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EC number: 205-811-5 | CAS number: 152-97-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- June to Aug 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 005
- Report date:
- 2005
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1997
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 6-fluoro-11,21-dihydroxy-16- methylpregna-1,4-diene-3,20-dione
- Cas Number:
- 152-97-6
- Molecular formula:
- C22H29FO4
- IUPAC Name:
- 6-fluoro-11,21-dihydroxy-16- methylpregna-1,4-diene-3,20-dione
- Details on test material:
- - Name of test material (as cited in study report): fluocortolone (ZK 10445) batch containing 5% ZK 344937 as impurity
- Batch No.of fluocortolone (ZK 10445): 13070001
- Purity of fluocortolone (ZK 10445): 98.2 %
- Batch No.of impurity (ZK 344937): PA172QCB248
- Purity of impurity (ZK 344937): 98.5 %
Constituent 1
Method
- Target gene:
- Histidine gene locus
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 98
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 100
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1535
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1537
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 induced male rat liver S9 mix
Type and composition of metabolic activation system:
- source of S9: Liver homogenates (S9: 9000 x g fraction), derived from male Sprague-Dawley rats pre-treated with Aroclor 1254, was obtained from ICN/Cappel Pharmaceuticals, Inc., Aurora, Ohio, USA, [S9 batch number:7756F; protein content 35.0 mg/mL; EROD activity: 4593.2 pmoles 7-hydroxyresorufin/min/mg S9 protein].
- concentration or volume of S9 mix and S9 in the final culture medium: The components of the standard S9 mix were 8 mmol/L MgCl;, 33 mmol/L KCl, 5 mmol/L glucose-6-phosphate, 4 mmol/L NADP, 100 mmol/L sodium phosphate, pH 7.4 and S9.at a concentration of0.1 mL per mL of mix (standard plate incorporation and E.coli) and for the preincubation test S9 at a concentration of 0.3 mL per mL of mix (only S.typhimurium). - Test concentrations with justification for top dose:
- Concentrations of test item (fluocortolone containing 5% ZK 344937 as impurity): 0.01, 0.05, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0 mg/plate (all strains)
- Vehicle / solvent:
- DMSO
Controls
- Untreated negative controls:
- yes
- Remarks:
- phosphate buffer
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- sodium azide
- N-dimethylnitrosamine
- benzo(a)pyrene
- cyclophosphamide
- ethylmethanesulphonate
- other: anthracene-2-amine (with metabolic activation), N-methyl-N'-nitro-N-nitrosoguanidine (without metabolic activation), 4-nitro-o-phenylenediamine (without metabolic activation)
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate
- Number of independent experiments: two
METHOD OF TREATMENT/ EXPOSURE:
- Test substance added in agar (plate incorporation); preincubation
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method, e.g.: background growth inhibition - Rationale for test conditions:
- As recommended by the OECD test guideline
- Evaluation criteria:
- please refer to 'Statistics'
- Statistics:
- The arithmetic means of the number ofmutant colonies of the 3 parallel plates in the negative control groups were compared with those of the compound groups. However, in the tables the given mean values are rounded to the nearest integer. A positive response was considered if the number of revertants of the compound groups compared to the number of revertants of the negative group was reproducibly higher than 2-fold. A dose-dependent increase in the number of
revertants was also considered to indicate a mutagenic effect.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation method: growth inhibition at 1.0 mg/plate and above (+/- S9 mix) preincubation modification: growth inhibition at 0.1 mg/plate and above (- S9 mix) or at at 1.0 mg/plate and above (+ S9 mix)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation method: growth inhibition at 1.0 mg/plate and above (+/- S9 mix) preincubation modification: growth inhibition at 0.1 mg/plate and above (- S9 mix) or at at 1.0 mg/plate and above (+ S9 mix)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation method: growth inhibition at 1.0 mg/plate and above (+/- S9 mix) preincubation modification: growth inhibition at 0.1 mg/plate and above (- S9 mix) or at at 1.0 mg/plate and above (+ S9 mix)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation method: growth inhibition at 1.0 mg/plate and above (+/- S9 mix) preincubation modification: growth inhibition at 0.1 mg/plate and above (- S9 mix) or at at 1.0 mg/plate and above (+ S9 mix)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- plate incorporation method: growth inhibition at 1.0 mg/plate and above (+/- S9 mix) preincubation modification: growth inhibition at 0.1 mg/plate and above (- S9 mix) or at at 1.0 mg/plate and above (+ S9 mix)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
Any other information on results incl. tables
None of the six tester strains showed increased reversion to prototrophy with the test item (fluocortolone containing 5% ZK 344937 as impurity) at the doses tested in the two tests (direct plate incorporation procedure and preincubation modification), either in the absence or presence of S9 mix.
Precipitates in the agar were observed at the maximum dose of 5.0 mg per plate.
In the direct plate incorporation assay with Salmonella typhimurium growth inhibition of the background lawn was observed from 1.0 mg per plate onwards without or with metabolic activation. When using the preincubation modification growth inhibition of the background lawn was observed in the experiments with Salmonella typhimurium from 0.1 mg per plate onwards without metabolic activation and from 1.0 mg per plate onwards with metabolic activation.
No growth inhibition ofthe background lawn was observed in the experiments with Escherichia coli WP2uvrA.
Negative controls and positive controls with known mutagens (anthracene-2-amine, benzo[a]pyrene, cyclophosphamide, ethyl methanesulfonate, N-methyl-N' -nitro-N-nitrosoguanidine, 2-nitro-9H-fluorene, 4-nitro-o-phenylenediamine, sodium azide, N-nitrosodimethylamine) produced the expected distinct increase in the number of revertant colonies.
