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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
Study period:
2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guidline study performed under GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

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

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of (1RS,2RS,7SR,8RS,9E)-9-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane and (1RS,2RS,7SR,8RS,9Z)-9-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane and (1RS,2SR,7SR,8SR,10E)-10-Ethylidene-3-oxatricyclo[6.2.1.0(2,7)] undecane and (1RS,2SR,7SR,8SR,10Z)-10-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane
Molecular formula:
C12H18O
IUPAC Name:
Reaction mass of (1RS,2RS,7SR,8RS,9E)-9-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane and (1RS,2RS,7SR,8RS,9Z)-9-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane and (1RS,2SR,7SR,8SR,10E)-10-Ethylidene-3-oxatricyclo[6.2.1.0(2,7)] undecane and (1RS,2SR,7SR,8SR,10Z)-10-Ethylidene-3-oxatricyclo [6.2.1.0(2,7)] undecane
Test material form:
gas under pressure: refrigerated liquefied gas
Details on test material:
- Physical state: liquid
- Storage condition of test material: Approximately 4 degrees C in the dark under nitrogen

Method

Species / strainopen allclose all
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9
Test concentrations with justification for top dose:
Preliminary test: 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 ug/plate
Experiment 1 (Range-finding test):
- Salmonella strains: 5, 15, 50, 150, 500, 1500, 5000 ug/plate
- E.coli strain: 15, 50, 150, 500, 1500, 5000 ug/plate
Experiment 2 (main test):
- Salmonella strains: 5, 15, 50, 150, 500, 1500 ug/plate
- E.coli strain: 15, 50, 150, 500, 1500, 5000 ug/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test material was immiscible in sterile distilled water at 50 mg/ml but was fully miscible in
dimethyl sulphoxide at the same concentration in solubility checks performed in-house. Dimethyl
sulphoxide was therefore selected as the vehicle.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
True negative controls:
yes
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-aminoanthracene; benzo(a)pyrene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours at 37 degrees C

NUMBER OF REPLICATIONS: triplicate

Evaluation criteria:
A test material will be considered non-mutagenic (negative) in the test system if the above criteria are not met. Although most experiments will give clear positive or negative results, in some instances the data generated will prohibit a definitive judgement about the test material activity. Results ofthis type will be reported as equivocal

Results and discussion

Test resultsopen allclose all
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
not specified
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
Vehicle controls validity:
not specified
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

Table 1. Range finding study

Range-finding test with and without S9
  Number of revertants (mean number of colonies per plate)
S9 Mix Test substance concentration (ug/plate) TA100 Mean TA1535 Mean WP2uvrA- Mean TA98 Mean TA1537 Mean
- 0 80 77 12 15 19 22 37 24 11 8
70 18 21 19 3
80 15 27 16 9
- 5 73 74 16 18 N/T  N/T 12 15 8 6
69 15  N/T 21 5
80 23 N/T  13 5
- 15 65 79 12 8 26 24 29 25 9 7
84 4 22 19 4
89 8 24 26 7
- 50 82 78 12 11 18 21 14 18 7 8
67 12 20 13 10
86 8 26 26 7
- 150 67 83 16 17 20 21 15 17 11 10
89 19 24 13 8
92 15 19 23 12
- 500 67* 69 11* 11 21 25 10 13 4 6
81* 10* 36 16 4
58* 13* 19 14 9
- 1500 59* 62 0* 0 15* 15 12* 9 0* 0
65* 0* 14* 7* 0*
62* 0* 15* 7* 0*
- 5000 0* 0 0* 0 18* 15 0* 0 0* 0
0* 0* 15* 0* 0*
0* 0* 11* 0* 0*
                       
+ 0 90 79 14 14 29 26 16 17 11 10
64 13 26 18 8
84 15 24 16 11
+ 5 82 88 5 7 N/T N/T  13 24 9 8
97 4  N/T 27 10
84 13  N/T 31 5
+ 15 81 84 7 7 18 21 21 22 5 5
95 5 22 22 9
75 10 24 24 1
+ 50 86 90 11 8 25 29 29 22 7 7
98 2 30 26 5
87 11 32 10 9
+ 150 87 75 15 13 18 24 19 24 12 10
64 14 27 21 11
73 10 26 31 8
+ 500 77 66 4 7 31 31 20 22 3 8
60 5 33 22 8
60 11 30 23 12
+ 1500 70* 74 5* 5 21 20 16* 18 5* 4
77* 0* 23 24* 3*
76* 9* 16 15* 4*
+ 5000 0* 0 0* 0 22* 19 0* 0 0* 0
0* 0* 27* 0* 0*
0* 0* 7* 0* 0*
                       
