Mixture of mono-, di- and triesters obtained with isooctadecanoic acid and hydrogenated castor oil

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-Study according to international guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

2000/32/EEC, Annex4D Ames test

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Salacos HCISV-L

IUPAC Name:

Salacos HCISV-L

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

Details on test material:

- Name of test material (as cited in study report): Salacos HCISV-L
- Substance type: organic
- Physical state: not stated
- Analytical purity: not stated
- Lot/batch No.: 40314
- Expiration date of the lot/batch: MAy 02, 2003
- Stability under test conditions: not stated
- Storage condition of test material: room temperature

Method

Target gene:

for S. typhimurium: his-
for E. Coli: trp-

Species / strain

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:

S9 liver fractions of rats, induced with phenobarbital/»- naphtoflavone.

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 33 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 33 ... 5000 µg/plate

Vehicle / solvent:

Solvent: Ethanol

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine, 2-Aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation; plate-incorporation

DURATION
- Preincubation period: 60 min
- Exposure duration: 48 h
- Expression time (cells in growth medium): none
- Selection time (if incubation with a selection agent): none
- Fixation time (start of exposure up to fixation or harvest of cells): none

SELECTION AGENT (mutation assays): none
SPINDLE INHIBITOR (cytogenetic assays): none
STAIN (for cytogenetic assays): none

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: not relevant

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth/number of colonies

Evaluation criteria:

The Salmonella typhimurium and Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
- regular background growth in the negative and solvent control
- the spontaneous reversion rates in the negative and solvent control are in the range of the laboratories historical data
- the positive control substances should produce a significant increase in mutant colony frequencies

Statistics:

except for calculation of mean values, no statistics performed

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

for Strain TA 1535 at 2500 and 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

It can be stated that during the dscribed mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
Therefore, Salacos HCISV-L is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.

Executive summary:

This study was performed to investigate the potential of Salacos HCISV-L to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 , and the Escherichia coli strain WP2 uvrA.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations:

33; 100; 333; 1000; 2500; and 5000 µg/plate

The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without metabolic activation in both independent experiments.

A minor toxic effect, evident as a reduction in the number of revertants, was observed in strain TA 1535 at 2500 and 5000 µg/plate without S9 mix in experiment I. No toxic effects were observed in the remaining strains and in experiment II.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Salacos HCISV-L at any dose level, neither in the presence nor absence of metabolic activation (S9 -mix). There was also no tendency of higher gene mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.