1,4:3,6-dianhydro-2,5-bis-O-(diphenoxyphosphoryl)-D-glucitol

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:

2010.06.01 - 2010-07-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (GLP)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Adopted on July 21, 1997

Deviations:

yes

Remarks:

Only 2-AA used as positive control in the presence of S9 mix

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology, BASF SE

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,4:3,6-dianhydro-2,5-bis-O-(diphenoxyphosphoryl)-D-glucitol

EC Number:

935-211-5

Cas Number:

1305113-15-8

Molecular formula:

C30H28O10P2

IUPAC Name:

1,4:3,6-dianhydro-2,5-bis-O-(diphenoxyphosphoryl)-D-glucitol

Constituent 2

Reference substance name:

phosphoric acid 6-(diphenoxy-phosphoryloxy)-hexahydro-furo[3,2-b]furan-3-yl ester diphenyl ester

IUPAC Name:

phosphoric acid 6-(diphenoxy-phosphoryloxy)-hexahydro-furo[3,2-b]furan-3-yl ester diphenyl ester

Details on test material:

Test substance: Isosorbid-O,O’-bis(diphenylphosphorsäureester); Lab test substance number: 10/0230-1
Physical state, appearance: solid, white
Batch identification: 9731/09/009 V1
Purity/composition: > 99% (w/w)
The test substance was homogen and stable throughout the study period
- Storage condition of test material: room temperature.

Method

Target gene:

his (salmonella strains), trp (e. coli)

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):

not applicable

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

not applicable

Metabolic activation:

with and without

Metabolic activation system:

liver S9-mix of phenobarbital (i.p.) and β-naphthoflavone (orally) induced rats

Test concentrations with justification for top dose:

0; 33; 100; 333; 1 000; 2 500 and 5 000 μg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2-AA), dissolved in DMSO, 2.5 μg/plate for the strains TA100, TA98, TA1357, TA1535, 60 μg/plate for E. coli WP2 uvrA

Remarks:

with metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 5 μg/plate dissolved in DMSO for strains TA 1535, TA 100

Remarks:

without metabolic activation

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine (NOPD), 10 μg/plate dissolved in DMSO for strain TA 98

Remarks:

without metabolic activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without metabolic activation

Migrated to IUCLID6: 100 μg/plate dissolved in DMSO for strain TA 1537

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

without metabolic activation

Migrated to IUCLID6: 5 μg/plate dissolved in DMSO for E. coli WP2 uvrA

Details on test system and experimental conditions:

Standard plate test and preincubation Test. 3 test plates per dose or per control.
Standard plate test (plate incorporation method) based on the method of Ames et al.
Preincubation Test procedure is based on the method described by Yahagi et al. and Matsushima et al.
After incubation at 37°C for 48 – 72 hours in the dark, the bacterial colonies (his+/trp+ revertants) are counted.
Determination of rate of induced back mutations of several bacteria mutants from histidine auxotrophy (his-/trp-) to histidine prototrophy (his+/trp+).

Evaluation criteria:

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

The experiment is considered valid if:
• The number of revertant colonies in the negative controls was within or nearby the range of the historical negative control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control substances both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
• The titer of viable bacteria was ≥ 10exp8/ml

The test substance is considered positive 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

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Weak bacteriotoxic effect (slight decrease in the number of his+ revertants) at 5000 μg/plate.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No relevant increase in the number of his+ or trp+ revertants was ovserved with or without metabolic activation in non of the tested strains.
No test substance precipitation was found.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative