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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:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2002-04-15 to 2002-06-13
Reliability:
2 (reliable with restrictions)
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 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

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium 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: water

- Justification for choice of solvent/vehicle: none given in report
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Water
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
Remarks:
Water
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
Remarks:
Water
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
Remarks:
Water
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
Remarks:
Water
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
Remarks:
Water
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 agar (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

DETERMINATION OF CYTOTOXICITY

- Method: relative total growth; background lawn assessment
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.
Statistics:
MANN and WHITNEY and Spearman’s rank.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: S. typhimurium TA 1535, TA 1537, TA 98, TA 102 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
TA 1537 without activation only: 5000 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 (TA 100 – MA) Number of revertants per plate (2 plates)

 

TA100

Conc.
(µg/plate)

Plate 1

Plate 2

Cytotoxic
(yes/no)

0*

135

150

No

0.316

123

122

No

1

123

108

No

3.16

109

124

No

10

115

125

No

31.6

148

114

No

100

108

115

No

316

105

121

No

1000

109

105

No

3160

106

150

No

5000

118

125

No

*Solvent control: Water

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*

35.3

32.7

No

133

137.3

No

293.3

280.7

No

100

26.3

39

No

125.3

140.7

No

181.3

325.7

No

316

36.7

32.7

No

136.7

141

No

290.3

317

No

1000

37

43.7

No

139.3

144.3

No

298

319.7

No

3160

30.7

31.3

No

140.3

137.7

No

308.7

315.3

No

5000

37.3

48.7

No

164

143.7

No

317.7

333

No

Positive control

1170.3

1110

No

1139.3

1241.3

No

1094

1066.7

No

*Solvent control: Water

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*

13.3

17.7

No

7.3

8

No

100

13.7

18.3

No

6

6

No

316

16.7

17

No

6.7

5.3

No

1000

13.7

14.7

No

7

5

No

3160

13.7

14

No

7

7.3

No

5000

15

16

No

5

6.7

No

Positive control

736

921.3

No

885

893.7

No

*Solvent control: Water

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*

30.3

24.3

No

124

135

No

285.7

280.3

No

100

29.7

35.7

No

114.7

132

No

310.3

300

No

316

31

38.3

No

119.3

123.3

No

312.3

309

No

1000

28

34.3

No

120.7

136

No

320.3

296.7

No

3160

31.3

35

No

124.7

136

No

301

289

No

5000

27.7

33.3

No

111.3

143

No

245.7

298

No

Positive control

831.1

1110.7

No

1218

1195.3

No

1150.3

1161.7

No

*Solvent control: Water

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*

16

14

No

8

6

No

100

15

11.3

No

4.3

5

No

316

13.3

14.3

No

5

7.3

No

1000

16.3

13

No

5

7

No

3160

17

14

No

6.7

5.7

No

5000

12

16.3

No

4

11.7

Yes

Positive control

884

888.3

No

645.3

627

No

*Solvent control: Water

Applicant's summary and conclusion

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

In a valid and reliable study conducted in accordance with OECD 471 and under GLP, no mutagenic effect was observed for the test substance tested up to cytotoxic concentration in any of the test strains in a plate incorporation study with and without metabolic activation. The result was confirmed in a pre-incubation study. It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the test.