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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
23-Aug-1996 to 07-Oct-1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to an appropriate OECD test guideline, and under GLP, with acceptable restrictions. The restrictions were that the strains used did not comply with the current guideline

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(no TA102 or E. coli WP2 uvrA, 2-AA only as positive control with S9)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Kalcol 2098 (chemical name: lauryl alcohol)
- Substance type: clear, colourless liquid
- Physical state: liquid
- 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.: 3397
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: shielded from light, not necessarily in darkness
- Other:
Date received: 05-Aug-1996

Method

Target gene:
histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: histidine deficient
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
other: histidine deficient
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S9 .
Test concentrations with justification for top dose:
0.5 (only TA1538 and 1537), 1.5, 5, 15, 50, 150 and 500 (only TA98, 100, 1535) µg/plate (based on a preliminary screening test)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
3 µg/plate (TA100), 5 µg/plate (TA1535), without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
80 µg/plate (TA1537), without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylene daimine
Remarks:
5 µg/plate (TA1538), without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
0.2 µg/plate (TA98), without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
1 µg/plate (TA100), 2 µg/plate (TA1535 and TA 1537), 0.5 µg/plate (TA1538 and TA98), with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar

DURATION
- Exposure duration: 48 hours at 37 deg C

NUMBER OF REPLICATES: triplicates
Evaluation criteria:
Considered positive if there is concentration-related and statistically significant increase in reverse mutation rate in one or more bacterial strains at sub toxic dose levels. To be considered negative, the number of induced revertants should be <2-fold  the number of spontaneous revertants and concentrations should extend to the limits of solubility or toxicity up to a maximum of 5000 µg/plate.
Statistics:
Dunnetts linear regression method.

Results and discussion

Test resultsopen allclose all
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:
cytotoxicity
Remarks:
50-150 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
50 µg/plate
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: An oily precipitate was observed at 1500 ug/plate and above in the preliminary toxicity assay, this did not interfere with the scoring of revertant colonies and was not reported when this concentration was tested in the repeat study.
- Other confounding effects: no data

COMPARISON WITH HISTORICAL CONTROL DATA: No

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- With and without metabolic activation: The test material exhibited a visible reduction in background lawn at and above 150 ug/plate in all the strains
tested. Strains TA1538, 1537 and 1535 also showed a reduction in background lawn at 50 ug/plate. This indicates that the material was tested to a 
toxic level.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

STATISTICAL RESULTS: No statistically significant increase in reverse mutation rate at any dose level tested with or without metabolic activation.

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

Concentrationµg/plate        TA 100       TA 1535        TA 1538          TA 98        TA 1537
 - MA  + MA  - MA  + MA  - MA   + MA  - MA  + MA  - MA  + MA
0 113 111 23 18 13 26 27 31 7 9
0.5 N/T N/T N/T N/T 14 N/T N/T N/T 7  N/T
1.5 113 111 25 15 15 25 25 30 7 10
5 115 105 21 12 15 24 23 30 7 9
15 111 107 23 12 16 27 27 29 8 10
50 88 92 26 15 9 28 25 25 6 9
150 48 72 24 11 0 11 14 24 0 5
500 0 27 0 7 N/T 0 7 18 N/T 0
Positive control 529 1001 136 232 685 513 185 842 879 309

N/T Not tested

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

 Concentration µg/plate TA 100 TA 1535 TA 15381 TA 98 TA 1537
-MA +MA -MA +MA -MA +MA -MA +MA -MA +MA
0 86 99 15 13 14 22 21 36 9 11
0.5 96 102 13 N/T 11 2.5 20 N/T 6 N/T
1.5 92 100 13 16 12 22 19 36 8 10
5 93 100 13 15 12 20 20 35 9 9
15 93 101 14 11 15 25 22 30 6 12
50 68 99 12 11 11 21 18 37 7 12
150 25* 70 6* 12 0* 19 9* 28 0* 10
500 N/T N/T N/T 8 N/T N/T N/T 13 N/T 1
1500 N/T N/T N/T 2* N/T N/T N/T 0* N/T 0*
Positive control 449 1008 293 210 708 442 134 602 1011 337

* Very thin or absent background lawn

N/T Not tested

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

In a reliable study, the C12 alcohol Kalcohl 2098 did not increase the reverse mutation rate in histidine dependent bacterial strains of Salmonella typhimurium in the presence or absence of metabolic activation. The material was tested to cytotoxic concentrations. The study was performed in compliance with GLP. It is concluded that dodecan-1-ol is negative for the induction of mutations in bacteria under the conditions of the test.