Paraffin oils, sulfochlorinated, saponified

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jan - Feb 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

E. coli WP2 strain or S. typhimurium TA 102 were not investigated.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

E. coli WP2 strain or S. typhimurium TA 102 were not investigated.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Paraffin oils, sulfochlorinated, saponified

EC Number:

269-144-1

EC Name:

Paraffin oils, sulfochlorinated, saponified

Cas Number:

68188-18-1

Molecular formula:

typical example: C15H31Na03S

IUPAC Name:

n-C14-C17 alkanes, secondary monosulphonic acids, sodium salts

impurity 1

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

impurity 2

Reference substance name:

Sodium chloride

EC Number:

231-598-3

EC Name:

Sodium chloride

Cas Number:

7647-14-5

Molecular formula:

ClNa

IUPAC Name:

sodium chloride

impurity 3

Reference substance name:

Glycerides, mixed decanoyl and octanoyl

EC Number:

277-452-2

EC Name:

Glycerides, mixed decanoyl and octanoyl

Cas Number:

73398-61-5

IUPAC Name:

73398-61-5

impurity 4

Reference substance name:

Sodium hydroxide

EC Number:

215-185-5

EC Name:

Sodium hydroxide

Cas Number:

1310-73-2

Molecular formula:

HNaO

IUPAC Name:

sodium hydroxide

Test material form:

solid

Specific details on test material used for the study:

Information from study report:
- Name of test material: Emulgator E30, Fest
- CAS number: 5896-54-8
The CAS number in report (5896-54-8) is misleading, since this CAS number does only represent 1-Penta-decanesulfonic acid, sodium salt (1:1), but it is confirmed that the substance used is correctly assigned to CAS 68188-18-1.
- Appearance/Further information: The test substance was supplied as white wax-like leaf-let and was received at the test institute on March 06, 1991.
- Batch No. of test material: 210291
- Purity: 95 %
- Analytical reference: TGL 39237
- Solubility and stability of the test substance in the solvent/vehicle: The substance is known to be stable in water.

Method

Target gene:

Histidine locus

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:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S9 preparation from the liver of Aroclor 1254 induced Sprague-Dawley rats.

Test concentrations with justification for top dose:

For initial test and independent repeat: 5, 1.5, 0.5, 0.15, 0.05, 0.015, 0.005 mg/plate with and without metabolic activation

Vehicle / solvent:

Water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene 10 µg/plate (all strains)

Remarks:

The positive controls 2-nitrofluorene, Na-azide and 9-aminoacridine were used without S9 mix; the positive control 2-aminoanthracene was used with S9 mix.

Details on test system and experimental conditions:

METHOD OF APPLICATION: Plate Incorporation Test
For each concentration and tester strain three parallel plates were used.

DETERMINATION OF CYTOTOXICITY
- Method: background growth

Evaluation criteria:

The threshold value given in the following tables is the number of revertants (average number of revertant colonies from test compound) for which X² is >/= 6.6 with the average number of spontaneous revertant colonies being the corresponding solvent control. If the number of revertants with the test compound would be greater than the calculated threshold value the number of revertants with the test substance would be significantly greater than the spontanous rate in the solvent control (error probability p

Statistics:

The statistical significance of the increase in the mutation frequency is examined in the X²-test with respect to its statistical significance. A statistically significant dose-related increase in the number of revertants occurs if several dilutions of the test compound are significant greater than the average number of spontaneous revertant colonies.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

Mutagenicity of the test material was not detected, neither in the initial experiment in concentrations up to and including 5 mg/plate, nor in the independent repetition with up to and including 0.5 mg/plate, both conducted with and without a metabolic activating system.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the first test toxic effects were exhibited when the highest concentration of 5 mg/plate was applied. The tester strains gave reduced numbers of revertants compared to the untreated control however, accompanied by a scattered background growth of the inoculated bacteria. Growth of strain TA 1538 was completely inhibited by a concentration of 0.5 mg/plate. Strain TA 1537 showed the same effect with a concentration of 1.5 mg/plate.
Therefore the highest concentration of the test substance was 0.5 mg/plate in the repetition experiments either with or without metabolic activation.

Applicant's summary and conclusion

Executive summary:

A bacterial reverse mutation assay (Ames test) according to OECD TG 471 was carried out with and without a metabolic activating system with the 5 tester strains S. thyphimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100, however no E. coli WP2 strain or S. typhimurium TA 102 was investigated. One independent repetition of the test was performed. Mutagenicity of the test material was not detected. Toxicity to S. typhimurium could be observed at the higher concentrations.