Alcohols, C12-14

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

other: published data

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(no TA102 or E. coli WP2 uvrA; 2-AA only positive control with S9)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Hexadecan-1-ol

EC Number:

253-149-0

EC Name:

Hexadecan-1-ol

Cas Number:

36653-82-4

Molecular formula:

C16H34O

IUPAC Name:

hexadecan-1-ol

Test material form:

solid: compact

Details on test material:

- Name of test material (as cited in study report): Kalcohl 6098 (1-hexadecanol)- Substance type: white granular solid- Physical state: solid- Analytical purity: no data- Impurities (identity and concentrations): no data- Composition of test material, percentage of components: no data- Isomers composition: no data- Purity test date: no data- Lot/batch No.: 2439- Expiration date of the lot/batch: no data- Stability under test conditions: no data- Storage condition of test material: room temperature in the dark

Method

Target gene:

histidine

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor 1254-induced male rat livers

Test concentrations with justification for top dose:

50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: not stated

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9, 3 µg/plate for TA100, 5 µg/plate for TA1535

Positive control substance:

other: 1103

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9, 80 µg/plate for TA1537

Positive control substance:

other: 28

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9, 5 µg/plate for TA1538

Positive control substance:

other: 1342 4-nitro-o-phenylenediamine

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

without S9, 0.2 µg/plate for TA98

Positive control substance:

other: 25

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

with S9, all strains, 0.5, 1 or 2 µg/plate

Positive control substance:

other: 1342 2-aminoanthracene

Details on test system and experimental conditions:

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

DURATION
- Preincubation period: not applicable
- Exposure duration: 48 hours

NUMBER OF REPLICATES: duplicate test, each performed with triplicate plates

DETERMINATION OF CYTOTOXICITY
- Method: growth of bacterial lawn

Evaluation criteria:

For a substance to be considered positive, it should have induced a concentration-related and statistically significant increase in mutation rate in one or more starins of bacteria in the presence and/or absence of S9 in both experiments at sub-toxic concentrations. To be considered negative the number of induced revertants compared to spontaneous revertants should be less than two-fold at each concentration employed.

Statistics:

Methods recommended by the United Kingdom Environmental Mutagen Society and normally Dunnett's method of linear regression

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:

no cytotoxicity

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Water solubility: no data
- Precipitation: at 5000 mg/plate, but this did not interfere with scoring of revertant colonies
- Other confounding effects: no data

RANGE-FINDING/SCREENING STUDIES: yes, non-toxic up to 5000 ug/plate in TA100

COMPARISON WITH HISTORICAL CONTROL DATA: vehicle control results said to be "within the normal range"

ADDITIONAL INFORMATION ON CYTOTOXICITY: no further data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

STATISTICAL RESULTS: Dunnetts test was used and showed no statistically significant differences between test and control plates.

Table 1 Experiment 1 Revertants per plate (mean of 3 plates)

Concentration       µg/plate	TA 100	TA 100	TA 1535	TA 1535	TA 1538	TA 1538	TA 98	TA 98	TA 1537	TA 1537
	- MA	+ MA	- MA	+ MA	- MA	+ MA	- MA	+ MA	- MA	+ MA
0	107	99	33	19	15	21	19	34	8	12
50	91	85	34	12	14	25	24	34	11	10
150	92	89	27	15	17	28	25	35	11	11
500	88	86	23	15	20	29	25	43	10	13
1500	95	98	30	16	16	19	25	40	9	13
5000	95	95	29	18	16	26	23	39	6	12
Positive control	419	672	137	171	544	235	148	236	277	209

 

Table 2 Experiment 2 Revertants per plate (mean of 3 plates)

Concentration       µg/plate	TA 100	TA 100	TA 1535	TA 1535	TA 1538	TA 1538	TA 98	TA 98	TA 1537	TA 1537
	- MA	+ MA	- MA	+ MA	- MA	+ MA	- MA	+ MA	- MA	+ MA
0	90	113	20	11	14	11	17	27	7	12
50	82	111	21	12	19	14	18	26	8	9
150	87	102	25	12	10	13	17	32	9	8
500	81	107	14	15	13	13	17	27	8	7
1500	89	104	19	14	8	9	17	24	9	10
5000	74	90	15	11	8	11	17	28	9	10
Positive control	530	600	171	195	589	196	152	305	496	206

 

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results :negative

In a reliable study, performed according to OECD guideline 471, the C16 alcohol Kahlcol 6098 did not increase the reverse mutation rate in histidine dependent bacterial strains of Salmonella typhimurium in the presence or absence of metabolic activation at concentrations up to 5000 µg/plate. This concentration was not cytotoxic.