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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
publication
Title:
Mutagenicity of 42 Chemicals in Salmonella
Author:
Zeiger, E.
Year:
1990
Bibliographic source:
Environmental and Molecular Mutagenesis 16 (18): 32-54

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
strain with A/T base pair as primary mutation site missing
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): n-butyl chloride

Method

Target gene:
His Operon
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 100, TA 1535, TA 97, TA 98
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
two metabolic activation systems were used separately: 1.) Aroclor induced Sprague Dawley rat liver S9-mix; 2.) Aroclor induced Syrian hamster liver S9-mix; 30 % and 10% S9 in the S9 mixture)
Test concentrations with justification for top dose:
preincubation method: 100, 333, 1000, 3333, 6666 and 10000 μg/plate
vapour desiccator method: 0.1, 0.5, 1, 2, 3 and 4 mL/chamber
Vehicle / solvent:
preincubation method: DMSO
Controls
Untreated negative controls:
yes
Remarks:
vapour desiccator method
Negative solvent / vehicle controls:
yes
Remarks:
preincubation method
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: with metabolic activation: 2-aminoanthracene for all strains; without metabolic activation: sodium azide for TA 100 and TA1535, 9-aminoacridine for TA 97 and 4-nitro-o-phenylenediamine for TA 98.
Details on test system and experimental conditions:
METHOD OF APPLICATION:
Preincubation method: Cells and test compound or DMSO were incubated for 20 minutes at 37 °C in the presence of either S9-mix or buffer. After the addition of soft agar, the contents of each tube was poured onto minimal medium, and the plates were incubated at 37°C for 48 hours [Haworth et al. 1983]
Vapour desiccator method: S9 or buffer was incorporated into the top agar and poured onto the plate. The lids of the plates were removed and the plates were stacked on a perfoated porcelain plate in a 9 L glass desiccator jug containing a magnetic stirring bar. A measured volume of test chemical, in liquid form, was introduced into a watch glass suspended below the porcelain plate, and the desiccator was sealed and placed on a magnetic stirrer in a 37 °C incubator. After 24 h the plates were removed from the desiccator and incubated at 37°C in air for an additional 24 h.


NUMBER OF REPLICATIONS:
vapour desiccator method: 2
preincubation method: 1





Evaluation criteria:
A chemical was judged mutagenic or weakly mutagenic if it gave a reproducible dose-related response over the solvent control in replicate trials, and judged questionable if the results of individual trials were not reproducible or if only single doses produced increase in his+ revertants in repeat trials. Chemicals were judged nonmutagenic, if they do not meet the criteria for mutagenic or questionable response.

Results and discussion

Test results
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
in preincubation and vapour desiccator method
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
from 3333 µg/plate and at 4 mL/chamber
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

Preincubation method

number of revertants: mean value of negative control

number of revertants: mean value of positive control

max. number of revertants: mean value of test material [µg/plate]

without S9-mix

 

 

TA 1535

 14 ± 0.9

 958 ± 34.3

 17 ± 1.5 [100]

TA 100

 119 ± 7.4

 955 ± 37.8

 135 ± 9.9 [100]

TA 97

 206 ± 9.5

 742 ± 42.2

 213 ± 4.6 [100]

TA 98

 36 ± 4.0

 677 ± 20

 36 ± 1.8 [100]

with S9-mix (30% HLI)

 

 

TA 1535

 10 ± 1.9

607 ± 16.8

13 ± 0.9 [1000]

TA 100

 163 ± 12.0

837 ± 39.5

176 ± 5.8 [1000]

TA 97

 212 ± 8.1

436 ± 2.2

230 ± 2.9 [100]

TA 98

 45 ± 3.3

770 ± 11.3

43 ± 5.2 [1000]

with S9-mix (30% RLI)

 

 

TA 1535

 18 ± 3.0

105 ± 3.6

18 ± 0.7 [1000]

TA 100

 175 ± 0.9

440 ± 26.1

170 ± 7.9 [1000]

TA 97

 230 ± 5.8

442 ± 3.5

233 ± 1.7 [333]

TA 98

 42 ± 6.5

168 ± 3.5

49 ± 3.2 [100]

Vapour desiccator method

number of revertants: mean value of negative control

number of revertants: mean value of positive control

number of revertants: mean value of test material [ml/chamber]

without S9-mix

 

 

TA 1535

 14 ± 1.2

1671 ± 41.8

16 ± 2.7 [0.1]

TA 100

 144 ± 8.6

1569 ± 21.7

147 ± 28.3 [0.1]

TA 97

 208 ± 17.2

978 ± 54.7

198 ± 15.3 [0.5]

TA 98

34 ± 2.8

557 ± 17.4

43 ± 3.8 [1.0]

with S9-mix (30% HLI)

 

 

TA 1535

11 ± 4.2

459 ± 21.2

20 ± 0.9 [0.5]

TA 100

165 ± 6.0

843 ± 24.8

158 ± 9.7 [1.0]

TA 97

140 ± 6.8

564 ± 15.9

172 ± 8.5 [1.0]

TA 98

22 ± 3.3

657 ± 47.6

42 ± 4.7[1.0]

with S9-mix (30% RLI)

 

 

TA 1535

 19 ± 3.2

90 ± 6.4

20 ± 2.5 [1.0]

TA 100

 142 ± 9.5

463 ± 58

145 ± 10.5 [0.5]

TA 97

166 ± 20.1

461 ± 10.7

186 ± 9.7 [0.5]

TA 98

 33 ± 4.8

120 ± 7.5

39 ± 4.1 [1.0]

HLI: aroclor 1254-induced hamster liver S9

RLI: aroclor 1254-induced rat liver S9

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative