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EC number: 239-590-1 | CAS number: 15541-60-3
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Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
MPP was Negative in the Ames test and in vitro chromosome aberration tests.
Link to relevant study records
- 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:
- 7 January to 9 February 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP study conducted in accordance with an internationally recognised method
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- histidine synthesis
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction
- Test concentrations with justification for top dose:
- Preliminary toxicity test: 0, 0.15, 0.5, 1.5, 5. 15, 50, 500, 1500, 5000 µg/plate
Range finding test: 0, 50, 150, 500, 1500, 5000 µg/plate
Main test: 0, 50, 150, 500, 1500, 5000 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Remarks:
- 2 µg/plate for WP2uvrA-, 3 µg/plate for TA100 and 5 µg/plate for TA1535
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- 80 µg/plate for TA1537
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- 0.2 µg/plate for TA98
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- In the presence of S9 only
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- In the presence of S9 only
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium; in agar (plate incorporation)
DURATION
- Preincubation period: not applicable
- Exposure duration: 48 hours (all tests)
NUMBER OF REPLICATIONS: 3
NUMBER OF CELLS EVALUATED:
DETERMINATION OF CYTOTOXICITY
- Method: number of revertant colonies - Evaluation criteria:
- The test material may be considered positive in this test system if it induces a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.
- Statistics:
- Dunnett's method of linear regression to determine significance of increase in revertant count
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Remarks:
- The test material was non-toxic to the strains of bacteria used (TAl 00 and WP2uvrA-).
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
None
RANGE-FINDING/MUTATION TEST
The test material caused no visible reduction in the growth of the bacterial background lawn at any dose level. The test material was, therefore, tested up to the maximum recommended dose level of 5000 µg/plate. No test material precipitate was observed on the plates at any of the doses
tested in either the presence or absence of S9-mix.
No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.
COMPARISON WITH HISTORICAL CONTROL DATA:
The vehicle (dimethyl sulphoxide) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the 89-mix were validated. - Remarks on result:
- other: strain/cell type: Salmonella uvrB-, Escherichia coli (WP2uvrA-)
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
The test material was considered to be non-mutagenic under the conditions of this test. - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 10 September 2010 to 2 February 2011
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- lymphocytes:
- Details on mammalian cell type (if applicable):
- Obtained from two healthy non-smoking donors
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9
- Test concentrations with justification for top dose:
- First test:
In the absence of S9 mix - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 µg/mL.
In the presence of S9 mix (2% v/v) - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 µg/mL.
Second test:
In the absence of S9 mix - 21 hour continuous treatment: 68.4, 114 and 190 µg/mL.
In the presence of S9 mix (5% v/v) - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 µg/mL. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: culture medium
- Justification for choice of solvent/vehicle: prior to commencing testing, the solubility of the test substance in solvents compatible with the test system was assessed. MPP was found to form a dosable fine suspension in culture medium at 0.38 mg/mL. On dosing a 0.38 mg/mL fine suspension at 50% v/v (2500 μL per 5 mL culture) into culture medium, giving a final concentration of 190 μg/mL, no precipitate (visible by eye) was observed. - Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- In the absence of S9 mix
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- In the presence of S9 mix
- 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
First test
In the absence of S9 mix - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 μg/mL.
In the presence of S9 mix (2% v/v) - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 μg/mL.
Second test
In the absence of S9 mix - 21 hour continuous treatment: 68.4, 114 and 190 μg/mL.
In the presence of S9 mix (5% v/v) - 3 hour treatment, 18 hour recovery: 68.4, 114 and 190 μg/mL.
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): 10% Giemsa, prepared in buffered water (pH 6.8).
NUMBER OF CELLS EVALUATED: metaphase analysis 100, mitotic index 1000
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index;
OTHER EXAMINATIONS:
- Determination of polyploidy: yes - Evaluation criteria:
- An assay is considered to be acceptable if the negative and positive control values lie within the current historical control range.
The test substance is considered to cause a positive response if the following conditions are met:
i) Statistically significant increases (P<0.01) in the frequency of metaphases with aberrant chromosomes (excluding gaps) are observed at one or more test concentration.
ii) The increases exceed the negative control range of this laboratory, taken at the 99% confidence limit.
iii) The increases are reproducible between replicate cultures.
iv) The increases are not associated with large changes in pH, osmolality of the treatment medium or extreme toxicity.
v) Evidence of a concentration-related response is considered to support the conclusion.
A negative response is claimed if no statistically significant increases in the number of aberrant cells above concurrent control frequencies are observed, at any concentration. - Species / strain:
- lymphocytes:
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH:
- Effects of osmolality:
- Evaporation from medium:
- Water solubility:
- Precipitation:
- Other confounding effects:
RANGE-FINDING/SCREENING STUDIES:
COMPARISON WITH HISTORICAL CONTROL DATA:
ADDITIONAL INFORMATION ON CYTOTOXICITY: - Remarks on result:
- other: other: derived from human blood
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
It is concluded that MPP has shown no evidence of causing an increase in the frequency of structural chromosome aberrations in this in vitro cytogenetic test system, under the experimental conditions described.
Referenceopen allclose all
Toxicity:
In the absence of S9 mix following 3 hour treatment, MPP caused no significant reduction in the mitotic index at 190 μg/mL, compared to the solvent control value. As this was the maximum achievable solubility within this test system, the concentrations selected for the metaphase analysis were 68.4, 114 and 190 μg/mL.
In the presence of S9 mix (2% v/v final concentration) following 3 hour treatment, MPP caused no significant reduction in the mitotic index at 190 μg/mL, compared to the solvent control value. As this was the maximum achievable solubility within this test system, the concentrations selected for the metaphase analysis were 68.4, 114 and 190 μg/mL.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
In vitro genotoxicity:
A bacterial reverse mutation assay (Ames test, SafePharm, 2004) has been undertaken following OECD/EU test methods. Experiments were performed both in the absence and presence of metabolic activation at concentrations up to 5000 µg/plate. The substance did not induce reverse mutation in Salmonella typhimurium or Escherichia coli.
The ability to cause chromosomal damage has been investigated (HLS, 2011) in cultured human lymphocytes, in vitro in the absence and presence of S9 metabolic activation according to OECD/EU test methods. Two independent experiments were performed, in the first the cells were treated for 3 hours in the presence and absence of S9 metabolism and harvested after 18 hours. A second experiment was performed as for the first test except that cells in the absence of S9 mix were subject to continuous 21-hour treatment. MPP did not induce chromosomal aberrations.
Justification for classification or non-classification
Negative in the Ames test and in vitro chromosome aberration and gene mutation tests.
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