Registration Dossier
Registration Dossier
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EC number: 204-000-3 | CAS number: 112-72-1
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutation (Bacterial reverse mutation assay / Ames test): negative with and without activation in S. typhimurium strains TA 98, TA100, TA1535, TA1537 and TA 1538 (similar to OECD Test Guideline 471) (Safepharm Laboratories, 1996). For completeness, a 5th-strain bacterial reverse mutation test is commissioned with the registered substance and will be conducted according to OECD Test Guideline 471 and in compliance with GLP.
Cytogenicity in mammalian cells: the related substance C12 and 13 alcohols; linear and monobranched, type 2 was negative in CHO cells (OECD Test Guideline 473) (Sasol, 1998)
Cytogenicity in mammalian cells: the related substance docosan-1-ol was negative with and without activation in Chinese hamster ovary cells (similar to OECD Test Guideline 473) (Iglesias, 2002b).
An in vitro micronucleus test is commissioned with the registered substance and will be conducted according to OECD Test Guideline 487 and in compliance with GLP.
Mutagenicity in mammalian cells: the related substance docosa-1-ol was negative with and without activation in Chinese hamster lung V79 cells (similar to OECD Test Guideline 476) (Iglesias, 2002b).
Mutagenicity in mammalian cells: the related substance Fatty alcohol blend (containing 40.77% C8 and 55.3% C10): negative with and without activation in L5178Y mouse lymphoma cells (similar to OECD Test Guideline 476) (Inveresk, 1992).
An in vitro mammalian cell gene mutation tests using the thymidine kinase gene is commissioned with the registered substance and will be conducted according to OECD Test Guideline 490 and in compliance with GLP.
Link to relevant study records
- 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.
- 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
- Target gene:
- histidine
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- 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 - 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
- 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
- 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
- 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
- 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.
- 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
- 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
- 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
- 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
- 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
- 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.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1998
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- 1997
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor induced rat liver S9
- Test concentrations with justification for top dose:
- Test 1: 0.1 - 500 µg/ml; Test 2: with S9 1 -50 µg/ml, without S9 0.5 - 20 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: none given in report. Standard solvent - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- without activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with activation
- Details on test system and experimental conditions:
- ACTIVATION: 1 ml Aroclor induced rat liver S9 mix, NADP as cofactor
METHOD OF APPLICATION: in medium
DURATION
Exposure duration: Test 1: +S9 3 hours, -S9 18 hours, Test 2: +S9 3 hours; -S9 18 or 32 hours
- Expression time (cells in growth medium): 18 hours
- Fixation time (start of exposure up to fixation or harvest of cells): Test 1 18 hours; Test 2 18 or 32 hours
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (final concentration 0.1 mM)
TAIN (for cytogenetic assays): Giesma
NUMBER OF REPLICATIONS: Duplicate cultures, independent repeat assay
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; other: observation of culture
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
- Other: interstitial deletions - Evaluation criteria:
- CRITERIA FOR EVALUATING RESULTS: The test is considered positive if the aberration frequency of at least one concentration is significantly above concurrent control frequencies.
- Statistics:
- Fisher's exact probability test (two-sided)
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- +S9 40 µg/ml; -S9 15 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not valid
- Positive controls validity:
- valid
- Additional information on results:
- GENOTOXIC EFFECTS: - With and without metabolic activation:
There were no statistically significant increase in total numbers of chromosome aberrations at any dose level tested
There was no increase in the incidence of polyploids or endoreduplicates.
PRECIPITATION CONCENTRATION: 125 µg/ml.
MITOTIC INDEX: The mitotic index was measured on 1000 cells and was always >40% of control levels and usually >50% for the dose levels which were scored for chromosome aberrations.
STATISTICAL RESULTS: Fischers exact probability test (two-sided) did not indicate any significant difference between test and control groups. - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- Alcohols, C12-13-branched and linear has been tested for clastogenicity in a valid study conducted according to OECD Test Guideline 473 and in compliance with GLP in CHO K1 cells. The test substance did not increase the incidence of chromosome aberrations in Chinese hamster ovary cells at dose levels up to cytotoxic concentrations in the presence or absence of metabolic activation. The result of the repeat experiment confirmed that of the initial assay. Appropriate positive and solvent controls were added and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations under the conditions of this test.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- not stated
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- without detailed documentation
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Principles of method if other than guideline:
- Well-conducted study according to protocol very similar to OECD guideline 473
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- not applicable
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: minimum essential medium
- Properly maintained: no data
- Periodically checked for Mycoplasma contamination: no data
- Periodically checked for karyotype stability: no data
- Periodically "cleansed" against high spontaneous background: no data - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver microsomal fraction from male rats prepared according to Ames et al., 1977
- Test concentrations with justification for top dose:
- 0.6, 10.0 and 20.0 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: solubility - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without metabolic activation
- Positive control substance:
- ethylmethanesulphonate
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- with metabolic activation
- Positive control substance:
- cyclophosphamide
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: not applicable
- Exposure duration: 4 hours
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): 7 and 24 (or 28) hours at 20 µg/ml, 18 hours at 0.6, 10 and 20 µg/ml
SPINDLE INHIBITOR (cytogenetic assays): Colcemid, 0.2 µg/ml
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2 cultures per concentration
NUMBER OF CELLS EVALUATED: 100 per slide, 200 per concentration
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
- Evaluation criteria:
- To be considered positive, either a statistically significant, concentration-related increase in the number of structural chromosome aberrations, or a statistically significant positive response at one of the concentrations
- Statistics:
- Chi-squared test performed for cells with aberration (excluding gaps)
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: presumably >20 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- 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: insoluble
- Precipitation: no data
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES: yes, but no data presented
COMPARISON WITH HISTORICAL CONTROL DATA: no data
ADDITIONAL INFORMATION ON CYTOTOXICITY: at 20 µg/ml, mitotic index not reduced, plating efficiency not reduced - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- In a reliable study, according to a protocol that is similar to OECD 473, behenyl alcohol (C22) did not increase the incidence of chromosome aberrations in Chinese hamster V79 cells in the presence or absence of metabolising fraction at concentrations up to 20 µg/ml. There was no evidence of cytotoxicity at this dose level.
