Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

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
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Report date:
1986

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Life Science Research Ltd.
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2-methylbut-3-yn-2-ol
EC Number:
204-070-5
EC Name:
2-methylbut-3-yn-2-ol
Cas Number:
115-19-5
Molecular formula:
C5H8O
IUPAC Name:
2-methylbut-3-yn-2-ol
Details on test material:
- Name of test material (as cited in study report): P0073

Method

Target gene:
The characteristics of the individual strains are as follows:
TA 1535 - contains a histidine missense mutation but is also deficient in a DNA repair system (uvr B) and has a defective lipopolysaccharide coat on the cell wall. It is reverted by many agents causing base-pair substitutions, but is not sensitive to frameshift mutagens.
TA 1537 - bears a histidine frameshift mutation. Like TA 1535, it is defective in a DNA repair system and lipopolysaccharide coat. It is sensitive to agents causing frameshift mutations involving insertion or deletion of a single base-pair.
TA 1538 - contains another histidine frameshift mutation. Again it has a defective DNA repair system and lipopolysaccharide coat. It is reverted by agents causing deletion of two adjacent base-pairs (double frameshift mutations).
TA 100 - is the same as TA 1535 but contains a resistance transfer factor conferring ampicillin resistance and increasing sensitivity to some mutagens (plasmid pKM 101). In addition to base-pair substitutions, it is also able to detect certain frameshift mutagens.
TA 98 - is TA 1538 with the addition of the pKM 101 plasmid. It is reverted by a variety of mutagens, but not by simple alkylating agents causing base-pair substitutions.
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
- Type and identity of media: Cultures of all organisms were prepared by overnight incubation of freshly inoculated nutrient broth (Oxoid No.2).
Species / strain / cell type:
S. typhimurium TA 1538
Details on mammalian cell type (if applicable):
- Type and identity of media: Cultures of all organisms were prepared by overnight incubation of freshly inoculated nutrient broth (Oxoid No.2).
Metabolic activation:
with and without
Metabolic activation system:
Liver microsomal preparation (S9 mix)
Test concentrations with justification for top dose:
Series of eight different concentrations of test material from 2.5 ug to 5 mg per plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: distilled water
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
2 ug/plate TA1535
Positive control substance:
other: 2-acetylaminofluorene
Remarks:
5 ug/plate TA 1535
Positive control substance:
9-aminoacridine
Remarks:
50 ug/plate TA 1537
Positive control substance:
2-nitrofluorene
Remarks:
5 ug/plate TA 1538, TA 98
Positive control substance:
benzo(a)pyrene
Remarks:
5 ug/plate TA 1537, TA 1538, TA 100, TA 98
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Expression time (cells in growth medium): 48 hours

NUMBER OF REPLICATIONS: 3 on two separate occasions.

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: thinning of the background lawn
Evaluation criteria:
Increases in revertant colony numbers.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Slight reduction in background lawn
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Slight reduction in background lawn
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: Preliminary toxicity test for dose selection using test strain TA 98.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

No increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to test item at levels from 50 to 5000 ug per plate. Thus, the test item was considered non mutagenic under the conditions of the test.
Executive summary:

An in vitro study on bacteria was carried out according to EU method B.13/14 and OECD guideline 471. The test itemwas examined for mutagenic activity in five histidine-dependent auxotrophs of Salmonella typhimurium,strains TA 98, TA 1538, TA 100, TA 1535 and TA 1537, using pour-plate assays. Each test, in each strain, was conducted on two separate occasions and run in triplicate. The studies, which were conducted in the absence and presence of an activating system derived from rat liver (5-9 mix), employed a range of levels of test item from 50 to 5000 ug per plate, selected following a preliminary toxicity test in strain TA 98, and included solvent (distilled water) controls with and without S-9 mix. No increases in reversion to prototrophy were obtained with any of the five bacterial strains at the compound levels tested, either in the presence or absence of S-9 mix. Marked increases in the number of revertant colonies were induced by the known mutagens benzo[a]pyrene, 2-nitrofluorene, 2-aminoanthracene, 9-aminoacridine and sodium azide when examined under similar conditions. It was concluded that the test item was devoid of mutagenic activity under the conditions of the test.