Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

This study was performed with a structural analogue substance according to GLP and the methods applied are fully compliant with OECD TG 471.

It was concluded that with and without addition of S9 mix as the external metabolizing system, the test material was not mutagenic under the experimental conditions described.

Link to relevant study records

Referenceopen allclose all

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:
April 18 - 29, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Target gene:
Salmonella: histidine operon
E coli. tryptophane operon
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Details on mammalian cell lines (if applicable):
- Type and identity of media:
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
S9 mix from Aroclor 1254 pretreated rats
Test concentrations with justification for top dose:
1st series: 5, 1.58, 5, 15.8, 50, 158, 500 µg/plate
2nd series: 15.8, 50, 158, 500, 1580 µg/plate
Vehicle:
H2O
Negative controls:
yes
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Sodium azide, 2-Aminoanthracene, 9-Aminoacridine, Daunomycin, 4-Nitroquinoline-N-oxide
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 3-5 hours
- Exposure duration: about 2 days


SELECTION AGENT (mutation assays): Histidine, Tryptophane

NUMBER OF REPLICATIONS:
Negative controls 6
Test material 3
Positive controls 3


NUMBER OF CELLS EVALUATED: about 1e9


DETERMINATION OF CYTOTOXICITY
- Method: other: background cytotox


Evaluation criteria:
- valid assay (cf. laboratory historical data)
- no or weak increase in revertant colonies = negative
- clear, dose dependent (over at least two concentrations), and reproducible increase in revertant colonies= positve
Statistics:
not applied
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no
- Effects of osmolality: no
- Evaporation from medium: no
- Water solubility: ok
- Precipitation: yes, at 1580 µg/plate with and without metabolic activation
- Other confounding effects: no

Summary Table 1stseries

Metabolic

Activation

Test

Material

Concentr.

[µg/plate]

 

Revertants per plate (Mean ± SD)

 

 

 

 

 

 

TA 98

TA 100

TA 1535

TA 1537

WP2 uvrA

 

 

 

 

 

 

 

 

 

Without Activation

H2O

 

 

31 ± 8

101 ± 12

24 ± 5

26 ± 8

38 ± 2

 

 

 

 

 

 

 

 

Test item

0.500

 

25 ± 5

115 ± 6

22 ± 3

27 ± 10

38 ± 3

 

1.58

 

27 ± 4

107 ± 16

22 ± 6

31 ± 3

34 ± 7

 

5.00

 

22 ± 8

113 ± 4

29 ± 8

20 ± 2

38 ± 6

 

15.8

 

22 ± 4

116 ± 16

22 ± 1

28 ± 2

34 ± 11

 

50.0

 

18 ± 1

102 ± 13

21 ± 4

32 ± 3

38 ± 5

 

158

 

22 ± 3

110 ± 13

21 ± 2

24 ± 6

31 ± 6

 

500

 

20 ± 1

112 ± 15

20 ± 6

21 ± 8

29 ± 4

 

 

 

 

 

 

 

 

DAUN

1.00

 

201 ± 19

 

 

 

 

NaN3

2.00

 

 

1290 ± 94

815 ± 14

 

 

9-AA

50.0

 

 

 

 

1601 ± 111

 

NQO

2.00

 

 

 

 

 

2043 ± 102

 

 

 

 

 

 

 

 

 

With

Activation

H2O

 

 

32 ± 4

131 ± 22

27 ± 9

25 ± 4

34 ± 13

 

 

 

 

 

 

 

 

Test item

0.500

 

22 ± 2

141 ± 7

23 ± 6

17 ± 6

44 ± 6

 

1.58

 

35 ± 10

156 ± 7

25 ± 5

15 ± 2

36 ± 4

 

5.00

 

19 ± 4

146 ± 7

26 ± 10

19 ± 8

48 ± 5

 

15.8

 

23 ± 8

132 ± 19

29 ± 6

12 ± 5

40 ± 7

 

50.0

 

19 ± 2

124 ± 6

20 ± 3

15 ± 7

41 ± 3

 

158

 

23 ± 9

137 ± 2

23 ± 2

13 ± 5

34 ± 8

 

500

 

18 ± 4

120 ± 7

17 ± 6

14 ± 3

32 ± 9

 

 

 

 

 

 

 

 

2-AA

2.00

 

252 ± 89

971 ± 105

 

 

 

2-AA

5.00

 

 

 

274 ± 24

333 ± 114

 

2-AA

10.0

 

 

 

 

 

314 ± 99

 

 

 

 

 

 

 

 

 

Key to Positive Controls

 

 

 

NaN3

2-AA

9-AA

DAUN

NQO

Sodium azide

2-Aminoanthracene

9-Aminoacridine

Daunomycin

4-Nitroquinoline-N-oxide

 

 

Summary Table 2ndseries

 

Metabolic

Activation

Test

Material

Concentr.

[µg/plate]

 

Revertants per plate (Mean ± SD)

 

 

 

 

 

 

TA 98

TA 100

TA 1535

TA 1537

WP2 uvrA

 

 

 

 

 

 

 

 

 

Without Activation

H2O

 

 

35 ± 6

119 ± 13

28 ± 2

23 ± 4

26 ± 3

 

 

 

 

 

 

 

 

Test item

15.8

 

31 ± 5

116 ± 7

34 ± 10

21 ± 2

18 ± 5

 

50.0

 

27 ± 3

125 ± 15

25 ± 7

23 ± 3

19 ± 8

 

158

 

31 ± 6

120 ± 6

20 ± 3

30 ± 1

17 ± 5

 

500

 

26 ± 4

119 ± 7

31 ± 9

22 ± 6

19 ± 3

 

1580

 

27 ± 10S E

128 ± 5S E

27 ± 4S E

25 ± 4S E

17 ± 2S E

 

 

 

 

 

 

 

 

DAUN

1.00

 

373 ± 39

 

 

 

 

NaN3

2.00

 

 

870 ± 41

648 ± 34

 

 

9-AA

50.0

 

 

 

 

519 ± 235

 

NQO

2.00

 

 

 

 

 

724 ± 62

 

 

 

 

 

 

 

 

 

With

Activation

H2O

 

 

35 ± 9

128 ± 13

28 ± 7

20 ± 7

35 ± 9

 

 

 

 

 

 

 

 

Test item

15.8

 

36 ± 2

122 ± 17

23 ± 4

21 ± 6

29 ± 3

 

50.0

 

34 ± 6

128 ± 2

25 ± 4

20 ± 3

40 ± 2

 

158

 

31 ± 7

114 ± 6

25 ± 2

19 ± 8

42 ± 9

 

500

 

33 ± 8

139 ± 4

25 ± 2

22 ± 6

38 ± 3

 

1580

 

36 ± 10S E

126 ± 13S E

22 ± 6S E

19 ± 2S E

40 ± 1S E

 

 

 

 

 

 

 

 

2-AA

2.00

 

188 ± 38

 

 

 

 

2-AA

5.00

 

 

1189 ± 26

 

 

 

2-AA

10.0

 

 

 

188 ± 11

269 ± 24

132 ± 3

 

 

 

 

 

 

 

 

 

 

Key to Positive Controls

Key to Plate Postfix Codes

 

 

NaN3

2-AA

9-AA

DAUN

NQO

Sodium azide

2-Aminoanthracene

9-Aminoacridine

Daunomycin

4-Nitroquinoline-N-oxide

S

E

Plated as suspension

Precipitation until end of experiment

 

Conclusions:
With and without addition of S9 mix as the external metabolizing system, the test material was not mutagenic under the experimental conditions described.
Executive summary:

This study was performed according to GLP and the methods applied are fully compliant with OECD TG 471. With and without addition of S9 mix as the external metabolizing system, the test material was not mutagenic under the experimental conditions described.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
For this endpoint information from a structural similar compound is available. This study for this similar compound was performed according to GLP and the methods applied are fully compliant with OECD TG 471. See chapter 13 report for a more detailed justification.
Reason / purpose:
read-across source
Related information:
Composition 1
Test material information:
Composition 1
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Based on the data obtained from the structural analogue substance, the test item is not classified for mutagenicity according to Regulation (EC) No 1272/2008.

Justification for classification or non-classification