- Executive summary:
In an in vitro chromosome aberration study, Chinese hamster lung fibroblasts (V79) were incubated with 0.6, 10.0 and 20.0 µg/ml of test material dissolved in ethanol for 4 hours, with and without metabolic activation.
The test substance did not increase the incidence of chromosome aberrations in Chinese hamster V79 cells in the presence or absence of metabolic activation when tested up to limit concentration. There was no evidence of cytotoxicity at this dose level. The study was comparable to guideline without detailed documentation (publication). It is considered that read across to the registered substance is valid and scientifically justifiable.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor induced rat liver S9
- Test concentrations with justification for top dose:
- Toxicity assay: 0.4, 4.3, 43.2, 432, 4320 µg/ml; Mutagenicity assay: 9.4, 18.8, 37.5, 75, 150, 300 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO was used in toxicity assay, acetone in mutagenicity assay
- Justification for choice of solvent/vehicle: due to impurity peaks in the chromatograms, solvent was changed to acetone. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 3-methylcholanthrene
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- ACTIVATION: 1.0 ml S9 mix containing 10% S9 and cofactors NADP and glucose-6-phosphate added to give final volume of 10 ml
METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: none
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 11-14 days
SELECTION AGENT (mutation assays): trifluorothymidine
NUMBER OF REPLICATIONS: duplicate cultures, independent repeat experiment
DETERMINATION OF CYTOTOXICITY
- Method: other: cloning efficiency - Evaluation criteria:
- A substance was considered positive if there was an increase of at least 1.7 fold in at least one of the highest doses was significant and associated with an increase in mutant numbers and/or an upward trend in the remaining doses, in two experiments under the same activation conditions.
- Statistics:
- Statistical evaluation was performed if marginal responses were recorded.
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 43.2 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Conclusions:
- Fatty alcohol blend has been tested according to a protocol that is similar to OECD 476 and under GLP. No increase in the mutant frequency was observed with or without metabolic activation in either the initial or repeat experiment up to cytotoxic concentrations. Solvent and positive controls gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.
- Executive summary:
Fatty alcohol blend has been tested according to a protocol that is similar to OECD 476 and under GLP. No increase in the mutant frequency was observed with or without metabolic activation in either the initial or repeat experiment up to cytotoxic concentrations. Solvent and positive controls gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.
The in vitro and in vivo data available for members of the category and supporting substances indicate that the C6-24 alcohols are not genotoxic. In addition, the category of LCAAs under consideration does not contain any structural elements that are of concern for potential mutagenic activity.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- no data
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Principles of method if other than guideline:
- Well-conducted study according to a protocol very similar to OECD guideline 476
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HGPRT
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: no data
- Properly maintained: no data
- Periodically checked for Mycoplasma contamination: no data
- Periodically checked for karyotype stability: no data
- Periodically "cleansed" against high spontaneous background: no data - Metabolic activation:
- with and without
- Metabolic activation system:
- no data, but for Ames test, liver microsomal fractions from male rats prepared by "established methods"
- Test concentrations with justification for top dose:
- 2.0, 7.5, 15.0, and 20.0 ug/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol, final concentration in culture medium <=1% v/v
- Justification for choice of solvent/vehicle: solubility - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without metabolic activation
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- 1.0 ug/ml
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- with metabolic activation
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Remarks:
- 15.4 ug/ml
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: not applicable
- Exposure duration: 4 hours
- Expression time (cells in growth medium): no data
- Selection time (if incubation with a selection agent): no data
- Fixation time (start of exposure up to fixation or harvest of cells): no data
SELECTION AGENT (mutation assays): thioguanine
NUMBER OF REPLICATIONS:
- 2 independent experiments, both with and without metabolic activation
NUMBER OF CELLS EVALUATED: no data
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- To be considered positive, statistically significant concentration-related increase in mutant frequency, or a reproducible and statistically significant positive response for at least one concentration
- Statistics:
- no data
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: presumably >20 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- 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: insoluble
- Precipitation: no data
- Other confounding effects: no data
RANGE-FINDING/SCREENING STUDIES: yes, but no data presented
COMPARISON WITH HISTORICAL CONTROL DATA: no data
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- With metabolic activation: mean relative cell survival over the test concentrations ranged from 89.1% (20 ug/ml) to 93.8% (15 ug/ml).
- Without metabolic activation: mean relative cell survival ranged from 96% (15 ug/ml) to 120.2 % (20 ug/ml). - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- In a reliable study, behenyl alcohol (C22) did not increase the gene mutation rate in Chinese hamster V79 cells in the presence or absence of metabolic activation at concentrations up to 20 ug/ml. It is concluded that the test substance is negative for mutagenicity in mammalian cells under the conditions of this test.
Referenceopen allclose all
Table 1:Number of revertants per plate (mean of 3 plates) for Test 1
Conc. |
[TA 100] |
[TA 1535] |
[TA 1538] |
[TA 98] |
[TA 1537] |
||||||||||
-MA |
+MA |
Cytotoxic |
- MA |
+MA |
Cytotoxic |
- MA |
+MA |
Cytotoxic |
-MA |
+ MA |
Cytotoxic |
-MA |
+ MA |
Cytotoxic |
|
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. |
[TA 100] |
[TA 1535] |
[TA 1538] |
[TA 98] |
[TA 1537] |
||||||||||
-MA |
+MA |
Cytotoxic |
-MA |
+MA |
Cytotoxic |
-MA |
+MA |
Cytotoxic |
-MA |
+MA |
Cytotoxic |
-MA |
+MA |
Cytotoxic |
|
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
Chromosome aberration assay: Test 1
Treatment time 18 hrs |
||||||
Treatment |
Activation |
Concentration µg/ml |
Number of cells |
Total - gaps |
Total + gaps |
|
Negative control DMSO |
-MA |
0 |
200 |
0 |
4 |
|
Test substance
|
-MA |
2.5 |
200 |
0 |
6 |
|
-MA |
5 |
200 |
0 |
6 |
||
-MA |
10 |
200 |
2 |
8 |
||
Positive control Mitomycin C |
-MA |
0.025 |
200 |
48 |
48 |
|
Treatment time 3 hrs |
||||||
Treatment |
Concentration µg /ml |
Number of cells |
Total - gaps |
Total + gaps |
||
Negative control DMSO |
+MA |
0 |
200 |
0 |
2 |
|
Test substance
|
+MA |
10 |
200 |
1 |
6 |
|
+MA |
20 |
200 |
1 |
5 |
||
+MA |
30 |
200 |
1 |
5 |
||
Positive control Cyclophosphamide |
+MA |
3.75 |
200 |
48 |
48 |
|
Chromosome aberration assay: Test 2
Treatment |
-MA |
Concentration µg /ml |
Number of cells |
Total - gaps |
Total + gaps |
Treatment time 18 hrs |
|||||
Negative control DMSO |
-MA |
0 |
200 |
1 |
5 |
Test substance
|
-MA |
5 |
200 |
2 |
8 |
-MA |
7.5 |
200 |
0 |
5 |
|
-MA |
10 |
200 |
1 |
4 |
|
Positive control Mitomycin C |
-MA |
0.025 |
200 |
54 |
54 |
Treatment time 3 hrs, incubation 18 hours |
|||||
Treatment |
Concentration µg /ml |
Number of cells |
Total - gaps |
Total + gaps |
|
Negative control DMSO |
+MA |
0 |
200 |
0 |
8 |
Test substance
|
+MA |
10 |
200 |
1 |
2 |
+MA |
20 |
200 |
1 |
7 |
|
+MA |
30 |
200 |
1 |
7 |
|
Positive control Cyclophosphamide |
+MA |
3.75 |
200 |
105 |
105 |
Treatment time 32 hrs |
|||||
Treatment |
Concentration µg /ml |
Number of cells |
Total - gaps |
Total + gaps |
|
Negative control DMSO |
-MA |
0 |
200 |
0 |
8 |
Test substance |
-MA |
10 |
200 |
0 |
7 |
Treatment time 3 hrs, incubation 32 hours |
|||||
Treatment |
Concentration µg /ml |
Number of cells |
Total - gaps |
Total + gaps |
|
Negative control DMSO |
+MA |
0 |
200 |
2 |
3 |
Test substance |
+MA |
30 |
200 |
0 |
5 |
Table 1 Cytogenicity: 7 hour fixation. Aberrations in 200 cells
Activation |
Concentration µg/ml |
Percent aberrant cells |
||
incl gaps |
excl gaps |
exchanges |
||
Without |
0* |
4.0 |
1.5 |
0 |
20 |
2.5 |
0.5 |
0 |
|
With |
0* |
4.0 |
1.5 |
0 |
20 |
7.0 |
2.5 |
0 |
|
* Solvent control with ethanol
** Only 100 cells counted for positive controls
Table 2 Cytogenicity: 18 hour fixation. Aberrations in 200 cells
Activation |
Concentration µg/ml |
Percent aberrant cells |
||
incl gaps |
excl gaps |
exchanges |
||
Without |
Negative control |
5.5 |
1.5 |
0 |
0* |
4.0 |
1.5 |
0.5 |
|
0.6 |
4.5 |
2.0 |
0 |
|
10 |
4.0 |
1.0 |
0.5 |
|
20 |
3.0 |
0.5 |
0 |
|
Positive control** |
12.0 |
9.0 |
4.0 |
|
With |
Negative control |
2.5 |
1.5 |
0 |
0* |
2.5 |
1.5 |
0.5 |
|
0.6 |
5.5 |
3.0 |
0.5 |
|
10 |
4.0 |
2.5 |
0 |
|
20 |
4.0 |
2.5 |
0.5 |
|
Positive control** |
16.0 |
13.0 |
5.5 |
|
* Solvent control with ethanol
** Only 100 cells counted for positive controls
Table 3 Cytogenicity: 18 hour fixation. Aberrations in 200 cells
Activation |
Concentration µg/ml |
Percent aberrant cells |
||
incl gaps |
excl gaps |
exchanges |
||
Without |
0* |
6.0 |
2.5 |
0.5 |
20 |
3.5 |
2.0 |
0 |
|
With |
0* |
1.0 |
0.5 |
0 |
20 |
4.0 |
2.5 |
0.5 |
|
* Solvent control with ethanol
** Only 100 cells counted for positive controls
Table 1 Experiment 1 Mutant frequency (average of 3 plates per culture)
Concentration µg/ml |
Relative total growth % |
Mean mutant count (MC) |
Mutant fraction x 10¿¿ |
Increase over control |
||||
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
|
Solvent control |
81 |
92 |
19 |
24 |
27 |
36 |
-
|
- |
112 |
103 |
32 |
25 |
35 |
36 |
|||
102 |
101 |
25 |
31 |
30 |
35 |
|||
104 |
103 |
26 |
27 |
31 |
32 |
|||
Positive control |
75 |
34 |
182 |
151 |
264 |
299 |
8.6 |
7.6 |
63 |
38 |
150 |
135 |
268 |
235 |
|||
9.4 |
129 |
85 |
17 |
30 |
21 |
36 |
1.0 |
0.9 |
100 |
88 |
29 |
18 |
39 |
26 |
|||
18.8 |
111 |
92 |
27 |
21 |
31 |
25 |
1.2 |
0.7 |
109 |
71 |
39 |
19 |
44 |
24 |
|||
37.5 |
72 |
92 |
18 |
24 |
25 |
34 |
0.9 |
1.0 |
79 |
71 |
27 |
27 |
33 |
35 |
|||
75 |
- |
- |
NP |
NP |
- |
- |
- |
- |
- |
- |
NP |
NP |
- |
- |
|||
150 |
- |
- |
NP |
NP |
- |
- |
- |
- |
- |
- |
NP |
NP |
- |
- |
|||
300 |
- |
- |
NP |
NP |
- |
- |
- |
- |
- |
- |
NP |
NP |
- |
|
|||
NP = Not plated, too toxic for assessment
Table 2 Experiment 2 Mutant frequency (average of 3 plates per culture)
Concentration µg/ml |
Relative total growth % |
Mean mutant count (MC) |
Mutant fraction x 10¿¿ |
Increase over control |
||||