Positive controls ENNG ENNG ENNG 4NQO 9AA
Concentration (ug/plate) 3 5 2 0.2 80
without S9 306 320 136 221 293 300 141 160 2435 1859
317 257 343 159 1364
338 269 265 181 1778
Positive controls 2AA 2AA 2AA BP 2AA
Concentration (ug/plate) 1 2 10 5 2
With S9 1767 1794 276 252 582 647 431 356 472 473
2105 278 697 307 492
1510 201 662 329 456

Table 2. Main study

Main test with and without S9
  Number of revertants (mean number of colonies per plate)
S9 Mix Test substance concentration (ug/plate) TA100 Mean TA1535 Mean WP2uvrA- Mean TA98 Mean TA1537 Mean
- 0 112 109 16 21 15 14 21 26 6 7
102 32 17 38 2
11 16 11 18 13
- 5 90 84 10 14 N/T 30 27 2 5
73 18 33 6
88 15 17 8
- 15 111 107 23 17 21 19 20 19 8 8
99 14 15 19 3
110 15 21 18 14
- 50 91 90 13 24 16 19 23 20 7 9
81 27 21 15 8
87 32 19 21 13
- 150 82 93 17 18 20 19 7 9 14 7
93 18 22 15 3
105 19 15 6 4
- 500 72* 68 0* 0 22 16 0* 0 1* 3
68* 0* 9 0* 3*
64* 0* 16 0* 4*
- 1500 0* 0 0* 0 16* 16 0* 0 0* 0
0* 0* 17* 0* 0*
0* 0* 14* 0* 0*
- 5000 N/T N/T 0* 0 N/T N/T
0*
0*
                       
+ 0 76 75 8 9 28 26 28 26 10 19
77 8 23 23 6
71 11 26 26 8
+ 5 70 78 10 9 24 29 24 29 4 18
70 8 31 31 4
95 10 31 31 7
+ 15 78 71 9 9 19 23 19 23 10 14
66 8 25 25 8
68 9 25 25 7
+ 50 74 74 8 9 38 34 38 34 10 13
70 8 24 24 11
79 10 40 40 13
+ 150 65 60 9 7 28 22 28 22 5 18
54 6 18 18 4
61 6 19 19 7
+ 500 58* 55 9* 10 16* 16 16* 16 5* 16
57* 13* 18* 18* 9*
50* 7* 14* 14* 8*
+ 1500 0* 0 0* 0 0* 0 0* 0 0* 0
0* 0* 0* 0* 0*
0* 0* 0* 0* 0*
+ 5000 N/T N/T 0* 0 N/T N/T
0*
0*
                       
Positive controls ENNG ENNG ENNG 4NQO 9AA
Concentration (ug/plate) 3 5 2 0.2 80
without S9 431 400 338 348 650 663 395 396 1489 1276
333 334 709 419 1183
437 373 629 373 1157
Positive controls 2AA 2AA 2AA BP 2AA
Concentration (ug/plate) 1 2 10 5 2
With S9 867 747 277 261 537 503 398 333 251 377
505 236 468 283 530
870 269 503 319 351

BP: Benzo( a )pyrene

2AA: 2-Aminoanthracene

ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine

4NQO: 4-Nitroquinoline-l-oxide

9AA: 9-Aminoacridine

N/T: Not tested at this dose level

*Partial absence ofbacterial background lawn

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the conditions of this study, the material is considered to be non mutagenic.
Executive summary:

The study was performed to the requirements of OECD Guideline 471 and EU Method B13/14 under GLP conditions, to evaluate the potential mutagenicity of the test substance in a bacterial reverse mutation assay using S.typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli strain WP2 uvrA in both the presence and absence of S-9 mix. A preliminary test was performed to determine the test concentrations for main study. The test material caused a visible reduction in the growth of the bacterial background lawn to all of the tester strains, initially at and above 500 ug/plate. The test material was, therefore, tested up to the maximum recommended dose level of 5000 ug/plate or its toxic limit depending on strain type. In the main study, the plate incorporation method was used and the test substance was evaluated at a concentration of up to 5000 µg/plate. Positive controls appropriate for each strain, in the presence and absence of S9 -mix, were included. The test substance did not induce any significant, reproducible increases in the observed number of revertant colonies in any of the strains tested, either in the presence or absence of S9-mix. The sensitivity of the test system and the metabolic activity of the S9-mix were demonstrated by the increases in the numbers of revertant colonies induced by the positive control substances. It was concluded that, under the conditions of this assay, the test substance gave a negative, i.e. non-mutagenic response in S.typhimurium strains TA98, TA100, TA1535, TA1537 and E.coli strain WP2 uvrA in the presence and absence of S-9 mix