Reaction mass of 1-(allyloxy)dodecan-2-ol and 1-(allyloxy)tetradecan-2-ol

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:

07 Aug - 30 Aug 2001

Reliability:

2 (reliable with restrictions)

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:

other: Test guideline of Industrial Safety and Health Act

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Duplicate plating was used without scientifically justifications. 2-Aminoanthracene was used as the positive control with the metabolic activation but the metabolic activation by S9-mix was not confirmed. The main test was not repeated.

GLP compliance:

yes

Remarks:

according to Japanese GLP standard

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1-(allyloxy)dodecan-2-ol and 1-(allyloxy)tetradecan-2-ol

EC Number:

944-498-6

Molecular formula:

Not applicable

IUPAC Name:

Reaction mass of 1-(allyloxy)dodecan-2-ol and 1-(allyloxy)tetradecan-2-ol

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL- Lot/batch No.of test material: 000830STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL- Storage condition of test material: at room temperature in the dark- Solubility and stability of the test substance in the solvent/vehicle: insoluble in water, soluble in DMSOTREATMENT OF TEST MATERIAL PRIOR TO TESTING- Final dilution of a dissolved stock liquid: 50 mg/mL in DMSO

Method

Target gene:

trp and his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

other: E. Coli WP2 uvr A-

Metabolic activation:

with and without

Metabolic activation system:

Purcahsed S9, prepared from the livers of rats treated with phenobarbital and 5-6-benzoflavone.

Test concentrations with justification for top dose:

0.625, 1.25, 2.5, 5, 10, and 20 μg/plate without metabolic activation: TA100, TA1535, TA98 and TA1537 tested up to cytotoxic concentrations2.5, 5, 10, 20, 39, and 78 μg/plate with metabolic activation: TA100, TA1535, TA98 and TA1537 tested up to cytotoxic concentrations10, 20, 39, 78, 156, and 313 μg/plate with and without metabolic activation: WP2 uvrA- tested up to the cytotoxic concentration

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO - Justification for choice of solvent/vehicle: due to the limited solubility of the test substance in water, DMSO was selected as the vehicle

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

9-aminoacridine

sodium azide

N-ethyl-N-nitro-N-nitrosoguanidine

other: Furylfuramide for TA100 and TA98 without S9 mix,

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubationDURATION- Preincubation period: 20 minutes- Exposure duration: 48 hNUMBER OF REPLICATIONS: 2 replicates in each test

Evaluation criteria:

A test substance was considered positive in the Ames test if: a) a more than 2-fold increase in the number of revertants was observedb) the increase was dose-related, and c) the positive result was reproducible.A test substance was considered negative in the Ames test if none of the above criteria were met under all experimental conditions.

Statistics:

Mean values were calculated.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Remarks:

, TA100, TA1535, TA1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 10 μg/plate without S9 mix in TA 100, TA 1535, and TA 1537; starting at 20 μg/plate without S9 mix in TA98, starting at 78 μg/plate with S9 mix in TA100, TA1535, TA98 and TA1537.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Remarks:

