Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 May - 05 June 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report Date:
2019

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted 21 July 1997
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
30 May 2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed

Method

Target gene:
his operon (for S. typhimurium strains) and trp operon (for E. coli strain)
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats treated with phenobarbital/beta-naphthoflavone
Test concentrations with justification for top dose:
First experiment: 3, 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate with and without metabolic activation (tested up to the recommended maximum concentration)
Second experiment: 33, 100, 333, 1000, 2500 and 5000 µg/plate with and without metabolic activation (tested up to the recommended maximum concentration)
Vehicle / solvent:
- Vehicle used: deionized water
- Justification for choice of vehicle: The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-o-phenylene-diamine, (4-NOPD), 2-Aminoanthracene (2-AA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar plate incorporation (first experiment); preincubation (second experiment)

DURATION
- Preincubation period: 1 h
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: bacterial growth inhibition, reduction in the number of spontaneous revertants (below a factor of 0.5) or a clearing of the bacterial background lawn.


Rationale for test conditions:
In the pre-experiment the concentration range of the test item was 3 – 5000 µg/plate. The pre-experiment is reported as experiment I. Since no relevant toxic effects were observed
Evaluation criteria:
- A test item is considered as a mutagen if a biologically relevant increase in the number of revertants of twofold or above (strains TA 98, TA 100, and WP2 uvrA) or threefold or above (strains TA 1535 and TA 1537) the spontaneous mutation rate of the corresponding solvent control is observed.
- A dose dependent increase is considered biologically relevant if the threshold is reached or exceeded at one or more concentrations.
- An increase of revertant colonies equal or above the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
- A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant
Statistics:
Mean numbers of the revertant colonies of each concentration of each test system was calculated and compared to that of the solvent control.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
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
True negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
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
True negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
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
True negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium 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
True negative controls validity:
valid
Positive controls validity:
valid
Key result
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
True negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
No relevant effect on bacterial growth was observed in any strain at any test concentration.

Any other information on results incl. tables

Table 1: Summary Results Experiment I (plate incorporation)

 

 

Test

Group

Dose Level

(per plate)

Revertant Colony Counts (Mean ±SD)

 

 

 

 

 

 

 

 

 

 

 

TA 1535

TA 1537

TA 98

TA 100

WP2 uvrA

 

 

 

 

 

 

 

 

Without Activation

Deionised water

 

16 ± 4

10 ± 3

28 ± 2

106 ± 9

50 ± 3

Untreated

 

11 ± 4

11 ± 1

33 ± 6

100 ± 8

55 ± 7

Test item

3 µg

13 ± 4

12 ± 2

29 ± 12

94 ± 13

47 ± 18

 

10 µg

16 ± 2

11 ± 3

34 ± 10

104 ± 19

49 ± 10

 

33 µg

16 ± 4

10 ± 1

34 ± 8

93 ± 16

52 ± 3

 

100 µg

15 ± 5

10 ± 3

30 ± 3

99 ± 8

52 ± 15

 

333 µg

12 ± 2

6 ± 2

34 ± 5

92 ± 11

58 ± 10

 

1000 µg

11 ± 1

12 ± 4

32 ± 3

98 ± 12

53 ± 10

 

2500 µg

10 ± 1

11 ± 2

34 ± 11

93 ± 14

49 ± 9

 

5000 µg

16 ± 3

11 ± 1

37 ± 10

88 ± 3

57 ± 17

NaN3

10 µg

1119 ± 35

 

 

1542 ± 75

 

4-NOPD

10 µg

 

 

405 ± 30

 

 

4-NOPD

50 µg

 

73 ± 10

 

 

 

MMS

2.0 µL

 

 

 

 

963 ± 28

 

 

 

 

 

 

 

 

With Activation

Deionised water

 

11 ± 5

16 ± 4

35 ± 6

114 ± 12

55 ± 10

Untreated

 

16 ± 4

16 ± 1

40 ± 12

116 ± 14

62 ± 1

Test item

3 µg

9 ± 5

15 ± 4

45 ± 6

98 ± 13

51 ± 6

 

10 µg

11 ± 3

17 ± 7

36 ± 8

118 ± 16

54 ± 6

 

33 µg

11 ± 4

16 ± 4

40 ± 12

115 ± 10

56 ± 11

 

