Cyclohexane, oxidized, non-acidic by-products, distn. residues

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: Guideline study with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Remarks:

(BASF AG, Department of Experimental Toxicology and Ecology)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Cyclohexane, oxidized, non-acidic by-products, distn. residues

EC Number:

271-816-4

EC Name:

Cyclohexane, oxidized, non-acidic by-products, distn. residues

Cas Number:

68609-04-1

IUPAC Name:

(1s,4s)-cyclohexane-1,4-diol (cyclohexyloxy)cyclohexane 2-butylcyclohexan-1-one 8-oxatricyclo[7.4.0.0^{2,7}]trideca-1(9),2(7)-diene cyclohexane cyclohexane-1,3-diol cyclohexanol phenol hydrate

Details on test material:

- Name of test material (as cited in study report): ANONE-OIL SPLIT K1
- Physical state: brown, vey viscous, homogeneous liquid
- Analytical purity: not reported
- Lot/batch No.: 96075-3
- Stability under test conditions: stability of the test substance was proven for a time frame of 5 months
- Storage condition of test material: room temperature

The test substance is a complex mixture of a large number of individual components. Main component is dicyclohexyl ether (9.4%).
Contents as determined by GC:
2g/100g H2O
49% components >1% (peak area)

The identity of the mixture was confirmed by IR- and H-NMR-spectroscopy.

Method

Target gene:

histidine and tryptophan auxotrophy

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S-9 mix

Test concentrations with justification for top dose:

Doses standard plate test: 0, 20, 100, 500, 2500 and 5000 µg/plate;
Doses preincubation test: 0, 20, 100, 500, 1000 and 2000 µg/plate (Salmonella strains) 0, 20, 100, 500, 2500 and 5000 µg/plate (E. coli)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene for TA100, TA98, TA1535, TA1537 and WP2uvrA

Remarks:

with S9

Controls 2

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: N-methyl-N-nitro-N-nitrosoguanidine for TA100 and TA1535; 4-nitro-o-phenylendiamine for TA98; 9-aminoacridine for TA1537; N-ethyl-N-nitro-N-nitrosoguanidine for WP2uvrA

Remarks:

without S9

Details on test system and experimental conditions:

- The standard plate test (SPT) was performed according to the method of Ames et al. (1973, 1975) and the Preincubation test (PIT) according to the methods of Yahagi et al. (1977) and Matsushima et al. (1980).
- 3 test plates per dose

Evaluation criteria:

A substance to be characterized as positive has to fulfill thefollowing requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

≥ 2500 µg/plate (SPT), ≥ 500 µg/plate (PIT)

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: a test substance precipitation was found at 5000 ug/plate

Remarks on result:

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

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard Plate Test: Revertants/plate (mean ± SD)

Dose (µg/plate)	TA 98		TA 100		TA 1535		TA 1537		WP2uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	28±8	42±4	128±5	128±4	22±2	21±1	10±2	11±1	29±2	38±2
20	26±2	41±5	108±8	107±10	22±4	20±4	10±2	11±1	36±2	40±2
100	22±2	39±4	103±8	101±17	20±3	18±2	9±2	8±2	33±3	38±6
500	20±2	31±4	100±13	102±12	22±2	14±2	8±2	8±2	34±1	36±4
2500	18±3	21±2	29±8	24±2	13±2	12±3	3±2	5±2	31±3	33±3
5000	9±3p	12±2p	8±1b/p	5±2b/p	3±1p	4±1p	1±1b/p	1±1b/p	25±4p	26±1p
PC	1311±269	1200±55	1562±84	1490±171	1685±36	150±19	742±65	168±22	536±23	262±16

Preincubation Test: Revertants/plate (mean ± SD)

Dose (µg/plate)	TA 98		TA 100		TA 1535		TA 1537		WP2uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	30±6	35±3	123±6	149±8	21±2	22±4	11±1	12±1	31±3	42±3
20	34±5	36±6	121±13	139±19	17±2	16±3	11±3	14±3	34±3	31±5
100	28±4	34±4	121±9	156±2	17±2	17±0	11±2	9±2	33±3	26±4
500	22±5b	33±4	106±16	143±5	14±2b	12±4	3±2b	8±1b	36±4	24±5
1000	20±3b	31±10b	90±5b	108±17b	12±2b	10±2b	2±1b	9±2b	-	-
2000	13±2b	23±4b	64±7b	92±2b	6±2b	11±3b	0b	6±2b	-	-
2500	-	-	-	-	-	-	-	-	31±3	20±2
5000	-	-	-	-	-	-	-	-	22±4b/p	22±2b/p
PC	817±39	638±75	1202±28	782±83	1326±29	145±14	626±92	124±12	524±24	223±5

b: reduced background growth, p- precipitation, PC- positive control