Reaction mass of 2-Hexyl-1-dodecanol and 2-Octyl-1-decanol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 21 to 22, 2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: well documented study performed according to OECD guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, C16-20-branched

EC Number:

274-758-8

EC Name:

Alcohols, C16-20-branched

Cas Number:

70693-04-8

Molecular formula:

not applicable, UVCB

IUPAC Name:

Alcohols, C16-20-branched

Details on test material:

- Name of test material (as cited in study report): ISOFOL 18T
- Substance type: pure active substance
- Physical state: colourless liquid
- Lot/batch No.: 0183/MA
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: no data

Method

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:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

liver homogenate from Arochlor-1254 induced rats

Test concentrations with justification for top dose:

0.05, 0.1, 0.5, 1.0 and 5.0 mg/plate

Vehicle / solvent:

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

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Sodium azide (50 µg/ml) for Salmonella typhimurium TA100 and TA 1535; 2-Nitrofluorene (50 µg/ml) for Salmonella typhimurium TA98; 9-Aminoacridine (1000 µg/ml) for Salmonella typhimurium TA1537; Methyl methanesulfonate (1% v/v) for E. coli WP2 uvrA

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (10 µg/ml) for Salmonella typhimurium TA 100, TA98, TA1535 and TA1537; 2-Aminoanthracene (100 µg/ml) for E. coli strain WP2 uvrA)

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48-72 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- test on cytotoxicity only on Salmonella thyphimurium strain TA 100 and E. Coli WP2 uvrA in a preliminary test at concentrations of 0.05, 0.1, 0.5, 1.0 and 5.0 mg/plate; evaluation of toxicity based on 1. revesion frequency, 2. viability and 3. integrity of the background

OTHER EXAMINATIONS:
Other:
- verification of genetic markers of test strains; histidine requirement, crystal violet sensitivity, ampicillin resistence, ultraviolet sensitivity and spontaneous reversion rates for Salmonella typhimurium; tryptophan requirment and sensitivity to Mitomycin C and ampicillin resistance for E. coli strain
- Determination of the exact cell concentration at the initiation of the test by a % transmittance reading at 650 nm and verification by plate count determination

Evaluation criteria:

A positive response is considered if there is a statistically significant difference (at p ¿ 0.05) in the number of revertants between the solvent control and the test article; and either strain TA98 or TA100 responds to a dose that produces a mean reversion frequency of the corresponding solvent control plates, or if strains TA 1535, TA1537, WP2 uvrA respond to a dose producing a threefold increase in the mean reversion frequency. In addition, the response must be dose-dependent or increasing concentrations of the test article must show increasing mean reversion frequencies. In evaluating the results, consideration is given to the degree of toxicity exhibited by the dose causing the twofold/threefold or greater increase in reversion frequency and the magnitude of the increase in reversion frequency.

A negative response is considered if there is no statistically significant difference (at p=0.05) in the number of revertants between the solvent control and the test article. A response is considered negative if strains TA98 and TA100 respond to a dose that produces mean reversion frequencies that are less than twice that of the mean reversion frequencies of the corresponding solvent control plates; strains TA1535, TA1537, WP2 uvrA respond to a dose producing reversion frequencies that are less than three times that of the corresponding solvent control plates, and there is no evidence of a dose-dependent response.

A response is considered equivocal if it does not fulfil the criteria of either a negative or a positive response and/or the Study Director does not consider the response to be either positive or negative.

Statistics:

One Way Analysis of Variance

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
preliminary screening test with 0.05, 0.1, 0.5, 1.0 and 5.0 mg/plate on tester strains Salmonella typhimurium TA100 and E. coli WP2 uvra. The test article was found to be non cytotoxic at these dose levels for both stains.

