Hexanedioic acid, polymer with 2,2'-oxybis[ethanol]

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: Study conducted according to OECD guideline 471 and GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

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:

Polyesterol 90212

IUPAC Name:

Polyesterol 90212

Constituent 2

Reference substance name:

Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol]

EC Number:

618-460-1

Cas Number:

9010-89-3

Molecular formula:

(C6 H10 O4 . C4 H10 O3)x

IUPAC Name:

Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol]

Test material form:

other: liquid

Details on test material:

Stability of the test substance in the vehicle DMSO was not determined analytically.

COMPOSITION:
ca. 99% (w/w) Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] / Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] / 9010-89-3
ca. 2% 2,3,7,8-tetrahydro-2,5,8-benzotrioxacycloundecine-1,9-dione / 1,4,7-Trioxacyclotridecane-8,13-dione / 1,4,7-Trioxacyclotridecane-8,13-dione / 6607-34-7
ca. 10 ppm tin bis(2-ethylhexanoate) / Bis(2-ethylhexanoyloxy)tin / Hexanoic acid, 2-ethyl, tin(2+) salt (2:1) / 301-10-0
ca. 10 ppm titanium tetrabutanolate / titanium tetrabutanolate / tetrabutyl titanate / 5593-70-4

Method

Target gene:

his-, trp-

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:

S9 liver fraction from male Wistar rats, induced by 80 mg/kg b.w. phenobarbital (i.p.) and beta-naphthoflavone (oral) on three consecutive days. Cofactors in S9 mix: MgCl2 (8 mM), KCl (33 mM), G6P (5mM), NADP (4 mM), phosphate buffer (15 mM, pH 7.4).

Test concentrations with justification for top dose:

0, 33, 100, 333, 1000, 2500, 5000 ug/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Remarks:

Sterility control: Soft agar plates with S9 mix, buffervehicle or test substance but without tester strain.

Negative solvent / vehicle controls:

yes

Remarks:

With and without S9 mix, contains vehicle at the same concentration and volume for all tester strains.

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

other: 2-aminoanthracene, N-methyl-N'-nitro-N-nitrosoguianidine, 4-nitro-o-phenylenediamine

Remarks:

With S9 mix: 2-aminoanthracene; without Sß mix: N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitro-o-phenylenediamine, 9-aminoacridine, 4-nitroquinoline-N-oxide

Details on test system and experimental conditions:

Assessment of mutagenicity in bacteria was performed as two independent experiments, a standard plate test and a preincubation test.

STANDARD PLATE TEST
Test substance, bacterial culture (Salmonella typhimurium or E. coli) and, if applicable, S9 mix, were added to soft agar heated up to 42-45 °C. Samples were mixed and immediately poured onto Vogel-Bonner (Salmonella typhimurium) or minimal (E. coli) agar plates. After incubation at 37 °C for 48-72 hours in the dark, his+ (Salmonella typhimurium) or trp+ (E. coli) revertants were counted.

PREINCUBATION TEST
Test substance, bacterial suspension and, if applicable, S9 mix were incubated on a shaker at 37 °C for 20 minutes. Subsequently, soft agar was added. Samples were mixed and immediately poured onto agar plates. After incubation at 37 °C for 48-72 hours in the dark, bacterial colonies were counted.

REPLICATES
3 test plates per dose or per control

Evaluation criteria:

The experiment was considered valid if the number of revertant colonies in the negative and in the positive control group (+/-S9) were within the range of the respective historical controls (or above, in the case of positive control substances), if the titer of viable bacteria was >= 10^8/ml, and if the sterility controls showed no signs of bacterial contamination.
A substance was considered mutagenic if there was 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 with or without S9 mix.
A substance was considered non-mutagenic if the number of revertants for all tester strains were wthin the historical negative control range under all conditions in at least two independent experiments.

Statistics:

3 plates per dose group or per control were evaluated.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

No mutagenicity detected in preincubation test or standard plate test.

Cytotoxicity / choice of top concentrations:

other: occasionally observed depending on the strain and test conditions from about 2500 ug/plate onward.

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No substance precipitation was found with and without S9 mix.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the experimental conditions chosen, the test substance is not mutagenic in the bacterial reverse mutation assay in the absence and the presence of metabolic activation.

Executive summary:

The test substance Hexanedioic acid oligomeric reaction products with 2,2'-oxybis[ethanol] was tested for mutagenicity in the bacterial reverse mutation assay in the absence and presence of metabolic activation. The test was conducted as two independent experiments: a standard plate test and a preincubation test. In both experiments, test substance concentrations ranging from 33 to 5000 ug/plate were tested. Bacteriotoxicity was observed depending on the strain and the test conditions from about 2500 ug/plate onward. No precipitation of the test substance was found with and without S9 mix. The test substance did not cause a relevant increase in the number of his+ or trp+ revertants in the standard plate test or the preincubation test either in the presence or the absence of S9 mix. Based on these results, it can be concluded that the test substance is not mutagenic in the bacterial reverse mutation assay under the experimental conditions chosen.