3-[[5-methyl-2-(1-methylethyl)cyclohexyl]oxy]propane-1,2-diol

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:

21 December 1998 to 21 January 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was conducted in 1998 to 1999 in accordance with OECD Guideline 471/472 on the appropriate strains of bacteria (S. typhimuriumTA1535, TA100, TA1537 and TA98 as well as E. coli WP2uvrA in both the presence and absence of metabolic activation. Whilst not conducted to GLP the report does include a signature page confirming that the report accurately reflects the proceedings of the experiment signed by the laboratory manager. Neither the batch details nor the purity of the substance are reported. Concurrent controls were run as a part of the experiment. Six dose levels were tested during the study, however the experiments were run on duplicate rather than triplicate plates and whilst the study result in an unequivocally negative result, the experiment was not duplicated and no justification for this was given in the report. Despite its basic nature and some deviations from the approved guidelines the study can be considered to be reliable with restrictions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Plates were tested in duplicate not triplicate and the negative result was not confirmed

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Deviations:

yes

Remarks:

Plates were tested in duplicate not triplicate and the negative result was not confirmed

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-[[5-methyl-2-(1-methylethyl)cyclohexyl]oxy]propane-1,2-diol

EC Number:

289-296-2

EC Name:

3-[[5-methyl-2-(1-methylethyl)cyclohexyl]oxy]propane-1,2-diol

Cas Number:

87061-04-9

Molecular formula:

C13H26O3

IUPAC Name:

3-{[5-methyl-2-(propan-2-yl)cyclohexyl]oxy}propane-1,2-diol

Test material form:

liquid

Method

Target gene:

Histidine synthesis in S. typhimurium strains, and tryptophan synthesis in the E. coli strain tested.

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media: Nutrient Broth No. 2 (Oxoid)

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

- Type and identity of media: Nutrient Broth No. 2 (Oxoid)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

Range finding test: 0, 0.305, 1.22, 4.88, 19.5, 78.1, 313, 1250 and 5000 µg/plate
Main test: 0, 39.1, 78.1, 156, 313, 625 and 1250 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 2-(2-furyl)-3- (5-nitro-2-furyl) acrylamide; 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

NUMBER OF REPLICATIONS: Performed in duplicate

DETERMINATION OF CYTOTOXICITY
- Method: assessment of the bacterial lawn growth

Evaluation criteria:

Plates were assessed for a significant increase in the number of revertant colonies.

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:

cytotoxicity

Remarks:

Observed at the two highest doses tested

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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:

Observed at the two highest doses tested

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
Based on the results of the range finding test, the maximum concentration selected for the main test was 1250 µg/plate as the test material caused a reduction in the growth of the bacterial lawn at concentrations above this dose.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Dose response curves for the number of revertants both with and without metabolic activation are presented in the attached figures.

Table 1: Results of the main test

Metabolic activation	Test material concentration	Replicate	Number of revertant colonies per plate
			Base-pair substitution			Frameshift
			TA 100	TA 1535	WP-2uvrA	TA 98	TA 1537
- S9 Mix	Solvent control	R1	148	9	17	16	9
		R2	156	7	17	12	14
		Mean	(152)	(8)	(17)	(14)	(12)
	39.1	R1	138	11	14	14	13
		R2	160	13	15	17	6
		Mean	(149)	(12)	(15)	(16)	(10)
	78.1	R1	159	6	14	17	13
		R2	155	11	18	15	11
		Mean	(157)	(9)	(16)	(16)	(12)
	156	R1	152	6	16	22	6
		R2	152	8	22	14	9
		Mean	(152)	(7)	(19)	(18)	(8)
	313	R1	151	9	17	7	7
		R2	166	8	19	7	8
		Mean	(159)	(9)	(18)	(7)	(8)
	625	R1	62*	6*	13*	0*	2*
		R2	82*	4*	10*	5*	2*
		Mean	(72)	(5)	(12)	(3)	(2)
	1250	R1	0*	0*	0*	0*	0*
		R2	0*	0*	0*	0*	0*
		Mean	(0)	(0)	(0)	(0)	(0)
+ S9 Mix	Solvent control	R1	173	9	19	24	12
		R2	165	9	19	23	11
		Mean	(169)	(9)	(19)	(24)	(12)
	39.1	R1	152	8	16	31	11
		R2	167	8	1	28	11
		Mean	(160)	(8)	(16)	(30)	(11)
	78.1	R1	158	13	30	33	9
		R2	198	16	11	26	13
		Mean	(178)	(15)	(21)	(30)	(11)
	156	R1	152	12	23	31	19
		R2	164	16	26	30	9
		Mean	(158)	(14)	(25)	(31)	(14)
	313	R1	170	3	14	19	16
		R2	163	8	11	28	15
		Mean	(167)	(6)	(13)	(24)	(16)
	625	R1	150*	7*	18*	19*	9*
		R2	128	5*	23*	18*	4*
		Mean	(117)	(6)	(21)	(19)	(7)
	1250	R1	122*	7*	12*	15*	3*
		R2	125	4*	8*	20*	1*
		Mean	(124)	(6)	(10)	(18)	(2)
Positive controls - S9 Mix	Positive control substance		AF-2	NaN3	AF-2	AF-2	9-AA
	Concentration (µg/plate)		0.01	0.5	0.01	0.1	80
		R1	486	488	93	264	259
		R2	516	491	85	263	285
		Mean	(501)	(490)	(89)	(264)	(272)
Positive controls + S9 Mix	Positive control substance		2-AA	2-AA	2-AA	2-AA	2-AA
	Concentration (µg/plate)		1.0	2.0	10	0.5	2.0
		R1	826	286	1292	239	116
		R2	867	311	1369	255	112
		Mean	(847)	(299)	(1331)	(247)	(114)
AF-2 = 2-(2-furyl)-3- (5-nitro-2-furyl) acrylamide NaN3= sodium azide 9-AA = 9-aminoacridine 2-AA = 2 -aminoanthracene *Inhibition of bacterial growth

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative Both with and without metabolic activation

Under the conditions of the test, the test material was found to be negative for mutagenicity in both the S. typhimurium and E. coli strains tested in the presence and absence of metabolic activation.

Executive summary:

The mutagenicity of the test material was assessed using a bacterial reverse mutagenicity assay (Ames test). The study was performed in compliance with the current OECD guidelines 471 and 472 with a few minor deviations from the standard protocol. Under the conditions of the test, the test material was found to be negative in all strains tested both with and without metabolic activation.