Glycerol, ethoxylated, esters with acrylic acid

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

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

Test material information

Constituent 1

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:

Glycerol, ethoxylated, esters with acrylic acid

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