Glycerol, propoxylated, esters with acrylic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

19 May 2000

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Version / remarks:

August 1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycerol, propoxylated, esters with acrylic acid

EC Number:

500-114-5

EC Name:

Glycerol, propoxylated, esters with acrylic acid

Cas Number:

52408-84-1

Molecular formula:

(C3H6O)m(C3H6O)n(C3H6O)oC12H14O6

IUPAC Name:

Poly[oxy(methyl-1,2-ethanediyl)], .alpha.,.alpha.',.alpha.''-1,2,3- propanetriyltris[.omega.-[(1-oxo-2-propenyl)oxy]]-

Test material form:

liquid

Specific details on test material used for the study:

Batch No.: GK0561/158
Storage: Room temperature
Stability: guaranteed until June 8, 2005

Method

Target gene:

his operon

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
- source of S9 : Aroclor 1254 induced rat liver
- method of preparation of S9 mix: a sufficient amount of S-9 fraction was thawed at room temperature and 1 volume of S-9 fraction is mixed with 9 volumes of S-9 supplement (cofactors). The concentrations of the cofactors in the S-9 mix are:
8 mM MgCl2
33 mM KCI
5 mM Glucose-6-phosphatase
4 mM NADP
15 mM Phosphate buffer (pH 7.4)
The phosphate buffer (6) is prepared by mixing an Na2HPO4 solution with an NaH2PO4 solution in a ratio of about 4 :1.
- S9 in the final culture medium : 0.5 mL
- quality controls of S9: To demonstrate the efficacy of the S-9 mix in this assay, the S-9 batch was characterized with benzo(a)pyrene

Test concentrations with justification for top dose:

20 µg - 5 000 µg/plate (Standard plate test)
20 µg - 5 000 µg/plate (Preincubation test)

Vehicle / solvent:

- Vehicle used: DMSO
- Justification for choice of vehicle:Due to the limited solubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.

Controls

Untreated negative controls:

yes

Remarks:

Sterility and Vehicle controls

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene; N-methyl-N'-nitro-N-nitrosoguanidine; 4-nitro-o-phenylendiamine; 9-a minoacridine (AAC); 4-nitroquinoline-N-oxide

Details on test system and experimental conditions:

METHOD OF APPLICATION: in suspension (soft agar;amino acid solution;test solution or vehicle;fresh bacterial culture; S-9 mix or phosphate buffer were mixed and poured onto agar plates)

DURATION (Preincubation and Standardtest were performed)
- Preincubation period: 20 min
- Exposure duration: 48h-72h
- Fixation time (start of exposure up cell count): 48h-72h

SELECTION AGENT: histidine

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth (number of revertants were counted)

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.

A test substance is generally considered nonmutagenic in this test if:
-The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Toxicity detected by a
- decrease in the number of revertants
- clearing or diminution of the background lawn (= reduced his- or trp- background growth)
- reduction in the titer

Results and discussion

Test results

Test results 1

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

slight decrease in the number of revertantswas observed depending on the strain and test conditions at doses ≥ 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

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

Remarks:

slight decrease in the number of revertantswas observed depending on the strain and test conditions at doses ≥ 5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation and time of the determination: No precipitation was found.

STUDY RESULTS
- Concurrent vehicle negative and positive control data : Please refer to Table 1 to 10.

Ames test:
- Signs of toxicity : A slight decrease in the number of revertants was occasionally observed in the standard plate test depending on the strain and test conditions at 5 000 pg/plate. In the preincubation assay a weak bacteriotoxicity (slight decrease in the number of revertants and / or a slight reduction in the titer) was observed depending on the strain and test conditions at doses > 2500 pg/plate.
- Individual plate counts: Please refer to Table 1 to 10
- Mean number of revertant colonies per plate and standard deviation : Please refer to Table 1 to 10

HISTORICAL CONTROL DATA
- Positive historical control data: Please refer to Table 11 to 15
- Negative historical control data: Please refer to Table 16 to 20