100 µg

10 ± 5

16 ± 1

40 ± 6

110 ± 6

56 ± 16

 

333 µg

13 ± 4

15 ± 2

43 ± 9

94 ± 12

58 ± 8

 

1000 µg

9 ± 3

20 ± 6

38 ± 3

96 ± 14

59 ± 7

 

2500 µg

10 ± 1

14 ± 6

41 ± 15

105 ± 26

61 ± 4

 

5000 µg

10 ± 4

18 ± 4

40 ± 7

85 ± 5

53 ± 5

2-AA

2.5 µg

325 ± 24

417 ± 10

3168 ± 188

2727 ± 207

 

2-AA

10.0 µg

 

 

 

 

306 ± 12

 

 

 

 

 

 

 

 

NaN3

2-AA

4-NOPD

MMS

sodium azide

2-aminoanthracene

4-nitro-o-phenylene-diamine

methyl methane sulfonate

 

Table 2: Summary Results Experiment II (pre incubation)

Metabolic

Activation

Test

Group

Dose Level

(per plate)

Revertant Colony Counts (Mean ±SD)

 

 

 

 

 

 

 

 

 

 

 

TA 1535

TA 1537

TA 98

TA 100

WP2 uvrA

 

 

 

 

 

 

 

 

Without Activation

Deionionised water

 

17 ± 6

15 ± 6

30 ± 4

102 ± 16

52 ± 9

Untreated

 

15 ± 3

18 ± 1

36 ± 3

113 ± 16

55 ± 4

Test item

33 µg

13 ± 3

14 ± 4

34 ± 9

122 ± 14

60 ± 7

 

100 µg

15 ± 4

16 ± 7

41 ± 2

108 ± 11

56 ± 10

 

333 µg

15 ± 0

15 ± 6

32 ± 8

106 ± 17

64 ± 5

 

1000 µg

14 ± 6

16 ± 3

31 ± 10

113 ± 9

61 ± 4

 

2500 µg

13 ± 4

17 ± 3

33 ± 2

116 ± 5

58 ± 9

 

5000 µg

16 ± 6

13 ± 3

33 ± 9

103 ± 11

56 ± 7

NaN3

10 µg

1290 ± 5

 

 

1603 ± 111

 

4-NOPD

10 µg

 

 

447 ± 10

 

 

4-NOPD

50 µg

 

86 ± 4

 

 

 

MMS

2.0 µL

 

 

 

 

760 ± 31

 

 

 

 

 

 

 

 

With Activation

Deionionised water

 

16 ± 4

17 ± 6

44 ± 8

110 ± 9

61 ± 4

Untreated

 

10 ± 2

15 ± 2

46 ± 6

112 ± 3

63 ± 9

Test item

33 µg

13 ± 1

16 ± 4

41 ± 14

108 ± 2

64 ± 17

 

100 µg

15 ± 7

16 ± 5

47 ± 6

103 ± 6

68 ± 9

 

333 µg

15 ± 6

15 ± 4

44 ± 5

110 ± 9

60 ± 3

 

1000 µg

12 ± 3

19 ± 4

48 ± 4

102 ± 1

61 ± 1

 

2500 µg

14 ± 2

16 ± 6

45 ± 3

95 ± 11

54 ± 6

 

5000 µg

13 ± 4

16 ± 4

47 ± 8

110 ± 3

59 ± 7

2-AA

2.5 µg

308 ± 13

265 ± 6

2865 ± 116

3148 ± 186

 

2-AA

10.0 µg

 

 

 

 

366 ± 1

 

 

 

 

 

 

 

 

NaN3

2-AA

4-NOPD

MMS

sodium azide

2-aminoanthracene

4-nitro-o-phenylene-diamine

methyl methane sulfonate

 

Table 3: Individual Results of Experiment I (Without metabolic activation)

Strain

Compound

Dose level per plate

Mean revertants per plate

Standard Deviation

Ratio treated / solvent

Individual revertant

colony counts

TA 1535

Test item

3 µg

13.3

3.8

0.8

15, 16, 9

 

 

10 µg

15.7

1.5

1.0

16, 14, 17

 

 

33 µg

15.7

4.2

1.0

19, 17, 11

 

 

100 µg

14.7

5.1

0.9

9, 19, 16

 

