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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:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restriction - Neither TA 102 nor E. Coli are included in the panel of cells tested. - No data on GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(neither TA 102 nor E. Coli are included in the panel of cells tested. Second positive control substance besides 2-AA for tests with S9 mix not included).
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3,4-Dichloroaniline-6-sulfonic acid
EC Number:
700-413-2
Cas Number:
6331-96-0
Molecular formula:
C6 H5 Cl2 N O3 S
IUPAC Name:
3,4-Dichloroaniline-6-sulfonic acid

Method

Target gene:
Histidine operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S-9 fraction mixed with a series of cofactors (MgCl2, KCl, Glu-6-p, NADP, phosphate buffer (NaH2PO4, Na2HPO2.2H2O)
Test concentrations with justification for top dose:
20, 100, 500, 2,500 and 5,000 µg/plate.
Vehicle / solvent:
- Vehicle: DMSO.
- Justification for choice of vehicle: no data.
Controls
Untreated negative controls:
yes
Remarks:
(sterility control)
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: (+ S-9): 2-aminoanthracene, 2-AA; (- S-9): N-methyl-N'-nitro-N-nitrosoguanidine, MNNG; 4-nitro-o-phenylene-diamine, 4-NOPD, 9 -aminoacridine chloride monohydrate.
Remarks:
10 µg 2-AA/plate (all strains); 5 µg MNNG/plate (TA 1535 and TA 100); 10 µg 4-NOPD/plate (TA 98), 100 µg 9-AA-Cl (TA 1537).
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation test) (experiment 1) and pre incubation test (experiment 2).

DURATION
- Exposure duration: 48 hours at 37°C.
- Preincubation period: 20 min

NUMBER OF REPLICATIONS: 2 experiments; 3 cultures/experiment.

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of spontaneous revertants or a clearing of the bacterial background lawn.
Evaluation criteria:
In general, a test chemical is considered positive if the following criteria are met:
- Doubling of the spontaneous mutation rate (control).
- Dose-response relationship.
- Reproducibility of the results
Statistics:
No statistical evaluation needed be performed.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Standard plate test (TA 100 and TA 1537 only; at 5000 µg/plate only), Preincubation test (all strains; ≥ 2500 µg/plate)
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test item was observed at 5,000 µg/plate in the preincubation test and with S9 mix only.

RANGE-FINDING/SCREENING STUDIES:
- no data
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Table of results (plate incorporation test); without metabolic activation

 

 

 

TA 98

TA 100

TA 1535

TA 1537

 

Concentration (µg/plate)

S9

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

test subst.

control

no

20

 

110

 

15

 

11

 

20

no

21

1.1

104

0.9

18

1.2

9

0.8

100

no

21

1.1

113

1.0

15

1.0

9

0.8

500

no

19

1.0

101

0.9

15

1.0

11

1.0

2,500

no

20

1.0

96

0.9

15

1.0

10

0.9

5,000

no

23

1.2

64

0.6

12

0.8

7

0.6

 

 

 

 

 

 

 

 

 

 

 

 

control

no

20

 

110

 

15

 

11

 

 

positive control

no

931

46.6

2050

18.6

1797

119.8

517

47.0

 

Table 2: Table of results (plate incorporation test); with metabolic activation 

 

 

 

TA 98

TA 100

TA 1535

TA 1537

 

Concentration (µg/plate)

S9

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

test subst.

control

yes

36

 

118

 

15

 

11

 

20

yes

32

0.9

126

1.1

16

1.1

16

1.5

100

yes

34

0.9

117

1.0

17

1.1

15

1.4

500

yes

31

0.9

113

1.0

15

1.0

14

1.3

2,500

yes

24

0.7

113

1.0

15

1.0

11

1.0

5,000

yes

21

0.6

124

1.1

17

1.1

11

1.0

 

 

 

 

 

 

 

 

 

 

 

 

control

yes

36

 

118

 

15

 

11

 

 

positive control

yes

1357

37.7

958

8.1

137

9.1

122

11.1

 

Table 3: Table of results (Preincubation test); without metabolic activation

 

 

 

TA 98

TA 100

TA 1535

TA 1537

 

Concentration (µg/plate)

S9

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

test subst.

control

no

21

 

104

 

14

 

8

 

20

no

16

0.8

88

0.8

12

0.9

9

1.1

100

no

11

0.5

83

0.8

13

0.9

6

0.8

500

no

16

0.8

91

0.9

13

0.9

6

0.8

2,500

no

B

-

B

-

13B

0.9

B

-

5,000

no

0

0.0

0

0.0

0

0.0

0

0.0

 

 

 

 

 

 

 

 

 

 

 

 

control

no

21

 

104

 

14

 

8

 

 

positive control

no

893

42.5

866

8.3

982

70.1

371

46.4

 

Table 4: Table of results (Preincubation test); with metabolic activation 

 

 

 

TA 98

TA 100

TA 1535

TA 1537

 

Concentration (µg/plate)

S9

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

X (n=3)

revertant factor

test subst.

control

yes

38

 

121

 

14

 

10

 

20

yes

34

0.9

117

1.0

15

1.1

9

0.9

100

yes

38

1.0

106

0.9

14

1.0

9

0.9

500

yes

38

1.0

94

0.8

16

1.1

9

0.9

2,500

yes

24

0.6

74

0.6

11

0.8

8

0.8

5,000

yes

0Bp

0.0

28p

0.2

11p

0.8

0Bp

0.0

 

 

 

 

 

 

 

 

 

 

 

 

control

yes

38

 

121

 

14

 

10

 

 

positive control

yes

448

11.8

587

4.9

184

13.1

102

10.2

 

B: reduced His- Background

P: precipitation

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative