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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:
2002-04-03 to 2002-05-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
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): silan GF 18
- Substance type: monoconstituent substance
- Physical state: liquid
- Analytical purity: 98.4 %
- Purity test date:
- Lot/batch No.: KG 02846
- Expiration date of the lot/batch: 2004-01-01
- Stability under test conditions: stability in solvent: at least a week excluding humidity
- Storage condition of test material: at room temperature, no direct sunlight

Method

Target gene:
histidine
Species / strain
Species / strain / cell type:
other: 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:
100-5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: none given in report
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 100 and TA 1535 without metabolic 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 metabolic 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 metabolic 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 metabolic 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 metabolic 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 metabloic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation;
DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours

NUMBER OF REPLICATIONS: 3 plates per concentration and 2 independent experiments

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: other: condition of bacterial lawn
Evaluation criteria:
A response is considered positive if there is a statistically significant dose-related increase in the number of revertants relative to the solvent control, by at least 2-fold (TA 98, 100 and 102) or 3-fold (TA 1535 or 1537), and that this is reproducible and revertants are confirmed by streaking to histidine-free agar plates.
Statistics:
Mann and Whitney U-test and Spearman's rank correlation coefficient.

Results and discussion

Test results
Species / strain:
other: TA 98, TA 100, TA 102, TA 1535, TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 1537 µg/plate and above
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
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 Plate incorporation test average number of revertants per plate (mean of 3 plates)

 

Concentration (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

-MA

+MA

-MA

+MA

-MA

+MA

-MA

+MA

-MA

+MA

5000

35.3

34.3

179.7

157.3

150.7

151.7

0.0*

0.0*

4.0*

3.7*

3160

31.7

26.3

134.0

137.3

172.3

120

39.7*

0.0*

2.7*

5.7*

1000

40.3

19.0

122.7

128.0

161.7

155.7

22.7

0.0*

5.0

5.3*

316

32.0

27.3

121.3

166.0

172.3

202

28.0

1.3

2.7

5.3

100

46.7

28.3

109.3

159.7

197.3

240.7

22.7

37.7

4.7

4.3

Negative control 100 µL/plate

32.0

36

119.0

153.0

220.7

292.0

16.3

27.7

5.3

3.7

Positive control

817.0

333

447.3

491.7

1231.0

905.7

957.3

841.3

290.3

290.7

 * = scarce background lawn

Table 2 Preincubation test average number of revertants per plate (mean of 3 plates)

Concentration (µg/plate)

TA 98

TA 100

TA 102

TA 1535

TA 1537

-MA

+MA

-MA

+MA

-MA

+MA

-MA

+MA

-MA

+MA

5000

0.0*

0.0*

31.0*

72.7*

1.3*

0.0*

0.0*

0.0*

0.0*

0.0*

3160

10.3*

0.0*

65.3*

80.3*

0.0*

0.0*

0.0*

7.0*

0.0*

2.3*

1000

10.7*

50.0*

93.0*

116.3*

74.3*

0.0*

0.0*

18.7*

0.0*

0.0*

316

12.3

25.7

119.7

135

280

340.7

3.3

20.7

3

13.3

100

19.7

29.7

102.3

132.3

275.3

370.7

11

17.3

3.7

12.3

Negative control 100 µl/plate

32.3

35

119.7

126

246.7

364.7

12

18.7

6.3

14.7

Positive control

303.3

313

464

477

1131

1153.7

891

984.3

286.3

288.7

* = scarce background lawn

Applicant's summary and conclusion

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

(3-chloropropyl)triethoxysilane has been tested in a valid and reliable study according to OECD 471 and under GLP. No increase in the number of revertants was observed in any strain with or without activation in either the initial plate incorporation assay or the subsequent preincubation assay. The solvent and positive controls gave the expected results. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.