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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:
02 December 2013 to 17 December 2013
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:
2013
Report Date:
2014

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: clear liquid
Details on test material:
- Name of test material (as cited in study report): 3-PPB

- Substance type: organic
- Physical state: clear liquid
- Analytical purity: 99.1%

- Purity test date:
- Lot/batch No.: EH101912A
- Expiration date of the lot/batch: 19 October 2014

- Storage condition of test material: Room temperature under nitrogen
- Other:

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: compound insoluble in water
Controls
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:
- Exposure duration: 72h
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS: 3

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:

OTHER:
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 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
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 3-PPB 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 3-PPB. No precipitate was observed on any plates containing 3-PPB. 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 3-PPB at any concentration up to and including 5000 μg/plate in either the presence or absence of S9 mix.