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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 15 to May 16, 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in 2006 according to OECD Method 471 and EU Annex V test B 13 and B14 and in accordance with GLP. Study material is well characterized.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Test material is a white solid which was received at testing laboratory on January 11, 2006 and stored at room temperature in the dark.

Method

Target gene:
four histidine-requiring strains and one tryptophan-requiring strain
Species / strainopen allclose all
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver post mitochondrial fraction (rat-liver S-9)
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver post mitochondrial fraction (rat-liver S-9)
Test concentrations with justification for top dose:
Range finding experiment and main experiment carried out at concentrations from 15 to 5000 ug/plate for salmonella and 50 to 50000 ug/plate for e coli strain. For the main study doses of 15, 150 ,500, 1500 and 5000 ug/plate for salmonella and 50, 150, 500, 1500 and 5000 ug/plate for e coli strain where used. An additional dose had been added to main test to allow for test material induced toxicity.
Vehicle:
Test solutions were prepared by dissolving BMS 587319-03 in sterile anhydrous grade dimethyl sulphoxide (DMSO).
Controlsopen allclose all
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO)
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
Migrated to IUCLID6: use strain TA98 without S-9)
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Migrated to IUCLID6: use strain TA100, TA 1535 without S-9)
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Migrated to IUCLID6: (use strain TA 1537 without S-9)
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
Migrated to IUCLID6: use strain TA98 with S-9
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
Migrated to IUCLID6: use strain WP2uvrA without S-9
Negative controls:
yes
Remarks:
(see solvent controls)
Solvent controls:
yes
Remarks:
treatment with solvent DMSO
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
use strain TA100,TA1535,TA1537, WP2uvrA with S-9
Details on test system and conditions:
A preliminary range finding experiment was conducted with strain TA100 and WP2uvrA at 10 concentrations from 15 to 5000 ug/plate plus negative (solvent) controls. There was a minor error in were the plates were left in incubator for 18 hours longer but no variations were noted and they were deemed acceptable. No evidence of toxicity was observed following this treatment for WP2uvrA but weakened lawns for TA100 at high doses of 5000 ug/plate were noted. This test was acceptable.For experiment 1 and 2 the Experiment 2 involved the five strains with and without metabolic activation using the same concentrations as preliminary but adding a dose for samonella strains to allow for test material induced toxicity. Normal plating treatment procedures followed. No evidence of toxicity was observed following this treatment. Negative and positive control treatments were included for all strains in both experiments. The mean numbers of revertant colonies on negative control plates were all acceptable.
Evaluation criteria:
Acceptance criteria for validity of assay: the mean negative control counts fell within normal ranges, the positive control chemical induced clear increases in revertant numbers confirming discrimination between different strains and an active S-9 preparation, and no more than 5 % of the plates were lost through contamination or some other unforeseen event. Evaluation criteria: test article would be considered mutagenic if: the assay was valid, and an increase in frequency in revertant colonies in excess of 2 or 3 fold depending on the tester strain was noted verses the concurrent solvent controls.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Remarks:
>5000 ug/plate
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Remarks:
(> 5000 ug/plate)
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without metabolic activation

It was concluded that BMS 587319-03 did not induce mutation in four histidine-requiring strains of Salmonella typhimurium and one tryptophan-requiring strain of e-coli when tested under conditions of this study. These conditions included treatment concentrations up to 5000 ug/plate in the absence and in the presence of a rat liver metabolic activation system (S-9).