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Toxicological information

Genetic toxicity: in vitro

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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:
25 June to 1 October 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to approved guidelines and in compliance with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Copy of certificate of compliance from UK GLP Monitoring Authority included in report
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Propane-1,2-diyl dibenzoate
EC Number:
242-894-7
EC Name:
Propane-1,2-diyl dibenzoate
Cas Number:
19224-26-1
Molecular formula:
C17H16O4
IUPAC Name:
2-(benzoyloxy)propyl benzoate
Constituent 2
Reference substance name:
1,2-propan-diyl-dibenzoate
IUPAC Name:
1,2-propan-diyl-dibenzoate
Constituent 3
Reference substance name:
K-FLEX* PG
IUPAC Name:
K-FLEX* PG
Test material form:
other: clear light yellow liquid
Details on test material:
- Name of test material (as cited in study report): Kalama® K-FLEX® PG
- Substance type: Plasticizer
- Physical state: Colourless to pale yellow liquid
- Analytical purity: 97.1% propylene glycol esters
- Lot/batch No.: KAKPG43301
- Expiration date of the lot/batch: 05 March 2016
- Storage condition of test material: Ambient temperature

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: compound insoluble in water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
2-nitrofluorene
sodium azide
benzo(a)pyrene
other: 2-Aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk

DURATION
- Preincubation period: 30 minutes (second test only)
- Exposure duration: 72h
- Expression time (cells in growth medium): not applicable to this test
- Selection time (if incubation with a selection agent): selection agent not used
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable to this test

SELECTION AGENT (mutation assays): not applicable to this test
SPINDLE INHIBITOR (cytogenetic assays): not applicable to this test
STAIN (for cytogenetic assays): not applicable to this test

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED: not applicable to this test

DETERMINATION OF CYTOTOXICITY
- Method: observed as a reduction in revertant colony numbers


OTHER EXAMINATIONS:
None
Evaluation criteria:
If exposure to a test substance produces a reproducible increase in revertant colony numbers of at least twice (three times in the case of strains TA1535 and TA1537) that of the concurrent vehicle controls, with some evidence of a positive concentration-response relationship, it is considered to exhibit mutagenic activity in this test system.

If exposure to a test substance does not produce a reproducible increase in revertant colony numbers, it is considered to show no evidence of mutagenic activity in this test system. No statistical analysis is performed.

If the results obtained fail to satisfy the criteria for a clear “positive” or “negative” response, even after additional testing, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers. The statistical procedures used are those described by Mahon et al (1989) and are usually Dunnett’s test followed, if appropriate, by trend analysis. Biological importance will be considered along with statistical significance. In general, treatment-associated increases in revertant colony numbers below two or three times those of the vehicle controls (as described above) are not considered biologically important. It should be noted that it is acceptable to conclude an equivocal response if no clear results can be obtained.
Statistics:
The mean number and standard deviation of revertant colonies were calculated for all groups. The “fold-increases” relative to the vehicle controls were calculated in order to compare the means for all treatment groups with those obtained for the vehicle control groups.

Results and discussion

Test resultsopen allclose all
Key result
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:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Toxicity was seen in strain WP2 uvrA (pKM101) following exposure to PGDB at 5000 µg/plate in the first test. No signs of toxicity were observed with any other strains following exposure to PGDB in either of the tests.
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
It was concluded that PGDB showed no evidence of mutagenic activity in this bacterial system under the test conditions employed.
Executive summary:

No evidence of toxicity was obtained following exposure to PGDB. No precipitate was observed on any plates containing PGDB. A maximum exposure concentration of 5000 μg/plate was, therefore, selected for use in the second test. No substantial increases in revertant colony numbers over control counts were obtained with any of the tester strains following exposure to PGDB at any concentration up to and including 5000 μg/plate in either the presence or absence of S9 mix.