 

333 µg

12.0

2.0

0.7

12, 14, 10

 

 

1000 µg

10.7

0.6

0.7

11, 11, 10

 

 

2500 µg

10.3

0.6

0.6

10, 11, 10

 

 

5000 µg

16.3

3.2

1.0

15, 20, 14

 

Deionised water

 

16.3

4.0

 

14, 21, 14

 

Untreated Control

 

11.0

4.0

 

7, 15, 11

 

 

 

 

 

 

 

TA 1537

Test item

3 µg

12.3

1.5

1.3

14, 12, 11

 

 

10 µg

11.0

2.6

1.1

10, 9, 14

 

 

33 µg

9.7

1.2

1.0

9, 11, 9

 

 

100 µg

9.7

2.5

1.0

12, 7, 10

 

 

333 µg

6.3

2.3

0.7

5, 9, 5

 

 

1000 µg

12.0

4.4

1.2

7, 14, 15

 

 

2500 µg

11.3

2.3

1.2

10, 10, 14

 

 

5000 µg

10.7

0.6

1.1

11, 11, 10

 

Deionised water

 

9.7

2.5

 

7, 12, 10

 

Untreated Control

 

10.7

1.2

 

10, 12, 10

 

 

 

 

 

 

 

TA 98

Test item

3 µg

29.0

12.1

1.0

22, 22, 43

 

 

10 µg

34.0

10.1

1.2

32, 25, 45

 

 

33 µg

34.3

8.3

1.2

37, 25, 41

 

 

100 µg

30.3

3.1

1.1

31, 27, 33

 

 

333 µg

34.3

5.1

1.2

30, 33, 40

 

 

1000 µg

31.7

2.9

1.1

30, 35, 30

 

 

2500 µg

34.3

10.7

1.2

32, 46, 25

 

 

5000 µg

37.3

10.0

1.3

26, 41, 45

 

Deionised water

 

28.0

2.0

 

26, 30, 28

 

Untreated Control

 

33.0

6.1

 

36, 26, 37

 

 

 

 

 

 

 

TA 100

Test item

3 µg

93.7

12.9

0.9

99, 79, 103

 

 

10 µg

104.0

19.2

1.0

113, 117, 82

 

 

33 µg

92.7

15.6

0.9

91, 109, 78

 

 

100 µg

99.3

7.6

0.9

96, 108, 94

 

 

333 µg

91.7

10.6

0.9

82, 103, 90

 

 

1000 µg

98.0

12.2

0.9

106, 84, 104

 

 

2500 µg

92.7

14.2

0.9

108, 80, 90

 

 

5000 µg

87.7

2.5

0.8

88, 85, 90

 

Deionised water

 

105.7

9.0

 

101, 116, 100

 

Untreated Control

 

100.3

7.8

 

94, 98, 109

 

 

 

 

 

 

 

WP2

Test item

3 µg

47.0

18.2

0.9

68, 37, 36

uvrA

 

10 µg

49.3

10.0

1.0

38, 53, 57

 

 

33 µg

52.0

2.6

1.0

49, 53, 54

 

 

100 µg

52.0

15.1

1.0

69, 47, 40

 

 

333 µg

57.7

10.1

1.2

47, 59, 67

 

 

1000 µg

53.3

10.0

1.1

57, 42, 61

 

 

2500 µg

49.0

8.5

1.0

41, 58, 48

 

 

5000 µg

56.7

16.8

1.1

53, 75, 42

 

Deionised water

 

50.0

2.6

 

47, 52, 51

 

Untreated Control

 

54.7

7.0

 

48, 54, 62

 

 

 

 

 

 

 

TA 1535

NaN3

10 µg

1119.3

35.2

68.5

1152, 1124, 1082

TA 1537

4-NOPD

50 µg

73.0

10.1

7.6

82, 62, 75

TA 98

4-NOPD

10 µg

405.3

29.8

14.5

425, 420, 371

TA 100

NaN3

10 µg

1542.0

75.4

14.6

1629, 1500, 1497

WP2 uvrA

MMS

2.0 µL

962.7

28.2

19.3

985, 972, 931

NaN3

4-NOPD

MMS

sodium azide

4-nitro-o-phenylene-diamine

methyl methane sulfonate

 

Table 4: Individual Results of Experiment I (With metabolic activation)

Strain

Compound

Dose level per plate

Mean revertants per plate

Standard Deviation

Ratio treated / solvent

Individual revertant

colony counts

TA 1535

Test item

3 µg

9.3

4.9

0.8

15, 6, 7

 

 

10 µg

11.3

2.5

1.0

11, 14, 9

 

 

33 µg

11.0

3.6

1.0

7, 14, 12

 

 

100 µg

9.7

4.6

0.9

7, 15, 7

 

 

333 µg

13.0

3.6

1.1

17, 10, 12

 

 

1000 µg

9.0

3.0

0.8

12, 6, 9

 

 

2500 µg

9.7

0.6

0.9

9, 10, 10

 

 

5000 µg

10.0

4.0

0.9

10, 14, 6

 

Deionised water

 

11.3

5.1

 

17, 10, 7

 

Untreated Control

 

15.7

4.2

 

17, 11, 19

 

 

 

 

 

 

 

TA 1537

Test item

3 µg

15.3

4.0

1.0

19, 16, 11

 

 

10 µg

17.0

7.0

1.1

9, 22, 20

 

 

33 µg

16.3

4.0

1.0

14, 21, 14

 

 

100 µg

16.0

1.0

1.0

17, 16, 15

 

 

333 µg

15.3

1.5

1.0

15, 14, 17

 

 

1000 µg

19.7

5.5

1.3

14, 25, 20

 

 

2500 µg

13.7

6.4

0.9

10, 10, 21

 

 

5000 µg

18.0

3.6

1.1

14, 19, 21

 

Deionised water

 

15.7

4.0

 

12, 20, 15

 

Untreated Control

 

15.7

1.2

 

17, 15, 15

 

 

 

 

 

 

 

TA 98

Test item

3 µg

45.0

6.1

1.3

41, 42, 52

 

 

10 µg

36.3

7.6

1.0

43, 38, 28

 

 

33 µg

39.7

12.4

1.1

32, 33, 54

 

 

100 µg

40.3

6.4

1.1

43, 45, 33

 

 

333 µg

42.7

9.3

1.2

47, 49, 32

 

 

1000 µg

37.7

2.5

1.1

38, 35, 40

 

 

2500 µg

41.0

14.7

1.2

58, 32, 33

 

 

5000 µg

40.0

7.0

1.1

47, 40, 33

 

Deionised water

 

35.3

5.9

 

31, 33, 42

 

Untreated Control

 

40.0

11.5

 

51, 28, 41

 

 

 

 

 

 

 

TA 100

Test item

3 µg

97.7

13.1

0.9

84, 99, 110

 

 

10 µg

117.7

15.6

1.0

134, 103, 116

 

 

33 µg

115.0

9.5

1.0

125, 106, 114

 

 

100 µg

109.7

5.5

1.0

110, 115, 104

 

 

333 µg

94.3

12.1

0.8

90, 85, 108

 

 

1000 µg

96.0

14.4

0.8

100, 108, 80

 

 

2500 µg

105.0

26.0

0.9

75, 121, 119

 

 

5000 µg

85.3

4.9

0.7

82, 91, 83

 

Deionised water

 

114.3

11.6

 

122, 120, 101

 

Untreated Control

 

116.0

13.7

 

113, 104, 131

 

 

 

 

 

 

 

WP2

Test item

3 µg

50.7

5.5

0.9

56, 51, 45

uvrA

 

10 µg

53.7

5.9

1.0

56, 47, 58

 

 

33 µg

56.0

11.4

1.0

64, 61, 43

 

 

100 µg

55.7

15.9

1.0

74, 46, 47

 

 

333 µg

58.0

7.5

1.1

51, 66, 57

 

 

1000 µg

58.7

6.7

1.1

53, 66, 57

 

 

2500 µg

60.7

3.5

1.1

64, 57, 61

 

 

5000 µg

53.0

5.0

1.0

48, 58, 53

 

Deionised water

 

54.7

9.8

 

49, 66, 49

 

Untreated Control

 

62.3

0.6

 

63, 62, 62

 

 

 

 

 

 

 

