3,7,11-Trimethyldodec-1-en-3-ol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Principles of method if other than guideline:

Ames II Assay according to Gee, P. et al. (1998) and on the "Users Manual" prepared by XENOMETRIX, Inc., Colorado, USA

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3,7,11-trimethyldodec-1-en-3-ol

EC Number:

609-221-2

Cas Number:

3625-46-5

Molecular formula:

C15H30O

IUPAC Name:

3,7,11-trimethyldodec-1-en-3-ol

Test material form:

liquid

Details on test material:

Tetrahydronerolidol (THNER)
CAS No. 3625-46-5
Batch No.: Anlage 067, 13 Jun 2001
Purity: 96 %

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: Anlage 067, 13.06.01
- Date of manufacture: June 2001

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: The stability of the test substance throughout the study has not been verified by reanalysis. The analytical investigations were carried out in the Analytical Departmetn, BASF AG (characterization)
- Stability of the test substance in the vehicle: The stability of the test substance in the vehicle DMSO and in water has not been determined analytically. Detailed information may be requested from the sponsor BASF AG.

Method

Target gene:

his

Species / strain

Species / strain 1

Species / strain / cell type:

other: Salmonella typhimurium TA Mix (Mixed strains TA 7001 - TA 7006) Liquid Fluctuation Test - Microtiter Version

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9-mix

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500 and 5000 µg/ml

Vehicle / solvent:

- Vehicle used: DMSO
- Justification for choice of vehicle: Due to the limited solubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2-AA); 5 µg/ml

Remarks:

with metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Mixture of 2-nitrofluorene (2-NF; 0.25 µg/ml) and 4-nitroquinoline-N-oxide (4-NQO; 0.0625 µg/ml)

Remarks:

without metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium (liquid fluctuation test with and without S-9 mix)


DURATION
- Exposure duration: 90 min, 37°C
- Selection time (if incubation with a selection agent): ca. 48, 37°C


SELECTION AGENT (mutation assays): histidine-deficient Ames II Reversion Indicator medium (Xenometrix)


NUMBER OF REPLICATIONS: Triplicate plates per dose, control chemical or vehicle


DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of positive wells, clearing or diminution of the background lawn (= reduced his' background growth) leading from turbid to non-turbid purple wells

OTHER: Acceptance criteria
Generally, the experiment is considered valid if the following criteria are met:
- the number of revertant wells in the negative controls was within the normal range of the historical control data for each tester strain
- the sterility controls revealed no indication of bacterial contamination
- the positive control articles both with and without S-9 mix induced a significant
increase in the number of positive wells within the range of the historical control data

Evaluation criteria:

EVALUATION CRITERA
- The test chemical is considered positive in this assay if the following criteria are met:
a dose-related increase in the number of positive wells by a factor of about 2 (calculated primarily on the basis of baseline data) in at least one tester strain either without S-9 mix or after adding a metabolizing system.
- A test substance is generally considered non-mutagenic in this test if:
the number of revertant wells for all tester strains were within the historical negative control range under all experimental conditions

ACCEPTANCE CRITERIA
Generally, the experiment is considered valid if the following criteria are met :
• The number of revertant wells in the negative controls was within the normal range of the historical control data for each tester strain
• The sterility controls revealed no indication of bacterial contamination .
• The positive control articles both with and without S-9 mix induced a significant increase in the number of positive wells within the range of the historical control data

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

other: Salmonella typhimurium TA Mix (Mixed strains TA 7001 - TA 7006) Liquid Fluctuation Test - Microtiter Version

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found

Any other information on results incl. tables

Result:

Dose µg/plate	metabolic activation	TA98			TA Mix (Mixed strains TA 7001 - TA 7006)
		Plate 1	Plate 2	Plate 3	Plate 1	Plate 2	Plate 3
DMSO	-	2	3	3	1	1	1
4	-	3	1	0	0	1	1
20	-	1	0	0	0	0	0
100	-	0	1	2	1	1	0
500	-	1	3	0	0	0	2
2500	-	2	2	1	1	0	1
5000	-	5	0	1	0	1	1
4-NQO + 2-NF	-	30	20	22	28	23	25
DMSO	+	3	2	5	0	1	2
4	+	6	1	1	1	2	0
20	+	0	3	1	2	1	1
100	+	1	0	5	0	0	1
500	+	3	2	1	0	0	0
2500	+	1	1	3	1	0	0
5000	+	3	4	3	1	0	1
2-AA	+	48	48	48	29	27	24

Applicant's summary and conclusion

Conclusions:

Thus, under the experimental conditions chosen here, it is concluded that the test substance is not a mutagenic agent in the Ames II Assay (Salmonella typhimurium reverse mutation assay) .

Executive summary:

The test substance was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several strains of Salmonella typhimurium in a modified version of the traditional Ames test, i .e. the Ames II Assay (microtiter version) .

STRAINS: TA 98, TA Mix (Mixed strains TA 7001 - TA 7006)

DOSE RANGE: 4 µg - 5,000 µg/ml

TEST CONDITIONS: Liquid fluctuation test both with and without metabolic activation (Aroclor-induced rat liver S-9 mix)

SOLUBILITY: No precipitation of the test substance was found

TOXICITY: No bacteriotoxic effect was observed

MUTAGENICITY: An increase in the number of positive wells (his+ revertants) was not observed either without S-9 mix or after the addition of a metabolizing system