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
|
Solvent control |
94 |
99 |
25 |
35 |
28 |
43 |
- |
- |
113 |
110 |
30 |
41 |
26 |
43 |
|||
92 |
99 |
29 |
31 |
28 |
37 |
|||
- |
93 |
- |
30 |
- |
37 |
|||
Positive control |
84 |
20 |
152 |
107 |
182 |
315 |
7.5 |
7.7 |
83 |
21 |
172 |
122 |
221 |
298 |
|||
10 |
103 |
- |
31 |
NP |
33 |
- |
1.3 |
- |
98 |
- |
32 |
NP |
39 |
- |
|||
20 |
128 |
- |
30 |
NP |
29 |
- |
1.1 |
- |
86 |
- |
28 |
NP |
29 |
- |
|||
30 |
93 |
90 |
21 |
27 |
26 |
30 |
1.1 |
0.9 |
91 |
79 |
29 |
40 |
32 |
39 |
|||
40 |
57 |
90 |
34 |
38 |
48 |
42 |
1.3 |
0.9 |
84 |
94 |
20 |
29 |
22 |
33 |
|||
50 |
32 |
91 |
20 |
31 |
26 |
31 |
1.0 |
0.8 |
29 |
93 |
21 |
32 |
30 |
36 |
|||
60 |
- |
71 |
NP |
36 |
- |
36 |
- |
0.8 |
- |
56 |
NP |
24 |
- |
28 |
|||
70 |
- |
37 |
NP |
22 |
- |
25 |
- |
0.6 |
- |
23 |
NP |
25 |
- |
26 |
|||
80 |
- |
- |
NP |
NP |
- |
- |
- |
- |
- |
- |
NP |
NP |
- |
- |
|||
NP = Not plated: 3 highest dose levels, too toxic for assessment
Table 1 Results of mutagenicity in V79 cells (mean of 2 cultures)
Concentration µg/ml |
Mean relative cell survival (%) |
Mean mutants per culture |
Mutant colonies per 10 E06 cells |
|||
-MA |
+MA |
-MA |
+MA |
-MA |
+MA |
|
Negative |
105.1 |
98.2 |
2.7 |
4.8 |
8.65 |
47.4 |
0* |
100 |
100 |
4.5 |
2.1 |
14.7 |
8.75 |
2 |
101.3 |
92.55 |
4.1 |
6.3 |
12.5 |
21.65 |
7.5 |
102.4 |
93.6 |
5.4 |
4.9 |
16.1 |
15.75 |
15 |
96.0 |
93.8 |
3.4 |
1.7 |
12.3 |
6.35 |
20 |
120.2 |
89.1 |
4.3 |
4.4 |
17.9 |
16.95 |
Positive control |
67.3 |
104.6 |
156.7 |
39.1 |
1143.7 |
163.4 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Mouse micronucleus study: the related substance dodecan-1-ol was
negative in mice after oral administration (gavage) (OECD Test Guideline
474) (Henkel, 1992).
Mouse micronucleus study: the related substance docosan-1-ol was
negative after oral administration (similar to OECD Test Guideline 474)
(Iglesias, 2002b).
Micronucleus study in mice: the related substance Fatty alcohol blend (containing 40.77% C8 and 55.3% C10) was negative after oral administration (OECD Test Guideline 474) (Inveresk, 1992).
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 26 November 1991 to 11 February 1332
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- only 1000 PCE per animal were scored for micronuclei
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- CD-1
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River (UK) Limited, Manston Road, Kent
- Age at study initiation: 5-7 weeks
- Weight at study initiation: 27-30 g (males) 18-23 g (females)
- Assigned to test groups randomly: yes
- Fasting period before study: no information
- Housing: individually in polypropylene/stainless steel cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 10 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19
- Humidity (%): 38
- Air changes (per hr): no information
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle(s)/solvent(s) used: corn oil
- Justification for choice of solvent/vehicle: none given; standard vehicle
- Concentration of test material in vehicle: sufficient to give required dose in appropriate volume of vehicle
- Amount of vehicle (if gavage or dermal): 10 mg/ml/day - Duration of treatment / exposure:
- Animals were dosed on three consecutive days.
- Frequency of treatment:
- daily
- Post exposure period:
- 96 hours
- Dose / conc.:
- 500 mg/kg bw/day (nominal)
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- Dose / conc.:
- 2 000 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 5 (positive control, low and mid dose) or 10 (vehicle control, high dose)
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- - Positive control substance: cyclophosphamide
- Justification for choice of positive control(s): none given - standard positive control
- Route of administration: no information
- Doses / concentrations: 40 mg/ kg bw - Tissues and cell types examined:
- Bone marrow
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: no deaths occurred in preliminary toxicity assay
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Animals dosed at 0, 24 and 48 hours; samples taken at 72 and 96 hours
DETAILS OF SLIDE PREPARATION: Air dried slides were fixed in methanol then stained with 1% May-Grunwald for 5 minutes then counterstained in 15% Giesma for 15 minutes
METHOD OF ANALYSIS: 1000 PCE scored for micronuclei; PCE/NCE ratio was determined for 300 cells, using x 1000 oil immersion objective - Evaluation criteria:
- An increase in micronucleus frequency of greater than 0.28%.
- Statistics:
- No statistical evaluation described.
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Conclusions:
- Fatty alcohol blend has been tested according to OECD 474 and under GLP. Male and female mice were dosed with 500, 1000 and 2000 mg/kg bw. No increase in the number of micronucleated PCE was observed (1000 PCE scored per animal). It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the test. No toxicity to bone marrow or general toxicity was observed.