Direct plate incorporation test on Salmonella typhimurium strain TA98 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 17 | 19 | 2 | 33 | 34 | 2 | 1.0 | 1.0 |
|
| 19 |
|
| 36 |
|
|
|
|
|
| 21 |
|
| 32 |
|
|
|
|
Phosphate buffer | 50 µL | 19 | 21 | 2 | 40 | 40 | 7 | 1.1 | 1.2 |
|
| 22 |
|
| 47 |
|
|
|
|
|
| 22 |
|
| 34 |
|
|
|
|
ZK10445/ZK344937 | 0.1 mg | 15 | 13 | 3 | 41 | 33 | 7 | 0.7 | 1.0 |
|
| 10 |
|
| 31 |
|
|
|
|
|
| 15 |
|
| 28 |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 17 | 16 |
| 31 | 38 | 6 | 0.8 | 1.1 |
|
| 15 |
|
| 42 |
|
|
|
|
|
| 15 |
|
| 42 |
|
|
|
|
ZK10445/ZK344937 | 0.5 mg | 7 | 9 | 3 | 42 | 39 | 5 | 0.5 | 1.1 |
|
| 9 |
|
| 33 |
|
|
|
|
|
| 12 |
|
| 41 |
|
|
|
|
ZK10445/ZK34493 7 | 1.0 mg | 10 B | 11 | 3 | 29 | 34 | 5 | 0.6 | 1.0 |
|
| 14 B |
|
| 38 |
|
|
|
|
|
| 8 B |
|
| 36 |
|
|
|
|
ZK10445/ZK344937 | 2.5 mg | 7 B | 8 |
| 33 B | 37 | 4 | 0.4 | 1.1 |
|
| 8 B |
|
| 36 B |
|
|
|
|
|
| 8 B |
|
| 41 B |
|
|
|
|
ZK10445/ZK34493 7 | 5.0 mg | 1 mPB |
|
| 45 mPB | 40 | 5 | 0.0 | 1.2 |
|
| 1 mPB |
|
| 38 mPB |
|
|
|
|
|
| 0 mPB |
|
| 36 mPB |
|
|
|
|
Anthracene-2-amine | 2.5 µg | 12 | 12 | 5 | 709 | 778 | 60 | 0.6 | 23.1 |
|
| 8 |
|
| 822 |
|
|
|
|
|
| 17 |
|
| 802 |
|
|
|
|
Benzo[a]pyrene | 2.5 µg | 8 | 9 |
| 243 | 243 | 6 | 0.5 | 7.2 |
|
| 10 |
|
| 238 |
|
|
|
|
|
| 8 |
|
| 249 |
|
|
|
|
2-Nitrofluorene | 10.0 µg | 731 | 721 | 21 | 229 | 244 | 13 | 37.9 | 7.2 |
|
| 697 |
|
| 251 |
|
|
|
|
|
| 735 |
|
| 251 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Direct plate incorporation test on Salmonella typhimurium strain TA 100 | |||||||||
|
|
|
|
|
|
|
|
|
|
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 100 | 103 | 4 | 92 | 94 | 3 | 1.0 | 1.0 |
|
| 101 |
|
| 92 |
|
|
|
|
|
| 108 |
|
| 98 |
|
|
|
|
Phosphate buffer | 50µL | 101 | 107 | 6 | 90 | 95 | 5 | 1.0 | 1.0 |
|
| 109 |
|
| 96 |
|
|
|
|
|
| 112 |
|
| 99 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 100 | 87 | 11 | 86 | 89 | 2 | 0.8 | 0.9 |
|
| 81 |
|
| 90 |
|
|
|
|
|
| 80 |
|
| 90 |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 82 | 88 | 8 | 88 | 85 | 16 | 0.9 | 0.9 |
|
| 84 |
|
| 100 |
|
|
|
|
|
| 97 |
|
| 68 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 100 | 87 | 15 | 87 | 79 | 7 | 0.8 | 0.8 |
|
| 71 |
|
| 76 |
|
|
|
|
|
| 89 |
|
| 73 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 82 | 86 | 6 | 98 | 85 | 11 | 0.8 | 0.9 |
|
| 93 |
|
| 82 |
|
|
|
|
|
| 83 |
|
| 76 |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 80B | 88 | 12 | 106B | 94 | 12 | 0.9 | 1.0 |
|
| 102B |
|
| 94B |
|
|
|
|
|
| 83B |
|
| 83B |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 28 mPB | 25 | 4 | 47 mB | 46 | 8 | 0.2 | 0.5 |
|
| 21 mPB |
|
| 38 mPB |
|
|
|
|
|
| 25 mPB |
|
| 53 mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 112 | 114 | 10 | 939 | 953 | 38 | 1.1 | 10.1 |
|
| 105 |
|
| 996 |
|
|
|
|
|
| 125 |
|
| 924 |
|
|
|
|
Benzo[a]pyrene | 2.5µg | 83 | 83 | 0 | 478 | 445 | 29 | 0.8 | 4.7 |
|
| 83 |
|
| 425 |
|
|
|
|
|
| 83 |
|
| 433 |
|
|
|
|
Sodium azide | 5.0µg | 437 | 453 | 27 | 164 | 158 | 5 | 4.4 | 1.7 |
|
| 485 |
|
| 154 |
|
|
|
|
|
| 438 |
|
| 157 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
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|
Direct plate incorporation test on Salmonella typhimurium strain TA1535 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 15 | 17 | 3 | 16 | 14 | 3 | 1.0 | 1.0 |
|
| 21 |
|
| 14 |
|
|
|
|
|
| 15 |
|
| 11 |
|
|
|
|
Phosphate buffer | 50 µL | 21 | 19 | 4 | 12 | 16 | 3 | 1.1 | 1.2 |
|
| 21 |
|
| 18 |
|
|
|
|
|
| 14 |
|
| 18 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 26 | 22 | 5 | 11 | 12 | 5 | 1.3 | 0.9 |
|
| 24 |
|
| 8 |
|
|
|
|
|
| 16 |
|
| 18 |
|
|
|
|
ZK10445/ZK344937 | 0.25mg | 21 | 19 | 4 | 9 | 10 | 3 | 1.1 | 0.8 |
|
| 15 |
|
| 8 |
|
|
|
|
|
| 22 |
|
| 14 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 16 | 20 | 3 | 9 | 11 | 2 | 1.2 | 0.8 |
