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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
Study period:
2002-05-03 to 2002-08-01
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report Date:
2002

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid

Method

Target gene:
histidine operon
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:
Aroclor induced rat liver S9
Test concentrations with justification for top dose:
10, 31.6, 100, 316, 1000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Ethylene glycol dimethylether

- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 1535, TA 100 (without activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
TA 98 (without activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA 1537 (without activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
TA 102 (without activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-anthracene amide
Remarks:
TA 98, TA 102, TA 1537 (with activation)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
TA 100, TA 1535 (with activation)
Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium (plate incorporation); preincubation.

DURATION:

- Preincubation period: 60 minutes.

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

NUMBER OF REPLICATIONS: 3 plates for each test concentration; the initial plate incorporation assay was repeated using the pre-incubation method.

DETERMINATION OF CYTOTOXICITY:

- Method: Background lawn assessment, relative colony counts.

METABOLIC ACTIVATION:
Aroclor induced rat liver S9 was tested for protein and P-450 content. S9 mix contained 5% S9, and glucose-6-phosphate and NADP as co-factors. 0.5 ml S9 mix were added to 2 ml top agar, 0.1 ml test material and 0.1 ml cell suspension, giving a final concentration of approximately 1% S9.
Evaluation criteria:
A result is positive if the number of revertants is significantly increased compared with the solvent control to at least 2-fold of the solvent control for TA 98, TA 100 and TA 102 and 3-fold of the solvent control for TA 1535 and TA 1537 in both experiments.

Positive results have to be reproducible and the histidine independence of the revertants has to be confirmed by streaking on histidine-free agar plates.

Cytotoxicity is defined as a reduction in the number of colonies by >50% compared with the solvent control and/or a sparse background lawn.
Statistics:
MANN and WHITNEY and Spearman’s rank.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
316 - 1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
316 - 1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
316 - 1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
316 - 1000 μg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Results were within range of historical control data
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 2: Dose range-finding study Number of revertants per plate (2 plates)

 

TA100

Conc.
(µg/plate)

Plate 1

Plate 2

Cytotoxic
(yes/no)

0*

112

115

No

0.316

117

105

No

1

111

110

No

3.16

126

112

No

10

101

114

No

31.6

116

121

No

100

172

181

No

316

165

173

No

1000

132

154

Yes

3160

0

0

Yes

5000

0

0

Yes

*solvent control with Ethylene glycol dimethylether

Table 3: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

 

TA98

TA100

TA102

Conc.
(
µg/plate)

MA

+

MA

Cytotoxic
(yes/no)

 MA

+

MA

Cytotoxic
(yes/no)

MA

+

 MA

Cytotoxic
(yes/no)

0*

42

51.7

No

133

158

No

278

295

No

10

36.3

48.7

No

154

148

No

274.7

273.7

No

31.6

34.7

53

No

139

153.3

No

268.3

275

No

100

34.3

57

No

137.7

131.3

No

271.3

270

No

316

47.7

47.3

No

143.3

139.3

No

258

272.3

No

1000

31.3

40.3

Yes

131

160.7

Yes

286.7

267.3

Yes

Positive control

1145.7

1100

No

1194.3

1196.3

No

1320

1150.3

No

*solvent control with Ethylene glycol dimethylether

Table 3: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

 

TA1535

TA1537

Conc.µg/plate

— MA

+

 MA

Cytotoxic
(yes/no)

MA

+

 MA

Cytotoxic
(yes/no)

0*

13

15.3

No

6

4

No

10

13

12

No

3.7

4.7

No

31.6

14.7

14

No

3.3

5.7

No

100

14.3

13

No

3

2.7

No

316

11

13

No

3.7

4.7

No

1000

12

16.3

No

4.3

3.3

No

Positive control

1187

1189

No

1191.3

1241.7

No

*solvent control with Ethylene glycol dimethylether

Table 4: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

 

TA98

TA100

TA102

Conc.
µg/plate

MA

+

 MA

Cytotoxic
(yes/no)

MA

+

 MA

Cytotoxic
(yes/no)

 MA

+

 MA

Cytotoxic
(yes/no)

0*

37.3

30.7

No

143.7

154

No

280.3

290.3

No

10

31.7

30.3

No

187

160.7

No

276.3

283.7

No

31.6

38

35.7

No

171.7

144.7

No

288.7

286.3

No

100

35

36.3

No

178

152

No

283.3

281.7

No

316

0

36

Yes

162.7

144.7

No

297.3

265

Yes

1000

0

0

Yes

0

0

Yes

0

0

Yes

Positive control

894.3

983.7

No

1326.3

1333.3

No

1338.3

1344.3

No

*solvent control with Ethylene glycol dimethylether

Table 4: Experiment 2 Preincubation Number of revertants per plate (mean of 3 plates)

 

TA1535

TA1537

Conc.
µg/plate

MA

+

MA

Cytotoxic
(yes/no)

— MA

+

 MA

Cytotoxic
(yes/no)

0*

13.7

13

No

3

4.3

No

10

13.3

13.7

No

3

4.7

No

31.6

14.3

12

No

3

3.3

No

100

12.3

12.3

No

2.3

4.3

No

316

14.3

12

Yes

4

4.3

Yes

1000

0

12.7

Yes

0

0

Yes

Positive control

487.3

499.7

No

502

503

No

*solvent control with Ethylene glycol dimethylether

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without metabolic activation

[2-(perfluorohexyl)ethyl]dichloro(methyl)silane has been tested in a reliable study, conducted according to OECD 471 and in compliance with GLP, no test-substance related increase in the number of revertants was observed with and without metabolic activation when tested up to cytotoxic concentration in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. The result of the initial plate incorporation assay was confirmed in an independent experiment using the pre-incubation method. Appropriate positive and solvent controls were included and gave expected results. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.