Any other information on results incl. tables

Table 1: No. of revertants (mean values and standard deviations; MA = metabolic activation)
	S. typhimurium TA98
	- MA		+ MA
Concentration (mg/plate)	1. experiment	2. experiment	1. experiment	2. experiment
ethanol	45 ± 3.5	41 ± 2.1	47 ± 1.7	49 ± 4.2
0.05	47 ± 1.0	42 ± 1.5	46 ± 0.6	49 ± 7.2
0.1	44 ± 3.8	41 ± 6.7	43 ± 1.5	49 ± 6.4
0.5	40 ± 2.0	41 ± 4.5	46 ± 1.0	48 ± 7.5
1.0	44 ± 3.5	41 ± 0.0	44 ± 1.7	51 ± 3.8
5.0	47 ± 1.2	38 ± 2.6	44 ± 1.2	51 ± 5.5
positive control	1092 ± 42.1	1056 ± 48.0	831 ± 25.7	816 ± 96.0
	S. typhimurium TA100
	- MA		+ MA
Concentration (mg/plate)	1. experiment	2. experiment	1. experiment	2. experiment
ethanol	131 ± 6.1	124 ± 6.7	124 ± 4.0	127 ± 10.1
0.05	129 ± 0.6	125 ± 5.0	122 ± 2.1	123 ± 9.5
0.1	128 ± 1.7	129 ± 0.6	125 ± 1.2	121 ± 11.0
0.5	131 ± 6.4	125 ± 6.4	124 ± 4.0	126 ± 9.2
1.0	131 ± 4.6	127 ± 1.0	123 ± 4.9	124 ± 3.8
5.0	123 ± 4.4	117 ± 10.1	123 ± 6.1	121 ± 19.2
positive control	1287 ± 77.7	1125 ± 82.1	943 ± 26.6	961 ± 12.2
	S. typhimurium TA1535
	- MA		+ MA
Concentration (mg/plate)	1. experiment	2. experiment	1. experiment	2. experiment
ethanol	19 ± 2.1	23 ± 3.2	17 ± 2.1	14 ± 3.8
0.05	21 ± 8.5	22 ± 2.9	22 ± 4.0	13 ± 1.7
0.1	13 ± 1.2	25 ± 3.8	12 ± 0.6	15 ± 1.7
0.5	16 ± 0.6	26 ± 3.6	17 ± 3.2	16 ± 3.1
1.0	13 ± 3.2	25 ± 4.9	18 ± 3.8	15 ± 3.0
5.0	19 ± 4.0	23 ± 1.7	22 ± 1.7	21 ± 1.0
positive control	1205 ± 56.2	1190 ± 76.8	82 ± 5.7	88 ± 1.5
	S. typhimurium TA1537
	- MA		+ MA
Concentration (mg/plate)	1. experiment	2. experiment	1. experiment	2. experiment
ethanol	16 ± 1.2	16 ± 3.5	14 ± 2.0	18 ± 1.0
0.05	16 ± 2.9	19 ± 3.1	14 ± 3.1	18 ± 2.3
0.1	18 ± 1.0	16 ± 3.5	13 ± 2.5	18 ± 4.0
0.5	14 ± 4.0	15 ± 1.0	15 ± 2.0	17 ± 2.9
1.0	16 ± 2.6	19 ± 4.2	15 ± 2.1	16 ± 4.0
5.0	15 ± 3.0	15 ± 1.7	14 ± 1.5	13 ± 4.9
positive control	1291 ± 33.3	1257 ± 26.6	33 ± 3.5	34 ± 4.4
	E. coli WP2 uvrA
	- MA		+ MA
Concentration (mg/plate)	1. experiment	2. experiment	1. experiment	2. experiment
ethanol	30 ± 5.6	28 ± 3.5	32 ± 8.4	34 ± 11.9
0.05	27 ± 2.1	28 ± 1.7	34 ± 5.1	35 ± 8.7
0.1	27 ± 3.8	29 ± 3.5	31 ± 2.1	33 ± 4.5
0.5	28 ± 8.7	29 ± 6.2	41 ± 5.5	35 ± 7.2
1.0	23 ± 3.0	28 ± 1.5	33 ± 6.4	32 ± 2.9
5.0	25 ± 3.8	25 ± 7.6	34 ± 4.0	31 ± 9.5
positive control	477 ± 16.7	530 ± 62.0	96 ± 7.6	103 ± 2.6

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Based on the results of this study under the experimental conditions alcohols 16-20, branched do not induce gene mutations by base pair
changes or frameshifts in the genome and are therefore not mutagenic.

Executive summary:

Based on the results of this study under the experimental conditions alcohols 16-20, branched do not induce gene mutations by base pair

changes or frameshifts in the genome and are therefore not mutagenic.