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EC number: 270-470-1 | CAS number: 68441-66-7
- 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
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- 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:
- 02 Apr 2021 to 26 Apr 2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 021
- Report date:
- 2021
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- OECD Guideline 471. Genetic Toxicology: Bacterial Reverse Mutation Test. (Adopted 21 July 1997, Corrected 26 June 2020).
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- EC Guideline No. 440/2008. Part B: Methods for the Determination of Toxicity and other health effects, Guideline B.13/14: "Mutagenicity: Reverse Mutation Test using Bacteria”. Official Journal of the European Union No. L142, 31 May 2008.
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid
- EC Number:
- 270-470-1
- EC Name:
- Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid
- Cas Number:
- 68441-66-7
- Molecular formula:
- not available due to complexity of the substance
- IUPAC Name:
- Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid
- Test material form:
- liquid
- Details on test material:
- Identification: Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid
Batch (Lot) Number: 2019194336
Expiry date: 03 June 2022
Physical Description: Clear light yellow liquid
Purity/Composition: UVCB
Storage Conditions: At room temperature
Additional information
Test Facility test item number: 212207/A
Purity/Composition correction factor: No correction factor required
Test item handling: No specific handling conditions required
Information about the purity and composition of the test item is not available since the test item is an UVCB (Substance of Unknown or Variable composition, Complex Reaction Products or Biological Materials). Since a sample relevant for the purpose of this study was tested, it was concluded that the study integrity was not affected by the omission of this
information.
Constituent 1
- Specific details on test material used for the study:
- No further details specified in the study report.
Method
- Target gene:
- histidine and tryptophan
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Details on mammalian cell type (if applicable):
- Test System: Salmonella typhimurium bacteria and Escherichia coli bacteria
Rationale: Recommended test system in international guidelines (e.g. OECD, EC).
Source: Trinova Biochem GmbH, Germany [Master culture from Dr. Bruce N. Ames (TA1535, TA1537, TA98, TA100; and Master culture from The National Collections of Industrial and Marine Bacteria, Aberdeen, UK (WP2uvrA)]
The Salmonella typhimurium strains were checked at least every year to confirm their
histidine-requirement, crystal violet sensitivity, ampicillin resistance (TA98 and TA100),
UV-sensitivity and the number of spontaneous revertants.
The Escherichia coli WP2uvrA strain detects base-pair substitutions. The strain lacks an excision repair system and is sensitive to agents such as UV. The sensitivity of the strain to a wide variety of mutagens has been enhanced by permeabilization of the strain using
Tris-EDTA treatment (Ref.1). The strain was checked to confirm the tryptophan-requirement, UV-sensitivity and the number of spontaneous revertants at least every year.
Stock cultures of the five strains were stored in the ultra-low freezer set to maintain -150°C. - Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- Not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-Fraction
Rat liver microsomal enzymes (S9 homogenate) were obtained from Trinova Biochem GmbH, Giessen, Germany and were prepared from male Sprague Dawley rats that had been injected intraperitoneally with Aroclor 1254 (500 mg/kg body weight).
Each S9 batch was characterized with the mutagens benzo-(a)-pyrene (Sigma) and 2-aminoanthracene, which require metabolic activation, in tester strain TA100 at concentrations of 5 µg/plate and 2.5 µg/plate, respectively.
Preparation of S9-Mix
S9-mix was prepared immediately before use and kept refrigerated. S9-mix contained per 10 mL: 30 mg NADP (Randox Laboratories Ltd., Crumlin, United Kingdom) and 15.2 mg glucose-6-phosphate (Roche Diagnostics, Mannheim, Germany) in 5.5 mL Milli-Q water (Millipore Corp., Bedford, MA., USA); 2 mL 0.5 M sodium phosphate buffer pH 7.4; 1 mL 0.08 M MgCl2 solution (Merck); 1 mL 0.33 M KCl solution (Merck). The above solution was filter (0.22 µm)-sterilized. To 9.5 mL of S9-mix components 0.5 mL S9-fraction was added (5% (v/v) S9-fraction) to complete the S9-mix. - Test concentrations with justification for top dose:
- Dose-range Finding Test: Eight concentrations, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate were tested in triplicate.
In the first mutation experiment, the test item was tested up to concentrations of 5000 µg/plate.
In the second mutation experiment, the test item was tested up to concentrations of 5000 µg/plate.
The highest concentration of the test item used in the subsequent mutation assays was 5000 µg/plate. At least five different doses (increasing with approximately half-log steps) of the test item were tested in triplicate in each strain in the absence and presence of S9-mix. The first experiment was a direct plate assay and the second experiment was a pre-incubation assay. - Vehicle / solvent:
- The test item formed a clear colourless solution in ethanol.
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 2-nitrofluorene
- sodium azide
- methylmethanesulfonate
- other: ICR-191; 2-AA
- Details on test system and experimental conditions:
- Environmental conditions
All incubations were carried out in a controlled environment at a temperature of 37.0 ± 1.0°C (actual range 36.6 - 39.3°C). The temperature was continuously monitored throughout the experiment. Due to addition of plates (which were at room temperature) to the incubator or due to opening and closing the incubator door, temporary deviations from the temperature may occur. Based on laboratory historical data these deviations are considered not to affect the study integrity.
Experimental Design
Dose-range Finding Test
Selection of an adequate range of doses was based on a dose-range finding test with the strains TA100 and WP2uvrA, both with and without S9-mix. Eight concentrations, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate were tested in triplicate.
The highest concentration of the test item used in the subsequent mutation assays was
5000 µg/plate. At least five different doses (increasing with approximately half-log steps) of the test item were tested in triplicate in each strain in the absence and presence of S9-mix. The first experiment was a direct plate assay and the second experiment was a pre-incubation assay.
The negative control (vehicle) and relevant positive controls were concurrently tested in each strain in the presence and absence of S9-mix.
First Experiment: Direct Plate Assay
The above mentioned dose-range finding study with two tester strains is reported as a part of the direct plate assay. In the second part of this experiment, the test item was tested both in the absence and presence of S9-mix in the tester strains TA1535, TA1537 and TA98. Top agar in top agar tubes was melted by heating to 45 ± 2°C. The following solutions were successively added to 3 mL molten top agar: 0.1 mL of a fresh bacterial culture
(109 cells/mL) of one of the tester strains, 0.1 ml of a dilution of the test item in ethanol or control solution and either 0.5 ml S9-mix (in case of activation assays) or 0.5 mL 0.1 M phosphate buffer (in case of non-activation assays).
The ingredients were mixed on a Vortex and the content of the top agar tube was poured onto a selective agar plate. After solidification of the top agar, the plates were inverted and incubated in the dark at 37.0 ± 1.0°C for 48 ± 4 h. After this period revertant colonies (histidine independent (His+) for Salmonella typhimurium bacteria and tryptophan independent (Trp+) for Escherichia coli) were counted.
Second Experiment: Pre-Incubation Assay
The test item was tested both in the absence and presence of S9-mix in all tester strains. Top agar in top agar tubes was melted by heating to 45 ± 2°C. The following solutions were
pre-incubated for 30 ± 2 minutes by 70 rpm at 37 ± 1°C, either 0.5 mL S9-mix (in case of activation assays) or 0.5 mL 0.1 M phosphate buffer (in case of non-activation assays),
0.1 mL of a fresh bacterial culture (109 cells/mL) of one of the tester strains, 0.05 mL of a dilution of the test item in ethanol or control solution. After the pre-incubation period 3 mL molten top agar was added to the solutions. The ingredients were mixed on a Vortex and the content of the top agar tube was poured onto a selective agar plate. After solidification of the top agar, the plates were inverted and incubated in the dark at 37.0 ± 1.0°C for 48 ± 4 h. After this period revertant colonies (histidine independent (His+) for Salmonella typhimurium bacteria and tryptophan independent (Trp+) for Escherichia coli) were counted.
