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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 1996 - January 1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
Bacterial strains tested not in accordance with the current recommendations (TA1538 instead of TA102), less than five different analysable concentrations of the test substance used
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
liquid

Method

Target gene:
His gene
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 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix from Aroclor 1254-induced rat liver
Test concentrations with justification for top dose:
0.002, 0.002, 0.2, 2 and 20 µL test solution/plate (1st experiment)
0.02, 0.06, 0.2, 0.6 and 2 µL test solution/plate (2nd experiment)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: 50% aqueous solution of test substance used
Controls
Untreated negative controls:
yes
Remarks:
water
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene, 2-aminofluorene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: not applicable
- Exposure duration: incubation at 37°C for 2 days

SELECTION AGENT (mutation assays): 0.05 mM L-histidine

NUMBER OF REPLICATIONS: 3
Evaluation criteria:
A test material is considered mutagenic if there is a reproducibly increasing dose-response curve of induced revertant colonies for at least 3 test concentrations. The minimal criteria for a positive response are a 2- to 3-fold increase in the number of revertants (at least 15 colonies) over the spontaneous number for the TA1535, TA1537, TA1538 and TA98 strains, and a 50% increase for the TA100 strain. In addition, a positive response must not be observed only at concentrations near toxic dose levels.
Statistics:
Mean and standard deviation calculation

Results and discussion

Test resultsopen allclose all
Key result
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:
from and above 0.6 µL/plate in the absence of S9, from and above 2 µL/plate (0.6 µL/plate for TA100) in the presence of S9
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
from and above 0.6 µL/plate in the absence of S9, from and above 2 µL/plate in the presence of S9
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Concentration

(µL test solution)

TA98

TA100

TA1535

TA1537

TA1538

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

Mean

SD

Ratio vs. controls

0 (water)

18

1

1.0

27

7

1.0

169

10

1.0

175

13

1.0

11

4

1.0

13

1

1.0

13

2

1.0

12

5

1.0

9

2

1.0

14

2

1.0

0.002

19

3

1.0

28

9

1.1

152

14

0.9

165

29

0.9

12

2

1.2

11

2

0.9

14

5

1.1

12

4

0.9

12

3

1.3

19

3

1.4

0.02

22

2

1.2

23

6

0.9

173

8

1.0

162

5

0.9

14

1

1.3

10

3

0.8

16

6

1.2

16

2

1.3

8

3

0.9

17

3

1.2

0.2

19

6

1.1

27

2

1.0

148

33

0.9

196

24

1.1

14

3

1.3

16

1

1.3

12

4

0.9

18

6

1.4

15

2

1.6

22

7

1.6

2

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

20

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

0*

0

0.0

Positive controls

2-Nitrofluorene

565

21

30.8

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

1600

50

177.8

-

-

-

2-Aminofluorene

-

-

-

318

18

11.9

-

-

-

1175

65

6.7

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Sodium azide

-

-

-

-

-

-

526

27

3.1

-

-

-

439

12

41.2

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2-Aminoanthracene

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

177

7

13.6

-

-

-

145

10

11.7

-

-

-

1375

56

98.2

9-Aminoacridine

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

263

14

19.7

-

-

-

-

-

-

-

-

-

* Cytotoxicity

First experiment: Summary of numbers of revertants per plate

Applicant's summary and conclusion

Conclusions:
Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was not mutagenic in a bacterial reverse mutation assay up to cytotoxic concentrations, with and without metabolic activation.
Executive summary:

Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was tested in a bacterial reverse mutation (Ames) test using the Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100 strains. The bacterial strains were exposed on minimal agar plates (using a plate incorporation method) to a range of concentrations up to 20 µL of test solution per plate, both in the presence or absence of an exogenous metabolic activation system, consisting of S9 mix from Aroclor 1254 -induced rat liver. Two independent experiments were performed in triplicate.

In this Ames test, no significant increase in the mean number of revertants over the respective vehicle controls was observed in any of the bacterial strains tested, either in the presence or in the absence of metabolic activation, up to 0.2 or 0.6 µL test solution/plate depending on the strain. Cyotoxic effects were observed at higher dose levels.

Under the conditions of this assay, Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was not mutagenic up to cytotoxic concentrations.