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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:
05 Nov 1998 - 06 Apr 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
No historical positive control data.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report date:
1999

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted in 1997
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Dodecanoic acid, ester with 1,2-propanediol
EC Number:
253-462-2
EC Name:
Dodecanoic acid, ester with 1,2-propanediol
Cas Number:
37321-62-3
Molecular formula:
C15H30O3 C27H52O4
IUPAC Name:
Dodecanoic acid, ester with 1,2-propanediol

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Details on mammalian cell type (if applicable):
CELLS USED
- Source of cells: B. Ames Laboratory, University of California, Berkeley, USA
Genetic characteristics were checked for each frozen stock culture preparation
The number of colonies obtained in the absence of any treatment (spontaneous revertants) was compared versus historical control data for each culture.

MEDIA USED
- Type and identity of media including CO2 concentration if applicable: Vogel-Bonner medium.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: not stated
Metabolic activation:
with and without
Metabolic activation system:
Freeze dried S9 fraction from rat liver treated with a mixture of phenobarbital and methylcholanthrene, batch SL 63, IFFA-CREDO, France
Test concentrations with justification for top dose:
Preliminary experiment (TA 100): 52, 164, 512, 1600, 5000 µg/plate (0.52, 1.64, 5.12, 16, 50 mg/mL)
Precipitate in the range of 164 to 5000 µg/plate, cytotoxicity (reduction of bacterial lawn and/or decrease in the number of revertant colonies) at 52 to 5000 µg/plate observed.
Experiment 1: 1.7, 5.4, 17, 52, 164 µg/plate
Experiment 2: 9, 16, 29, 52, 92 µg/plate (top dose chosen to be just below precipitation)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO (Sigma, batch 38H3447)
- Justification for choice of solvent/vehicle: Test article was insoluble in water at 50 mg/mL. A clear solution was obtained with DMSO.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: Butyl hydroxyperoxide for TA102 and 2-Aminoanthracene for all strains with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation);
Cells were allowed to grow in nutrient broth for 6-16 h (37 degree Celsius) in incubator to the late exponential stationary or early stationary phase of growth (approx. 10E+08 or 10E+09 cells per mL). Optical density was used to check cell density.

- Exposure duration: 48 h minimum (37 degree Celsius incubator)

NUMBER OF REPLICATIONS:
2 experiment with triplicates each; one preliminary test with TA100 strain in triplicate

DETERMINATION OF CYTOTOXICITY
Reduction of bacterial lawn and/or decrease in the number of revertant colonies

Test article formulations were prepared on every treatment day and mixed with molten agar and bacterial culture with or without S9 mix.
Rationale for test conditions:
Top doses were chosen due to cytotoxicity and precipitation occurring at 164 µg/plate and upwards.
Evaluation criteria:
Individual plate counts
Statistics:
Standard deviation, variation. Dunnett's test was used to compare the counts at each dose level with the negative (vehicle) control.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
for Experiment 1 and 2
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
for Experiment 1 and 2
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
for Experiment 1 and 2
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
for Experiment 1 and 2
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
for Experiment 1 and 2
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: None
- Negative spontaneous revertants historical control data: from 1993 to 1197 on all five Salmonella strains with and without metabolic activation

ADDITIONAL INFORMATION ON CYTOTOXICITY/PRECIPITATION:
Preliminary experiment:
Cytotoxicity and precipitate at 164 µg/plate and upward.
Experiment 1:
Clear signs of cytotoxicity and precipitate at 164 µg/plate. Few signs of cytotoxicity, with a moderate degree of inter-strain reproducibility, mainly at 52 µg/plate.
Experiment 2:
No precipitate at tested dose levels up to 92 µg/plate. Few signs of cytotoxicity, with a moderate degree of inter-strain reproducibility, at 29 to 92 µg/plate.
Remarks on result:
other: without S9 significantly more revertant colonies at 1.7 µg/plate in Experiment 1

Applicant's summary and conclusion

Conclusions:
CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.