- Executive summary:
Fatty alcohol blend has been tested according to OECD 474 and under GLP. Male and female mice were dosed with 500, 1000 and 2000 mg/kg bw. No increase in the number of micronucleated PCE was observed (1000 PCE scored per animal). It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the test. No toxicity to bone marrow or general toxicity was observed.
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- not stated
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- Only 1000 erythrocytes were scored per animal, full experimental details were not reported, toxicity details were lacking. It was not compliant with GLP.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- (only 1000 PCEs per animal scored for micronuclei)
- Principles of method if other than guideline:
- Well-conducted study according to protocol very similar to OECD guideline 474
- GLP compliance:
- not specified
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: BRL Tierfarm Fullinsdorf, Switzerland
- Age at study initiation: >=10 weeks
- Weight at study initiation: no data
- Assigned to test groups randomly: no data
- Fasting period before study: 18 hours, but continued to receive water ad libitum
- Housing: Markrolon Type 1 cages with wire mesh tops and granulated soft wood bedding
- Diet (e.g. ad libitum): standard pellet diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: no data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): not regulated
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12
IN-LIFE DATES: no data - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle(s)/solvent(s) used: polyethylene glycol
- Justification for choice of solvent/vehicle: no data
- Concentration of test material in vehicle: no data [calculated: 5, 15 and 50 mg/ml]
- Amount of vehicle (if gavage or dermal): 10 ml/kg bw
- Lot/batch no. (if required): no data
- Purity: no data - Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: few details; test material suspended in vehicle
- Duration of treatment / exposure:
- single administration
- Frequency of treatment:
- single administration
- Post exposure period:
- none
- Dose / conc.:
- 50 mg/kg bw/day (nominal)
- Dose / conc.:
- 150 mg/kg bw/day (nominal)
- Dose / conc.:
- 500 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 6
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- cyclophosphamide
- Justification for choice of positive control(s): no data
- Route of administration: presumably oral gavage
- Doses / concentrations: 40 mg/kg bw - Tissues and cell types examined:
- bone marrow cells
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: based on previous study - 500 mg/kg bw estimated to be the "maximum attainable dose"
TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): 24, 48 and 72 hours after dosing
DETAILS OF SLIDE PREPARATION: femurs removed, marrow flushed out with foetal calf serum, cell suspension centrifuged and supernatant discarded, small drop of cell pellet spread on slide, air dried, stained with May-Grunwald, mounted; 1 slide/sample
METHOD OF ANALYSIS: 1000 polychromatic erythrocytes (PCEs) scored for micronuclei; polychromatic:normochromatic (PCE:NCE) ratio scored
OTHER: only 5/sex per dose level evaluated - Evaluation criteria:
- To be considered positive, either a statistically significant dose-related increase in the number of micronucleated PCEs or a reproducible, statistically significant positive response for at least one dose level
- Statistics:
- Mann-Whitney test
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- not specified
- Remarks:
- presumably toxic at >500 mg/kg bw
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: no data
- Solubility: no data
- Clinical signs of toxicity in test animals: no data
- Evidence of cytotoxicity in tissue analyzed: no data
- Rationale for exposure: no data
- Harvest times: no data
- High dose with and without activation: no data
- Other: presumably toxic above 500 mg/kg bw since this maximum dose was chosen for the main study on the basis of the results of the range-finding study
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): 0.03-0.09% for vehicle controls, 0.04-0.10% for test material treated, 0.71% for positive control
- Ratio of PCE/NCE (for Micronucleus assay): 1.05-1.27 for vehicle controls, 0.98-1.55 for test material treated, 0.93 for positive control
- Appropriateness of dose levels and route: appropriate (top dose was apparently the maximum tolerated dose, oral route relevant to humans)
- Statistical evaluation: no statistically significant increases in the frequency of micronuclei in mice treated with the test material; statistical significance not presented for positive control - Conclusions:
- In a reliable study, behenyl alcohol (C22) did not increase the incidence of micronuclei in mouse bone marrow cells after a single oral gavage dose of up to 500 mg/kg bw.
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 11-Feb-1992 to 27-Apr-1992
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- 1000 erythrocytes counted instead of 2000
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- other: albino mice, CFW 1
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Winkelmann
- Age at study initiation: 7-8 weeks
- Weight at study initiation: males 21-27 g, females 21-26 g
- Assigned to test groups randomly: yes, under following basis: allocated to treatment groups according to randomization table generated by computer programme or manually
- Fasting period before study: yes, overnight until 3-4 hours after dosing
- Housing: males, 1/cage, macrolon cages type I; females, <=3/cage, macrolon cages type II; filled with clean softwood bedding
- Diet (e.g. ad libitum): standard animal diet, Altromin No. 1314, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: >=6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25 +- 3 (occasionally 20-25)
- Humidity (%): 40 - 50 (occasionally 45-70)
- Air changes (per hr): no data, except "air-conditioned room"
- Photoperiod (hrs dark / hrs light): 12 / 12
IN-LIFE DATES (main study): From: 25-Feb-1992 To: 28-Feb-1992 - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle(s)/solvent(s) used: arachis oil
