Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 248-468-7 | CAS number: 27458-90-8
- 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
- Nanomaterial porosity
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutations
The potential of DI-tert-DODECYL DISULFIDE to induce a reverse mutation was evaluated in bacteria Salmonella typhimurium (Molinier, 1995). A preliminary toxicity test was performed to define the doses to be used for the mutagenicity study. The test substance was then tested in two independent tests, with or without a metabolic activation system, the S9 mix, prepared from a liver micrasomal fraction (S9) of rats induced with Araclor 1254. The tests were perf ormed according to the direct plate incorporation method except the second test with S9 mix, according to the preincubation method (one hour, 37°C). Five strains of bacteria Salmonella typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 102 were used. Each strain was exposed to five doses of the test substance (three plates/dose). After 48 to 72 hours of incubation at 37°C, the revertant colonies were scored. DI-tert-DODECYL DISULFIDE was dissolved in dimethylsulfoxide (DMSO). The number of revertants of the vehicle and positive controls was as specified in the acceptance criteria and within the range of the historical data. The top dose was selected according to the criteria specified in the international regulations. Since the test substance was non-toxic, freely soluble, the top dose was 5000 µg/plate. The selected range dose was: 312.5, 625, 1250, 2500 and 5000 µg/plate. The test substance did not induce any significant increase in the number of revertants, with or without S9 mix, in any of the five strains. Under these experimental conditions, DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this bacterial reverse mutation assay on Salmonella typhimurium.
The potential of DI-tert-DODECYL DISULFIDE to induce mutations at the TK (thymidine kinase) locus was evaluated in L5 l 78Y mouse lymphoma cells (Molinier, 1996). After a preliminary toxicity test, DI-tert-DODECYL DISULFIDE was tested in two independent experiments, with or without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Araclor 1254. Approximately 0.5.10e6 cells/ml in 20 ml culture medium with 5% horse serum were exposed to the test or control substances, in the presence or absence of S9 mix (final concentration of S9 fraction 2% ), for three hours at 37°C. Cytotoxicity was then determined using cloning efficiency (CE0) before expression of the mutant phenotype. Cells viability (using cloning efficiency CE1 ) and number of mutant clones (differentiating small and large colonies) were checked after the expression of the mutant phenotype. The test substance was dissolved in ethanol. The dose-levels for the positive contrals were as follows: without S9 mix: 25 µg/ml of methylmethane sulfonate, with S9 mix: 3 µg/ml of cyclophosphamide.
The cloning efficiencies CE0 and CE2 and mutation frequencies of the vehicle and positive controls were within the range of our historical data. The treament-levels were for both mutagenicity experiments, with and without S9 mix: 200, 400, 800 and 1600 µg/ml; 1600 µg/ml was the highest treatment-level above the limit of solubility in the final culture medium at the end of the treatment-period. The cloning efficiencies CE0 and CE2 were considered equivalent to that of the control cultures. DI-tert-DODECYL DISULFIDE did not induce any significant increase in the mutation frequency, with and without S9 mix. Under our experimental conditions, DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this mouse lymphoma assay.
Chromosomal aberrations
The potential of DI-tert-DODECYL DISULFIDE to induce chromosome aberrations was evaluated in cultured human lymphocytes (Molinier, 1996). The test substance, dissolved in ethanol, was tested in two independent tests, with or without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9) of rats induced with Aroclor 1254. For each culture, heparinised whole blood was added to culture medium containing a mitogen (phytohaemogglutinin) and incubated at 37°C in a humidified atmosphere of 5% CO2/95% air, for 48 hours.
First test: cells were then exposed to the test or control substances, with or without S9 mix, for three hours then rinsed. One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to black cells at the metaphase-stage of mitosis. Harvest time was 20 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times. As this test gives negative results, both with or without S9 mix, an additional test was performed as follows:
Second test:
. without S9 mix, cells were exposed continuously to the test or control substances.
. with S9 mix, cells were exposed to the test or control substances for three hours then rinsed.
One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to block cells at the metaphase-stage of mitosis. Harvest times were 20 hours and 44 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times and 24 hours after.
After hypotonic treatment (KCl 0.075 M), the cells were fixed in a methanol/acetic acid mixture (3/1; v/v), spread on glass slides and stained with Giemsa. All the slides were coded for scoring.
The doses of DI-tert-DODECYL DISULFIDE were as follows:
First test with and without S9 mix: 50, 100, 200, 400, 600 and 800 µg/rnl for treatment and 400, 600 and 800 µg/ml for chromosomal aberrations scoring. 800 µg/ml being the lowest precipitating dose.
Second test with and without S9 mix: 200, 400, 600 and 800 µg/ml for treatment and 400, 600 and 800 µg/ml for chromosomal aberrations scoring. The doses of the positive controls were 3 µg/ml mitomycin C (MMC) without S9 mix (3 hours of treatment) or 0.2 µg/ml (continuous treatment) and 50 µg/ml of cyclophosphamide (CPA) with S9 mix.
The frequency of cells with structural chromosome aberrations in the negative, vehicle and positive controls was as specified in the acceptance criteria and within the range of the historical data. The test substance did not induce any significant increase in the frequency of cells with structural chromosome aberrations, with or without S9 mix, for both of the two tests and for the two harvest times. Under these experimental conditions, DI-tert-DODECYL DISULFIDE did not induce chromosome aberrations in cultured human lymphocytes.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 26th May 1983 and revised Draft document of December 1994 .
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver micrasomal fraction (S9) of rats induced with Araclor 1254
- Test concentrations with justification for top dose:
- 312.5, 625, 1250, 2500 and 5000 µg/plate
- Vehicle / solvent:
- DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- mitomycin C
- other: 2-Anthramine
- Details on test system and experimental conditions:
- To assess the toxicity of the test substance to the bacteria, six doses (one plate/dose) were tested in the TA 98, TA 100 and TA 102 strains, with or without S9 mix.
The tests were peïf ormed according to:
. direct plate incorporation method (both tests without S9 mix, first test with S9 mix): 0.05 to 0.1 ml of the test substance solution, 0.5 ml of S9 mix when required and 0.1 ml of the strain were mixed with 2 ml of overlay agar containing traces of the relevant amino-acid and biotin and maintained at 45°C. After rapid homogenization, the mixture was spread out on a Petri plate containing minimum medium.
. preincubation rnethod (second test with S9 mix): 0.05 to 0.1 ml of the test substance solution, 0.5 ml of S9 mix and 0.1 ml of the strain were incubated for 60 minutes at 37°C prior adding the overlay agar and pouring onto the surface of a minimum agar plate.
After 48 to 72 hours of incubation at 37°C, revertants were scored with an automatic counter (Artek counter, model 880, O.S.I., 75015 Paris, France). - Evaluation criteria:
- Biological relevance of the results was considered first. In addition, the following criteria may be used as an aid for determining a positive response:
. a dose-related increase in the number of revertants,
and/or
. a reproducible increase in the number of revertants (i.e. a doubling in at least one strain when compared to that of the controls) for at least one of the doses. - Key result
- Species / strain:
- S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100 and TA 102
- 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
- Conclusions:
- DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this bacterial reverse mutation assay on Salmonella typhimurium.
- Executive summary:
The potential of DI-tert-DODECYL DISULFIDE to induce a reverse mutation was evaluated in bacteria Salmonella typhimurium. A preliminary toxicity test was performed to define the doses to be used for the mutagenicity study. The test substance was then tested in two independent tests, with or without a metabolic activation system, the S9 mix, prepared from a liver micrasomal fraction (S9) of rats induced with Araclor 1254. The tests were perf ormed according to the direct plate incorporation method except the second test with S9 mix, according to the preincubation method (one hour, 37°C). Five strains of bacteria Salmonella typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 102 were used. Each strain was exposed to five doses of the test substance (three plates/dose). After 48 to 72 hours of incubation at 37°C, the revertant colonies were scored. DI-tert-DODECYL DISULFIDE was dissolved in dimethylsulfoxide (DMSO). The number of revertants of the vehicle and positive controls was as specified in the acceptance criteria and within the range of the historical data. The top dose was selected according to the criteria specified in the international regulations. Since the test substance was non-toxic, freely soluble, the top dose was 5000 µg/plate. The selected range dose was: 312.5, 625, 1250, 2500 and 5000 µg/plate. The test substance did not induce any significant increase in the number of revertants, with or without S9 mix, in any of the five strains. Under these experimental conditions, DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this bacterial reverse mutation assay on Salmonella typhimurium.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Version / remarks:
- 4th April 1984 and revised Draft document of September 1995
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- other: In Vitro Mammalian Cell Gene Mutation Test
- Target gene:
- Thymidine kinase locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- L5178Y cells, originally obtained through ATCC, were kindly supplied by Dr. Oudelkhim-Diot
(Vitry-sur-Seine, France). - Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Araclor 1254
- Test concentrations with justification for top dose:
- 200, 400, 800 and 1600 µ/ml
1600 µg/ml was the highest treatment-level above the limit of solubility in the final culture medium at the end of the treatment-period. - Vehicle / solvent:
- ethanol
- 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:
- - Treatment
The day before treatment, the cells were seeded in 50 ml of RPMI 1640 medium containing 10% horse serum, L-Glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 µg/ml) and sodium pyruvate (200 µg/ml). The cells were then incubated at 37°C in a humidified atmosphere of 5% CO2/95% air. After 24-hour incubation, culture medium was removed and cells were counted.
Approximately 0.5 10e6 cells/ml in 20 ml culture medium with 5% horse serum, were exposed to the test or control substances, in the presence or absence of S9 mix (final concentration of S9 fraction: 2% ), for three hours at 37°C. The treatment medium was then removed and the cells were counted and seeded as follows:
Cytotoxicity plates
1.6 cells/well (one 96-well plate/culture = two plates/dose-level) to determine cytotoxicity using cloning efficiency (CE0 ). After 12 ± 1 days of incubation at 37°C, the clones were counted.
To enable the expression of the mutant phenotype, 2.10e5 cells/ml were reincubated at 37°C and replated on day 1 at 2.10e5 cells/ml. After two days, the cells were seeded as follows:
Viability plates
1.6 cells/well (one 96-well plate/culture = two plates/dose-level) to de fine the number of viable cells (CE,). After 12 ± 1 days of incubation at 37°C, the clones were counted.
Mutant plates
2000 cells/well (two 96-well plates/culture = four plates/dose-level) to select the TFT (trifluorothymidine resistant) mutant cells (for determination of CEmutant). After 12 ± 1 days of incubation at 37°C in the presence of 4 µg TFT/ml of culture medium, the clones were counted, differentiating small and large colonies:
- size of small colonies, < 25% of the diameter of the well,
- size of large colonies, > 25% of the diameter of the \vell.
- Preliminary toxicity test
To assess the cytotoxicity of the test substance, four dose-levels of DI-tert-DODECYL DISULFIDE (one culture/dose-level) were tested both with and without metabolic activation.
- Mutagenicity experiments In two independent experiments, four dose-levels of DI-tert-DODECYL DISULFIDE (two cultures/dose-level) were tested both with and without metabolic activation. In each experiment, the following controls were included using at least duplicate cultures:
vehicle controls: cultures treated with the vehicle (100 µl ethanol/20 ml culture medium)
positive controls: cultures treated with:
- MMS, in the absence of S9 mix,
- CPA, in the presence of S9 mix. - Evaluation criteria:
- A reproducible two-fold increase in the mutant frequency when compared with the vehicle controls, at any dose-level and/or evidence a dose-relationship were considered as a positive result. Reference to historical data, or other considerations of biological relevance might be also taken into account in the evaluation of the data obtained.
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- 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
- Positive controls validity:
- valid
- Additional information on results:
- PRELIMINARY TOXICITY TEST
The test substance was poorly soluble in the vehicle (ethanol), the limit of solubility in this vehicle being approximately 300 mg/ml. Consequently, with a maximum dose volume of 100 µl/20 ml culture medium, the highest treatment-level feasible was 1500 µg/ml; at this dose-level, a moderate emulsion was observed in the culture medium. The treatment-levels were: 150, 500, 1000 and 1500 µg/ml. No cytoxicity was noted after treatment, with and without S9 mix: the cloning efficiencies CE0 of treated cultures were considered equivalent to that of control cultures.
MUTAGENICITY EXPERIMENTS
The cloning efficiencies CE0 and CE2 and the mutation frequencies of the vehicle and positive controls were as specified in acceptance criteria and within the range of our historical data. The top dose was selected according to the criteria specified in the international regulations: since the test substance was non-toxic and poorly soluble, the top dose-level was a dose-level above the limit of solubility in the final culture medium at the end of the treatment period. Using an initial solution at 320 mg/ml vehicle and a treatment-volume of 100 µl/20 ml culture medium, the treament-levels were for both mutagenicity experiments with and without S9 mix: 200, 400, 800 and 1600 µg/ml. The cloning efficiencies CE0 and CE2 were considered equivalent to that of the control cultures: RCE0 was at least 72% and RCE2 67%. The test substance did not induce any significant increase in the mutation frequency, with and without S9 mix. - Conclusions:
- DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this mouse lymphoma assay
- Executive summary:
The potential of DI-tertDODECYL DISULFIDE to induce mutations at the TK (thymidine kinase) locus was evaluated in L5 l 78Y mouse lymphoma cells. After a preliminary toxicity test, DI-tert-DODECYL DISULFIDE was tested in two independent experiments, with or without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Araclor 1254. Approximately 0.5.10e6 cells/ml in 20 ml culture medium with 5% horse serum were exposed to the test or control substances, in the presence or absence of S9 mix (final concentration of S9 fraction 2% ), for three hours at 37°C. Cytotoxicity was then determined using cloning efficiency (CE0) before expression of the mutant phenotype. Cells viability (using cloning efficiency CE1 ) and number of mutant clones (differentiating small and large colonies) were checked after the expression of the mutant phenotype. The test substance was dissolved in ethanol. The dose-levels for the positive contrals were as follows: without S9 mix: 25 µg/ml of methylmethane sulfonate, with S9 mix: 3 µg/ml of cyclophosphamide.
The cloning efficiencies CE0 and CE2 and mutation frequencies of the vehicle and positive controls were within the range of our historical data. The treament-levels were for both mutagenicity experiments, with and without S9 mix: 200, 400, 800 and 1600 µg/ml; 1600 µg/ml was the highest treatment-level above the limit of solubility in the final culture medium at the end of the treatment-period. The cloning efficiencies CE0 and CE2 were considered equivalent to that of the control cultures. DI-tert-DODECYL DISULFIDE did not induce any significant increase in the mutation frequency, with and without S9 mix. Under our experimental conditions, DI-tert-DODECYL DISULFIDE did not show mutagenic activity in this mouse lymphoma assay.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- 26th May 1983 and revised draft document of September 1995
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- other: In Vitro Mammalian Chromosome Aberration Test
- Species / strain / cell type:
- primary culture, other: human lymphocytes
- Details on mammalian cell type (if applicable):
- human lymphocytes will corne from whole blood sampled into heparinised sterile tubes obtained from two healthy donors.
- Cytokinesis block (if used):
- colcemid
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix, prepared from a liver microsomal fraction (S9 fraction) of rats induced with Araclor 1254
- Test concentrations with justification for top dose:
- 50, 100, 200, 400, 600 and 800 µg/ml
The highest treatment-level feasible was 800 µg/ml; at this dose-level, a moderate emulsion was observed in the culture medium. - Vehicle / solvent:
- ethanol
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Details on test system and experimental conditions:
- - Treatment
At least three doses of the test substance (two cultures/dose) were tested, with or without metabolic activation. For treatment with S9 mix, 1.5% S9 (i.e. 15% S9 mix) is added to the culture medium. During each test, the following controls were made using at least duplicate cultures:
. negative control: untreated culture,
. vehicle control: culture treated with the vehicle (15 µl ethanol/5.5 ml culture medium),
. positive contrais: culture treated (only for the first harvest time) with:
- MMC, without S9 mix,
- CPA, with S9 mix.
