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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:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

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

Constituent 1
Chemical structure
Reference substance name:
Glycerol, ethoxylated, esters with acrylic acid
EC Number:
500-322-6
EC Name:
Glycerol, ethoxylated, esters with acrylic acid
Cas Number:
144086-03-3
Molecular formula:
UVCB substance
IUPAC Name:
[(uraniovanadio)methyl]borane
Details on test material:
- Name of test material (as cited in study report): Glycerin3EOTA
- Physical state: Beige, turbid liquid
- Analytical purity: >99%
- Lot/batch No.: GK0561/160
- Stability under test conditions: The stability of the test substance under storage conditions throughout the study period is guaranteed until June 15, 2005 as indicated by the sponsor, and the sponsor holds this responsibility
- Storage condition of test material: Room temperature

Method

Target gene:
S. typhimurium: his-
E. coli: trp-
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98; E. coli WP2 uvrA
Metabolic activation:
with and without
Metabolic activation system:
Liver S-9 mix from Aroclor 1254 induced male rats
Test concentrations with justification for top dose:
0; 20; 100; 500; 2500 and 5000 µg/plate in both the SPT and the PT
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: see below
Details on test system and experimental conditions:
Experiment 1: Standard plate test (SPT)
Test tubes containing 2 ml soft agar kept in a water bath at 45°C, and remaining components added in the following order:
0 .1 ml test solution or vehicle
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.

Experiment 2: Preincubation assay (PT)
0.1 ml test solution or vehicle, 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.

Experiment1 & 2:
In each experiment 3 test plates per dose or per control used; after incubation at 37°C for 48 hours in the dark, the bacterial colonies ( his+/trp+ revertants) are counted.
Positive control:
with metabolic activation: 2.5 (Salmonella) or 60 (E. coli) μg/plate 2-aminoanthracene for all strains;
without metabolic activation: 5 μg/plate N-methyl-N'-nitro-N-nitrosoguanidine for TA 100 and TA 1535, 10 μg/plate 4-nitro-o-phenylendiamine for TA 98, 100 μg/plate 9-aminoacridine for TA 1537 and 5 µg/plate 4-nitroquinoline-N-oxide for E. Coli WP2 uvrA, all substances were dissolved in DMSO.
The titer was determined and in regularly measurements the strain characteristics were checked. Sterility control was performed.
Evaluation criteria:
The test chemical is considered positive in this assay if the following criteria are met:
A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.
Statistics:
not performed

Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98; E. coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: >=2500 µg/plate in both the SPT and PIT; depending on strain and test condition
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
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 revertants in Controls maximum revertant factor dose dependency Assessment
TA 98 no 33 0.9 no negative
  yes 44 0.9 no negative
TA 100 no 103 1.1 no negative
  yes 114 1.2 no negative
TA 1535 no 18 1.1 no negative
  yes 18 1.0 no negative
TA 1537 no 9 1.4 no negative
  yes 11 1.1 no negative
WP2 uvr A no 37 1.1 no negative
  yes 37 1.1 no negative
Preincubation test (20 - 5000 µg/plate)
Strain Metabolic activation system mean revertants in Controls maximum revertant factor dose dependency Assessment
TA 98 no 26 1.0 no negative
  yes 31 0.9 no negative
TA 100 no 110 1.0 no negative
  yes 108 1.0 no negative
TA 1535 no 17 1.0 no negative
  yes 17 0.9 no negative
TA 1537 no 10 1.1 no negative
  yes 10 0.9 no negative
WP2 uvr A no 31 1.0 no negative
  yes 36 1.0 no negative

Applicant's summary and conclusion