- Justification for choice of solvent/vehicle: test material easily soluble at required concentration
- Concentration of test material in vehicle: not stated, but provided a dose level of 5000 mg/kg bw, so 500 mg/ml
- Amount of vehicle (if gavage or dermal): 10 ml/kg bw (main study), 20 ml/kg bw (range finding study)
- Lot/batch no. (if required): no data
- Purity: no data - Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: 500 mg/ml in arachis oil (main study)
- Duration of treatment / exposure:
- single administration
- Frequency of treatment:
- single administration
- Post exposure period:
- evaluated at 24, 48, 72 hours after administration
- Dose / conc.:
- 5 000 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 6
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- cyclophosphamide
- Justification for choice of positive control(s): not stated
- Route of administration: oral
- Dose: 20 mg/kg bw - Tissues and cell types examined:
- bone marrow
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: maximum tolerated dose, based on range-finding study (effects seen at 5000 mg/kg were piloerection only)
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): single administration, animals sacrificed 24, 48 and 72 hours after administration
DETAILS OF SLIDE PREPARATION: bone marrow collected from femurs, using foetal calf serum applied via a syringe, into a siliconised centrifuge tube; after centrifugation at 1000 rpm and removal of all but one drop of supernatant, cells of sediment carefully mixed; drop of cell suspension placed on clean, degreased microscope slide and immediately spread; 3 slides/animal; slides air dried at least overnight; stained with Giemsa; air dried and dipped in xylol for 3 minutes
METHOD OF ANALYSIS: 1 slide/animal chosen and given a random code; microscopic evaluation of slides from 5 males and 5 females per treatment group at 1000x magnification; number of micronucleated cells counted in 1000 polychromatic erythrocytes (PCEs)/animal; ratio of
polychromatic to normochromatic erythrocytes determined by counting and differentiating the first 1000 erythrocytes
OTHER: means and standard deviations calculated - Evaluation criteria:
- Statistically significant (p<0.05) increase in PCE compared to controls at any sampling time in either sex
Acceptability of test: positive controls induced statistically significant increase in frequency of micronucleated PCEs; solvent control incidence of micronuclei should reasonably fall within historical control range for the testing facility. - Statistics:
- Method used: Kastenbaum & Bowman
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- piloerection for 8 hours after administration; no mortality
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: 5000 mg/kg bw
- Solubility: used at 250 mg/ml
- Clinical signs of toxicity in test animals: piloerection
- Evidence of cytotoxicity in tissue analyzed: not examined
- Rationale for exposure: based on limit test in rats in which acute oral LD50 was >5000 mg/kg bw
- Harvest times: animals observed for 3 days
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no statistically significant increase in micronucleus frequency in any treatment group
- Ratio of PCE/NCE (for Micronucleus assay): treated groups similar to controls
- Appropriateness of dose levels and route: maximum tolerated dose of 5000 mg/kg bw used; guideline recommends maximum dose of 2000 mg/kg bw; oral route selected "taking into account the possible route of human exposure during manufacture, handling and use"
- Statistical evaluation: no statistically significant increases in micronuclei in treated groups of either sex; positive control produced a statistically significant increase in micronuclei
- Control incidence of micronuclei: not reported but presumably therefore within historical control range - Conclusions:
- Dodecan-1-ol has been tested a reliable study, conducted according to OECD guideline 474, no genotoxicity was seen in mice after a single oral dose of 5000 mg/kg bw. The study was performed in compliance with GLP.
Referenceopen allclose all
Table 1 Results of in vivo micronucleus study
Treatment mg/kg.bw /day |
Time of dosing (h) |
Time of sampling (h) |
Sex |
No. of surviving dosed mice |
Erythrocytes |
|||
Polychromatic cells (PCE) |
Mean PCE/NCE
|
|||||||
PCE Analysed |
No. of MN-PCE |
% MN-PCE |
||||||
Negative control |
0+24+48 |
72 |
M/F |
10 |
10000 |
14 |
0.14 |
0.93 |
96 |
M/F |
10 |
10000 |
10 |
0.10 |
1.02 |
||
Positive control |
0+24+48 |
72 |
M/F |
10 |
10000 |
150* |
1.50 |
0.46 |
500 |
0+24+48 |
72 |
M/F |
10 |
10000 |
9 |
0.09 |
1.00 |
1000 |
0+24+48 |
72 |
M/F |
10 |
10000 |
9 |
0.09 |
0.94 |
2000 |
0+24+48 |
72 |
M/F |
10 |
10000 |
8 |
0.08 |
0.86 |
96 |
M/F |
10 |
10000 |
24 |
0.24 |
0.90 |
||
PCE = Polychromatic erythrocytes
MN-PCE = Micronucleated PCE
MN-PCE = Micronucleated NCE
* = Positive response in PCE
Toxicity unclear, but possibly one male and one female mouse [per group?] died either spontaneously or due to gavage error.
Table 1 Results of micronucleus assay 24 hour sampling time
Treatment |
Suspending agent |
Low dose |
Mid dose |
High dose |
Concentration mg/kg bw |
0 |
40 |
50 |
150 |
Harvest time |
24 |
24 |
24 |
24 |
Micronucleated PCE (%) |
0.03 |
0.71 |
0.07 |
0.08 |
Ratio PCE/NCE |
1.27 |
0.93 |
0.98 |
1.07 |
Table 2 Results of micronucleus assay 48 hour sampling time
Treatment |
Suspending agent |
Test substance |
Test substance |
Test substance |
Concentration mg/kg bw |
0 |
50 |
150 |
500 |
Harvest time |
48 |
48 |
48 |
48 |
Micronucleated PCE (%) |
0.09 |
0.1 |
0.04 |
0.05 |
Ratio PCE/NCE |
1.05 |
1.06 |
1.01 |
1.23 |
Table 3 Results of micronucleus assay 72 hour sampling time
Treatment |
Suspending agent |
Low dose |
Mid dose |
High dose |
Concentration mg/kg bw |
0 |
50 |
150 |
500 |
Harvest time |
72 |
72 |
72 |
72 |
Micronucleated PCE (%) |
0.09 |
0.09 |
0.05 |
0.07 |
Ratio PCE/NCE |
1.41 |
1.33 |
1.55 |
1.46 |
The test substance did not increase the frequency of micronucleated erythrocytes or the PCE:NCE ratio in mice at any time interval after treatment (24, 48 or 72 hours) at dose levels up to 5000 mg/kg bw when compared to vehicle controls.
