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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report date:
1989

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(only four strains tested, 2-aminoanthracene was the sole indicator of the efficacy of the S9-mix)
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Alcohols, C13-15-branched and linear
EC Number:
287-625-4
EC Name:
Alcohols, C13-15-branched and linear
Cas Number:
85566-16-1
Molecular formula:
C13 H27 OH - C15 H31 OH
IUPAC Name:
Alcohols C13-C15 branched and linear, reaction products of C12-C14 olefines
Details on test material:
- Name of test material (as cited in study report): C13 - C15 - Alkohol
- Physical state: colorless liquid
- Analytical purity: 99.3% alcohols (0.5% low boiling substances, 0.2% high boiling substances)
- Composition of test material, percentage of components: 29.5% i-C13-alcohol, 33% n-C13-alcohol, 19% i-C15-alcohol, 17.8% n-C15-alcohol
- Lot/batch No.: from continuous production
- Sustance number: 88/577
- Storage: room temperature

Method

Target gene:
his-gene
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9-mix of Aroclor 1254 induced rat livers (Sprague-Dawley)
Test concentrations with justification for top dose:
20 - 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Complete solubility of the test substance in DMSO.
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see Details on test system and conditions for details
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and preincubation

Standard plate test:
The experimental procedure is based on the method of Ames et al ., 1975

Test tubes containing 2 ml portions of soft agar which consists of 100 ml agar (0.6% agar + 0.6% NaCl) and 10 ml amino acid solution (minimal amino acid solution for the determination of mutants: 0.5 mM histidine + 0.5 mM biotin) are kept in a water bath at 45°C, and the remaining components are added in the following order:

0.1 ml test solution
0.1 ml bacterial suspension
0.5 ml S-9 mix (in tests with metabolic activation)
or
0.5 ml phosphate buffer (in tests without metabolic activation)

After mixing, the samples are poured onto Vogel-Bonner agar plates (minimal glucose agar plates) within approx. 30 seconds.

Preincubation test:
The experimental procedure is based on the method described by Yahagi et al. (1977) and Matsushima et al. (1980).

0.1 ml test solution, 0.1 ml bacterial suspension and 0.5 ml S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the Vogel-Bonner agar plates within approx. 30 seconds.

Composition of the minimal glucose agar:
980 ml aqua dest.
20 ml Vogel-Bonner E medium
15 g Difco bacto agar
20 g D-glucose, monohydrate

After incubation at 37°C for 48 hours in the dark, the bacterial colonies (his+ revertants) are counted.

Positive controls:
The following positive control substances are used to check the mutability of the bacteria and the activity of the S-9 mix:

with S-9 mix:
10 µg 2-aminoanthracene (dissolved in DMSO) for the strains TA 100, TA 98, TA 1537 and TA 1535

without S-9 mix:
5 µg N-methyl-N-nitro-N-nitrosoguanidine (MNNG) (dissolved in DMSO) for the strains TA 100 and TA 1535,
10 µg 4-nitro-o-phenylendiamine (dissolved in DMSO) for the strain TA 98,
100 µg 9-aminoacridine chloride monohydrate (dissolved in DMSO) for the strain TA 1537

NUMBER OF REPLICATIONS: triplicate




Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control);
- dose-response relationship;
- reproducibility of the results;

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test (20 - 5000 µg/plate)
Strain Metabolic activation system mean his+/trp+revertant colonies (negative control) maximum revertant factor (conc. (µg/plate)) dose dependency Assessment maximum revertant factor (positive control)
TA 98 no 23 1.0 (20/500/2500/5000) no negative 29 (NPD)
  yes 34 1.1 (20/100) no negative 34.7 (2-AA)
TA 100 no 114 0.9 (100) no negative 14.8 (MNNG)
  yes 103 1.3 (100) no negative 17.2 (2-AA)
TA 1537 no 9 1.3 (20) no negative 47 (AAC)
  yes 12 0.9 (20/100/500) no negative 11.7 (2-AA)
TA 1535 no 15 1.0 (20) no negative 128.4 (MNNG)
  yes 15 1.2 (2500) no negative 10.5 (2-AA)
Preincubation test (20 - 5000 µg/plate)
Strain Metabolic activation system mean his+/trp+revertant colonies (negative control) maximum revertant factor (conc. (µg/plate)) dose dependency Assessment maximum revertant factor (positive control)
TA 98 no 24 1.1 (500) no negative 39.9 (NPD)
  yes 35 1.2 (500) no negative 26.5 (2-AA)
TA 100 no 110 1.1 (500) no negative 10.6 (MNNG)
  yes 125 1.1 (20) no negative 9.5 (2-AA)
TA 1537 no 8 1.2 (100) no negative 43.9 (AAC)
  yes 12 0.8 (20/100/2500) no negative 9.3 (2-AA)
TA 1535 no 19 1.2 (5000) no negative 59 (MNNG)
  yes 18 1.0 (20/5000) no negative 7.1 (2-AA)
2-AA = 2-aminoanthracene
NPD = N-nitro-o-phenylendiamine
MNNG = N-methyl-N-nitro-N-nitrosoguanidine
AAC = 9-aminoacridine chloride monohydrate

Applicant's summary and conclusion