Colony Counting
The revertant colonies were counted automatically with the Sorcerer Colony Counter. Plates with sufficient test item precipitate to interfere with automated colony counting were counted manually. Evidence of test item precipitate on the plates and the condition of the bacterial background lawn were evaluated when considered necessary, macroscopically and/or microscopically by using a dissecting microscope. - Rationale for test conditions:
- In accordance with test guidelines
- Evaluation criteria:
- A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at Charles River Den Bosch.
b) The selected dose-range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated.
A test item is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is not greater than two times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three times the concurrent control.
b) The negative response should be reproducible in at least one follow up experiment.
A test item is considered positive (mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is greater than two times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537 or TA98 is greater than three times the concurrent control.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment. - Statistics:
- No formal hypothesis testing was done.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- 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, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- First Experiment: Direct Plate Assay
The test item was initially tested in the tester strains TA100 and WP2uvrA as a dose-range finding test with concentrations of 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate in the absence and presence of S9-mix. Based on the results of the dose-range finding test, the following dose-range was selected for the mutation assay with the tester strains, TA1535, TA1537 and TA98 in the absence and presence of S9-mix: 17, 52, 164, 512, 1600 and
5000 μg/plate.
Precipitate
Precipitation of the test item on the plates was observed at concentrations of 1600 µg/plate and upwards in tester strains TA100 and WP2uvrA. The test item precipitated on the plates at the top dose of 5000 μg/plate in tester strains TA1535, TA1537 and TA98.
Toxicity
To determine the toxicity of the test item, the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies were observed.
No reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants were observed.
In strain TA100 (presence of S9-mix), fluctuations in the number of revertant colonies below the laboratory historical control data range were observed. However, since no dose-relationship was observed, these reductions are not considered to be caused by toxicity of the test item. It is more likely these reductions are caused by an incidental fluctuation in the number of revertant colonies.
Mutagenicity
In the direct plate test, no biologically relevant increase in the number of revertants was observed upon treatment with the test item under all conditions tested. In tester strains WP2uvrA (absence of S9-mix) and TA1537 (presence of S9-mix), the test item induced 2.1-fold and 4.0-fold increases in the number of revertant colonies compared to the solvent control, respectively. These increases were not dose related and well within the historical database. Therefore the observed increases were not considered biologically relevant.
Second Experiment: Pre-Incubation Assay
To obtain more information about the possible mutagenicity of the test item, a pre-incubation experiment was performed in the absence and presence of S9-mix. Based on the results of the first mutation assay, the test item was tested up to the dose level of 5000 µg/plate in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA.
Precipitate
The test item precipitated on the plates at dose levels of 1600 μg/plate and upwards in the absence of S9-mix and at the top dose level of 5000 µg/plate in the presence of S9-mix.
Toxicity
There was no reduction in the bacterial background lawn and no biologically relevant decrease in the number of revertants at any of the concentrations tested in all tester strains in the absence and presence of S9-mix.
In strain TA100 (presence of S9-mix), fluctuations in the number of revertant colonies below the laboratory historical control data range were observed. However, since no dose-relationship was observed, these reduction are not considered to be caused by toxicity of the test item. It is more likely these reductions are caused by an incidental fluctuation in the number of revertant colonies.
Mutagenicity
In the pre-incubation test, no biologically relevant increase in the number of revertants was observed upon treatment with the test item under all conditions tested. In tester strain TA1537 (absence of S9-mix), the test item induced a 5.0-fold increase in the number of revertant colonies compared to the solvent control. The increase was not dose related, well within the historical database and linked to a low solvent control. Therefore the observed increase was not considered biologically relevant.
Formulation Analysis
Accuracy
In the vehicle, no test item was detected.
The concentrations analyzed in the dose formulation samples were in agreement with target concentrations (i.e. mean accuracies between 90% and 110%).
Homogeneity
The dose formulation samples were homogeneous (i.e. coefficient of variation ≤ 10%).
Stability
Analysis of the dose formulation samples after storage yielded a relative difference of ≤ 10%, for the low samples. The dose formulation samples were found to be stable during storage at room temperature under normal laboratory light conditions for at least 4 hours. For the high samples, the relative difference was > 10% (i.e. -12%, see deviation).
Any other information on results incl. tables
Dose-Range Finding Test: Mutagenic Response of Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay
Direct Plate Assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with one Salmonella typhimurium and one Escherichia coli strain |
|||||||||||
|
TA100 |
WP2uvrA |
|
|
|
|
||||||
Without S9-mix |
||||||||||||
Positive control |
882 |
± |
54 |
|
1484 |
± |
44 |
|
|
|
|
|
Solvent control |
72 |
± |
10 |
|
10 |
± |
3 |
|
|
|
|
|
1.7 |
84 |
± |
10 |
|
14 |
± |
2 |
|
|
|
|
|
5.4 |
79 |
± |
7 |
|
12 |
± |
0 |
|
|
|
|
|
17 |
74 |
± |
16 |
|
16 |
± |
4 |
|
|
|
|
|
52 |
79 |
± |
11 |
|
10 |
± |
2 |
|
|
|
|
|
164 |
73 |
± |
10 |
|
15 |
± |
1 |
|
|
|
|
|
512 |
62 |
± |
14 |
NP |
15 |
± |
1 |
NP |
|
|
|
|
1600 |
76 |
± |
14 |
SP |
21 |
± |
1 |
SP |
|
|
|
|
5000 |
84 |
± |
3 |
nSP |
16 |
± |
4 |
nSP |
|
|
|
|
With S9-mix |
||||||||||||
Positive control |
718 |
± |
97 |
|
360 |
± |
14 |
|
|
|
|
|
Solvent control |
57 |
± |
13 |
|
11 |
± |
2 |
|
|
|
|
|
1.7 |
51 |
± |
3 |
|
16 |
± |
4 |
|
|
|
|
|
5.4 |
58 |
± |
23 |
|
18 |
± |
5 |
|
|
|
|
|
17 |
58 |
± |
11 |
|
20 |
± |
6 |
|
|
|
|
|
52 |
54 |
± |
3 |
|
19 |
± |
5 |
|
|
|
|
|
164 |
46 |
± |
11 |
|
15 |
± |
8 |
|
|
|
|
|
512 |
55 |
± |
7 |
NP |
16 |
± |
2 |
NP |
|
|
|
|
1600 |
63 |
± |
8 |
SP |
16 |
± |
4 |
SP |
|
|
|
|
5000 |
56 |
± |
8 |
nSP |
20 |
± |
9 |
nSP |
|
|
|
|
NP = No precipitate
SP = Slight precipitate
n = Normal bacterial background lawn
Experiment 1: Mutagenic Response of Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay
Direct Plate Assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium |
|||||||||||
|
TA1535 |
TA1537 |
TA98 |
|||||||||
Without S9-mix |
||||||||||||
Positive control |
910 |
± |
57 |
|
1092 |
± |
158 |
|
1498 |
± |
359 |
|
Solvent control |
9 |
± |
4 |
|
7 |
± |
4 |
|
19 |
± |
1 |
|
17 |
9 |
± |
5 |
|
4 |
± |
0 |
|
21 |
± |
6 |
|
52 |
7 |
± |
0 |
|
5 |
± |
2 |
|
17 |
± |
2 |
|
164 |
7 |
± |
7 |
|
7 |
± |
2 |
|
13 |
± |
2 |
|
512 |
11 |
± |
4 |
|
5 |
± |
2 |
|
20 |
± |
2 |
|
1600 |
6 |
± |
3 |
NP |
9 |
± |
5 |
NP |
18 |
± |
3 |
NP |
5000 |
13 |
± |
4 |
nSP |
4 |
± |
2 |
nSP |
17 |
± |
6 |
nSP |
With S9-mix |
||||||||||||
Positive control |
229 |
± |
27 |
|
241 |
± |
14 |
|
1209 |
± |
101 |
|
Solvent control |
10 |
± |
1 |
|
2 |
± |
2 |
|
24 |
± |
10 |
|
17 |
10 |
± |
3 |
|
3 |
± |
3 |
|
25 |
± |
8 |
|
52 |
9 |
± |
3 |
|
5 |
± |
2 |
|
19 |
± |
2 |
|
164 |
9 |
± |
6 |
|
5 |
± |
2 |
|
22 |
± |
8 |
|
512 |
11 |
± |
4 |
|
8 |
± |
4 |
|
23 |
± |
8 |
|
1600 |
12 |
± |
1 |
NP |
8 |
± |
5 |
NP |
22 |
± |
9 |
NP |
5000 |
14 |
± |
9 |
nSP |
4 |
± |
2 |
nSP |
26 |
± |
2 |
nMP |
MP = Moderate Precipitate
NP = No precipitate
SP = Slight precipitate
n = Normal bacterial background lawn
Experiment 2: Mutagenic Response of Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay
Pre-incubation Assay
Dose (µg/plate) |
Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium and one Escherichia coli strain. |
|||||||||||||||||||
|
TA1535 |
TA1537 |
TA98 |
TA100 |
WP2uvrA |
|||||||||||||||
Without S9-mix |
||||||||||||||||||||
Positive control |
967 |
± |
185 |
|
56 |
± |
11 |
|
1590 |
± |
84 |
|
782 |
± |
56 |
|
839 |
± |
129 |
|
Solvent control |
7 |
± |
0 |
|
1 |
± |
1 |
|
14 |
± |
10 |
|
107 |
± |
10 |
|
21 |
± |
6 |
|
17 |
10 |
± |
6 |
|
3 |
± |
2 |
|
17 |
± |
7 |
|
107 |
± |
12 |
|
23 |
± |
4 |
|
52 |
11 |
± |
1 |
|
3 |
± |
2 |
|
17 |
± |
5 |
|
100 |
± |
13 |
|
18 |
± |
1 |
|
164 |
9 |
± |
4 |
|
3 |
± |
2 |
|
15 |
± |
4 |
|
103 |
± |
8 |
|
20 |
± |
1 |
|
512 |
9 |
± |
2 |
NP |
3 |
± |
2 |
NP |
17 |
± |
10 |
NP |
110 |
± |
19 |
NP |
22 |
± |
8 |
NP |
1600 |
11 |
± |
1 |
SP |
5 |
± |
2 |
SP |
14 |
± |
3 |
SP |
122 |
± |
20 |
SP |
24 |
± |
9 |
SP |
5000 |
11 |
± |
3 |
nSP |
2 |
± |
1 |
nMP |
14 |
± |
2 |
nSP |
118 |
± |
9 |
nSP |
19 |
± |
8 |
nMP |
With S9-mix |
||||||||||||||||||||
Positive control |
112 |
± |
8 |
|
33 |
± |
7 |
|
367 |
± |
26 |
|
636 |
± |
151 |
|
345 |
± |
24 |
|
Solvent control |
12 |
± |
4 |
|
6 |
± |
2 |
|
23 |
± |
1 |
|
52 |
± |
11 |
|
24 |
± |
10 |
|
17 |
9 |
± |
2 |
|
4 |
± |
1 |
|
20 |
± |
3 |
|
57 |
± |
10 |
|
27 |
± |
5 |
|
52 |
8 |
± |
5 |
|
4 |
± |
1 |
|
27 |
± |
4 |
|
48 |
± |
3 |
|
27 |
± |
4 |
|
164 |
7 |
± |
2 |
|
2 |
± |
2 |
|
24 |
± |
1 |
|
43 |
± |
8 |
|
14 |
± |
2 |
|
512 |
11 |
± |
4 |
|
5 |
± |
2 |
|
19 |
± |
4 |
|
46 |
± |
7 |
|
32 |
± |
2 |
|
1600 |
11 |
± |
1 |
NP |
2 |
± |
2 |
NP |
18 |
± |
2 |
NP |
47 |
± |
11 |
NP |
21 |
± |
2 |
NP |
5000 |
16 |
± |
3 |
nSP |
2 |
± |
1 |
nMP |
24 |
± |
1 |
nSP |
57 |
± |
13 |
nSP |
27 |
± |
7 |
nSP |
MP = Moderate Precipitate
NP = No precipitate
SP = Slight precipitate
n = Normal bacterial background lawn
Historical Control data of the Solvent Control
|
TA1535 |
TA1537 |
TA98 |
TA100 |
WP2uvrA |
|||||
S9-mix |
- |
+ |
- |
+ |
- |
+ |
- |
+ |
- |
+ |
Range |
3 – 26 |
4 – 25 |
2 – 24 |
2 – 17 |
4 – 61 |
6 – 60 |
58 – 188 |
50 – 176 |
9 – 61 |
11 – 68 |
Mean |
9 |
10 |
5 |
5 |
14 |
18 |
109 |
101 |
24 |
29 |
SD |
3 |
3 |
2 |
2 |
5 |
6 |
19 |
21 |
9 |
10 |
Total number of plates |
2504 |
2484 |
2528 |
2493 |
2523 |
2528 |
2566 |
2508 |
2385 |
2361 |
SD = Standard deviation
Historical control data from experiments performed between Nov 2017 and Nov 2020.
