Acid chlorides, coco

Genetic toxicity: in vitro

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Acid chlorides, coco

EC Number:

269-133-1

EC Name:

Acid chlorides, coco

Cas Number:

68187-89-3

Molecular formula:

variable

IUPAC Name:

dodecanoyl chloride; tetradecanoyl chloride

Constituent 2

Reference substance name:

Fatty acids, coco, chlorinated

EC Number:

307-160-3

EC Name:

Fatty acids, coco, chlorinated

Cas Number:

97553-06-5

IUPAC Name:

97553-06-5

Constituent 3

Reference substance name:

Cocoyl chloride

IUPAC Name:

Cocoyl chloride

Constituent 4

Reference substance name:

96/239

IUPAC Name:

96/239

Details on test material:

- Name of test material (as cited in study report): Kokosfettsaeurechlorid)
- Batch No.: Vers. 148/U
- Physical state: clear liquid, yellowish
- Analytical purity: 99.5 % (by GC)
- Expiration date of the lot/batch: not reported
- Stability under test conditions: confirmed by the analytical report 98L00389
- Storage condition of test material: at room temperature under exclusion of moisture and oxygen (nitrogen atmosphere)

Method

Target gene:

his- (Salmonella typhimurium strains), trp- (E. coli strains)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S-9 mix

Test concentrations with justification for top dose:

20.0 μg - 5,000 µg/plate (SPT) in two trials: 20, 100, 500, 2500, 5000 µg/plate and 250, 500, 750, 1000 µg/plate
0.8 μg - 500 µg/plate (PIT)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: see free text

Details on test system and experimental conditions:

Experiment 1: Standard plate test (SPT)
Test tubes containing 2 mL soft agar kept in a water bath at 45°C, and remaining components added in the following order:
0.1 mL test solution or vehicle
0.1 mL bacterial suspension
0.5 mL S-9 mix (in tests with metabolic activation) or 0.5 mL phosphate buffer (in tests
without metabolic activation).
After mixing, the samples are poured onto Vogel-Bonner agar plates.

Experiment 2: Preincubation assay (PIT)
0.1 mL test solution or vehicle, 0.1 mL bacterial suspension and 0.5 mL S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 mL of soft agar is added and, after mixing, the samples are poured onto minimal agar plates.

Experiment1&2
In each experiment 3 test plates per dose or per control used; after incubation at 37°C for 48 - 72 hours in the dark, the bacterial colonies ( his+ and trp + revertants, respectively) are counted.

Positive control:
with metabolic activation: 2.5 μg/plate 2-aminoanthracene for each Salmonella strain; 60 µg/plate 2-aminoanthracene for the E. coli strain
without metabolic activation: 5 μg/plate N-methyl-N'-nitro-N-nitrosoguanidine for TA 100 and TA 1535, 10 μg/plate 4-nitro-o-phenylendiamine
for TA 98, 100 μg/plate 9-aminoacridine chloride monohydrate for TA 1537 and 10 µg/plate N-Ethyl-N-nitro-N-nitrosoguanidine for the E. coli strain. All substances were dissolved in DMSO.
The titer was determined and in regularly measurements the strain characteristics were checked. Sterility control was performed.

Evaluation criteria:

- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Statistics:

Mean and standard deviation calculated in result tables. No further data.

Results and discussion

Test results

Test results 1

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:

at doses >= 500 µg/plate in the SPT and at doses >= 100 µg/plate in the PIT

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

A test substance precipitation was found from about 1000 μg/plate onward.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test (20 - 5000 µg/plate)
Strain	Metabolic activation system	Replicates	max. revertant factor (Trial 20-5000 µg/plate)	max. revertant factor (Trial 250-1000 µg/plate)	dose dependency	Assessment
TA 98	no	3	1.0	0.9	no	negative
	yes	3	0.7	0.9	no	negative
TA 100	no	3	0.9	1.1	no	negative
	yes	3	0.9	0.9	no	negative
TA 1535	no	3	1.0	1.0	no	negative
	yes	3	0.9	0.9	no	negative
TA 1537	no	3	0.9	1.2	no	negative
	yes	3	1.0	0.9	no	negative
WP2 uvr A	no	3	1.1	not perf.	no	negative
	yes	3	0.9	not perf.	no	negative
Preincubation test (0.8 - 500 µg/plate)
Strain	Metabolic activation system	Replicates	maximum revertant factor	dose dependency	Assessment
TA 98	no	3	0.9	no	negative
	yes	3	0.9	no	negative
TA 100	no	3	1.0	no	negative
	yes	3	1.1	no	negative
TA 1535	no	3	0.9	no	negative
	yes	3	0.9	no	negative
TA 1537	no	3	0.9	no	negative
	yes	3	1.2	no	negative
WP2 uvr A	no	3	1.1	no	negative
	yes	3	0.9	no	negative

Applicant's summary and conclusion

Executive summary:

The study was performed according to OECD Guideline 471 and GLP requirements and is reliable with out any restriction. Cocoyl chloride was tested in the standard plate test (SPT) as well as in the preincubation assay (PIT) with and without metabolic activation (MA) in S. typhimurium TA98, TA100, TA1535 and TA1537 as well as in E. coli WP2uvrA at dose levels of 20 - 5000 µg/plate (SPT) or 0.8 - 500 µg/plate (PIT). No increase in the number of revertants was detected in any strain with and without MA. Vehicle controls and positive controls were valid. Cytotoxicity was found depending on the test method in concentration. Precipitation was seen in the SPT.

Conclusion:   

According to the results of the present study, the test substance Cocoyl chloride is not mutagenic in the Salmonella typhimurium/ Escherichia coli reverse mutation assay under the experimental conditions chosen here.