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EC number: 201-557-4 | CAS number: 84-74-2
Toxicological information
Genetic toxicity: in vitro
Currently viewing:
Administrative data
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: comparable to guideline study with acceptable restrictions
Data source
Referenceopen allclose all
- Reference Type:
- publication
- Title:
- Chromosome aberration and sister chromoatid exchange tests in Chinese hamster ovary cells in vitro III: Results with 27 chemicals
- Author:
- Gulati DK, Witt K, Anderson B
- Year:
- 1 989
- Bibliographic source:
- Environ. Mol. Mutagen. 13, 133-193
- Reference Type:
- publication
- Title:
- Tests for the induction of chromosomal aberrations and sister chromatid exchanges in cultured Chinese hamster ovary (CHO) cells.
- Author:
- Gulati DK, Sabharwal PS, Shelby MD
- Year:
- 1 985
- Bibliographic source:
- Progr. Mutat. Res. 5, 413-427
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- Bis(2-ethylhexyl) phthalate
- EC Number:
- 204-211-0
- EC Name:
- Bis(2-ethylhexyl) phthalate
- Cas Number:
- 117-81-7
- IUPAC Name:
- bis(2-ethylhexyl) phthalate
- Details on test material:
- - Name of test material (as cited in study report): Di-(2-ethylhexyl)phthalate DEHP
- Source : Fluka
- Purity : >99%
Batch 1 Lancaster Synthesis. Ltd. - DMSO (Ppt at >500 µg/ml)
Batch 2 Matheson Coleman & Bell 96% DMSO (Ppt at >500 µg/ml)
Batch 3 W. R. Grace & Co. 99.6% DMSO (Ppt at >500 µg/ml)
Constituent 1
Method
- Target gene:
- Chromosomal aberrations
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: McCoy's 5A medium (modified) supplemented with L-glutamine (2 mM), antibiotics, and 10% fetal bovine serum (FBS)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: no
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver fraction (S9) prepared from Aroclor 1254-induced male Sprague Dawley rats (Microbiological Associates, Bethesda, MD)
- Test concentrations with justification for top dose:
- 0, 5, 16, 50, 600, 1600, 2000, 3000, 4000, 5000 µg/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: mitomycin C (-S9) and cyclophosphamide (+S9)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium;
DURATION
- Preincubation period: 24h
- Exposure duration: 10-11h
- Harvest time: 12-13.5 h
For chemicals that caused cell cycle delay, harvest times were extended, generally in 5 h increments, with colcemid present for the last 2 h. For chromosomal aberrations tests, harvest times were similarly extended based on the observation of cell cycle delay in the sister chromatic exchange trials.
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (2-3h prior harvest)
STAIN (for cytogenetic assays): Giemsa 6%
NUMBER OF CELLS EVALUATED: 100 cells were scored for each dose in early studies and 200 cells per dose in later studies
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index;
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes - Statistics:
- The percentage of cells with aberrations was analyzed. Both the dose-response curve and individual dose points were statistically analyzed. A statistically significant (P < 0.003) trend test or a significantly elevated dose point (P < 0.05) was sufficient to indicate a chemical effect.
Results and discussion
Test results
- Species / strain:
- Chinese hamster Ovary (CHO)
- 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:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Downloaded from the NTP site:
http://ntp-apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=invitroca.cadata&study_no=923560&cas_no=117%2D81%2D7&endpointlist=CAB
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| In Vitro Cytogenetics - Chromosome Aberrations | ||||||||||||||||
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| Current Search Criteria | ||||||||||||||||
| Chemical Name: | Di(2-ethylhexyl) phthalate | |||||||||||||||
| CAS Number: | 117-81-7 | |||||||||||||||
| Study Type: | Chromosome Aberrations | |||||||||||||||
| Study ID: | 923560 | |||||||||||||||
| Table Instructions and Notes: | ||||||||||||||||
| Below is a summary of Chromosome Aberrations for the selected Study ID. Solvent control marked in white. | ||||||||||||||||
| Study Result: Negative | ||||||||||||||||
| Activation | Trial | Trial Call | ||||||||||||||
| No Activation | 1 | Negative | ||||||||||||||
| No Activation | 2 | Negative | ||||||||||||||
| Induced Rat Liver S9 | 3 | Negative | ||||||||||||||
| Induced Rat Liver S9 | 4 | Negative | ||||||||||||||
| Trial #:1 Activation: No Activation Date: 12/10/1982 Harvest Time: 13.5 hrs Trial Call: Negative | ||||||||||||||||
| Dose | Total Cells Examined | Total Aberrations | Complex Aberrations | Simple Aberrations | Other Abs. | |||||||||||
| µg/mL | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | ||||
| Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | |||||
| Cell | Abs. | Cell | Abs. | Cell | Abs. | Cell | Abs. | |||||||||
| Abs: Aberrations | ||||||||||||||||
| Vehicle Control: | Negative | 0 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Vehicle Control: | Dimethylsulfoxide | 0 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Positive Control: | Mitomycin C | 0.5 | 100 | 99 | 0.990 | 63.000 | 26 | 0.260 | 25.000 | 43 | 0.430 | 32.000 | 30 | 0.300 | 28.000 | |
| Test Chemical: | Di(2-ethylhexyl)phthalate (DEHP) | 50 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| 160 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 500 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 1600 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 5000 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| Trend: | 0.000 | 0.000 | 0.000 | |||||||||||||
| Probability: | 0.000 | 0.000 | 0.000 | |||||||||||||
| Trial #:2 Activation: No Activation Date: 12/21/1982 Harvest Time: 12.0 hrs Trial Call: Negative | ||||||||||||||||
| Dose | Total Cells Examined | Total Aberrations | Complex Aberrations | Simple Aberrations | Other Abs. | |||||||||||
| µg/mL | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | ||||
| Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | |||||
| Cell | Abs. | Cell | Abs. | Cell | Abs. | Cell | Abs. | |||||||||
| Abs: Aberrations | ||||||||||||||||
| Vehicle Control: | Negative | 0 | 100 | 2 | 0.020 | 2.000 | 1 | 0.010 | 1.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |
| Vehicle Control: | Dimethylsulfoxide | 0 | 100 | 2 | 0.020 | 2.000 | 2 | 0.020 | 2.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Positive Control: | Mitomycin C | 0.5 | 100 | 76 | 0.760 | 48.000 | 25 | 0.250 | 21.000 | 34 | 0.340 | 24.000 | 17 | 0.170 | 13.000 | |
| Test Chemical: | Di(2-ethylhexyl)phthalate (DEHP) | 2000 | 100 | 3 | 0.030 | 3.000 | 1 | 0.010 | 1.000 | 2 | 0.020 | 2.000 | 0 | 0.000 | 0.000 | |
| 3000 | 100 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |||
| 4000 | 100 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |||
| 5000 | 100 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |||
| Trend: | -1.064 | -2.126 | 0.308 | |||||||||||||
| Probability: | 0.856 | 0.983 | 0.379 | |||||||||||||
| Trial #:3 Activation: Induced Rat Liver S9 Date: 12/10/1982 Harvest Time: 12.0 hrs Trial Call: Negative | ||||||||||||||||
| Dose | Total Cells Examined | Total Aberrations | Complex Aberrations | Simple Aberrations | Other Abs. | |||||||||||
| µg/mL | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | ||||
| Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | |||||
| Cell | Abs. | Cell | Abs. | Cell | Abs. | Cell | Abs. | |||||||||
| Abs: Aberrations | ||||||||||||||||
| Vehicle Control: | Negative | 0 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Positive Control: | Cyclophosphamide | 50 | 100 | 74 | 0.740 | 51.000 | 16 | 0.160 | 15.000 | 40 | 0.400 | 32.000 | 18 | 0.180 | 17.000 | |
| Vehicle Control: | Dimethylsulfoxide | 0 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Test Chemical: | Di(2-ethylhexyl)phthalate (DEHP) | 50 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| 160 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 500 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 1600 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| 5000 | 100 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |||
| Trend: | 0.000 | 0.000 | 0.000 | |||||||||||||
| Probability: | 0.000 | 0.000 | 0.000 | |||||||||||||
| Trial #:4 Activation: Induced Rat Liver S9 Date: 12/21/1982 Harvest Time: 12.0 hrs Trial Call: Negative | ||||||||||||||||
| Dose | Total Cells Examined | Total Aberrations | Complex Aberrations | Simple Aberrations | Other Abs. | |||||||||||
| µg/mL | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | No. of | Abs | % Cells | ||||
| Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | Abs. | Per | With | |||||
| Cell | Abs. | Cell | Abs. | Cell | Abs. | Cell | Abs. | |||||||||
| Abs: Aberrations | ||||||||||||||||
| Vehicle Control: | Negative | 0 | 100 | 1 | 0.010 | 1.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| Positive Control: | Cyclophosphamide | 50 | 100 | 74 | 0.740 | 52.000 | 16 | 0.160 | 14.000 | 27 | 0.270 | 24.000 | 31 | 0.310 | 21.000 | |
| Vehicle Control: | Dimethylsulfoxide | 0 | 100 | 3 | 0.030 | 3.000 | 2 | 0.020 | 2.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |
| Test Chemical: | Di(2-ethylhexyl)phthalate (DEHP) | 1000 | 100 | 4 | 0.040 | 4.000 | 4 | 0.040 | 4.000 | 0 | 0.000 | 0.000 | 0 | 0.000 | 0.000 | |
| 2000 | 100 | 4 | 0.040 | 3.000 | 3 | 0.030 | 2.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |||
| 3000 | 100 | 2 | 0.020 | 2.000 | 1 | 0.010 | 1.000 | 1 | 0.010 | 1.000 | 0 | 0.000 | 0.000 | |||
| 4000 | 100 | 2 | 0.020 | 2.000 | 0 | 0.000 | 0.000 | 2 | 0.020 | 2.000 | 0 | 0.000 | 0.000 | |||
| Trend: | -0.768 | -1.634 | 0.909 | |||||||||||||
| Probability: | 0.779 | 0.949 | 0.182 | |||||||||||||
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
DEHP did not induce any noteworthy increase in the number of cells with structural chromosome aberration, both with and without S9 mix, in either experiment or at either harvest time. - Executive summary:
The potential of DEHP to induce structural chromosome aberrations in Chinese hamster ovary cells was evaluated with and without a metabolic activation system according a protocol similar to the OECD 473.
Cells were treated for about 10-11 h and colcemid was added 2-3 hr prior to tell harvest by mitotic shake-off.
The chromosome number was recorded for each cell and chromosome or chromatid type aberrations were classified into three categories: simple (breaks, fragments, double minutes), complex (interchanges, rearrangements), and other (pulverized, more than ten aberrations/cell).
Under these experimental conditions, DEHP did not induce any noteworthy increase in the number of cells with structural chromosome aberration, both with and without S9 mix, in either experiment or at either harvest time.
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