Tetradecanol

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:

19-Sep-1996 to 24-Sep-1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The restriction was that no cross linking strain was used, so does not comply with current guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

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 (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tetradecanol

EC Number:

204-000-3

EC Name:

Tetradecanol

Cas Number:

112-72-1

Molecular formula:

C14H30O

IUPAC Name:

tetradecan-1-ol

Test material form:

solid

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:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

Test 1: 15(-S9 only), 50, 150, 500, 1500, and 5000 µg/plate; Test 2: 50, 150, 500, 1500, and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Well-known solvent

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

TA100 (3 ug/plate) and TA1535 (5 ug/plate) without S9

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

TA1537 (8 ug/plate) without S9

Positive control substance:

9-aminoacridine

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

TA1538 (5 ug/plate) without S9

Positive control substance:

other: 4-nitro-o-phenylenediamine

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

TA98 (0.2 ug/plate) without S9

Positive control substance:

4-nitroquinoline-N-oxide

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

all strains (0.5, 1 or 2 ug/plate) with S9

Positive control substance:

other: 2-aminoanthracene

Details on test system and experimental conditions:

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

DURATION
- Exposure duration: approximately 48 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Thinning or absence of background lawn of non-revertant cells

OTHER: The mutation experiment was repeated on a separate day using fresh cultures and fresh test material formulations.

Evaluation criteria:

After incubation, numbers of revertant colonies were counted. For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of S9 in both experiments at sub-toxic dose levels. For a negative result the numbers of induced revertants should be less than two fold compared to controls.

Statistics:

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

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended 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: precipitation of test substance was observed at dose levels of 1500 ug/plate and above. Plates were counted manually at 1500 ug/plate and above.
- Other confounding effects: no data

RANGE-FINDING/SCREENING STUDIES: Strain TA 100 was exposed to the test substance at concentrations of 0, 50, 150, 500, 1500 and 5000 ug/plate in a preliminary toxicity study.

COMPARISON WITH HISTORICAL CONTROL DATA: no data

CYTOTOXIC CONCENTRATION: Slight cytotoxicity was indicated in a preliminary toxicity screen with TA100 at dose levels >= 500 ug/plate without 
metabolic activation. In the actual mutation study there was no evidence of cytotoxicity up to 5000 ug/plate with or without S9.

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table 1:Number of revertants per plate (mean of 3 plates) for Test 1

Conc. [µg /plate]	[TA 100]			[TA 1535]			[TA 1538]			[TA 98]			[TA 1537]
	-MA	+MA	Cytotoxic (yes/no)	- MA	+MA	Cytotoxic (yes/no)	- MA	+MA	Cytotoxic (yes/no)	-MA	+ MA	Cytotoxic (yes/no)	-MA	+ MA	Cytotoxic (yes/no)
0*	72	116	no	14	15	no	14	25	no	20	29	no	6	9	no
15	71	-	no	12	-	no	11	-	no	18	-	no	7	-	no
50	65	112	no	12	15	no	11	27	no	17	24	no	6	12	no
150	64	98	no	12	12	no	12	22	no	18	35	no	7	12	no
500	57	89	no	12	14	no	11	28	no	21	27	no	8	10	no
1500	55	72	no	12	17	no	11	22	no	19	27	no	7	9	no
5000	52	74	no	12	11	no	12	22	no	23	30	no	10	7	no
Positive control	597	831	no	348	195	no	636	488	no	217	485	no	550	282	no

*solvent control with DMSO

 

Table 2: Number of revertants per plate (mean of 3 plates) for Test 2

Conc. [µg /plate]	[TA 100]			[TA 1535]			[TA 1538]			[TA 98]			[TA 1537]
	-MA	+MA	Cytotoxic (yes/no)	-MA	+MA	Cytotoxic (yes/no)	-MA	+MA	Cytotoxic (yes/no)	-MA	+MA	Cytotoxic (yes/no)	-MA	+MA	Cytotoxic (yes/no)
0*	86	85	no	19	12	no	11	16	no	15	24	no	7	9	no
50	67	83	no	15	13	no	10	14	no	19	28	no	7	8	no
150	70	66	no	11	9	no	9	13	no	20	29	no	5	7	no
500	66	71	no	17	10	no	9	14	no	15	26	no	6	11	no
1500	66	72	no	9	11	no	7	12	no	13	20	no	5	8	no
5000	66	58	no	9	10	no	6	13	no	12	23	no	7	9	no
Positive control	459	1005	no	211	208	no	524	344	no	175	345	no	762	266	no

*solvent control with DMSO

 

 

Applicant's summary and conclusion

Conclusions:

In a reliable study, conducted in accordance with OECD guideline 471 and under GLP, the C14 alcohol Kalcol 4098 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. The top concentration was not cytotoxic. It is concluded that the test substance is not mutagenic to bacteria under the conditions of the test.

Executive summary:

In the bacterial mutagenicity study, the ability of tetradecan-1 -ol to induce mutations in bacteria was tested.

In test I, 50, 150, 500, 1500, and 5000 µg/plate of test material were applied to S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538 without metabolic activation. In test II, the same concentrations of the test material were applied to

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 1538, in the presence and absence of metabolic activation system. The exposure duration was approximately 48 hours.

After incubation, numbers of revertant colonies were counted. For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of metabolic activation in both experiments at sub-toxic dose levels. For a negative result the numbers of induced revertants should be less than two fold compared to controls.

The study reports tetradecan-1 -ol to be not mutagenic to S. typhimurium when tested up to limit concentration. The study was conducted according to an appropriate OECD test guideline, with acceptable restriction. The restriction was that no cross linking strain was used, so does not comply with current guidelines. It was compliant with GLP.