2-(2-butoxyethoxy)ethyl methacrylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15.12.1998-01.09.1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

OECD 471, GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

EEC Annex V Test B14 (1993)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-(2-butoxyethoxy)ethyl methacrylate

EC Number:

230-813-8

EC Name:

2-(2-butoxyethoxy)ethyl methacrylate

Cas Number:

7328-22-5

Molecular formula:

C12H22O4

IUPAC Name:

2-(2-butoxyethoxy)ethyl 2-methylprop-2-enoate

Test material form:

liquid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from male Sprage Dawley rats

Test concentrations with justification for top dose:

range finder: 8, 40, 200, 1000 and 5000 µg/plate
Experiment 1: 8, 40, 200, 1000 and 5000 µg/plate
Experiment 2: : 51.2; 128; 320; 800; 2000 and 5000 µg/plate

Vehicle / solvent:

Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Controls 2

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Controls 3

Positive controls:

yes

Positive control substance:

sodium azide

Controls 4

Positive controls:

yes

Positive control substance:

9-aminoacridine

Controls 5

Positive controls:

yes

Positive control substance:

other: Glutaraldehyd

Controls 6

Positive controls:

yes

Positive control substance:

other: 2-amonianthracene

Details on test system and experimental conditions:

The test article was completely soluble in the aqueous assay system at all concentrations treated, in each of the experiments performed.
Negative (solvent) and positive control treatments were included for all strains in both experiments. The mean numbers of revertant colonies on negative control plates all fell within acceptable ranges, and were significantly elevated by positive control treatments.

Range-finder:
TA100 using final concetrations of Butyldiglycol methacrylate at 8, 40, 200, 1000 and 5000 µg/plate, plus negative (solvent) and positive controls with and without metabolic activation.

No evidence of toxicity (as would normally be indicated by a thinning of the background bacteriallawn and/or areduction in revertant numbers) was observed following any of these range-finder treatments. These results were considered acceptable for mutagenicity assessment and
were therefore used to provide the TAl00 mutagenicity data for Experiment 1.

Main Experiment:
Experiment 1
TA98, TA102, TA1535, TA1537 inal concetrations of Butyldiglycol methacrylate at 8, 40, 200, 1000 and 5000 µg/plate, plus negative (solvent) and positive controls with and without metabolic activation. Following this experiment, evidence of toxicity (manifest as slight thinning of the
background bacterial lawn) was observed solely with strains TA98 and TA102 in the absence of S-9 at the maximum per dose.

Experiment 2:
TA98, TA100, TA1535, TA1537, TA102
Test concentrations: 51.2; 128; 320; 800; 2000 and 5000 µg/plate plus negative (solvent) and positive controls with and without metabolic activation.
Treatments in the presence of S9 in experiment 2 included a pre-incubation step for 1 hour at 37 °C before addition of molten agar at 46 °C.

Evaluation criteria:

Acceptance criteria
The assay was considered valid if the following criteria were met:
1) the mean negative control counts fell within the normal ranges as defined in Appendix 3
2) the positive control chemicals induced clear increases in revertant numbers confirming discrimination between different strains, and an
active S-9 preparation
3) no more than 5% of the plates were lost through contamination or some other unforeseen event.

On one experimental occasion, the solvent control counts for one strain were outside of the laboratory historical control ranges. Although these counts failed to meet Acceptance criteria 1), these data were considered valid .

Evaluation criteria:
The test article was considered to be mutagenic if:
1) the assay was valid (see acceptance criteria)
2) a dose related and reproducible increase in the number of revertants was observed, or a significant* and reproducible increase in the
number of revertants was induced at one or more test concentration.
* An increase in revertant numbers was considered to be significant if the number of revertant colonies was at least two times the mean negative control counts in strains TA98, TAl00 and TA 102 , or three times in strains TA1535 and TA1537.

Statistics:

According to the OECD guideline 471, a statistical analysis of the data is not mandatory.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Butydiglycol methacrylate did not induce mutation in the Salmonella typhimurium strains TA98, TAl00, TA1535, TA1537 and
TA 102 up to a concentration of 5000 µg/plate, in the absence and in the presence of a rat liver metabolic activation system (S-9).

Applicant's summary and conclusion

Conclusions:

Butydiglycol methacrylate did not induce mutation in the Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and
TA 102 up to a concentration of 5000 µg/plate, in the absence and in the presence of a rat liver metabolic activation system (S-9).

Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471 Salmonella . typhimurium TA98, TA100, TA1535, TA1537 and TA102 strains were exposed to Butyldiglycol methacrylate in DMSO at concentrations up to 5000 µg/plate in the presence and absence of mammalian metabolic activation. The test was performed as pre-incubation test with 1 hour pre-incubation.  The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.  It was concluded that Butyldiglycol methacrylate did not induce mutation in the Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and TA102, under the conditions employed in this study with test concentrations up to 5000 µg/plyte, in the absence and presence of rat liver metabolic activation system (S-9).

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