Historical Control Data of the Positive Control Items
|
TA1535 |
TA1537 |
TA98 |
TA100 |
WP2uvrA |
|||||
S9-mix |
- |
+ |
- |
+ |
- |
+ |
- |
+ |
- |
+ |
Range |
107 – 1530 |
78 – 1481 |
64 – 1475 |
52 – 1843 |
379 – 2118 |
265 – 2077 |
452 – 1993 |
397 – 2666 |
93 – 1999 |
109 – 1968 |
Mean |
972 |
289 |
818 |
293 |
1252 |
945 |
865 |
1432 |
1196 |
421 |
SD |
170 |
108 |
370 |
142 |
251 |
378 |
181 |
398 |
526 |
186 |
Total number of plates |
2360 |
2365 |
1979 |
2384 |
2451 |
2379 |
2382 |
2387 |
2269 |
2267 |
SD = Standard deviation
Historical control data from experiments performed between Nov 2017 and Nov 2020.
Applicant's summary and conclusion
- Conclusions:
- In conclusion, based on the results of this study it is concluded that Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.
- Executive summary:
The objective of this study was to determine the potential of Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid and/or its metabolites to induce reverse mutations at the histidine locus in several strains of Salmonella typhimurium (S.typhimurium; TA98, TA100, TA1535, and TA1537), and at the tryptophan locus of Escherichia coli (E. coli) strain WP2uvrA in the presence or absence of an exogenous mammalian metabolic activation system (S9).
The test was performed in two independent experiments, at first a direct plate assay was performed and secondly a pre-incubation assay.
The study procedures described in this report were based on the most recent OECD and EC guidelines.
Batch 2019194336 of the test item was a clear light yellow liquid. The vehicle of the test item was ethanol.
The concentrations analyzed in the dose formulation samples were in agreement with target concentrations (i.e. mean accuracies between 90% and 110%). The dose formulation samples were homogeneous (i.e. coefficient of variation ≤ 10%). The low dose formulation samples were found to be stable during storage at room temperature under normal laboratory light conditions for at least 4 hours. For the high samples, the relative difference was > 10% (i.e. -12%, seedeviation).
In the dose-range finding study, the test item was initially tested up to concentrations of 5000 µg/plate in the strains TA100 and WP2uvrA in the direct plate assay. The test item precipitated on the plates at dose levels of 1600 μg/plate and upwards. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed. Results of this dose-range finding test were reported as part of the first mutation assay.
In the first mutation experiment, the test item was tested up to concentrations of 5000 µg/plate in the strains TA1535, TA1537 and TA98. The test item precipitated on the plates at the top dose of 5000 μg/plate. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed.
In the second mutation experiment, the test item was tested up to concentrations of 5000 µg/plate in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA in the pre-incubation assay. The test item precipitated on the plates at dose levels of 1600 μg/plate and upwards in the absence of S9-mix and at the top dose level of 5000 µg/plate in the presence of S9-mix. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed.
In this study, acceptable responses were obtained for the negative and strain-specific positive control items indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
The test item did not induce a biologically relevant, dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in a follow-up experiment.
In conclusion, based on the results of this study it is concluded that Decanoic acid, mixed esters with dipentaerythritol, octanoic acid and valeric acid is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.
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