Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on the available information, the test item is not mutagenic in the Ames test under the selected experimental conditions.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 Mar 2000 to 19 Dec. 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Name of the test substance used in the study report: Neozapon Gelb R Stuecke ber. 100%
- Purity: Ca. 96.6%

performed under old CAS 73297-13-9
new CAS 85029-58-9, same structure
Target gene:
S. typhimurium: his
E. coli: trp
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced rat liver S-9 mix resp. uninduced hamster liver S-9 mix
Test concentrations with justification for top dose:
20 µg - 5000 µg/plate (Standard Plate Test)
4 µg - 2,500 µg/plate (Prival Preincubation Test)
Vehicle / solvent:
- Vehicle: DMSO
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
congo red
other: 2-aminoanthracene (2-AA), benzidine, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylendiamine (NOPD)
Details on test system and experimental conditions:
Ames Standard Plate Test and Prival Preincubation Test
Evaluation criteria:
The test chemical is considered positive in this assay if the following criteria are met :
A dose-related and reproducible increase in the number of revertant colonies, i .e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system .
A test substance is generally considered nonmutagenic in this test if:
The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out
independently of each other.
Statistics:
no statistics, standard deviation calculated
Key result
Species / strain:
S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
A bacteriotoxic effect was observed under all test conditions
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
A bacteriotoxic effect was observed under all test conditions
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
An increase in the number of his+ or trp+ revertants was not observed either in the Ames standard plate test or in the Prival preincubation test without S-9 mix or after the addition of a metabolizing system.

TOXICITY
A weak bacteriotoxic effect (slight decrease in the number of revertants, slight reduction in the titer) was observed in the Ames standard plate test using the Salmonella strains depending on the strain and test conditions from about 2,500 pg/plate onward . In the Prival preincubation assay bacteriotoxicity (reduced his- or trp- background growth, decrease in the number of revertants, reduction in the titer) was observed depending on the strain and test conditions from about 100 pg - 500 pg/plate onward .
SOLUBILITY
Test substance precipitation was found from about 500 pg/plate onward .

Table: Standard Test (average result of 3 plates, respectively)

Concentration [µg/plate]

TA 1535

TA 1537

TA 98

TA 100

E. coli WP2 uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Solvent control (DMSO)

18

19

10

11

31

38

114

128

36

36

20

20

17

9

10

21

30

96

116

25

45

100

18

18

8

12

20

43

91

113

29

43

500

17

15

5

11

20

25

98

86

25

34

2500

11

11

4

5

13

16

58

24

25

37

5000

11

7

3

1

12

5

62

16

25

35

Positive control

647

165

601

103

1081

580

619

722

550

225

Table: Preincubation test (average result of 3 plates, respectively)

Concentration [µg/plate]

TA 1535

TA 1537

TA 98

TA 100

E. coli WP2 uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Solvent control (DMSO)

17

17

9

13

25

35

108

109

36

29

Test item

 

4

19

17

8

12

21

35

105

105

37

25

20

16

16

5

11

20

30

60

89

33

23

100

13

16

4

10

15

25

56

81

27

27

500

12

11

1

5

12

20

31

59

26

25

2500

8

6

2

1

10

9

49

14

24

19

Positive control

 

MNNG

1093

 

 

 

 

 

1190

 

 

 

2-AA

 

178

 

104

 

561

 

622

 

220

AAC

 

 

632

 

 

 

 

 

 

 

NOPD

 

 

 

 

990

 

 

 

 

 

CONGOR.

 

 

 

 

 

370

 

 

 

 

BENZID

 

 

 

 

 

579

 

 

 

 

4-NQO

 

 

 

 

 

 

 

 

519

 

MNNG: N-methyl-N'-nitro-N-nitrosoguanidine

2 -AA: 2-aminoanthracene

AAC: 9-aminoacridine

NOPD: 4 -nitro-o-phenylendiamine

4 -NQO: 4 -Nitroquinoline-N-oxide

BENZID: Benzidine

CONGOR.: Congo red

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Genetic toxicity in vitro:

In a reverse gene mutation assay in bacteria (Ames-test), strains TA 1535, TA 100, TA 1537 and TA 98 of Salmonella typhimurium and strain WP2 uvrA of E. coli were exposed to the test substance at concentrations of 20, 100, 500, 2500 and 5000 µg/plate (BASF, 2000). Standard plate test and preincubation test both were conducted with and without metabolic activation (Aroclor induced rat liver S-9 mix). The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background. Based on the available information, the test substance is not mutagenic in the Ames test under the selected experimental conditions.

Justification for classification or non-classification

Based on the available information classification for genetic toxicity is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.

Categories Display