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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:
2005-10-18 to 2005-11-17
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report date:
2005

Materials and methods

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

Test material

Constituent 1
Chemical structure
Reference substance name:
3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine
EC Number:
219-588-7
EC Name:
3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine
Cas Number:
2469-55-8
Molecular formula:
C10H28N2OSi2
IUPAC Name:
3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine
Constituent 2
Reference substance name:
3,3’-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine
IUPAC Name:
3,3’-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine
Test material form:
other: liquid

Method

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:
Phenobarbital and Beta-Naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
See table 1
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO

- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 100, TA 1535 (without activation)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylene-diamine
Remarks:
TA 98, TA 1537 (without activation)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
TA 102 (without activation)
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
TA 1537, TA 102 (with activation)
Details on test system and experimental conditions:
ACTIVATION: The S9 supernatant was mixed S9 cofactor solution to result in a final protein concentration of 36 mg/mL in the cell cultures. The cofactors were: 8 mM MgCl2, 33 mM KCl, 5 mM Glucose-6-phosphate, 5 mM NADP in 100 mM sodium-ortho-phosphat-buffer pH 7.4.
METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION

- Preincubation period: 60 minutes

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

NUMBER OF REPLICATIONS: 3 plates for each test concentration

DETERMINATION OF CYTOTOXICITY
- Method: Background lawn assessment, revertant colony counts
Evaluation criteria:
Mean values of the spontanious reversion frequency are within historical control data range.

Positive controls should show a distinct enhancement of revertant rates over the control plate.

A test system is considered as mutagenic if there is a clear and dose-related increase in the number of revertants and / or a biologically relevant positive response for at least one of the dose groups in at least one tester strain with or without metabolic activation.

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 100 and TA 102 and 3-fold of the solvent control for TA 98, TA 1535 and TA 1537.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
500 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
300 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
800 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
300 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
1000 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
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 -2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
100 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
800 - 2500 ug/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
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: No mutagenic potential

Any other information on results incl. tables

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

TA 98

TA 100

Concentration (μg/Plate)

Plate 1

- MA

Plate 2

+ MA

Cytotoxic (Yes/No)

Plate 1

- MA

Plate 2

+ MA

Cytotoxic (Yes/No)

0*

1

1

No

1

1

No

3.16

1

0.8

No

1

1

No

10

0.9

0.9

No

1

1.3

No

31.6

0.8

1.2

No

1.1

1.3

No

100

1.1

1.2

No

1

1.2

No

316

1.1

1.2

No

1

1

Yes

1000

0.3

0.8

Yes

0

1.8

Yes

2500

0

0

Yes

0

0

Yes

5000

0

0

Yes

0

0

Yes

Positive Control

26.9

19.1

No

9.9

13

No

*solvent control with DMSO

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

 

TA 98

TA 100

TA 102

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

MA

+ MA

Cytotoxic
(yes/no)

MA

+ MA

Cytotoxic
(yes/no)

Negative control

31

45

No

116

104

No

210

235

No

0*

26

40

No

101

95

No

167

180

No

3.16

26

32

No

104

99

No

165

225

No

10

23

35

No

102

118

No

171

225

No

31.6

21

47

No

107

125

No

146

226

No

100

27

50

No

100

110

No

88

191

Yes

316

27

50

No

98

99

Yes

24

142

Yes

1000

7

33

Yes

2

172

Yes

0

48

Yes

2500

0

0

Yes

0

0

Yes

0

0

Yes

Positive control

690

762

No

1001

1234

No

1934

564

No

*solvent control with DMSO

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

 

TA 1535

TA 1537

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

— MA

+ MA

Cytotoxic
(yes/no)

Negative control

13

8

No

12

12

No

0*

7

11

No

15

9

No

3.16

12

11

No

11

17

No

10

13

12

No

14

15

No

31.6

13

13

No

15

17

No

100

9

9

No

16

19

No

316

5

12

Yes

10

27

Yes

1000

0

3

Yes

0

4

Yes

2500

0

0

Yes

0

0

Yes

Positive control

660

87

No

233

56

No

*solvent control with DMSO

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

 

TA 98

TA 100

TA 1535

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

MA

+ MA

Cytotoxic
(yes/no)

MA

+ MA

Cytotoxic
(yes/no)

Negative control

30

37

No

107

127

No

11

9

No

0*

25

40

No

111

113

No

6

9

No

50

17

-

No

109

-

No

9

-

No

100

31

34

No

98

99

No

13

9

No

200

32

49

No

96

97

No

7

10

No

300

25

48

No

118

102

Yes

7

10

Yes

500

22

40

Yes

75

88

Yes

2

10

Yes

800

23

41

Yes

18

86

Yes

0

11

Yes

1200

8

25

Yes

0

134

Yes

0

6

Yes

1600

0

19

Yes

0

192

Yes

1

0

Yes

2000

-

16

Yes

-

46

Yes

-

0

Yes

Positive control

1000

2114

No

1010

2545

No

720

125

No

*solvent control with DMSO

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

 

TA 1537

 Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

Negative control

13

11

No

0*

14

8

No

50

9

-

No

100

14

14

No

200

11

17

No

300

9

21

No

500

8

16

Yes

800

9

8

Yes

1200

0

13

Yes

1600

0

2

Yes

2000

-

0

Yes

Positive control

202

214

No

*solvent control with DMSO

 

TA 102

Conc.
(
µg/plate)

— MA

+ MA

Cytotoxic
(yes/no)

Negative control

220

308

No

0*

206

238

No

25

168

-

No

50

151

-

No

100

129

217

No

200

79

222

Yes

300

39

207

Yes

500

0

123

Yes

800

0

81

Yes

1200

-

52

Yes

Positive control

1891

871

No

*solvent control with DMSO

Applicant's summary and conclusion

Conclusions:
In a highly reliable test, conducted under OECD 471, with GLP, no mutagenic effect was observed for the test substance tested up to cytotoxic concentration in any of the test strains in two independent experiments without and with metabolic activation. The test substance is non-mutagenic in test strains used.