reaction products of 1,4-butanediol with 1,3-chloro-2,3-epoxypropane, esters with prop-2-enoic 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:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted 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:

adopted May 2000

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:

Reaction products of butane-1,4-diol and 1-chloro-2,3-epoxypropane, esters with acrylic acid

EC Number:

701-230-0

Cas Number:

52408-42-1

Molecular formula:

C16H26O8

IUPAC Name:

Reaction products of butane-1,4-diol and 1-chloro-2,3-epoxypropane, esters with acrylic acid

Test material form:

liquid

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media: 8 g Difco nutrient broth + 5 g NaCI/liter
- Properly maintained: yes

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

- Type and identity of media: 8 g Difco nutrient broth + 5 g NaCI/liter
- Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 rat liver induced S9 mix

Test concentrations with justification for top dose:

20µg, 100µg, 500µg, 2500µg, 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: low water solubility of test substance, DMSO had been demonstrated to be suitable in bacterial lreverse mutation tests and historical control data are available

Controls

Controls 1

Untreated negative controls:

other: sterility control

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Remarks:

with S9

Controls 2

Untreated negative controls:

other: sterility contol

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for TA 1535, TA 100; 4-nitro-o-phenylendiamine (NOPD) for TA 98; 9-aminoacridine (AAC) for TA 1537; 4-nitroquinoline-N-oxide (4-NQO) for E . coli WP2 uvrA

Remarks:

without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; with and without preincubation

DURATION
- Preincubation period: 20min
- Exposure duration: 48h,72h

SELECTION AGENT (mutation assays): lack of tryptophane in soft agar

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn (=reduced his- or trp- background
growth ); reduction in the titer

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.

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 .

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

slight at >= 2500µg/ml (standard) or >= 500 - 2500µg/ml (preincubation)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

slight at >= 2500µg/ml (standard) or >= 500 - 2500µg/ml (preincubation)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

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:

cytotoxicity

Remarks:

slight at >= 2500µg/ml (standard) or >= 500 - 2500µg/ml (preincubation)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

slight at >= 2500µg/ml (standard) or >= 500 - 2500µg/ml (preincubation)

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No increase in the number of his + or trp+ revertants in both, the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system.
Cytotoxicity was observed in Salmonella strains in the preincubation assay at and above 500 µg/plate - 2500 µg/plate depending on strain.

Applicant's summary and conclusion

Conclusions:

Thus, under the experimental conditions chosen here, it is concluded that the test substance is not a mutagenic agent in a bacterial reverse mutation test.

Executive summary:

In a gene mutation assay in bacteria (Ames) according to OECD471 and GLP (2002), 1,4 -butanediylbis[oxy(2 -hydroxy-3,1 -propanediyl)] diacrylate did not lead to an increase in the number of his + or trp+ revertants in both, the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system. Concentrations up to 5000µg/plate were tested in Salmonella strains TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli 1412 uvrA. Cytotoxicity was observed in Salmonella strains in the preincubation assay at and above 500µg/plate - 2500µg/plate depending on strain. S9 fraction was prepared from aroclor 1254 induced rat liver.