6,10-dimethylundecan-2-one

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:

2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

2000

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology BASF AG

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

6,10-dimethylundecan-2-one

EC Number:

216-509-8

EC Name:

6,10-dimethylundecan-2-one

Cas Number:

1604-34-8

Molecular formula:

C13H26O

IUPAC Name:

6,10-dimethylundecan-2-one

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Tetrahydrogeranylaceton
- Substance No.: 01/0275-1
- Batch-Nr.: B 633
- Purity: 97.5 area% (GC)
- Date of production: 17 Oct 2000
- Substance type: Liquid / colourless - clear

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: BASF, Batch No. B 633
- Purity: 97.5 %

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refrigerator
- Appearance, consistency: colorless liquid

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: 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.

Method

Target gene:

his, trp

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9-mix

Test concentrations with justification for top dose:

SPT: 20, 100, 500, 2500 and 5000 µg/plate;
PIT: 4, 20, 100, 500 and 2500 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Due to the Iimited 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

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

N-ethyl-N-nitro-N-nitrosoguanidine

other: 4-nitro-o-phenylendiamine (NOPD)

Remarks:

Positive control substance with S9-mix: 2-aminoanthracene (2-AA); 2.5 µg/plate, dissolved in DMSO, strains: TA1535, TA100, TA1537, TA98; 60 µg/plate, dissolved in DMSO, strain: E.coli WP2 uvrA

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 20 min, 37°C
- Exposure duration: 48 - 72 h, 37°C

NUMBER OF REPLICATIONS: 3 test plates per dose or per control

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

The test chemical is considered positive in this assay if the following criteria are met:
- A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontanous mutation rate in at least one tester strain eitherwithout S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if:
- The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out
independently of each other

Generally, the experiment is considered valid if the following criteria are met:
- The number of revertant colonies in the negative controls was within the normal range of the historical data for each tester strain.
- The sterility controls revealed no indication of bacterial contamination.
- The positive control articles both with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical control data.
- The titer of viable bacteria was >= 10^9/mL.

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:

other: SPT: >= 500 µg/plate, PIT >= 500 µg/plate

Vehicle controls validity:

valid

Untreated negative 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

Remarks:

PIT >=2 500 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY:
A weak bacteriotoxic effect (slight decrease in the number of revertants and/or reduction in the titer) was observed in the standard plate test depending on the strain and test conditions from about 500 µg - 2500 µg/plate onward.
In the preincubation assay bacteriotoxicity (reduced background growth, slight decrease in the number of revertants, slight reduction in the titer) was observed depending on the strain and test conditions at doses of about >= 500 µg/plate.

Solubility:
No test substance precipitation was found.

Any other information on results incl. tables

Results:

Standard plate test:

Dose µg/plate	metabolic activation	TA98			TA100			TA1535			TA1537			E.coli WP2 uvr A
		Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3
0	+	35	40	36	141	123	139	17	19	22	11	12	10	27	29	27
20	+	30	27	32	113	119	120	17	16	19	13	11	7	24	27	28
100	+	26	24	23	124	103	114	16	14	16	10	8	7	26	22	33
500	+	24	20	20	109	98	93	15	15	15	10	8	7	27	28	23
2500	+	16	21	18	57	43	73	16	20	12	6	7	5	17	21	20
5000	+	16	10	19	32	53	36	14	12	10	5	4	5	18	20	17
2.5 µg 2-Aminoanthracene	+	651	570	570	649	638	636	202	213	207	99	89	103
60 µg 2-Aminoanthracene	+													200	216	222
0	-	24	28	29	102	104	111	19	16	17	7	10	10	30	25	27
20	-	25	22	18	107	111	106	17	15	19	7	3	9	26	21	26
100	-	23	24	19	110	122	111	18	14	19	6	9	5	21	19	22
500	-	19	21	20	93	88	97	16	13	16	7	7	8	25	21	26
2500	-	14	17	15	98	81	71	14	16	14	7	9	5	21	20	19
5000	-	14	13	11	54	44	59	10	13	11	5	5	4	19	21	20
5 µg MNNG	-				683	633	513	632	572	652
10 µg 4-Nitro-o-phenylendiamin	-	722	682	703
100 µg 9-amninoacridine	-										411	420	463
5 µg 4-nitroquinoline-N-oxide	-													460	473	409

Preincubation test:

Dose µg/plate	metabolic activation	TA98			TA100			TA1535			TA1537			E.coli WP2 uvra
		Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3	Trial 1	Trial 2	Trial 3
0	+	32	34	44	103	108	104	16	18	20	10	10	11	25	33	31
4	+	30	36	33	107	107	104	13	19	17	11	12	11	24	30	22
20	+	39	29	33	125	109	100	15	16	13	10	11	9	30	27	22
100	+	32	30	31	78	66	94	15	10	10	11	14	9	24	31	27
500	+	24	31	30	90	96	72	12	10	14	10B	8B	6B	39	31	22
2500	+	22B	19B	21B	69B	54B	47B	11B	9B	12B	8B	6B	4B	24B	17B	16B
2.5 µg 2-Aminoanthracene	+	736	822	804	739	682	622	109	117	122	113	147	128
60 µg 2-Aminoanthracene	+													287	244	261
0	-	36	38	36	116	105	128	20	16	19	12	12	7	26	27	31
4	-	33	46	34	102	105	110	16	15	16	16	9	13	28	29	26
20	-	27	29	29	98	97	125	15	20	16	8	7	9	21	27	29
100	-	28	31	24	79	89	74	20	18	15	6	8	6	24	27	22
500	-	24	29	24	90	66	66	16	13	15	2B	4B	7B	23	26	20
2500	-	10B	21B	20B	66B	67B	64B	10B	10B	14B	0B	0B	0B	22B	23B	17B
5 µg MNNG	-				557	687	637	991	873	962
10 µg 4-Nitro-o-phenylendiamin	-	936	956	982
100 µg 9-amninoacridine	-										413	460	449
5 µg 4-nitroquinoline-N-oxide	-													425	557	529

B: reduced backround growth

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions chosen in this study, it is concluded that the test substance is not a mutagenic agent in a bacterial reverse mutation test.

Executive summary:

The test substance was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial strains, i.e. Salmonella typhimurium and Escherichia coli, in a reverse mutation assay.

- Strains: TA 1535, TA 100, TA 1537, TA 98 and E.coli WP2 uvrA

- Dose range: 20 µg - 5.000 µg/plate (SPT), 4 µg - 2.500 µg/plate (PIT)

- Test conditions: Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (Aroclor-induced rat liver S9 mix)

- Solubility: No precipitation of the test substance was found.

- Toxicity: A bacteriotoxic effect was observed under all test conditions

- Mutagenicity: An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S9 mix or after the addition of a metabolizing system