Mean values per group in the micronucleus test with 1-Dodecanol
a) Number of micronucleated cells per 1000 polychromatic erythrocytes (PCE)
b) Ratio of polychromatic to normochromatic erythrocytes (PCE/NCE)
Treatment group; (sampling time) |
Species and sex |
Dose mg/kg |
Micronucleated cells 1000 PCE |
Ratio of PCE/NCE |
||
Mean |
Range |
Mean |
Range |
|||
Negative control (24 hours) arachis oil |
male mice |
10 ml/kg |
3.60 |
0 - 9 |
1.11 |
0.80 - 1.31 |
female mice |
10 ml/kg |
2.00 |
0 - 4 |
1.34 |
1.02 - 1.07 |
|
Positve control (24 hours) cyclophosphamide |
male mice |
20 |
13.40 |
10 - 16 |
1.21 |
0.90 - 1.72 |
female mice |
20 |
10.80 |
7 - 14 |
0.95 |
0.67 - 1.28 |
|
1-Dodecanol (Lorol C12-99)
|
|
|
|
|
|
|
Limit dose (24 hours) |
male mice |
5000 |
2.60 |
0 - 5 |
1.08 |
0.94 - 1.26 |
female mice |
5000 |
2.40 |
2 - 3 |
1.01 |
0.90 - 1.18 |
|
Limit dose (48 hours) |
male mice |
5000 |
3.00 |
1 - 4 |
0.89 |
0.48 - 1.16 |
female mice |
5000 |
2.00 |
0 - 5 |
1.18 |
0.90 - 1.68 |
|
Limit dose (72 hours) |
male mice |
5000 |
2.60 |
2 - 4 |
1.65 |
0.91 - 2.14 |
female mice |
5000 |
1.60 |
0 - 4 |
1.33 |
1.08 - 1.55 |
|
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Reliable information on the genetic toxicity of tetradecan-1-ol was available for bacterial mutagenicity, however the study was performed with 4 strains and the fifth strain S. typhimurium TA102 or E.coli WP2 uvrA or E.coli WP2 uvrA (pKM101) is missing (Safepharm Laboratories, 1996b). For endpoints where no information was available, key studies were chosen from studies on closely related linear or branched alcohols of similar chain length. The choice of key study was based on reliability and similarity of chain length. The data available from standard in vitro and in vivo genetic toxicity assays for all related substances show no evidence of mutagenic potential.
A full discussion of the Category and considerations of RAAF Assessment Entities can be found in the Human Health Alcohols C6-24 Category report (PFA, 2021).
Tetradecan-1-ol has been tested for mutagenicity to bacteria in a reliable study, conducted according to OECD Test Guideline 471 and in compliance with GLP (Safepharm Laboratories, 1996b). No evidence of a test-substance related increase in the number of revertants was observed with or without metabolic activation in Salmonella typhimuriumstrains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 in the initial or repeat experiments up to limit concentration. Appropriate positive and solvent controls were included and gave the expected results. It is concluded that the test substance is negative for mutagenicity to bacterial under the conditions of the test
Fatty alcohol blend (containing 40.77% C8 and 55.3% C10) has been tested for mutagenicity to bacteria, in a study which was conducted according to OECD Test Guideline 471 and in compliance with GLP. No evidence of a test-substance related increase in the number of revertants was observed with or without metabolic activation in Salmonella typhimuriumstrains TA 1535, TA 1537, TA 98, TA 100 and TA 1538 in the initial or repeat experiments up to cytotoxic concentration. Appropriate positive and solvent controls were included and gave the expected results. It is concluded that the test substance is negative for mutagenicity to bacterial under the conditions of the test (Inveresk, 1992).
Alcohols, C12-13-branched and linear has been tested for clastogenicity in a valid study conducted according to OECD Test Guideline 473 and in compliance with GLP in CHO K1 cells (Sasol, 1998). The test substance did not increase the incidence of chromosome aberrations in Chinese hamster ovary cells at dose levels up to cytotoxic concentrations in the presence or absence of metabolic activation. The result of the repeat experiment confirmed that of the initial assay. Appropriate positive and solvent controls were added and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations under the conditions of this test.
Docosan-1-ol has been tested for clastogenicity in a valid study conducted according to OECD Test Guideline 473 without information on GLP compliance in Chinese hamster lung fibroblasts (V79) (Iglesias, 2002b). The test substance did not increase the incidence of chromosome aberrations in Chinese hamster lung fibroblasts (V79) at dose levels up to cytotoxic concentrations in the presence or absence of metabolic activation. Appropriate solvent, negative and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations under the conditions of this test.
Docosan-1-ol has been tested for mutagenicity in Chinese hamster lung fibroblasts (V79) cells according to OECD Test Guideline 476 and in compliance with GLP (Iglesias, 2002). No test-substance induced increase in the number of mutations was observed when tested with or without metabolic activation up to cytotoxic concentrations. Appropriate solvent, negative and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the study
Fatty alcohol blend (containing 40.77% C8 and 55.3% C10) has been tested for mutagenicity in mouse lymphoma L5178Y cells according to OECD Test Guideline 476 and in compliance with GLP. No test-substance induced increase in the number of mutations was observed when tested with or without metabolic activation up to cytotoxic concentrations. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the study (Inveresk, 1992).
Fatty alcohol blend has been tested according to OECD Test Guideline 474 and under GLP (Inveresk, 1992). No evidence for a test substance induced increase in the incidence of micronucleated PCE was observed (1000 PCE scored per animal). Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the test. No toxicity to bone marrow or general toxicity was observed.
Dodecan-1-ol has been tested for the induction of micronuclei in mice according to OECD Test Guideline 474 and in compliance with GLP. No evidence for a test substance induced increase in the incidence of micronucleated normochromatic erythrocytes in mice bone marrow. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance does not cause damage to chromosomes under the conditions of the test (Henkel, 1992).
Docosan-1-ol has been tested for the induction of micronuclei in mice according to OECD Test Guideline 474 but without information on GLP compliance. No evidence for a test substance induced increase in the incidence of micronucleated normochromatic erythrocytes in mice bone marrow. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance does not cause damage to chromosomes under the conditions of the test Iglesias, 2002b).
