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Toxicological information

Genetic toxicity: in vitro

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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:
2000-05-09 to 2000-05-19
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:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1997
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital and Naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
50-5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO

- Justification for choice of solvent/vehicle: It is assumed by the reviewer that the solvent was chosen for its solubility properties and relative non-toxicity to bacteria
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
water
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:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 100 (without activation)
Untreated negative controls:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
TA 102 (without activation)
Untreated negative controls:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 1535 (without activation)
Untreated negative controls:
yes
Remarks:
water
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:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
WP2 uvrA (without activation)
Untreated negative controls:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoflouorene
Remarks:
All strains (with activation)
Untreated negative controls:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
All strains (with activation)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

ACTIVATION:

DURATION

- Preincubation period: 30 minutes

- Expression time (cells in growth medium): 72 hours

NUMBER OF REPLICATIONS: 3 plates for each test concentration

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth; background lawn assessment
Evaluation criteria:
For a test compound or a test substance to be considered positive, it must (in two independent experiments) cause at least a doubling in the mean revertants per plate of at least one tester strain. The increase must be accompanied by a dose response towards increasing concentrations of the test article. Single increases in revertant frequencies, which are not dose-related and not reproducible in two independent tests, are considered non-relevant. If however these increases do occur in both tests, this will be taken as an indication of a mutagenic effect.

A reduction in the number of spontaneous revertants, and / or a thinning of the background lawn is indicative of toxicity.

Results and discussion

Test resultsopen allclose all
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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: precipitation noted at 1600 and 5000 µg/plate

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: Experiment 1 Plate incorporation Number of revertants per plate

 

TA 98

TA 100

TA 102

Conc.
(
µl/ml)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

Negative Control

13

13

No

129

237

No

380

434

No

0*

15

9

No

104

2219

No

329

351

No

50

18

2

No

103

237

No

398

312

No

160

16

7

No

111

233

No

388

308

No

500

16

5

No

130

225

No

350

347

No

1600

22

7

No

135

223

No

345

298

No

5000

13

7

No

138

199

No

349

268

No

Positive control

100

2216

No

587

1637

No

1205

837

No

Positive control 2

-

1606

No

-

1524

No

-

522

No

*solvent control with DMSO

Table 2: Experiment 1 Plate incorporation Number of revertants per plate

 

TA 1535

TA 1537

WP2 uvrA

Conc.
(
µl/ml)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

Negative Control

12

15

No

13

16

No

12

12

No

0*

14

8

No

12

15

No

9

11

No

50

13

7

No

14

17

No

11

15

No

160

11

7

No

9

12

No

9

14

No

500

13

9

No

11

16

No

8

12

No

1600

13

10

No

17

11

No

11

14

No

5000

10

10

No

11

9

No

9

12

No

Positive control

576

31

No

43

69

No

185

40

No

Positive control 2

-

142

No

-

116

No

-

20

No

*solvent control with DMSO

Table 3: Experiment 2 Pre incubation Number of revertants per plate

 

TA 98

TA 100

TA 102

Conc.
(
µl/ml)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

Negative Control

26

30

No

196

197

No

406

365

No

0*

23

33

No

181

182

No

350

361

No

50

31

34

No

182

185

No

391

360

No

160

30

31

No

195

179

No

361

336

No

500

37

31

No

196

174

No

374

359

No

1600

27

35

No

200

175

No

399

334

No

5000

27

28

No

191

200

No

385

277

No

Positive control

137

1966

No

680

1331

No

1390

1173

No

Positive control 2

-

1113

No

-

1064

No

-

471

No

*solvent control with DMSO

Table 3: Experiment 2 Pre incubation Number of revertants per plate

 

TA 1535

TA 1537

WP2 uvrA

Conc.
(
µl/ml)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

MA

+

MA

Cytotoxic
(yes/no)

Negative Control

15

11

No

6

10

No

13

11

No

0*

16

9

No

9

9

No

10

13

No

50

13

10

No

8

7

No

13

17

No

160

16

11

No

9

15

No

12

18

No

500

11

10

No

6

10

No

14

16

No

1600

12

11

No

11

13

No

8

16

No

5000

12

12

No

9

14

No

12

12

No

Positive control

555

24

No

39

78

No

222

52

No

Positive control 2

-

128

No

-

71

No

-

22

No

*solvent control with DMSO

Applicant's summary and conclusion

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

Triethoxy(3-thiocyanatopropyl)silane has been tested in a reliable study conducted in accordance with OECD 471 (1997) and in compliance with GLP using Salmonella typhimurium strains TA 98, 100, 1535, 1537 and 1538, and E.coli WP2uvrA. No evidence of test substance induced increase in the number of revertants was observed up to limit concentrations in any of the bacteria tested without and with metabolic activation in either the initial plate incorporation assay or the repeat experiment using the preincubation method. Appropriate untreated, solvent and positive controls were included and gave expected results. It is concluded that the test substance is non-mutagenic under the conditions of the test.