Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
other information
Study period:
Oct 2002 to Aug 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well reported GLP Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
ZK 5668
IUPAC Name:
ZK 5668
Details on test material:
- Name of test material (as cited in study report): ZK 5668
- Batch No.: 14610801

Method

Target gene:
Histidine gene locus
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
phenobarbital/beta-naphthoflavone induced liver S9-mix
Test concentrations with justification for top dose:
1,4,6-Trienol: first experiment: eight concentrations from 3 to 5000 µg/plate; second experiment 10 concentrations from 0.3 to 5000 µg/plate
Sodium azide: 10 µg/plate
4-Nitro-o-phenylenediamine: 10 µg/plate (TA 98) or 50 µg/plate (TA 1537)
Methyl methane sulfonate: 4 µl/plate
2-Aminoanthracene: 2.5 µg/plate (10 µg/plate in TA 102)






Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
DMSO or water deionised
Positive controls:
yes
Positive control substance:
other: without metabolic activation: Sodium azide, 4-Nitro-o-phenylenediamine, Methyl methane sulfonate; with metabolic activation: 2-Aminoanthracene

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Precipitation of the test item was observed without metabolic activation in strains TA 1537 and TA 102 at 5000 µg/plate in the first experiment, and from 1000 µg/plate up to 5000 µg/plate in the second experiment.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with ZK 5668 at any concentration level, neither in the presence nor absence of metabolic activation (S9 mix). However, in the first experiment an increase in revertant colony numbers was observed in strain TA 102 in the absence of metabolic activation, already beginning at the lowest concentration but with no dose dependent increase. The threshold of two times the number of the corresponding solvent control was reached at 100 µg/plate. In order to check this questionable effect an additional experiment was performed with an adjusted concentration range of 0.3 up to 5000 µg/plate. No increase in revertant colony numbers was observed in this additional experiment and the effects observed in the first experiment were judged to be based on biologically

irrelevant fluctuations in the number of colonies.

Applicant's summary and conclusion

Executive summary:

1,4,6 -Trienol (ZK 5668) was examined in two experiments for mutagenic activity up to 5000 µg/plate in the five histidine-dependent Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 with and without metabolic activation.

No cytotoxic effects were seen up to 5000 µg/plate. Precipitation of the test item was observed without metabolic activation in strains TA 1537 and TA 102 at 5000 µg/plate in the first experiment, and from 1000 µg/plate up to 5000 µg/plate in the second experiment.

There was no evidence for a mutagenic activity of 1,4,6 -Trienol, when tested up to the maximum recommended dose level of 5000 µg/plate in the absence and presence of S9 mix.