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EC number: 640-964-5 | CAS number: 218451-68-4
Toxicological information
Genetic toxicity: in vitro
Currently viewing:
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:
- October - November 1996
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 996
- Report date:
- 1997
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Reaction products of coconut oil with polyethyleneglycol and 2-ethyl-2-(hydroxymethyl)propane-1,3-diol
- EC Number:
- 640-964-5
- Cas Number:
- 218451-68-4
- Molecular formula:
- not applicable because UVCB
- IUPAC Name:
- Reaction products of coconut oil with polyethyleneglycol and 2-ethyl-2-(hydroxymethyl)propane-1,3-diol
- Details on test material:
- - Name of test material (as cited in study report): Marlowet 4753
- Physical state: slightly yellow liquid, containing flakes and crystals
- Lot/batch No.: 01/96
- Expiration date of the lot/batch: no data
- Other:
Date of production: 14th week 1996
Stability: > 1 year
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 or Phenobarbital/beta-Naphthoflavone induced rat liver S9 mix
- Test concentrations with justification for top dose:
- Plate incorporation test:
50 / 160 / 500 / 1600 / 5000 µg/plate with all strains with and without S9 mix
30 / 100 / 300 / 100 / 3000 µg/plate, repetition with strain TA 98 without S9 mix (due to failure of positive control and toxicity)
10 / 30 / 100 / 300 / 100 µg/plate, repetition with strain TA 1537 with S9 mix (due to toxicity)
1 / 3 / 10 / 30 / 100 µg/plate, repetition with strain 1537 without S9 mix (due to failure of positive control and toxicty)
Preincubation test:
10 / 30 / 100 / 300 / 1000 µg/plate with strain TA 98 with and without S9 mix
30 / 100 / 300 / 100 / 3000 µg/plate with strain TA 100 with and without S9 mix
16 / 50 / 160 / 500 / 1600 µg/plate with strain TA 1535 with and without S9 mix
10 / 30 / 100 / 300 / 1000 µg/pklate with strain TA 1537 with S9 mix
0.1 / 0.3 / 1 / 3 / 10 µg/plate with strain TA 1537 without S9 mix - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: not mentioned
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- water
- Negative solvent / vehicle controls:
- yes
- Remarks:
- acetone (25 µL/plate)
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Nitrofluorene (2.5 µg in DMSO/plate) for the strain TA 98; Sodium azide (5.0 µg in aqua bidest./plate) for strain TA 100; Sodium azide (2.5 µg in aqua bidest./plate) for strain TA 1535; 9-Aminoacredine (25 µg in DMSO/plate) for strain TA 1537
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- acetone (25 µL/plate)
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Aminoanthracene (2.5 µg in DMSO/plate) with all strains
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation) and preincubation, performed in two independent tests
DURATION
- Preincubation period: 30 minutes
- Exposure duration: 72 hours
NUMBER OF REPLICATIONS: 3 per concentration
DETERMINATION OF CYTOTOXICITY
- Method: examination of bacterial background lawn (reduced or absent), a reduction in the number of spontaneous revertants was jugded as also indicative of toxicity
OTHER EXAMINATIONS:
- Other: Determination of the frequency of induced or spontaneous reversion to histidine independence with negative controls (H2O), solvent controls (acetone), test substance concentrations and positive controls; determination of the titers of overnight cultures
OTHER: none - Evaluation criteria:
- Criteria for determination of a valid test:
- in the the solvent control, each tester strain culture must exhibit a characteristic meean number of spontanious revertants
- to ensure that appropriate numbers of bacteria are plated, overnight culture titers must be in excess of 10E8 bacteria/mL - Statistics:
- no statistics performed
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- MA, resp. at 5000 µg/plate with S9 in the plate incorporation test
- 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:
- at 5000 µg/plate with and without S9 in the plate incorporation test
- 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:
- ambiguous
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 5000 µg/plate with and without S9 in the plate incorporation test
- 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:
- 1600 µg/plate and above with S9, resp. 100 µg/plate and above without S9 in the plate incorporation test
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not measured
- Effects of osmolality: not applicable
- Evaporation from medium: not applicable
- Water solubility: not mentioned
- Precipitation: no
- Other confounding effects: none
RANGE-FINDING/SCREENING STUDIES: not performed
COMPARISON WITH HISTORICAL CONTROL DATA: not performed
ADDITIONAL INFORMATION ON CYTOTOXICITY: In the plate incorporation test, treatment with 50 - 5000 µg/plate was associated with excess toxicity to strain TA 1537. This part of the test was repeated with lower test subtance concentrations. In the preincubation test,all strains were exposed to lower, toxicity adjusted test substance concentrations. - Remarks on result:
- other: strain/cell type: Salmonella typhimurium
Any other information on results incl. tables
Table #1a: Plate incorporation test: Number of revertants per plate (mean of 3 plates) |
|||||||||||||||
| Concentration µg/plate |
Strain TA 98 | Strain TA 100 | Strain TA 1535 | Strain TA 1537 | |||||||||||
| + S9 mix | - S9 mix | Cytotoxicity | + S9 mix | - S9 mix | Cytotoxicity | + S9 mix | - S9 mix | Cytotoxicity | + S9 mix | - S9 mix | Cytotoxicity | ||||
| Neg. control | 40 ± 13 | 26 ± 3 | no | 155 ± 35 | 145 ± 11 | no | 11 ±1 | 9 ± 2 | no | 17 ± 2 | 22 ± 4 | no | |||
| Solvent control | 41 ± 5 | 24 ± 5 | no | 135 ± 31 | 156 ± 11 | no | 10 ± 1 | 7 ± 2 | no | 16 ± 10 | 16 ± 3 | no | |||
| 50 | 34 ± 10 | 19 ± 2 | no | 128 ± 6 | 108 ± 21 | no | 12 ± 3 | 7 ± 4 | no | 20 ± 1 | 16 ± 3 | no | |||
| 160 | 31 ± 3 | 22 ± 5 | no | 143 ± 41 | 125 ± 14 | no | 15 ± 6 | 8 ± 1 | no | 13 ± 3 | 5 ± 1 | +S9: no, -S9: yes | |||
| 500 | 47 ± 8 | 16 ± 7 | no | 137 ± 5 | 124 ± 12 | no | 13 ± 3 | 7 ± 2 | no | 13 ± 2 | 5 ± 3 | +S9: no, -S9: yes | |||
| 1600 | 39 ± 9 | 13 ± 6 | no | 148 ± 18 | 113 ± 6 | no | 7 ± 2 | 4 ± 0 | no | 5 ± 2 | 2 ± 0 | yes | |||
| 5000 | 36 ± 5 | 7 ± 5 | yes | 118 ± 6 | 107 ± 11 | yes | 3 ± 1 | 28 ± 7 | yes | 5 ± 2 | 3 ± 2 | yes | |||
| Pos. control | 623 ± 37 | 67 ± 12 | no | 947 ± 150 | 586 ± 19 | no | 312 ± 52 | no | 52 ± 10 | 32 ± 14 | no | ||||
| Solvent: Acetone | |||||||||||||||
Table #1b: Repetition of Plate incorporation test: Number of revertants per plate (mean of 3 plates) |
|||||||||||||||
| Concentration µg/plate |
Strain TA 98 | Strain TA 1537 | |||||||||||||
| + S9 mix | - S9 mix | Cytotoxicity | + S9 mix | - S9 mix | Cytotoxicity | ||||||||||
| Neg. control | not tested | 15 ± 3 | no | 16 ± 4 | 19 ± 4 | no | |||||||||
| Solvent control | 14 ± 4 | no | 14 ± 4 | 14 ± 6 | no | ||||||||||
| 1 | not tested | not tested | 13 ± 8 | -S9: no | |||||||||||
| 3 | 8 ± 5 | -S9: no | |||||||||||||
| 10 | 19 ± 6 | 7 ± 2 | no | ||||||||||||
| 30 | 16 ± 9 | no | 15 ± 2 | 6 ± 3 | no | ||||||||||
| 100 | 12 ± 3 | no | 21 ± 7 | 4 ± 2 | + S9: no, -S9: yes | ||||||||||
| 300 | 16 ± 4 | no | 15 ± 5 | not tested | +S9: no | ||||||||||
| 1000 | 10 ± 2 | no | 9 ± 3 | +S9: no | |||||||||||
| 3000 | 8 ± 3 | yes | not tested | ||||||||||||
| Pos. control | 147 ± 5 | no | 62 ± 5 | 70 ± 18 | no | ||||||||||
| Solvent: Acetone | |||||||||||||||
Table #2a: Preincubation test: Number of revertants per plate (mean of 3 plates) |
|||||||||||||||
| Strain TA 98 | Strain TA 100 | Strain TA 1535 | Strain TA 1537 | ||||||||||||
| Concentration µg/plate |
+ S9 mix | - S9 mix | Cytotoxicty | Concentration µg/plate |
+ S9 mix | - S9 mix | Cytotoxicity | Concentration µg/plate |
+ S9 mix | - S9 mix | Cytotoxicity | Concentration µg/plate |
+ S9 mix | - S9 mix | Cytotoxicity |
| Neg. control | 26 ± 6 | 16 ± 5 | no | Neg. control | 204 ± 20 | 158 ± 14 | no | Neg. control | 19 ± 7 | 14 ± 4 | no | Neg. control | 45 ± 5 | 22 ± 2 | no |
| Solvent control | 25 ± 3 | 22 ± 6 | no | Solvent control | 194 ± 27 | 155 ± 11 | no | Solvent control | 15 ± 4 | 9 ± 2 | no | Solvent control | 44 ± 3 | 18 ± 1 | no |
| 10 | 31 ± 6 | 22 ± 5 | no | 30 | 177 ± 7 | 135 ± 16 | no | 16 | 16 ± 4 | 17 ± 3 | no | 0.1 | not tested | 20 ± 2 | S9: no |
| 30 | 31 ± 7 | 19 ± 3 | no | 100 | 187 ± 6 | 163 ± 19 | no | 50 | 21 ± 3 | 12 ± 3 | no | 0.3 | 13 ± 3 | S9: no | |
| 100 | 35 ± 5 | 17 ± 3 | no | 300 | 185 ± 22 | 170 ± 12 | no | 160 | 17 ± 4 | 10 ± 2 | no | 1 | 24 ± 5 | S9: no | |
| 300 | 39 ± 3 | 19 ± 6 | no | 1000 | 215 ± 8 | 160 ± 7 | no | 500 | 17 ± 8 | 11 ± 4 | no | 3 | 24 ± 3 | S9: no | |
| 1000 | 30 ± 7 | 15 ± 2 | no | 3000 | 200 ± 16 | 148 ± 12 | no | 1600 | 12 ± 1 | 10 ± 3 | no | 10 | 52 ± 5 | 26 ± 6 | no |
| Pos.control | 1523 ± 75 | 149 ± 9 | no | Pos. control | 1877 ± 44 | 599 ± 18 | no | Pos.control | 225 ± 20 | 352 ± 15 | no | 30 | 47 ± 7 | not tested | +S9 : no |
| 100 | 48 ± 8 | +S9 : no | |||||||||||||
| 300 | 49 ± 2 | +S9 : no | |||||||||||||
| 1000 | 48 ± 12 | +S9 : no | |||||||||||||
| Pos. Control | 179 ± 8 | 52 ± 3 | no | ||||||||||||
Applicant's summary and conclusion
- Conclusions:
- In the plate incorporation test without metabolic activation with strain TA 1535 treatment with 5000 µg/plate Coconut oil, reaction product with polyethylene glycol and trimethylolpropane resulted in an increased revertant frequency (factor 4.2) but accompanied with toxicity. Therefore, this mutagenic response was jugded by the author of the study to be of no relevance. In the preincubation test, all tester strains were exposed to lower, toxicity adjusted test compound concentratons. With the above mentioned exception, treatment with the test compound did not result in a significant increase in the revertant frequency of the tester strains. All four tester strains exhibited a positive mutagenic response with the positive controls tested both with and without metabolic activaton by S9 mix. Negative (solvent) controls were also tested with each strain, and the mean numbers of spontaneous revertants were considered acceptable.
Coconut oil, reaction products with polyethylene glycol and trimethylolpropane was jugded not to be a bacterial mutagen. - Executive summary:
Coconut oil, reaction products with polyethylene glycol and trimethylolpropane was tested for its ability to induce reverse mutation in an in vitro bacterial system.
Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 were treated with the test compound by the Ames test plate incorporation as well as the preincubation method. Five dose levels covering a total range between 0.1 and 5000 µg/plate, in triplicate both with and without the addition of a metabolising system (Aroclor 1254 or Phenobarbital/beta-Naphthoflavone induced rat liver S9 mix) were employed.
All four bacterial strains exhibited mutagenic responses to the appropriate positive control substance. Solvent controls were also tested with each strain and the mean numbers of spontaneous revertants were in an acceptable range.
A reproducible mutagenic activity of the test compound to any of the tester strains TA 98, TA 100, TA 1535 or TA 1537 was not observed with and without metabolic activation.
It is therefore concluded, that Coconut oil, reaction products with polyethylene glycol and trimethylolpropane is not a bacterial mutagen.
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