TA 1535

2-AA

2.5 µg

325.0

23.6

28.7

345, 299, 331

TA 1537

2-AA

2.5 µg

416.7

9.7

26.6

406, 419, 425

TA 98

2-AA

2.5 µg

3168.3

188.3

89.7

3143, 2994, 3368

TA 100

2-AA

2.5 µg

2726.7

207.1

23.8

2910, 2768, 2502

WP2 uvrA

2-AA

10.0 µg

306.3

11.6

5.6

312, 293, 314

2-AA

2-aminoanthracene

 

 

Table 5: Individual Results of Experiment II (Without metabolic activation)

Strain

Compound

Dose level per plate

Mean revertants per plate

Standard Deviation

Ratio treated / solvent

Individual revertant

colony counts

TA 1535

Test item

33 µg

12.7

3.2

0.7

14, 9, 15

 

 

100 µg

15.0

4.0

0.9

11, 15, 19

 

 

333 µg

15.0

0.0

0.9

15, 15, 15

 

 

1000 µg

13.7

5.5

0.8

10, 20, 11

 

 

2500 µg

13.0

3.6

0.8

12, 17, 10

 

 

5000 µg

15.7

6.0

0.9

15, 22, 10

 

Deionised water

 

17.0

6.2

 

22, 19, 10

 

Untreated Control

 

14.7

2.5

 

12, 15, 17

 

 

 

 

 

 

 

TA 1537

Test item

33 µg

14.0

3.6

1.0

10, 15, 17

 

 

100 µg

16.0

6.6

1.1

9, 22, 17

 

 

333 µg

14.7

5.5

1.0

9, 15, 20

 

 

1000 µg

16.3

2.5

1.1

19, 16, 14

 

 

2500 µg

17.3

2.9

1.2

14, 19, 19

 

 

5000 µg

12.7

3.2

0.9

9, 15, 14

 

Deionised water

 

14.7

6.0

 

14, 9, 21

 

Untreated Control

 

17.7

1.2

 

17, 19, 17

 

 

 

 

 

 

 

TA 98

Test item

33 µg

34.0

8.5

1.1

25, 42, 35

 

 

100 µg

41.3

1.5

1.4

40, 43, 41

 

 

333 µg

31.7

8.1

1.0

41, 26, 28

 

 

1000 µg

31.3

9.8

1.0

20, 37, 37

 

 

2500 µg

33.3

1.5

1.1

33, 32, 35

 

 

5000 µg

33.3

9.3

1.1

36, 41, 23

 

Deionised water

 

30.3

4.0

 

28, 35, 28

 

Untreated Control

 

35.7

2.5

 

36, 33, 38

 

 

 

 

 

 

 

TA 100

Test item

33 µg

122.0

13.9

1.2

115, 138, 113

 

 

100 µg

107.7

11.0

1.1

114, 114, 95

 

 

333 µg

105.7

16.7

1.0

119, 87, 111

 

 

1000 µg

113.3

8.6

1.1

104, 121, 115

 

 

2500 µg

116.3

5.0

1.1

121, 111, 117

 

 

5000 µg

103.3

11.2

1.0

113, 106, 91

 

Deionised water

 

102.0

16.1

 

85, 104, 117

 

Untreated Control

 

113.3

16.2

 

132, 104, 104

 

 

 

 

 

 

 

WP2

Test item

33 µg

60.3

7.0

1.2

53, 67, 61

uvrA

 

100 µg

56.0

10.0

1.1

66, 46, 56

 

 

333 µg

64.0

5.0

1.2

59, 64, 69

 

 

1000 µg

60.7

4.2

1.2

64, 56, 62

 

 

2500 µg

57.7

9.3

1.1

62, 64, 47

 

 

5000 µg

55.7

6.5

1.1

62, 56, 49

 

Deionised water

 

51.7

9.1

 

45, 48, 62

 

Untreated Control

 

54.7

4.0

 

59, 54, 51

 

 

 

 

 

 

 

TA 1535

NaN3

10 µg

1290.0

5.2

75.9

1287, 1296, 1287

TA 1537

4-NOPD

50 µg

86.0

3.6

5.9

87, 89, 82

TA 98

4-NOPD

10 µg

447.3

10.2

14.7

443, 440, 459

TA 100

NaN3

10 µg

1603.0

110.7

15.7

1710, 1489, 1610

WP2 uvrA

MMS

2.0 µL

760.3

30.7

14.7

785, 726, 770

NaN3

4-NOPD

MMS

sodium azide

4-nitro-o-phenylene-diamine

methyl methane sulfonate

 