|
| 22 |
|
| 12 |
|
|
|
|
|
| 22 |
|
| 13 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 23 | 22 | 3 | 6 | 11 | 4 | 1.3 | 0.8 |
|
| 25 |
|
| 13 |
|
|
|
|
|
| 19 |
|
| 13 |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 15 | 15 | 2 | 14 | 12 | 2 | 0.9 | 0.9 |
|
| 17 |
|
| 12 |
|
|
|
|
|
| 13 |
|
| 11 |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 10 mPB | 13 | 3 | 11 mPB | 10 |
| 0.7 | 0.7 |
|
| 13 mPB |
|
| 9 mPB |
|
|
|
|
|
| 15 mPB |
|
| 9 mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 15 | 19 | 4 | 113 | 107 | 8 | 1.1 | 7.8 |
|
| 20 |
|
| 98 |
|
|
|
|
|
| 23 |
|
| 110 |
|
|
|
|
Cyclophosphamide | 400.0 µg | 36 | 35 | 7 | 364 | 365 |
| 2.0 | 26.7 |
|
| 41 |
|
| 366 |
|
|
|
|
|
| 27 |
|
| 364 |
|
|
|
|
Sodium azide | 5.0µg | 505 | 498 | 22 | 114 | 133 | 21 | 29.3 | 9.7 |
|
| 515 |
|
| 156 |
|
|
|
|
|
| 473 |
|
| 129 |
|
|
|
|
|
|
|
|
|
|
|
|
|
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|
|
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|
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|
Direct plate incorporation test on Salmonella typhimurium strain TA1538 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 9 | 12 | 3 | 26 | 23 | 5 | 1.0 | 1.0 |
|
| 12 |
|
| 17 |
|
|
|
|
|
| 14 |
|
| 26 |
|
|
|
|
Phosphate buffer | 50 µL | 8 | 8 |
| 27 | 28 | 2 | 0.7 | 1.2 |
|
| 9 |
|
| 27 |
|
|
|
|
|
| 7 |
|
| 31 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 11 | 8 | 3 | 17 | 22 | 7 | 0.7 | 1.0 |
|
| 5 |
|
| 30 |
|
|
|
|
|
| 9 |
|
| 19 |
|
|
|
|
ZK10445/ZK344937 | 0.25mg | 8 | 7 |
| 22 | 18 | 6 | 0.6 | 0.8 |
|
| 7 |
|
| 22 |
|
|
|
|
|
| 7 |
|
| 11 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 8 | 7 | 2 | 17 | 18 |
| 0.6 | 0.8 |
|
| 8 |
|
| 19 |
|
|
|
|
|
| 4 |
|
| 18 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 8 | 7 | 2 | 25 | 25 |
| 0.6 | 1.1 |
|
| 9 |
|
| 25 |
|
|
|
|
|
| 5 |
|
| 26 |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 2 mB | 2 |
| 27 | 24 | 3 | 0.2 | 1.0 |
|
| 2 mB |
|
| 21 |
|
|
|
|
|
| 3 mB |
|
| 23 |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 0 mPB | 0 | 0 | 13 mPB | 16 | 4 | 0.0 | 0.7 |
|
| 0 mPB |
|
| 20 mPB |
|
|
|
|
|
| 0 mPB |
|
| 15 mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 4 | 7 | 4 | 615 | 621 | 19 | 0.6 | 27.0 |
|
| 6 |
|
| 606 |
|
|
|
|
|
| 12 |
|
| 643 |
|
|
|
|
2-Nitrofluorene | 10.0µg | 811 | 735 | 101 | 312 | 304 | 8 | 63.0 | 13.2 |
|
| 620 |
|
| 296 |
|
|
|
|
|
| 774 |
|
| 304 |
|
|
|
|
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|
|
Direct plate incorporation test on Salmonella typhimurium strain TA1537 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 6 | 9 | 3 | 10 | 12 | 2 | 1.0 | 1.0 |
|
| 9 |
|
| 14 |
|
|
|
|
|
| 11 |
|
| 12 |
|
|
|
|
Phosphate buffer | 50 µL | 8 | 8 |
| 14 | 11 | 3 | 1.0 | 0.9 |
|
| 9 |
|
| 9 |
|
|
|
|
|
| 8 |
|
| 11 |
|
|
|
|
ZK1044S/ZK344937 | 0.1mg | 6 | 8 | 2 | 10 | 11 | 4 | 0.9 | 0.9 |
|
| 9 |
|
| 8 |
|
|
|
|
|
| 9 |
|
| 15 |
|
|
|
|
ZK10445/ZK344937 | 0.25mg | 8 | 8 | 3 | 7 | 11 | 4 | 0.9 | 0.9 |
|
| 5 |
|
| 15 |
|
|
|
|
|
| 10 |
|
| 12 |
|
|
|
|
ZK! 0445/ZK344937 | 0.5mg | 5 | 6 | 3 | 8 | 8 |
| 0.7 | 0.7 |
|
| 9 |
|
| 9 |
|
|
|
|
|
| 4 |
|
| 7 |
|
|
|
|
ZK! 044S/ZK344937 | 1.0mg | 5B | 5 |
| 8B | 9 | 2 | 0.6 | 0.8 |
|
| 6B |
|
| 8B |
|
|
|
|
|
| 4B |
|
| 11B |
|
|
|
|
ZKI 0445/ZK344937 | 2.5mg | 1 mB | 1 |
| 4 mB | 5 | 1 | 0.1 | 0.4 |
|
| 1 mB |
|
| 5 mB |
|
|
|
|
|
| 0 mB |
|
| 5 mB |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 0 mPB | 0 | 0 | 5 mPB | 3 | 2 | 0.0 | 0.2 |
|
| 0 mPB |
|
| 1 mPB |
|
|
|
|
|
| 0 mPB |
|
| 2 mPB |
|
|
|
|
Anthracene-2-amine | 2.5mg | 8 | 7 |
| 101 | 98 | 8 | 0.8 | 8.1 |
|
| 7 |
|
| 103 |
|
|
|
|
|
| 7 |
|
| 89 |
|
|
|
|
4-NPDA | 10.0 µg | 63 | 66 | 7 | 17 | 21 | 5 | 7.6 | 1.8 |
|
| 74 |
|
| 26 |
|
|
|
|
|
| 60 |
|
| 20 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| |||||||||
Direct plate incorporation assay E.coli WP2uvrA |
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 19 | 22 | 3 | 29 | 31 | 3 | 1.0 | 1.0 |
|
| 23 |
|
| 34 |
|
|
|
|
|
| 25 |
|
| 29 |
|
|
|
|
Phosphate buffer | 50 µL | 22 | 25 | 4 | 32 | 33 | 2 | 1.1 | 1.1 |
|
| 30 |
|
| 31 |
|
|
|
|
|
| 23 |
|
| 35 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 25 | 21 | 4 | 21 | 28 | 8 | 0.9 | 0.9 |
|
| 20 |
|
| 27 |
|
|
|
|
|
| 18 |
|
| 36 |
|
|
|
|
ZKl 0445/ZK344937 | 0.25 mg | 23 | 25 | 3 | 31 | 29 | 2 | 1.1 | 1.0 |
|
| 29 |
|
| 27 |
|
|
|
|
|
| 24 |
|
| 30 |
|
|
|
|
ZKl 0445/ZK344937 | 0.5mg | 18 | 18 | 5 | 27 | 32 | 6 | 0.8 | 1.0 |
|
| 23 |
|
| 31 |
|
|
|
|
|
| 14 |
|
| 38 |
|
|
|
|
ZKl 0445/ZK344937 | 1.0mg | 29 | 24 | 5 | 20 | 28 | 7 | 1.1 | 0.9 |
|
| 20 |
|
| 29 |
|
|
|
|
|
| 22 |
|
| 34 |
|
|
|
|
ZK l 0445/ZK344937 | 2.5mg | 23 | 20 | 3 | 38 | 33 | 4 | 0.9 | 1.1 |
|
| 21 |
|
| 31 |
|
|
|
|
|
| 17 |
|
| 30 |
|
|
|
|
ZKl 0445/ZK344937 | 5.0mg | 15 mP | 16 | 2 | 26 mP | 25 | 2 | 0.7 | 0.8 |
|
| 15 m P |
|
| 23 mP |
|
|
|
|
|
| 18 mP |
|
| 2S mP |
|
|
|
|
Anthracene-2-amine | 10.0 µg | 25 | 23 | 2 | 155 | 192 | 34 | 1.0 | 6.3 |
|
| 21 |
|
| 200 |
|
|
|
|
|
| 22 |
|
| 221 |
|
|
|
|
EMS | 5.0µL | 612 | 572 | 75 | 490 | 553 | 62 | 25.6 | 18.0 |
|
| 619 |
|
| 614 |
|
|
|
|
|
| 48S |
|
| 555 |
|
|
|
|
MNNG | 5.0µg | 289 | 268 | 33 | 222 | 238 | 23 | 12.0 | 7.8 |
|
| 285 |
|
| 229 |
|
|
|
|
|
| 230 |
|
| 264 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain T A98 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 14 | 14 | 0 | 47 | 40 | 8 | 1.0 | 1.0 |
|
| 14 |
|
| 41 |
|
|
|
|
|
| 14 |
|
| 31 |
|
|
|
|
Phosphate buffer | 50 µL | 10 | 13 | 3 | 34 | 44 | 10 | 1.0 | 1.1 |
|
| 16 |
|
| 54 |
|
|
|
|
|
| 14 |
|
| 44 |
|
|
|
|
ZK!0445/ZK344937 | 0.1mg | 14 | 12 | 2 | 32 | 36 | 5 | 0.9 | 0.9 |
|
| 12 |
|
| 36 |
|
|
|
|
|
| 11 |
|
| 41 |
|
|
|
|
ZKl 0445/ZK344937 | 0.25mg | 11 | 10 | 2 | 37 | 33 | 4 | 0.7 | 0.8 |
|
| 7 |
|
| 30 |
|
|
|
|
|
| 11 |
|
| 33 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 3mB | 5 | 2 | 29 | 35 | 5 | 0.4 | 0.9 |
|
| 7 mB |
|
| 37 |
|
|
|
|
|
| 5 mB |
|
| 39 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 1022) B1) | 78 | 28 | 20B | 22 | 2 | 5.6 | 0.5 |
|
| 482) B1) |
|
| 24B |
|
|
|
|
|
| 852) B1) |
|
| 21B |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 782) B1) | 29 | 43 | 18B | 19 | 3 | 2.0 | 0.5 |
|
| 6 B1) |
|
| 17B |
|
|
|
|
|
| 2 B1) |
|
| 23B |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 1mPB1) | 1 | 1 | 15mPB | 17 | 9 | 0.0 | 0.4 |
|
| 0 mPB1) |
|
| 27mPB |
|
|
|
|
|
| 1 mPB1) |
|
| 10mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 13 | 13 | 4 | 244 | 245 | 13 | 1.0 | 6.2 |
|
| 17 |
|
| 259 |
|
|
|
|
|
| 10 |
|
| 233 |
|
|
|
|
Anthracene-2-amine | 3.5µg | 22 | 23 |
| 326 | 376 | 57 | 1.6 | 9.5 |
|
| 22 |
|
| 438 |
|
|
|
|
|
| 24 |
|
| 365 |
|
|
|
|
Benzo[a]pyrene | 2.5µg | 5 | 9 | 7 | 166 | 153 | 18 | 0.7 | 3.9 |
|
| 6 |
|
| 161 |
|
|
|
|
|
| 17 |
|
| 132 |
|
|
|
|
2-Nitrofluorene | 10.0 µg | 798 | 799 | 25 | 245 | 217 | 30 | 57.0 | 5.5 |
|
| 774 |
|
| 221 |
|
|
|
|
|
| 824 |
|
| 185 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA98 (a) | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 16 | 18 | 3 |
| / | / | 1.0 | / |
|
| 22 |
|
|
|
|
|
|
|
|
| 16 |
|
|
|
|
|
|
|
Phosphate buffer | 50 µL | 20 | 23 | 3 |
| / | / | 1.3 | / |
|
| 25 |
|
|
|
|
|
|
|
|
| 23 |
|
|
|
|
|
|
|
ZK!0445/ZK344937 | 0.05mg | 23 | 18 | 5 |
| / | / | 1.0 | / |
|
| 14 |
|
|
|
|
|
|
|
|
| 17 |
|
|
|
|
|
|
|
ZKl 0445/ZK344937 | 0.10mg | 10 | 13 | 3 |
| / | / | 0.7 | / |
|
| 15 |
|
|
|
|
|
|
|
|
| 15 |
|
|
|
|
|
|
|
ZK!0445/ZK344937 | 0.25mg | 20 | 15 | 5 |
| / | / | 0.8 | / |
|
| 14 |
|
|
|
|
|
|
|
|
| 11 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.5 mg | 4mB | 2 | 2 |
| / | / | 0.1 | / |
|
| 1mB |
|
|
|
|
|
|
|
|
| 2mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.75mg | 1mB | 1 | 2 |
| / | / | 0.1 | / |
|
| 0mB |
|
|
|
|
|
|
|
|
| 3mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 472)mB1) | 24 | 20 |
| / | / | 1.3 | / |
|
| 8mB1) |
|
|
|
|
|
|
|
|
| 172)mB1) |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 2.0mg | 0mB1) | 6 | 11 |
| / | / | 0.4 | / |
|
| 0mB1) |
|
|
|
|
|
|
|
|
| 192)mB1) |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 3.0mg | 12mB1) | 9 | 3 |
| / | / | 0.5 | / |
|
| 6mB1) |
|
|
|
|
|
|
|
|
| 10mB1) |
|
|
|
|
|
|
|
2-NF | 10.0 µg | 958 | 874 | 149 |
| / | / | 48.5 | / |
|
| 702 |
|
|
|
|
|
|
|
|
| 961 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA100 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 88 | 93 | 4 | 100 | 101 | 8 | 1.0 | 1.0 |
|
| 96 |
|
| 110 |
|
|
|
|
|
| 94 |
|
| 94 |
|
|
|
|
Phosphate buffer | 50 µL | 101 | 101 | 2 | 102 | 109 | 10 | 1.1 | 1.1 |
|
| 100 |
|
| 105 |
|
|
|
|
|
| 103 |
|
| 121 |
|
|
|
|
ZKl0445/ZK344937 | 0.1mg | 77 | 83 | 13 | 95 | 88 | 10 | 0.9 | 0.9 |
|
| 73 |
|
| 77 |
|
|
|
|
|
| 98 |
|
| 93 |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 74 | 75 | 2 | 110 | 98 | 12 | 0.8 | 1.0 |
|
| 77 |
|
| 86 |
|
|
|
|
|
| 74 |
|
| 98 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 69 | 41 | 24 | 98 | 91 | 7 | 0.4 | 0.9 |
|
| 24 |
|
| 84 |
|
|
|
|
|
| 30 |
|
| 91 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 5 mB | 11 | 5 | 90 | 82 | 20 | 0.1 | 0.8 |
|
| 15 mB |
|
| 59 |
|
|
|
|
|
| 12 mB |
|
| 97 |
|
|
|
|
ZK10445/ZK34493 7 | 2.5mg | 25 m B | 23 | 4 | 103B | 86 | 20 | 0.2 | 0.8 |
|
| 26 m B |
|
| 91B |
|
|
|
|
|
| 18 mB |
|
| 64B |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 18 mPB | 20 | 2 | 42mPB | 53 | 12 | 0.2 | 0.5 |
|
| 22 mPB |
|
| 66mPB |
|
|
|
|
|
| 20 mPB |
|
| 50mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 113 | 110 | 14 | 234 | 225 | 9 | 1.2 | 2.2 |
|
| 95 |
|
| 217 |
|
|
|
|
|
| 123 |
|
| 223 |
|
|
|
|
Anthracene-2-amine | 3.5µg | 76 | 88 | 21 | 281 | 322 | 40 | 1.0 | 3.2 |
|
| 77 |
|
| 361 |
|
|
|
|
|
| 112 |
|
| 325 |
|
|
|
|
DMNA | 5.0µL | 88 | 88 | 0 | 1561 | 1471 | 127 | 0.9 | 14.5 |
|
| 88 |
|
| 1526 |
|
|
|
|
|
| 88 |
|
| 1325 |
|
|
|
|
Sodium azide | 5.0µg | 498 | 486 | 12 | 130 | 128 | 3 | 5.2 | 1.3 |
|
| 485 |
|
| 124 |
|
|
|
|
|
| 475 |
|
| 130 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA100 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50µL | 104 | 102 | 3 |
| I | / | 1.0 |
|
|
| 98 |
|
|
|
|
|
|
|
|
| 104 |
|
|
|
|
|
|
|
Phosphate buffer | 50 µL | 90 | 99 | 8 |
| I | / | 1.0 |
|
|
| 102 |
|
|
|
|
|
|
|
|
| 105 |
|
|
|
|
|
|
|
ZK! 0445/ZK344937 | 0.01mg | 89 | 89 | 2 |
| I | / | 0.9 |
|
|
| 88 |
|
|
|
|
|
|
|
|
| 91 |
|
|
|
|
|
|
|
ZK10445/ZK34493 7 | 0.05mg | 86 | 90 | 4 |
| I | / | 0.9 |
|
|
| 90 |
|
|
|
|
|
|
|
|
| 94 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 91 | 79 | 16 |
| I | / | 0.8 | / |
|
| 61 |
|
|
|
|
|
|
|
|
| 84 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 52 | 66 | 14 |
| I | / | 0.6 |
|
|
| 65 |
|
|
|
|
|
|
|
|
| 80 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 16 mB | 19 | 6 |
| I | / | 0.2 |
|
|
| 26 mB |
|
|
|
|
|
|
|
|
| 15 mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 30 mB | 22 | 8 |
| I | / | 0.2 |
|
|
| 15 mB |
|
|
|
|
|
|
|
|
| 21 mB |
|
|
|
|
|
|
|
Sodium azide | 5.0µg | 726 | 690 | 41 |
| I | / | 6.8 |
|
|
| 699 |
|
|
|
|
|
|
|
|
| 646 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA1535 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 17 | 18 |
| 19 | 16 | 4 | 1.0 | 1.0 |
|
| 18 |
|
| 11 |
|
|
|
|
|
| 18 |
|
| 18 |
|
|
|
|
Phosphate buffer | 50 µL | 22 | 24 | 2 | 13 | 13 |
| 1.4 | 0.8 |
|
| 25 |
|
| 13 |
|
|
|
|
|
| 25 |
|
| 12 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 12 | 13 |
| 17 | 16 | 5 | 0.7 | 1.0 |
|
| 12 |
|
| 20 |
|
|
|
|
|
| 14 |
|
| II |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | II | 13 | 2 | 12 | 18 | 8 | 0.7 | 1.1 |
|
| 13 |
|
| 27 |
|
|
|
|
|
| 14 |
|
| 15 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 13B | 12 | 2 | 20 | 21 | 6 | 0.7 | 1.3 |
|
| 10B |
|
| 27 |
|
|
|
|
|
| 12B |
|
| 15 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 7B | 7 |
| 8B | 11 | 2 | 0.4 | 0.7 |
|
| 8B |
|
| 12B |
|
|
|
|
|
| 7B |
|
| 12B |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 6B | 8 | 2 | 7B | 7 | 3 | 0.5 | 0.4 |
|
| 9B |
|
| 4B |
|
|
|
|
|
| 9B |
|
| 9B |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 7 mPB | 5 | 2 | 4 mPB | 3 | 0.3 | 0.2 |
|
|
| 4 mPB |
|
| 4 mPB |
|
|
|
|
|
| 4 mPB |
|
| 2 mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 13 | 11 | 2 | 62 | 63 | 10 | 0.6 | 3.9 |
|
| 9 |
|
| 54 |
|
|
|
|
|
| 10 |
|
| 73 |
|
|
|
|
Anthracene-2-amine | 3.5µg | 9 | 10 |
| 69 | 62 | 6 | 0.5 | 3.9 |
|
| 10 |
|
| 59 |
|
|
|
|
|
| 10 |
|
| 57 |
|
|
|
|
Cyclophosphamide | 400.0 µg | 26 | 32 | 6 | 491 | 530 | 38 | 1.8 | 33.1 |
|
| 37 |
|
| 533 |
|
|
|
|
|
| 33 |
|
| 566 |
|
|
|
|
Sodium azide | 5µg | 431 | 423 | 26 | 52 | 39 | 12 | 24.0 | 2.4 |
|
| 445 |
|
| 37 |
|
|
|
|
|
| 394 |
|
| 28 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain T A1538 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 10 | 11 | 4 | 20 | 21 | 2 | 1.0 | 1.0 |
|
| 15 |
|
| 23 |
|
|
|
|
|
| 7 |
|
| 21 |
|
|
|
|
Phosphate buffer | 50 µL | 12 | 13 |
| 18 | 18 | 3 | 1.2 | 0.8 |
|
| 14 |
|
| 15 |
|
|
|
|
|
| 13 |
|
| 20 |
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 10 | 9 |
| 10 | 12 | 2 | 0.9 | 0.6 |
|
| 9 |
|
| 14 |
|
|
|
|
|
| 9 |
|
| 13 |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | II | 10 | 3 | 20 | 18 | 2 | 0.9 | 0.9 |
|
| 12 |
|
| 18 |
|
|
|
|
|
| 7 |
|
| 17 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 9 mB | 4 | 5 | 15 | 20 | 5 | 0.3 | 0.9 |
|
| 1 mB |
|
| 25 |
|
|
|
|
|
| 1 mB |
|
| 19 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 1 mB | 0 |
| 15 | 12 | 3 | 0.0 | 0.5 |
|
| 0 mB |
|
| 10 |
|
|
|
|
|
| 0 mB |
|
| 10 |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 0 mB | 0 | 0 | 17B | 14 | 4 | 0.0 | 0.7 |
|
| 0mB |
|
| 16B |
|
|
|
|
|
| 0 mB |
|
| 10B |
|
|
|
|
ZK10445/ZK34493 7 | 5.0mg | 0 mPB | 0 | 0 | 5mPB | 4 |
| 0.0 | 0.2 |
|
| 0 mPB |
|
| 3mPB |
|
|
|
|
|
| 0 mPB |
|
| 3mPB |
|
|
|
|
Anthracene-2-amine | 2.5µg | 6 | 7 | 1 | 198 | 192 | 9 | 0.7 | 9.0 |
|
| 8 |
|
| 182 |
|
|
|
|
|
| 8 |
|
| 197 |
|
|
|
|
Anthracene-2-amine | 3.5µg | 6 | 6 | 1 | 246 | 233 | 24 | 0.6 | 10.9 |
|
| 7 |
|
| 248 |
|
|
|
|
|
| 6 |
|
| 206 |
|
|
|
|
2-Nitrofluorene | 10.0 µg | 794 | 789 | 9 | 195 | 215 | 20 | 73.9 | 10.1 |
|
| 778 |
|
| 214 |
|
|
|
|
|
| 794 |
|
| 235 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA1538 (a) | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 12 | 13 | 1 |
| I | I | 1.0 | / |
|
| 14 |
|
|
|
|
|
|
|
|
| 13 |
|
|
|
|
|
|
|
Phosphate buffer | 50 µL | 9 | 9 | 1 |
| I | I | 0.7 | I |
|
| 9 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.01 mg | 11 | 9 | 2 |
| I | I | 0.7 | I |
|
| 7 |
|
|
|
|
|
|
|
|
| 9 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.05 mg | 18 | 12 | 6 |
| / | / | 0.9 | / |
|
| 11 |
|
|
|
|
|
|
|
|
| 6 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 5 B | 5 | 1 |
| / | / | 0.4 | / |
|
| 4 mB |
|
|
|
|
|
|
|
|
| 6 mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 11mB | 7 | 4 | / | I | 0.5 | I |
|
|
| 3mB |
|
|
|
|
|
|
|
|
| 6mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.5 mg | 19mB | 6 | 11 | / | I | 0.5 | / |
|
|
| 0mB |
|
|
|
|
|
|
|
|
| 0 mB |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 1.0 mg | 0 mB 0mB | 0 | 0 | I | / | 0.0 | / |
|
|
| 0 mB |
|
|
|
|
|
|
|
2-Nitrofluorene | l0.0 µg | 899 | 976 | 70 | I | / | 75.1 | / |
|
|
| 994 |
|
|
|
|
|
|
|
|
| l036 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA1537 | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 4 | 8 | 3 | 18 | 21 | 4 | 1.0 | 1.0 |
|
| 10 |
|
| 26 |
|
|
|
|
|
| 9 |
|
| 19 |
|
|
|
|
Phosphate buffer | 50 µL | 5 | 5 |
