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EC number: 294-436-0 | CAS number: 91722-33-7 A complex combination of organic compounds separated after condensation of the vapors from the destructive distillation of wood.
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
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:
- 07.05.2014 - 16.05.2014
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Strain: TA98Genotype: trp./his mutation his D3052, type of mutation: FrameshiftMutations: cell wall: rfa / DNA-repair: uvrBMain DNA target: GCStrain: TA100Genotype: trp./his mutation hisG46, type of mutation: Base pair substitutionMutations: cell wall: rfa / DNA-repair: uvrBMain DNA target: GCStrain: TA1535Genotype: trp./his mutation his hisG46 type of mutation: Base pair substitutionMutations: cell wall: rfa / DNA-repair: uvrBMain DNA target: GCStrain: TA1537Genotype: trp./his mutation his D3052, type of mutation: FrameshiftMutations: cell wall: rfa / DNA-repair: uvrBMain DNA target: GCStrain: WP2uvrAGenotype: trp./his mutation his trpE, type of mutation: Base pair substitutionMutations: cell wall: + / DNA-repair: uvrAMain DNA target: AT
- Species / strain / cell type:
- S. typhimurium TA 98
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- other: Mutation in hisD3052, rfa, uvrB and pKM101
- Species / strain / cell type:
- S. typhimurium TA 100
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- other: Mutation in hisG46, rfa, uvrB and pKM101
- Species / strain / cell type:
- S. typhimurium TA 1535
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- other: Mutation in hisG46, rfa and uvrB
- Species / strain / cell type:
- S. typhimurium TA 1537
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- other: Mutation in hisC3076, rfa and uvrB
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- other: Mutation in trpE and uvrA
- Metabolic activation:
- with and without
- Metabolic activation system:
- post mitochondrial supernatant (S9) prepared from livers of Phenobarbital/β-naphthoflavone-induced rats
- Test concentrations with justification for top dose:
- 5000; 1581; 500; 158; 50 and 15.8 µg/plate
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-1,2-phenylene-diamine (NPD)
- Remarks:
- Dose quantity/plate: 4 µg; Strain: Salmonella TA98
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Sodium azide (SAZ)
- Remarks:
- Dose quantity/plate: 2 µg; Strain: Salmonella TA100 and TA1535
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- Dose quantity/plate: 50 µg; Strain: Salmonella TA1537
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- Dose quantity/plate: 2 µL; Strain: E.coli WP2uvrA
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene (2AA)
- Remarks:
- Dose quantity/plate: 2 µg; Strain: all of Salmonella strains
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene (2AA)
- Remarks:
- Dose quantity/plate: 50 µg; Strain: E.coli strain
- Species / strain:
- S. typhimurium TA 98
- 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
- Species / strain:
- S. typhimurium 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
- Species / strain:
- S. typhimurium TA 1535
- 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
- Species / strain:
- S. typhimurium TA 1537
- 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
- Species / strain:
- E. coli WP2 uvr A
- 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
- Remarks on result:
- other: other: plate incorporation method and pre-incubation method
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- In conclusion, the test item Tar wood has no mutagenic activity on the applied bacterium tester strains under the test conditions used in this study.
- Executive summary:
The test item Tar wood was tested with regard to a potential mutagenic activity using the Bacterial Reverse Mutation Assay.
The experiments were carried out using histidine-requiring auxotroph strains of Salmonella typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the tryptophan-requiring auxotroph strain of Escherichia coli (Escherichia coli WP2 uvrA) in the presence and absence of a post mitochondrial supernatant (S9) prepared from livers of phenobarbital/β-naphthoflavone-induced rats.
The study included a Preliminary Solubility Test, a Preliminary Range Finding Test (Informatory Toxicity Test), an Initial Mutation Test (Plate Incorporation Test), and a Confirmatory Mutation Test (Pre-Incubation Test).
In the Range Finding Test as well as in the Initial Mutation Test the plate incorporation method was used.
Based on the results of the Solubility Test and the Range Finding Test the test item was dissolved in Dimethyl sulfoxide in a concentration of 100 mg/mL.
Based on the results of the preliminary Range Finding Test the following concentrations of the test item were prepared and used in the Initial and Confirmatory Mutation Tests:
5000; 1581; 500; 158; 50 and 15.8 μg/plate.
In the Initial and Confirmatory Mutation the test item concentrations, including the controls (untreated, vehicle and positive reference) were tested in triplicate.
No substantial, biological relevant increases were observed in revertant colony numbers of any of the five test strains following treatment with Tar wood at any concentration level, either in the presence or absence of metabolic activation (S9 Mix) in the performed experiments. Sporadic increases in revertant colony numbers compared to the vehicle control values were observed in both independently performed main experiments.
However, there was no tendency of higher mutation rates with increasing concentrations beyond the generally acknowledged border of biological relevance in the performed experiments. See details in the Section 9. RESULTS.
The highest revertant colony number increase over the spontaneous rate of the vehicle control plates was observed in the Confirmatory Mutation Test (Pre-Incubation Test) in S. typhimurium TA1537 at 158 μg/plate, without metabolic activation (-S9 Mix). The mutation rate was: 2.82*. This relatively high value was unique, remained within the laboratory’s historical control data range and remained below the genotoxicological threshold for being positive.
Inhibitory effect of the test item was obtained in the Confirmatory Mutation Test following the pre-incubation procedure. The inhibitory effect of the test item was indicated by lower revertant colony numbers than the revertant colony numbers of the vehicle controls (that were often below the corresponding historical control data ranges) and affected background lawn development: absent, reduced or slightly reduced background lawn.
The revertant colony numbers of vehicle control (Dimethyl sulfoxide, DMSO) plates with and without S9 Mix demonstrated the characteristic mean number of spontaneous revertants and were in line with the corresponding historical control data ranges.
The spontaneous revertant colony numbers of the DMSO vehicle control plates were slightly lower (the actual average number: 16) than the characteristic mean numbers agreed with the actual historical control data range (historical control range: 17-49) in the Initial Mutation Test in Salmonella typhimurium TA98 in the presence of metabolic activation (+S9 Mix). The lower counts were evaluated as acceptable without any influence on the final conclusion of the study.
The reference mutagen treatments (positive controls) showed the expected, biological relevant increases in induced revertant colonies in all experimental phases, in all tester strains.
The reported data of this mutagenicity assay show (see Appendix I to IV) that under the experimental conditions applied, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
* Mutation rate (MR): The mutation rate is the quotient of the mean revertant of test item treatment and the mean revertant of the vehicle control. In the case of S. typhimurium TA1537 a biologically relevant increase (positive result) is when the number of reversions is at least three times higher than the reversion rate of the vehicle control (Mutation rate ≥ 3.00).
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- yes
- Type of assay:
- comet assay
- Specific details on test material used for the study:
- Name: Scansmoke R909
Batch No: 03/2004 - Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- 3.7.2C cells
ATCC CRL-9518
lot 1661603 - Metabolic activation:
- with and without
- Metabolic activation system:
- S9
- Test concentrations with justification for top dose:
- Test 1
-S9: 0, 250, 400, 550, 650, 750 and 850 µg/mL
+S9: 0, 500, 750, 1000, 1200, 1300 and 1400 µg/mL
Test 2
-S9: 0, 250, 350, 450, 550, 600 and 650 µg/mL
+S9: 0, 500, 750, 1000, 1200, 1300 and 1400 µg/mL - Vehicle / solvent:
- - Vehicle used: RPMI5 medium
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- cyclophosphamide
- Details on test system and experimental conditions:
- DURATION
Test 1: Exposure during 3 h with and without metabolic activation
Test 2: Exposure during 3 h with and 24 h without metabolic activation - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with
- Genotoxicity:
- positive
- Remarks:
- dose associated at 450 to 650 µg/mL after 3 h treatment
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Positive controls validity:
- valid
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Remarks:
- dose associated at 1000 to 1400 µg/mL after 24 h treatment
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Positive controls validity:
- valid
Referenceopen allclose all
see attached report for more information
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Genetic toxicity in vivo
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- yes
- Type of assay:
- mammalian erythrocyte micronucleus test
- Specific details on test material used for the study:
- Name: Scansmoke R909
Batch No: 03/2004 - Species:
- mouse
- Strain:
- NMRI
- Details on species / strain selection:
- Strain: CRL: NMRI BR mice
- Sex:
- not specified
- Route of administration:
- oral: unspecified
- Vehicle:
- distilled water
- Dose / conc.:
- 500 mg/kg diet
- Dose / conc.:
- 1 000 mg/kg diet
- Dose / conc.:
- 2 000 mg/kg diet
- Control animals:
- yes
- Positive control(s):
- Cyclophosphamide (ip. administration)
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Executive summary:
No increases in the frequency of micronucleated polychromated erythrocytes (MCPE) in male and female mice at either 24 or 48 hours after treatment compared to the vehicle control.
Considerable differences in the ratio of polychromatic and normochromatic erythrocytes were not found after the treatment.
Biologically significant depression of PCE:NCE ratio was not observed in the study. The positive control showed significantly increased MCPE numbers compared to the control.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
The test item Tar wood was tested with regard to a potential mutagenic activity using the Bacterial Reverse Mutation Assay.
The experiments were carried out using histidine-requiring auxotroph strains of Salmonella typhimurium (Salmonella typhimurium TA98, TA100, TA1535 and TA1537), and the tryptophan-requiring auxotroph strain of Escherichia coli (Escherichia coli WP2 uvrA) in the presence and absence of a post mitochondrial supernatant (S9) prepared from livers of phenobarbital/β-naphthoflavone-induced rats.
The study included a Preliminary Solubility Test, a Preliminary Range Finding Test (Informatory Toxicity Test), an Initial Mutation Test (Plate Incorporation Test), and a Confirmatory Mutation Test (Pre-Incubation Test).
In the Range Finding Test as well as in the Initial Mutation Test the plate incorporation method was used.
Based on the results of the Solubility Test and the Range Finding Test the test item was dissolved in Dimethyl sulfoxide in a concentration of 100 mg/mL.
Based on the results of the preliminary Range Finding Test the following concentrations of the test item were prepared and used in the Initial and Confirmatory Mutation Tests:
5000; 1581; 500; 158; 50 and 15.8 μg/plate.
In the Initial and Confirmatory Mutation the test item concentrations, including the controls (untreated, vehicle and positive reference) were tested in triplicate.
No substantial, biological relevant increases were observed in revertant colony numbers of any of the five test strains following treatment with Tar wood at any concentration level, either in the presence or absence of metabolic activation (S9 Mix) in the performed experiments. Sporadic increases in revertant colony numbers compared to the vehicle control values were observed in both independently performed main experiments.
However, there was no tendency of higher mutation rates with increasing concentrations beyond the generally acknowledged border of biological relevance in the performed experiments. See details in the Section 9. RESULTS.
The highest revertant colony number increase over the spontaneous rate of the vehicle control plates was observed in the Confirmatory Mutation Test (Pre-Incubation Test) in S. typhimurium TA1537 at 158 μg/plate, without metabolic activation (-S9 Mix). The mutation rate was: 2.82*. This relatively high value was unique, remained within the laboratory’s historical control data range and remained below the genotoxicological threshold for being positive.
Inhibitory effect of the test item was obtained in the Confirmatory Mutation Test following the pre-incubation procedure. The inhibitory effect of the test item was indicated by lower revertant colony numbers than the revertant colony numbers of the vehicle controls (that were often below the corresponding historical control data ranges) and affected background lawn development: absent, reduced or slightly reduced background lawn.
The revertant colony numbers of vehicle control (Dimethyl sulfoxide, DMSO) plates with and without S9 Mix demonstrated the characteristic mean number of spontaneous revertants and were in line with the corresponding historical control data ranges.
The spontaneous revertant colony numbers of the DMSO vehicle control plates were slightly lower (the actual average number: 16) than the characteristic mean numbers agreed with the actual historical control data range (historical control range: 17-49) in the Initial Mutation Test in Salmonella typhimurium TA98 in the presence of metabolic activation (+S9 Mix). The lower counts were evaluated as acceptable without any influence on the final conclusion of the study.
The reference mutagen treatments (positive controls) showed the expected, biological relevant increases in induced revertant colonies in all experimental phases, in all tester strains.
.
Justification for classification or non-classification
The reported data of this mutagenicity assay show (see Appendix I to IV in the report) that under the experimental conditions applied, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.
In conclusion, the test item Tar wood has no mutagenic activity on the applied bacterium tester strains under the test conditions used in this study.
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