E. coli WP2 uvr A-

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

starting at 313 μg/plate +/- S9 in WP2 uvrA-

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Precipitation: no precipitationRANGE-FINDING/SCREENING STUDIES:A preliminary test was performed with test concentrations ranging from 0.625 - 5000 μg/plate with/without metabolic activation. No increase in the number of revertants was observed. Furthermore, cytotoxicity was observed at 10 μg/plate in TA100, TA1535, and TA1537 without metabolic activation, at 20 μg/plate in TA98 without metabolic activation, at 313 μg/plate in WP2urvA with and without metabolic activation, at 78 μg/plate in TA100, TA1535, TA98 and TA1537 with metabolic activation.HISTORICAL CONTROL DATA (means (means ± 2 standard deviation))- Positive historical control data: Without metabolic activation in TA100 by AF-2: 453 (378 - 528), TA1535 by NaN3: 346 (269 - 424), WP2urvA- by ENNG: 536 (362 - 710), TA98 by AF-2: 512 (417 - 607), and TA1537 by 9-AA: 197 (57 - 338)With metabolic activation by 2- AA in TA100: 656 (412 - 900), TA1535: 269 (224 - 314), WP2urvA-: 619 (476 - 763), TA98: 407 (295 - 520), and TA1537: 123 (91 - 155)- Negative (solvent/vehicle) historical control data:Without metabolic activation in TA100: 96 (76 - 116), TA1535: 9 (5 - 13), WP2urvA-: 20 (15 - 25), TA98: 17 (12 - 21), and TA1537: 7 (5 - 9)With metabolic activation in TA100: 106 (75 - 136), TA1535: 9 (4 - 14), WP2urvA-: 21 (16 - 27), TA98: 22 (16 - 28), and TA1537: 10 (6 - 13)

Any other information on results incl. tables

Table 1. Results for AHK without metabolic activation in the dose finding study

Without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate
		Base-pair substitution type			Frameshift type
		TA100	TA1535	WP2urvA-	TA98	TA1537
–	0	105	11	24	19	9
–	0.625	111	10	-	18	15
–	1.25	111	9	-	22	12
–	2.5	113	13	-	19	11
–	5	85	6	-	18	13
–	10	30*	0*	26	13	0*
–	20	0*	0*	27	11*	0*
–	39	-	-	25	-	-
–	78	-	-	24	-	-
–	156	-	-	26	-	-
–	313	-	-	30*	-	-
–	Positive controls	AF-2	NaN3	ENNG	AF-2	9-AA
	Mean No. of colonies/plate	526	421	648	583	171

 

Table 2. Results for AHK with metabolic activation in the dose finding study

With S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate
		Base-pair substitution type			Frameshift type
		TA100	TA1535	WP2urvA-	TA98	TA1537
+	0	125	8	21	23	13
+	2.5	102	9	-	24	16
+	5	106	6	-	20	14
+	10	103	6	25	22	18
+	20	102	7	36	18	20
+	39	118	9	35	20	15
+	78	79*	8*	32	20*	4*
+	156	-	-	21	-	-
+	313	-	-	23*	-	-
+	Positive controls	2-AA	2-AA	2-AA	2-AA	2-AA
	Mean No. of colonies/plate	847	314	762	518	154

 

Table 3. Results for AHK without metabolic activation in main study

Without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate
		Base-pair substitution type			Frameshift type
		TA100	TA1535	WP2urvA	TA98	TA1537
–	0	81	6	24	18	9
–	0.625	77	9	-	18	9
–	1.25	79	11	-	24	11
–	2.5	71	7	-	20	13
–	5	72	5	-	19	8
–	10	64*	0*	25	16	0*
–	20	0*	0*	24	10*	0*
–	39	-	-	24	-	-
–	78	-	-	22	-	-
–	156	-	-	27	-	-
–	313	-	-	16*	-	-
–	Positive controls	AF-2	NaN3	ENNG	AF-2	9-AA
	Mean No. of colonies/plate	455	417	606	487	166

Table 4. Test results for AKH with metabolic activation in the main study

With S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate
		Base-pair substitution type			Frameshift type
		TA100	TA1535	WP2urvA	TA98	TA1537
+	0	98	8	23	19	13
+	2.5	96	8	-	22	11
+	5	99	9	-	22	10
+	10	101	5	26	25	11
+	20	87	8	25	20	10
+	39	94	10	25	21	9
+	78	73*	7*	27	17*	4*
+	156	-	-	23	-	-
+	313	-	-	30*	-	-
+	Positive controls	2-AA	2-AA	2-AA	2-AA	2-AA
	Mean No. of colonies/plate	643	312	762	516	153

AF-2= Furylamide

NaN3 = sodium azido

ENNG = N-ethyl-N'-nitro-N'-nitrosogunanidine

9-AA = 9-aminoacridine

2AA = 2-aminoanthracene

* = growth inhibition