 males, oral administration
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide:
- Justification for choice of positive control(s): no data
- Route of administration: i.p. only
- Doses / concentrations: 25 mg/kg

Examinations

Tissues and cell types examined:
Bone marrow cells
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION:
i.p.: based on the results of the Sister Chromatid Exchange (SCE) dose-response study: one higher dose was selected, since chromosome aberration is less sensitive than SCE
oral/gavage: 1/3 of approx. oral LD50

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
- i.p. injection and gavage 22 h before colchicine, 24 hours before sacrifice
- colchicine (2 mg/kg) 2 h before sacrifice
- sacrifice and expelling of bone marrow 24 hours after treatment

DETAILS OF SLIDE PREPARATION:
- bone marrow chromosomes prepared according to Preston RJ et al (1987) Mutation Res. 198: 157-165
- staining of chromosomes on slide with Giemsa

METHOD OF ANALYSIS:
- 100 well-spread metaphase cells scored per animal
- mitotic indices (MI) calculated from 1000 cells / animal, expressed as percentage
- chromosome aberrations (CA) scored according to WHO method (Preston et al, see above)
- aberration frequencies of chromatid and chromosome type per cell calculated
- gaps recorded but not included in the frequency of aberrations per cell
Evaluation criteria:
Significant increase in chromosome aberration
Gaps recorded but not included in the frequency of aberrations per cell
Statistics:
Statistical calculations carried out from percentages of aberrant cells
Student's t-test, to compare results at each dose with control

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Remarks:
no mortality
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid

Any other information on results incl. tables

No significant increase in aberrations / cell and aberrant cells (%) for all doses tested, when compared to solvent control.

No significant difference in mitotic indices, when compared to control.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative