Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
September 12, 1988 – October 7, 1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test was conducted according to OECD Test Guideline No. 471, 1983, under GLP Standards, and QA. Chemical identity and purity of the test substance are not reported. One strain not tested.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report date:
1988

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1983
Deviations:
yes
Remarks:
Only four Salmonella test strains
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
yes
Remarks:
EU Method B.14 (Other effects-Mutagenicity: Salomonella typhimurium - Reverse Mutation Assay, 1984)
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Remarks:
Statement of Compliance
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of 3-[(diphenoxyphosphoryl)oxy]phenyl triphenyl 1,3-phenylene bis(phosphate) and tetraphenyl 1,3-phenylene bis(phosphate)
EC Number:
701-337-2
Cas Number:
not available
Molecular formula:
C30H24O8P2
IUPAC Name:
Reaction mass of 3-[(diphenoxyphosphoryl)oxy]phenyl triphenyl 1,3-phenylene bis(phosphate) and tetraphenyl 1,3-phenylene bis(phosphate)
Details on test material:
- Name of test material (as cited in study report): CR 733-S
- Physical state: no data
- Lot/batch No.: confidential
- Storage condition of test material: At room temperature in the dark

Method

Target gene:
His-gene: Amino acid histidine
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
other: S. typhimurium TA98 and TA100: pKM101. All strains: rfa, gal, chl, bio, uvrB
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9 induced by Aroclor 1254
Test concentrations with justification for top dose:
100, 333, 1000, 3330, and 5000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9-mix: TA1535: sodium azide; TA1537: 9-aminoacridine; TA98: daunomycine; TA100: methylmethanesulfonate. +S9-mix: all strains: 2-aminoanthracene.
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Selection time (if incubation with a selection agent): 48 hours

SELECTION AGENT (mutation assays): agar containing Histidine

NUMBER OF REPLICATIONS: 3 plates per concentration

NUMBER OF CELLS EVALUATED: 1 x 10*9 viable cells

DETERMINATION OF CYTOTOXICITY
- Method: Viability determination in non-selective agar: the percentage survival of the TA100 culture is determined by comparing the number of colonies on the solvent control plate with those on the plates containing the test substance, both in the absence and presence of S9-mix.

OTHER EXAMINATIONS:
Preliminary toxicity at 1.0, 3.3, 10, 33.3, 100, 333, 1000, 3330, and 5000 ug/plate.
Plate incorporation: independently repeated experiment.
Evaluation criteria:
A test substance is considered negative (not mutagenic) in the Ames test if:
a) The total number of revertants in any test strain at any concentration is not greater than two times the solvent control value, with or without metabolic activation.
b) The negative response should be reproducible in at least one independently repeated experiment.
A test substance is considered positive (mutagenic) in the Ames test if:
a) It induces at least a 2-fold, dose-related increase in the number of revertants with respect to the number induced by the solvent control in any of the test strains, either with or without metabolic activation. However, any mean plate count of less than 20 is considered to be not significant. If the test substance shows in the first test only a positive response at one or two concentrations, the assay is repeated with doses just below and exceeding those showing positive effects in the first test.
b) The positive response should be reproducible in at least one independently repeated experiment.
Statistics:
Mean and Standard Deviation

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: The test substance was not toxic towards the test strain TA100 in the preliminary toxicity test, both in the absence and presence of S9-mix. Therefore 5000 ug/plate was chosen as the top dose level in the mutation tests.

COMPARISON WITH HISTORICAL CONTROL DATA: The negative and strain-specific positive control values fell within the laboratory background historical ranges.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Not relevant

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

All bacterial strains showed negative responses over the entire dose range of the test substance. Based on the results of this study it is concluded that the test substance can be considered as not mutagenic in the Ames Salmonella/microsome assay.
Executive summary:

CR 733-S was tested in the Salmonella/microsome plate test at 100, 333, 1000, 3330, and 5000 ug/plate in the absence and presence of S9-mix. The test substance did not induce a dose-related increase in the number of revertant (His+) colonies in each of the four test strains (S. typhimurium TA1535, TA1537, TA98, and TA100). These results were confirmed in an independently repeated experiment. The test substance can, therefore, be considered as not mutagenic in this test system.