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EC number: 610-122-1 | CAS number: 433733-94-9
- 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
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- Nanomaterial specific surface area
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- Endpoint summary
- Stability
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- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Based on the results of this study it is concluded that LDT600 C3 is not mutagenic in the Salmonella typhimurium reverse mutation assay.
Diacetyl Thymidine (LDT600-C3) is not mutagenic in the TK mutation test system under the experimental conditions described in this report.
QSAR- in vitro gene mutation study in mammalian cells: negative
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 23.5.2009 - 29.5.2009
- 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
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his
- Species / strain / cell type:
- other: TA97, TA98, TA100, TA102 and TA1535
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver S9-mix
- Test concentrations with justification for top dose:
- 100, 333, 1000, 3330 and 5000 μg/plate
- Vehicle / solvent:
- - Vehicle/solvent used: dimethylsulfoxide (DMSO)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: without S9: sodium azide (TA1535), 2-nitrofluorene (TA98), methylmethanesulfonat (TA100), cumene hydroperoxide (TA102), ICR-191 (TA97); with S9: 1,8 dihydroxyanthraquinone (TA102), 2-aminoanthracene (all other strains)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: three replications in two independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: inspection of bacterial background lawn - Evaluation criteria:
- A test substance is considered positive in the test, if:
- The total number of revertants in tester strains TA100, TA97 and TA102 is greater than two times the concurrent control or the total number of revertants in tester strains TA1535 and TA98 is greater than three times the concurrent control.
- 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 independently repeated experiment. - Key result
- Species / strain:
- other: TA97, TA98, TA100, TA102 and TA1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Conclusions:
- All bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments.
The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
Based on the results of this study it is concluded that LDT600 C3 is not mutagenic in the Salmonella typhimurium reverse mutation assay. - Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- May 2019 - Aug 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)
- GLP compliance:
- yes
- Type of assay:
- bacterial forward mutation assay
- Target gene:
- Thymidine Kinase Gene
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- L5178Y/TK+/--3.7.2C mouse lymphoma cells.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver microsomal enzymes (S9 homogenate)
- Test concentrations with justification for top dose:
- In order to select appropriate dose levels for mutagenicity testing, cytotoxicity data were obtained by treating 8 x 106 cells (106 cells/mL for 3 hour treatment) or 6 x 106 cells (1.25 x 105 cells/mL for 24 hour treatment) with a number of test item concentrations increasing by approximately half log steps. The cell cultures for the 3 hour treatment were placed in sterile 30 mL centrifuge tubes, and incubated in a shaking incubator at 37.0 ± 1.0°C and 145 rpm. The cell cultures for the 24 hour treatment were placed in sterile 75 cm2 culture flasks at 37.0 ± 1.0°C. The test item was tested in the absence and presence of S9-mix.
The highest tested concentration was 2000 μg/mL exposure medium.
The surviving cells of the 3 hour treatment were subcultured twice to determine cytotoxicity.
After 24 hour of subculturing, the cells were counted and subcultured again for another 24 hours, after that the cells were counted. The surviving cells of the 24 hour treatment were subcultured once. After 24 hours of subculturing, the cells were counted. If less than 1.25 x 105 cells/mL were counted no subculture was performed. The suspension growth expressed as the reduction in cell growth after approximately 24 and 48 hours or only 24 hours cell growth, compared to the cell growth of the solvent control, was used to determine an appropriate dose-range for the mutagenicity tests. - Vehicle / solvent:
- dimethyl sulfoxide (DMSO)
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Test System L5178Y/TK+/--3.7.2C mouse lymphoma cells.
Rationale Recommended test system in international guidelines (e.g. OECD).
Source American Type Culture Collection, (ATCC, Manassas, USA) (2001).
All incubations were carried out in a humid atmosphere (80 - 100%, actual range 55 - 100%) containing 5.0 ± 0.5% CO2 in air in the dark at 37.0 ± 1.0°C (actual range 34.1 – 37.4°C).
Temperature and humidity were continuously monitored throughout the experiment. The CO2 percentage was monitored once on each working day. Temporary deviations from the temperature, humidity and CO2 percentage may occur due to opening and closing of the incubator door. Any variation to these conditions were evaluated and maintained in the raw data.
Stock cultures of the cells were stored in the freezer (-150°C). The cultures were checked for mycoplasma contamination. Cell density was kept below 1 x 106 cells/mL.
Per culture 8 x 106 cells (106 cells/mL for 3 hour treatment) or 6 x 106 cells (1.25 x 105 cells/mL for 24 hour treatment) were used. The cell cultures for the 3 hour treatment were placed in sterile 30 mL centrifuge tubes, and incubated in a shaking incubator at 37.0 ± 1.0°C and 145 rpm. The cell cultures for the 24 hour treatment were placed in sterile 75 cm2 culture flasks at 37.0 ± 1.0°C. Solvent and positive controls were included and the solvent control
was tested in duplicate. In the first experiment, cell cultures were exposed for 3 hours to Diacetyl Thymidine (LDT600-C3) in exposure medium in the absence and presence of S9-mix. In the second experiment, cell cultures were exposed to Diacetyl Thymidine (LDT600-C3) in exposure medium for 24 hours in the absence of S9-mix.
For the 3 hour treatment, cell cultures were exposed to the test item in exposure medium in the absence as well as in the presence of S9-mix. After exposure, the cells were separated from the treatment solutions by 2 centrifugation steps (216 g, 5 min). The first centrifugation step was followed by removal of the supernatant and resuspension of the cells in Hanks’ balanced salt solution and after the second centrifugation step the cells were resuspended in 50 mL growth medium (R10-medium).
For the 24 hour treatment, cell cultures were exposed to the test item in exposure medium in the absence of S9-mix. After exposure, the cells were separated from the treatment solutions by 2 centrifugation steps (216 g, 5 min). The first centrifugation step was followed by removal of the supernatant and resuspension of the cells in Hanks’ balanced salt solution and after the second centrifugation step the cells were resuspended in 20 mL growth medium
(R10-medium). The cells in the final suspension were counted with the coulter particle counter. - Evaluation criteria:
- A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls (CEday2) is between 65 and 120% in order to have an acceptable number of surviving cells analyzed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50 per 106 survivors and ≤ 170 per 106 survivors.
c) The suspension growth (SG) over the 2-day expression period for the solvent controls should be between 8 and 32 for the 3 hour treatment, and between 32 and 180 for the 24 hour treatment.
d) The positive control should demonstrate an absolute increase in the total mutation frequency, that is, an increase above the spontaneous background MF (an induced MF (IMF)) of at least 300 x 10-6. At least 40% of the IMF should be reflected in the small colony MF. And/or, the positive control has an increase in the small colony MF of at least 150 x 10-6 above that seen in the concurrent solvent control (a small colony IMF of 150 x 10-6).
All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented. - Statistics:
- In addition to the criteria stated below, any increase of the mutation frequency should be
evaluated for its biological relevance including comparison of the results with the historical
control data range.
The global evaluation factor (GEF) has been defined by the IWGT as the mean of the
negative/solvent MF distribution plus one standard deviation. For the micro well version of
the assay the GEF is 126.
A test item is considered positive (mutagenic) in the mutation assay if it induces a MF of
more than MF(controls) + 126 in a dose-dependent manner. An observed increase should be
biologically relevant and will be compared with the historical control data range.
A test item is considered equivocal (questionable) in the mutation assay if no clear conclusion
for positive or negative result can be made after an additional confirmation study.
A test item is considered negative (not mutagenic) in the mutation assay if: none of the tested
concentrations reaches a mutation frequency of MF(controls) + 126. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- 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 applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Remarks on result:
- no mutagenic potential (based on QSAR/QSPR prediction)
- Conclusions:
- Diacetyl Thymidine (LDT600-C3) is not mutagenic in the TK mutation test system under the experimental conditions described in this report.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Study period:
- January 2020
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- results derived from a valid (Q)SAR model and falling into its applicability domain, with limited documentation / justification
- Justification for type of information:
- The prediction was considered satisfactory because of the structural similarity and similar mechanism of action of the target chemical and the analogs.
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The chemical was entered in the QSAR Toolbox by SMILES. The identified structure of the chemical was
retained and subsequently added to the data matrix. After selecting the endpoint of interest
(genotoxicity), in the data module, the relevant databases were searched for experimental data. No
experimental data was found. The profiling module was used to identify the main characteristics of the
chemical. In category definition, the chemical profiler (OECD HPV chemical categories) was used to
categorize the chemical. In the data filling, in vitro genotoxicity test results with L5178Y mouse
lymphoma cells was selected because there was sufficient analog data available to be used to fill the
data gap. The data gap filling was initiated by read-across, as it is the appropriate data gap-filling
method for qualitative endpoints such as genotoxicity. The results were subsequently refined by DNA
binding by OECD, protein binding alerts for chromosomal aberration by OASIS, DNA alerts for CA and
MNT by OASIS and organic functional groups (US EPA). The prediction was considered satisfactory
because of the structural similarity and similar mechanism of action of the target chemical and the
analogs. - GLP compliance:
- not specified
- Target gene:
- QSAR
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not applicable
- True negative controls validity:
- not applicable
- Positive controls validity:
- not applicable
- Remarks on result:
- not measured/tested
- Conclusions:
- in vitro gene mutation study in mammalian cells: negative
Referenceopen allclose all
The test substance did not induce a significant dose-related increase in the number of revertant colonies in each of the five tester strains (TA1535, TA97, TA98, TA100 and TA102) both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently repeated experiment.
Database(s) used:
- ECHA CHEM
Category boundaries (applicability domain):
- Active descriptor(s) range:
- log Kow: from -0.38 to 0.23 target chemical is in domain
- Response range:
- in vitro gene mutation study in mammalian cells: negative (x9) (from Negative to Negative)
Profilers:
- Tertiary Amines (OECD HPV Chemical Categories) (primary grouping)
target chemical is in domain
- DNA binding by OECD (subcategorization)
target chemical is in domain
- Protein binding alerts for Chromosomal aberration by OASIS (subcategorization)
target chemical is in domain
- DNA alerts for CA and MNT by OASIS (subcategorization)
target chemical is in domain
- Organic functional groups (US EPA) (subcategorization)
target chemical is in domain
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
All bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments.
The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
Based on the results of this study it is concluded that LDT600 C3 is not mutagenic in the Salmonella typhimurium reverse mutation assay.
In the absence of S9-mix, Diacetyl Thymidine (LDT600-C3) did not induce a biologically
relevant increase in the mutation frequency in the first experiment. This result was confirmed
in a repeat experiment with modification in the duration of treatment.
In the presence of S9-mix, Diacetyl Thymidine (LDT600-C3) did not induce a biologically
relevant increase in the mutation frequency.
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