Discussion of trends in the Category of C6-24 linear and essentially-linear aliphatic alcohols (LCAA):
The in vitro and in vivo data available for members of the category and supporting substances indicate that the C6-24 alcohols are not genotoxic. In addition, the category of LCAAs under consideration does not contain any structural elements that are of concern for potential mutagenic activity (Ashby and Tenant, 1991). Furthermore, primary LCAAs (linear and branched) in the range C1 to C5 do not have a mutagenic potential (Bevan, 2001; OECD SIDS butan-1-ol, 2001). Moreover, in a review by WHO-JECFA a series of 22 saturated aliphatic branched-chain primary LCAAs and the corresponding aldehydes and acids in the range C4 to C8 showed no activity in a battery of in vitro and in vivo mutagenicity tests (WHO, 1999). On this basis it is concluded that the category of LCAAs does not have a mutagenic potential and that read-across within the category can be justified. Where data gaps exist, the gap is filled by read-across from reliable evidence within the C6-24 Alcohols Category, where possible using interpolation between at least two reliable studies using higher and lower carbon number test substances.
Conclusion: The category C6-24 LCAAs do not have a genotoxic potential.
Genetic toxicity of LCAAs
|
CAS |
CHEMICAL NAME |
Bacterial mutagenicity |
Bacterial mutagenicity |
Mammalian cytogenicity |
Mammalian cytogenicity |
Mamalian mutagenicity |
Mamalian mutagenicity |
In vivo studies |
In vivo studies |
|
|
|
Result (Rel.) |
Reference |
Result (Rel.) |
Reference |
Result (Rel.) |
Reference |
Result (Rel.) |
Study Type*(Ref) |
C6 |
111-27-3 |
Hexan-1-ol |
Neg; (1) |
Henkel, 1990 |
|
|
|
|
|
|
C7, 8 and 9 |
|
Alcohols, C7-9 |
Neg. (1) |
Shell, 1996 |
|
|
|
|
|
|
C8 |
111-87-5 |
Octan-1-ol |
Neg; (2) |
Henkel, 1982a; Huntingdon Life Sciences, 1996k |
|
|
|
|
|
|
C8 |
104-76-7 |
2-ethylhexan-1-ol Supporting Substance |
Neg; (2) |
Kirby, 1983 |
|
|
Neg; (2) |
Kirby, 1983 |
Neg; (2) |
MN;Dom. Leth (Putman, 1983; WHO, 1993) |
C8-10 |
none |
Fatty alcohol blend (40.7% C8 and 55.3% C10) Supporting Substance |
Neg(2) |
Inveresk (1992) |
|
|
Neg (1) |
Inveresk (1992) |
Neg (1) |
Inveresk(1992) |
C10 |
112-30-1 |
Decan-1-ol |
Neg (4) 2 strains only
|
(Huntingdon Life Sciences, 1996l) |
|
|
|
|
|
|
C12 |
112-53-8 |
Dodecan-1-ol |
Neg. (1)l |
(Safepharm Laboratories, 1996a)Shimizu, 1985 |
|
|
|
|
Neg. (2) |
Micronucleus; (Henkel, 1992) |
C12 and 13 |
75782-87-5 |
Alcohols, C12-13 |
Neg (2, >80% lin) |
Sasol, 1980 |
|
|
|
|
|
|
C12 and 13 |
740817-83-8 |
Alcohols, C12-13-branched and linear |
Neg (1 50% lin), |
Sasol, 1998 |
Neg (1 (50% lin) |
Sasol, 1998 |
|
|
|
|
C12 |
67762-25-8 |
C12-18 Alcohols, Type B Supporting |
Neg (2)Ames |
Henkel 1982 |
|
|
|
|
|
|
C 12-15 |
90604-40-3 |
Alcohols, C12-15-branched and linear |
Neg (1) |
Corning Hazleton, 1996 |
|
|
|
|
|
|
C14 |
112-72-1 |
Tetradecan-1-ol |
Neg (1) |
Safepharm Laboratories, 1996b |
|
|
|
|
|
|
C16 |
36653-82-4 |
Hexadecan-1-ol |
Neg (1) |
Safepharm Laboratories, 1996c |
|
|
|
|
|
|
C16 |
36653-82-4 |
Hexadecan-1-ol |
Neg. (2) |
Henkel, 1981 |
|
|
|
|
|
|
C16 |
68002-94-8 |
C16-18 and C18 Unsaturated Supporting |
Neg. Ames (2) |
Banduhn, 1989) |
|
|
|
|
|
|
C18 |
112-92-5 |
Octadecan-1-ol |
Neg (1)
|
Safepharm Laboratories, 1996d
|
|
|
|
|
Neg (2) MN |
Hachiya, 1982 |
C18 |
112-92-5 |
Octadecan-1-ol |
Neg(2) |
Henkel, 1981 |
|
|
|
|
|
|
C18 |
97552-91-5 |
C18-22 Alcohol Supporting |
Neg. Ames (2) |
Banduhn 1995 |
|
|
|
|
|
|
C22 |
661-19-8 |
Docosan-1-ol |
Neg (2),
|
Iglesias, 2002b, Thompson, 1997 |
Neg (2), |
Iglesias, 2002b |
Neg (2) |
Iglesias, 2002b |
Neg (2) |
Micronucleus Iglesias, 2002bª |
C24-32 |
|
D-002*** Supporting substance |
|
|
|
|
|
|
Neg (4) |
MN; Dom. Leth.Rodeiro 1998a |
* MN: Mouse bone marrow micronucleus test; Dom. Leth. Mouse Dominant Lethal test; UDS: Unscheduled DNA Synthesis assay
** Tested in S. typhimurium TA 98 and TA100, only.
***Mixture of very long chain fatty alcohols from hydrolysed bees wax
References:
Ashby, J., Tennant, R.W., 1991. Definitive relationships among chemical structure, carcinogenicity, and mutagenicity for 301 chemicals tested by the US NTP. Mutation Research 257, 229–306.
WHO, 1999. Technical Report Series 884 Evaluation of certain food additives and contaminants. 49th Report of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), Geneva.
Justification for classification or non-classification
Based on the available data, tetradecan-1ol does not require classification for genetic toxicity according to Regulation (EC) No 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