| 10 | 11 | 2 | 0.7 | 0.5 |
|
| 5 |
|
| 11 |
|
|
|
|
|
| 6 |
|
| 13 |
|
|
|
|
ZK1 0445/ZK344937 | 0.1mg | 5 | 7 | 2 | 22 | 19 | 4 | 0.9 | 0.9 |
|
| 7 |
|
| 14 |
|
|
|
|
|
| 8 |
|
| 21 |
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 2 mB | 3 | 2 | 14 | 13 | 2 | 0.4 | 0.6 |
|
| 3 mB |
|
| 11 |
|
|
|
|
|
| 5 mB |
|
| 13 |
|
|
|
|
ZK10445/ZK344937 | 0.5mg | 1 mB | 10 | 13 | 23 | 14 | 8 | 1.3 | 0.7 |
|
| 25mB |
|
| 9 |
|
|
|
|
|
| 3 mB |
|
| 10 |
|
|
|
|
ZK10445/ZK344937 | 1.0mg | 1 m B | 0 |
| 13 | 12 | 2 | 0.0 | 0.6 |
|
| 0 mB |
|
| 13 |
|
|
|
|
|
| 0 mB |
|
| 10 |
|
|
|
|
ZK10445/ZK344937 | 2.5mg | 0 mB | 0 |
| 6B | 9 | 3 | 0.0 | 0.4 |
|
| l mB |
|
| 8B |
|
|
|
|
|
| 0 mB |
|
| 12B |
|
|
|
|
ZK10445/ZK344937 | 5.0mg | 2 mPB | 1 |
| 0mPB | 2 | 2 | 0.1 | 0.1 |
|
| 0 mPB |
|
| 2mPB |
|
|
|
|
|
| 0 mPB |
|
| 4mPB |
|
|
|
|
Anthracene-2-amine | 5.0µg | 9 | 9 | 0 | 58 | 59 | 4 | 1.2 | 2.8 |
|
| 9 |
|
| 56 |
|
|
|
|
|
| 9 |
|
| 63 |
|
|
|
|
4-NPDA | 10.0 µg | 67 | 57 | 12 | 24 | 26 | 2 | 7.4 | 1.2 |
|
| 43 |
|
| 28 |
|
|
|
|
|
| 60 |
|
| 26 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA1537 (a) | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 11 | 11 | 1 |
| / | / | 1.0 | / |
|
| 11 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
Phosphate buffer | 50µL | 10 | 9 | 1 |
| / | / | 0.9 | / |
|
| 8 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.01 mg | 9 | 10 | 1 |
| / | / | 0.9 | / |
|
| 10 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.05 mg | 5 | 5 | 1 |
| / | / | 0.4 | / |
|
| 5 |
|
|
|
|
|
|
|
|
| 4 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.1mg | 5 | 5 | 1 |
| / | / | 0.4 | / |
|
| 5 |
|
|
|
|
|
|
|
|
| 4 |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.25 mg | 4 mB | 51 | 75 |
| / | / | 4.8 | / |
|
| 1382)B1) |
|
|
|
|
|
|
|
|
| 11B1) |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 0.5 mg | 1mB | 53 | 50 |
| / | / | 5.0 | / |
|
| 1012)B1) |
|
|
|
|
|
|
|
|
| 582)B1) |
|
|
|
|
|
|
|
ZK10445/ZK344937 | 1.0 mg | 0mB1) | 24 | 42 |
| / | / | 2.3 | / |
|
| 0mB1) |
|
|
|
|
|
|
|
|
| 732)B1) |
|
|
|
|
|
|
|
4-NPDA | 10.0µg | 65 | 70 | 8 |
| / | / | 6.6 | / |
|
| 79 |
|
|
|
|
|
|
|
|
| 67 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Salmonella typhimurium strain TA1537 (d) | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
|
|
|
|
|
|
|
|
|
|
DMSO | 50 µL | 8 | 7 | 1 |
| / | / | 1.0 | / |
|
| 8 |
|
|
|
|
|
|
|
|
| 6 |
|
|
|
|
|
|
|
Phosphate buffer | 50 µL | 5 | 8 | 4 |
| / | / | 1.1 | / |
|
| 13 |
|
|
|
|
|
|
|
|
| 7 |
|
|
|
|
|
|
|
ZK10445 | 2.5µg | 7 | 9 | 2 |
| / | / | 1.2 | / |
|
| 9 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
ZK10445 | 5µg | 5 | 6 | 1 |
| / | / | 0.8 | / |
|
| 5 |
|
|
|
|
|
|
|
|
| 7 |
|
|
|
|
|
|
|
ZK10445 | 10µg | 9 | 10 | 1 |
| / | / | 1.3 | / |
|
| 10 |
|
|
|
|
|
|
|
|
| 10 |
|
|
|
|
|
|
|
ZK10445 | 25µg | 5 | 8 | 4 |
| / | / | 1.1 | / |
|
| 13 |
|
|
|
|
|
|
|
|
| 6 |
|
|
|
|
|
|
|
ZK10445 | 50µg | 7B | 4 | 3 |
| / | / | 0.5 | / |
|
| 1B |
|
|
|
|
|
|
|
|
| 3B |
|
|
|
|
|
|
|
ZK10445 | 100µg | 4B | 6 | 2 |
| / | / | 0.8 | / |
|
| 7B |
|
|
|
|
|
|
|
|
| 7B |
|
|
|
|
|
|
|
ZK10445 | 250µg | 612)B1) | 27 | 30 |
| / | / | 3.6 | / |
|
| 9 B1) |
|
|
|
|
|
|
|
|
| 10 B1) |
|
|
|
|
|
|
|
ZK10445 | 500µg | 0 B1) | 2 | 3 |
| / | / | 0.2 | / |
|
| 0 B1) |
|
|
|
|
|
|
|
|
| 5 B1) |
|
|
|
|
|
|
|
ZK10445 | 1000µg | 0 B1) | 0 | 0 |
| / | / | 0.0 | / |
|
| 0 B1) |
|
|
|
|
|
|
|
|
| 0 B1) |
|
|
|
|
|
|
|
4-NPDA | 10.0µg | 89 | 85 | 11 |
| / | / | 11.6 | / |
|
| 93 |
|
|
|
|
|
|
|
|
| 73 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Preincubation test on Escherichia coli WP2uvrA | |||||||||
|
| -S9 | M | +/-SD | +S9 | M | +/-SD | -S9 | +S9 |
DMSO | 50 µL | 25 | 24 | 3 | 39 | 36 | 3 | 1.0 | 1.0 |
|
| 27 |
|
| 34 |
|
|
|
|
|
| 21 |
|
| 34 |
|
|
|
|
Phosphate buffer | 50 µL | 32 | 31 | 1 | 48 | 47 | 2 | 1.3 | 1.3 |
|
| 31 |
|
| 44 |
|
|
|