Table 6: Individual Results of Experiment II (With metabolic activation)

Strain

Compound

Dose level per plate

Mean revertants per plate

Standard Deviation

Ratio treated / solvent

Individual revertant

colony counts

TA 1535

Test item

33 µg

12.7

1.2

0.8

12, 14, 12

 

 

100 µg

15.3

6.5

1.0

9, 15, 22

 

 

333 µg

15.3

5.5

1.0

10, 15, 21

 

 

1000 µg

11.7

3.1

0.7

9, 15, 11

 

 

2500 µg

14.0

2.0

0.9

16, 12, 14

 

 

5000 µg

13.0

3.6

0.8

17, 10, 12

 

Deionised water

 

16.0

3.6

 

19, 12, 17

 

Untreated Control

 

10.3

1.5

 

9, 12, 10

 

 

 

 

 

 

 

TA 1537

Test item

33 µg

15.7

4.0

0.9

20, 12, 15

 

 

100 µg

16.3

4.5

1.0

16, 21, 12

 

 

333 µg

14.7

4.0

0.9

11, 19, 14

 

 

1000 µg

19.0

3.6

1.1

20, 15, 22

 

 

2500 µg

15.7

6.0

0.9

22, 10, 15

 

 

5000 µg

15.7

4.2

0.9

11, 19, 17

 

Deionised water

 

17.0

6.0

 

17, 23, 11

 

Untreated Control

 

15.3

1.5

 

15, 17, 14

 

 

 

 

 

 

 

TA 98

Test item

33 µg

41.3

13.6

0.9

43, 27, 54

 

 

100 µg

47.3

6.0

1.1

48, 53, 41

 

 

333 µg

43.7

4.9

1.0

47, 38, 46

 

 

1000 µg

48.3

4.2

1.1

45, 47, 53

 

 

2500 µg

45.3

2.5

1.0

48, 45, 43

 

 

5000 µg

46.7

7.8

1.1

38, 53, 49

 

Deionised water

 

43.7

8.1

 

51, 45, 35

 

Untreated Control

 

46.0

6.1

 

42, 53, 43

 

 

 

 

 

 

 

TA 100

Test item

33 µg

108.3

2.1

1.0

110, 106, 109

 

 

100 µg

103.3

5.8

0.9

100, 100, 110

 

 

333 µg

109.7

9.1

1.0

120, 103, 106

 

 

1000 µg

101.7

1.2

0.9

103, 101, 101

 

 

2500 µg

95.3

11.0

0.9

104, 99, 83

 

 

5000 µg

110.0

2.6

1.0

113, 108, 109

 

Deionised water

 

110.0

8.9

 

113, 100, 117

 

Untreated Control

 

112.0

2.6

 

110, 111, 115

 

 

 

 

 

 

 

WP2

Test item

33 µg

64.3

16.6

1.1

66, 47, 80

uvrA

 

100 µg

68.0

8.7

1.1

78, 63, 63

 

 

333 µg

59.7

3.2

1.0

62, 61, 56

 

 

1000 µg

61.3

0.6

1.0

62, 61, 61

 

 

2500 µg

54.3

6.1

0.9

49, 53, 61

 

 

5000 µg

58.7

6.7

1.0

53, 66, 57

 

Deionised water

 

60.7

3.5

 

61, 64, 57

 

Untreated Control

 

63.3

8.7

 

61, 73, 56

 

 

 

 

 

 

 

TA 1535

2-AA

2.5 µg

307.7

13.4

19.2

323, 302, 298

TA 1537

2-AA

2.5 µg

264.7

5.8

15.6

268, 268, 258

TA 98

2-AA

2.5 µg

2864.7

116.5

65.6

2994, 2768, 2832

TA 100

2-AA

2.5 µg

3147.7

186.5

28.6

3039, 3363, 3041

WP2 uvrA

2-AA

10.0 µg

366.3

1.2

6.0

365, 367, 367

2-AA

2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Under the conditions of the conducted test the substance was not mutagenic in any of the five tester strains (TA 98, TA 100, TA 1535, TA 1537 and WP2 uvrA) tested with and without metabolic activation up to 5000 µg/plate.