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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD)

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
1989
Report date:
1989

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
2,5,6-trimethylcyclohex-2-en-1-one
EC Number:
243-473-0
EC Name:
2,5,6-trimethylcyclohex-2-en-1-one
Cas Number:
20030-30-2
Molecular formula:
C9H14O
IUPAC Name:
2,5,6-trimethylcyclohex-2-en-1-one
Details on test material:
- Name of test material (as cited in study report): 2,3,6-Trimethylcyclohexen-1-on
- ZST Test Substance No . : 87/818
- Analytical purity: ca. 96 %
- Storage condition of test material: room temperature

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S9 mix
Test concentrations with justification for top dose:
Standard plate test (SPT): 100, 500, 2500 and 5000 ug/plate
Preincubation test (PIT): 100, 500 and 2500 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene, 10 µg/plate, dissolved in DMSO, for the strains TA100, TA98, TA1537 and TA1535
Remarks:
with metabolic activation
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitrosoguanidine; 5 µg/plate, dissolved in DMSO, for strains TA100 and TA1535
Remarks:
without metabolic activation
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylendiamine, 10 µg/plate, dissolved in DMSO, for strain TA98
Remarks:
without metabolic activation
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without metabolic activation

Migrated to IUCLID6: 100 µg/palte, dissolved in DMSO, for strain TA1537
Details on test system and experimental conditions:
METHOD OF APPLICATION:
The experimental procedures are based on the methods described by Ames, (1973, 1975), Yahagi et al. (1977) and Matsushima et al. (1980).

1ST EXPERIMENT: in agar (plate incorporation) (SPT)

DURATION
- Exposure duration: 48 - 72 h, 37°C


2ND EXPERIMENT: preincubation (PIT)


DURATION
- Preincubation period: 20 min, 37°C
- Exposure duration: 48 - 72 h, 37°C

NUMBER OF REPLICATIONS: triplicates; 3 test plates per dose or per control


DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn (= reduced his- or trp- background growth), reduction in the titer
Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results
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:
depending on the strain and test conditions at doses > 2500 µg/plate .
Vehicle 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

Results of SPT:

Strain metabolic activation Experiment  controls µg substance / plate Evaluation
solvent positive 20 100 500 2500 5000
TA 1535 - 1 17 2100 15 17 16 11 11 -
- 2 12 2150 22 18 23 13 8 -
- 3 14 1950 20 14 10 18 7 -
+ 1 21 123 21 23 19 19 7 -
+ 2 19 159 17 18 20 20 8 -
+ 3 19 154 22 18 23 16 13 -
TA 1537 - 1 7 245 8 6 11 8 B -
- 2 8 372 5 7 7 6 B -
- 3 8 367 8 10 8 9 B -
+ 1 12 129 6 7 9 7 7 -
+ 2 9 117 9 14 11 9 8 -
+ 3 8 142 8 8 8 9 4 -
TA 98 - 1 21 575 20 21 23 22 13B -
- 2 22 647 19 23 26 21 10B -
- 3 24 652 26 18 20 16 13B -
+ 1 33 1310 42 33 36 32 28 -
+ 2 32 1180 31 49 38 36 28 -
+ 3 35 1060 31 35 41 38 33 -
TA 100 - 1 101 1820 106 97 120 98 60B -
- 2 97 1920 97 101 118 101 71B -
- 3 102 1950 120 98 101 91 70B -
+ 1 121 1890 135 122 111 102 97 -
+ 2 128 1830 105 133 103 101 84 -
+ 3 115 1870 119 133 128 114 88 -

B: reduced his-backround

Results of PIT:

mean number of revertant colonies / plate  
Strain metabolic activation Experiment  controls µg substance / plate Evaluation
solvent positive 4 20 100 500 2500
TA 1535 - 1 19 1050 21 17 13 18 13 -
- 2 12 1080 21 15 13 16 13 -
- 3 12 1100 13 15 13 14 13 -
+ 1 13 139 15 17 18 12 13 -
+ 2 16 132 19 18 20 12 13 -
+ 3 18 110 22 19 17 14 13 -
TA 1537 - 1 8 423 7 11 10 5 B -
- 2 8 413 9 7 8 3 B -
- 3 7 377 9 10 7 5 B -
+ 1 13 129 9 8 7 8 B -
+ 2 12 100 9 8 9 10 B -
+ 3 11 85 9 8 10 11 B -
TA 98 - 1 19 985 24 26 26 23 B -
- 2 22 1050 26 23 21 13 B -
- 3 20 1330 24 21 20 9 B -
+ 1 32 878 30 41 33 33 B -
+ 2 36 810 34 30 30 34 B -
+ 3 34 700 37 34 40 36 B -
TA 100 - 1 106 1160 128 115 110 93 B -
- 2 106 1170 107 127 86 89 B -
- 3 118 1100 106 107 103 112 B -
+ 1 112 1060 123 98 118 128 B -
+ 2 128 1150 104 106 107 134 B -
+ 3 112 920 115 126 133 124 B -

B: reduced his-backround

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

According to the results of the present study, the test substance 2,3,6-Trimethylcyclohexen- 1-on is not mutagenic in the Ames test under the experimental conditions chosen above.