1,4,7-trioxacyclotridecane-8,13-dione

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

Study period:

1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline Study (OECD), GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(July 21, 1997)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Remarks:

Department of toxicolgy, BASF AG

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3,7,8-tetrahydro-2,5,8-benzotrioxacycloundecine-1,9-dione

EC Number:

229-554-3

EC Name:

2,3,7,8-tetrahydro-2,5,8-benzotrioxacycloundecine-1,9-dione

Cas Number:

6607-34-7

Molecular formula:

C10H16O5

IUPAC Name:

1,4,7-Trioxacyclotridecane-8,13-dione

Details on test material:

- Name as cited in report: Adipic acid, cyclic ester with diethyleneglycol
- Analytical purity: ca. 90%
- Physical state: Solid/white crystals
- Storage: refrigerated
- Lot/batch No.: B 74
- Stablity: verified by reanalysis

Method

Target gene:

S. typhimurium: his-
E. coli: trp -

Species / strain

Species / strain / cell type:

other: TA 1535, TA 100, TA 1537, TA 98 and E . coli WP2 uvrA

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from Aroclor-induced liver of male Sprague-Dawley rat (200 - 300g).

Test concentrations with justification for top dose:

STANDARD PLATE TEST (SPT): 0; 20; 100; 500; 2,500 and 5,000 µg/plate
PREINCUBATION TEST (PIT): 0; 4; 20; 100; 500 and 2,500 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Remarks:

(sterility control)

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: With S9 Mix; 2-aminoanthracene (2-AA) and without S9 mix; N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylendiamine (NOPD), 9- aminoacridine (AAC), 4-nitroquinoline-N-oxide (4-NQO)

Remarks:

The stability of the selected positive controls was well-defined under the selected culture conditions, since they are well-established reference mutagens.

Details on test system and experimental conditions:

1). STANDARD PLATE TEST (SPT) (in agar incorporation method)
- Exposure duration: 48 - 72 h (in the dark) @ 37°C

2). PREINCUBATION TEST (PIT)
- Preincubation period: 30 min
- Exposure duration: 48 - 72 h (in the dark) @ 37°C


DETERMINATION OF CYTOTOXICITY
Toxicity was detected by a
- decrease in the number of reve rtants
- clearing or diminution of the background lawn (= reduced his" or trp" background growth)
- reduction in the titer

Evaluation criteria:

The test chemical was considered positive in this assay if 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, was seen. A test substance was 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

Species / strain:

other: TA 1535, TA 100, TA 1537, TA 98 and E . coli WP2 uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 500 µg

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
A slight decrease in the number of revertants was observed in the standard plate test depending on the strain and test conditions from about 500 µg - 2,500 µg/plate onward. In the preincubtion assay, a weak bacteriotoxicity (reduced background growth, slighty decrease in the number of revertants and/or slight reduction in the titer) was observed depending on the strain and test conditions from about 500 pg/plate onward.

Any other information on results incl. tables

Standard Plate Test
Strain	Metabolic activation system	Replicates	Revertant factor range	dose dependency	Assessment
TA 98	no	3	0.5 - 0.9	no	negative
	yes	3	0.5 - 0.8	no	negative
TA 100	no	3	0.7 - 0.9	no	negative
	yes	3	0.5 - 0.9	no	negative
TA 1535	no	3	0.4 - 0.9	no	negative
	yes	3	0.4 - 0.9	no	negative
TA 1537	no	3	0.4 - 1.0	no	negative
	yes	3	0.4 - 0.7	no	negative
WP2uvrA	no	3	0.7 - 0.8	no	negative
	yes	3	0.8 - 1.0	no	negative

Preincubation Test
Strain	Metabolic activation system	Replicates	Revertant factor range	dose dependency	Assessment
TA 98	no	3	0.6 - 0.9	no	negative
	yes	3	0.6 - 0.8	no	negative
TA 100	no	3	0.9 - 1.0	no	negative
	yes	3	0.8 - 1.0	no	negative
TA 1535	no	3	0.6 - 0.9	no	negative
	yes	3	0.5 - 0.9	no	negative
TA 1537	no	3	0.6 - 0.9	no	negative
	yes	3	0.6 - 0.9	no	negative
WP2uvrA	no	3	0.9 - 1.0	no	negative
	yes	3	0.7 - 0.9	no	negative