|
|
| 31 |
|
| 48 |
|
|
|
|
ZK344937/10445 | 0.1mg | 28 | 24 | 5 | 34 | 40 | 6 | 1.0 | 1.1 |
|
| 18 |
|
| 42 |
|
|
|
|
|
| 26 |
|
| 45 |
|
|
|
|
ZK344937/10445 | 0.25mg | 28 | 26 | 2 | 38 | 35 | 3 | 1.1 | 1.0 |
|
| 25 |
|
| 32 |
|
|
|
|
|
| 25 |
|
| 36 |
|
|
|
|
ZK344937/10445 | 0.5mg | 26 | 29 | 4 | 49 | 43 | 6 | 1.2 | 1.2 |
|
| 33 |
|
| 44 |
|
|
|
|
|
| 29 |
|
| 37 |
|
|
|
|
ZK344937/10445 | 1.0mg | 19 | 21 | 10 | 25 | 24 | 5 | 0.9 | 0.7 |
|
| 32 |
|
| 28 |
|
|
|
|
|
| 13 |
|
| 19 |
|
|
|
|
ZK344937/10445 | 2.5mg | 16 | 17 | 1 | 27 | 27 | 1 | 0.7 | 0.8 |
|
| 18 |
|
| 28 |
|
|
|
|
|
| 16 |
|
| 26 |
|
|
|
|
ZK344937/10445 | 5.0mg | 10mP | 14 | 3 | 25 | 27 | 5 | 0.6 | 0.7 |
|
| 15mP |
|
| 32 |
|
|
|
|
|
| 16mP |
|
| 23 |
|
|
|
|
Anthracene-2-amine | 50.0µg | 70 | 63 | 10 | 151 | 159 | 9 | 2.6 | 4.4 |
|
| 51 |
|
| 169 |
|
|
|
|
|
| 67 |
|
| 156 |
|
|
|
|
EMS | 5.0µL | 1550 | 1475 | 65 | 1682 | 1667 | 73 | 60.6 | 46.7 |
|
| 1437 |
|
| 1588 |
|
|
|
|
|
| 1438 |
|
| 1731 |
|
|
|
|
MNNG | 5.0µg | 489 | 451 | 46 | 89 | 111 | 34 | 18.5 | 3.1 |
|
| 400 |
|
| 151 |
|
|
|
|
|
| 463 |
|
| 94 |
|
|
|
|
M: Mean +/-SD: Standard-deviation p: Precipitation +S9: With S9 mix -S9: Without S9 mix B: Background lawn reduced C: Contamination /: Not tested Quotient= Mean revertants (test substance) Mean revertants (solvent) 1) : Background lawn was extremely or totally suppressed 2) : Counts include, however, small "pinpoint" colonies which could not be unequivocally distinguished from normal colonies |
Applicant's summary and conclusion
- Conclusions:
- The mutagenic potential of the test item (fluocortolone batch containing 5% of ZK 344937 as impurity) was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 as well as the E. coli strain WP2 uvrA in the presence and absence of S9 mix according to OECD TG 471. Evidence of mutagenic activity was not seen up to the maximum recommended dose level of 5.0 mg/plate. No substantial increases in revertant colony numbers of any of the six tester strains were observed at any dose level in the presence and absence of metabolic activation. Precipitation was observed at the maximum dose level of 5.0 mg/plate. Therefore, the test item was considered to be non-mutagenic in the Salmonella typhimurium reverse mutation assay (plate incorporation method and preincubation modification).
- Executive summary:
In a reverse gene mutation assay in bacteria according to OECD guideline 471 (1997), Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, TA1538 and E.coli strain WP2uvrA were exposed to Fluocortolon in DMSO in concentrations of 0.01, 0.05, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0 mg/plate in the absence and presence of mammalian metabolic activation (rat liver S9 mix) in the plate incorporation assay and with the preincubation method. Cytotoxicity was observed from 1.0 mg/plate upwards (in the refinement only with S9 mix).Without metabolic activation cytotoxicity appeared at 0.1 mg/plate in the refinement.
Precipitation was observed at the highest concentration (5.0 mg/plate). The positive controls induced the appropriate responses in the corresponding strains. The mean numbers of revertant colonies in the negative controls were within the ranges of the historical control data.
There was no evidence of an increase in the number of revertant colonies that exceeded twice background in any of the tester strains (TA98, TA 100, TA1538, TA1535, TA1537 or E.coli WP2uvrA) examined at dose levels up to 5.0 mg/plate in the absence and presence of a metabolic activation source (S9). Therefore, test substance was considered to be non-genotoxic (non-mutagenic) under the conditions employed (preincubation assay).
There was no evidence of induced mutant colonies over background.
Under the conditions of the study, the test substance was negative for mutagenic potential.
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