Alcohols, C12-15-branched and linear

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1993

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, C12-16

EC Number:

272-490-6

EC Name:

Alcohols, C12-16

Cas Number:

68855-56-1

IUPAC Name:

tetradecan-1-ol

Method

Target gene:

Histidine operon.

Species / strain

Species / strain / cell type:

S. typhimurium, other: strains TA 98, TA 100, TA 1535, TA 1537, and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

1.25 to 5000 µg/plate (200 µg/plate for TA 1537)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: none given in report

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100 and TA 1535 without metabolic activation

Migrated to IUCLID6: 2 µg/plate

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene, 5 µg/plate

Remarks:

TA 1537 without metabolic activation

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 98, TA 100 with metabolic activation

Migrated to IUCLID6: 50 µg/plate

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 without metabolic activation

Migrated to IUCLID6: 50 µg/plate

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: glutaraldehyde 25 µg/plate

Remarks:

TA 102 without metabolic activation

Details on test system and experimental conditions:

ACTIVATION: The activity of the post-mitochondrial fraction was checked by the supplier. S9 mix contained 10% S9 fraction. 0.5 ml S9 mix was added to 2.5 ml of agar.

METHOD OF APPLICATION: in agar (plate incorporation); preincubation (Experiment 2 with metabolic activation)

DURATION
- Preincubation period: 1 h
- Exposure duration: 3 days
- Expression time (cells in growth medium): 3 days

SELECTION AGENT (mutation assays): histidine-deficient agar

NUMBER OF REPLICATIONS: Triplicate plates, experiment repeated

DETERMINATION OF CYTOTOXICITY
- Method: other: condition of bacterial lawn and/or reduction in the number of revertants.

Evaluation criteria:

Considered to be mutagenic if test was valid and reproducible and Dunnetts test gave a positive response (p<0.01) with a significant dose correlation.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 10 µg/plate (TA 1537)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

PRECIPITATION CONCENTRATION: Precipitation observed at 1000 µg/plate and above.

CYTOTOXIC CONCENTRATION:
- With and without metabolic activation: There was variability in the toxic response of the various tester strains to Dobanol 25. As a result, in the second experiment various additional dose levels were added. Strain TA 1537 was particularly sensitive and slight toxicity was observed at 10 µg/plate. There was no obvious toxicity with TA 1535 up to 1000 µg/plate although slight toxic effects may have been obscured by precipitation at this level and above. Toxic effects were seen in TA98 and 100 with S9 at 250 µg/plate and with TA102 at 62.5 µg/plate +S9 and at 250 µg/plate -S9. It was concluded that a sufficient number of dose levels were assessed where toxicity did not confound the result.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1 Experiment 1 Plate incorporation: Number of revertants per plate (mean of 3 plates)

Strain	TA98	TA100	TA1535	TA1537 *	TA102	TA98	TA100	TA1535	TA1537	TA102
Dose	- MA					+MA
0	11.8	88.8	10.6	6.2	317.2	23.2	98.6	17.6	14	448
0.32	NT	NT	NT	8.7	NT	NT	NT	NT	NT	NT
1.6	NT	NT	NT	8	NT	NT	NT	NT	NT	NT
8	13.7	78.7	16	8	293.3	28	103	12.3	12.3	399.3
40	13	93	13.7	8.3	338.7	22.7	107.3	14.3	11.3	301
200	10.3	65	13.3	9	289	19.7	95	15	8.7	304.7
1000	10	56.7	13.7	NT	246.7	15	78	13.7	9.3	266.7
5000	7	48.7	11	NT	195	13.7	79	11.3	7.7	221
Positive control	1289.7	637.7	454	603.3	811.3	1154.3	1573	NT	NT	NT

NT Not tested

Table 2 Experiment 2 Plate incorporation, without activation: Number of revertants per plate (mean of 3 plates)

Strain	TA98	TA100	TA1535	TA1537 *	TA102 *
Dose	- MA
0	15.6	85.4	6.8	10	325.6
62.5	13.3	60.7	9	(2.5) 9.7	(12.5) 292.3
125	12.3	76.7	8.7	(5) 7	(25) 280
250	11.7	55.3	7.7	(10) 9	(50)  292.7
500	9	63.3	6.7	(20) 11.3	(100} 295.3
1000	5.7	57.7	9	(40) 5.7	(200) 253.7
Positive control	1231.3	615.3	474.7	610	1002.7

Figures in brackets indicate doses where these differ from column 1

NT Not tested

Table 3 Experiment 2 Pre-incubation, with activation: Number of revertants per plate (mean of 3 plates)

 

Strain	TA98 *	TA100 *	TA1535	TA1537 *	TA102 *
Dose	+ MA
0	18	106.4	12.8	12	344
6.25	NT	NT	NT	NT	368
12.5	19.3	109	(62.5) 15	(1.25) 7	340
25	24	97	(125) 19.3	(2.5) 5	317
50	21.3	100.7	(250) 9	(5) 11.7	327
100	16.7	99	(500) 16.7	(10) 8	304
200	12.7	92.3	(1000) 11.7	(20) 6	NT
Positive control	620.3	470.7	NT	NT	NT

Figures in brackets indicate doses where these differ from column 1

NT Not tested

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

In a valid and reliable study according to OECD TG 171 and under GLP, C12-16 alcohol Dobanol 25 did not increase the reverse mutation rate in histidine dependent bacterial strains of Salmonella typhimurium in the presence or absence of metabolic activation at dose levels including those which showed some degree of cytotoxicity. The results of the initial plate incorporation experiment were confirmed in the independent repeat experiment which used the plate incorporation method without metabolic activation and the pre-incubation method with metabolic activation. It is concluded that the test substance is negative for mutagenicity to bacteria.