For each culture, 0.5 ml of heparinised whole blood was added to 5 ml of RPMI 1640 medium containing 20% fetal calf serum, L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (100 µg/ml) and 3.6% of a mitogen (phytohaemagglutinin, PHA, to stimulate the lymphocytes to divide) in a tube. The tubes were then placed at 37°C for 48 hours. First test Cells were then exposed to the test or control substances, with or without S9 mix, for three hours then rinsed. One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to block cells at the metaphase-stage of mitosis. Harvest time was 20 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times. As this test gives negative results, both with or without S9 mix, an additional test was performed as follows:·
Second test
. without S9 mix, cells were exposed continuously to the test or control substances
. with S9 mix, cells were exposed to the test or control substances for three hours then rinsed.
One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to block cells at the metaphase-stage of mitosis. Harvest times were 20 hours and 44 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times and 24 hours after.
After hypotonie treatment (KCI 0.075 M), the cells were fixed in a methanol/acetic acid mixture (3/1; v/v), spread on glass slides and stained with Giemsa. All the slides were coded for scoring. - Evaluation criteria:
- Biological relevance of the results was considered first. In addition, the following criteria may be used as an aid for deterrnining a positive response: a reproducible and statistically significant increase in the frequency of cells with structural chromosome aberrations for at Ieast one of the doses and one of the two harvest times.
- Statistics:
- For each test and for each harvest time, the frequency of cells with structural chromosome aberrations (excluding gaps) in treated cultures was compared to that of the control cultures. The comparison was performed using the X² test, in which p = 0.05 was used as the lowest level of significance.
- Species / strain:
- primary culture, other: human lymphocytes
- 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:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- First test:
The test substance was poorly soluble in the vehicle (ethanol), the limit of solubility in this vehicle being approximately 300 mg/ml. Consequently, with a maximum dose volume of 15µl/5.5ml culture medium, the highest treatment-level feasible was 800 µg/ml; at this dose-level, however, a moderate emulsion was observed in the culture medium.
The treatment-levels were: 50, 100, 200, 400, 600, 800 µg/ml. No cytotoxicity was noted after treatment, with or without S9 mix: the mitotic index of treated cultures was considered equivalent to that of contrai cultures. The top dose-level for scoring was selected according to the criteria specified in the international regulations: since the test substance was non-toxic, poorly soluble, the top dose was a dose above the limit of solubility in the final culture medium at the end of the treatment period: 800 µg/ml. Therefore, chromosome aberrations were scored on the slides corresponding to the following three doses: 400, 600, 800 µg/ml.
The test substance did not induce chromosome aberrations both with and without S9 mix after three hours of exposure. Therefore a second test was performed using a continuous exposure without S9 mix until harvest and a later harvest time both with and without S9 mix.
Second test:
The doses for treatment were: 200, 400, 600, 800 µg/ml. The mitotic index of treated cultures was considered equivalent to that of the controls. Therefore the slides corresponding to the three following doses were scored for chromosome aberrations: 400, 600, 800 µg/ml. The test substance did not induce chromosome aberrations both with and without S9 mix for both harvest times. The frequencies of cells with structural chromosome aberrations of the negative, vehicle and positive controls were as specified in acceptance criteria and within the range of our historical data for both tests and both harvest times. - Conclusions:
- DI-tert-DODECYL DISULFIDE did not induce chromosome aberrations in cultured human lymphocytes
- Executive summary:
The potential of DI-tertDODECYL DISULFIDE to induce chromosome aberrations was evaluated in cultured human lymphocytes. The test substance, dissolved in ethanol, was tested in two independent tests, with or without a metabolic activation system, the S9 mix, prepared from a liver microsomal fraction (S9) of rats induced with Aroclor 1254. For each culture, heparinised whole blood was added to culture medium containing a mitogen (phytohaemogglutinin) and incubated at 37°C in a humidified atmosphere of 5% CO2/95% air, for 48 hours.
First test: cells were then exposed to the test or control substances, with or without S9 mix, for three hours then rinsed. One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to black cells at the metaphase-stage of mitosis. Harvest time was 20 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times. As this test gives negative results, both with or without S9 mix, an additional test was performed as follows:
Second test:
. without S9 mix, cells were exposed continuously to the test or control substances.
. with S9 mix, cells were exposed to the test or control substances for three hours then rinsed.
One and a half hours before harvest, each culture was treated with a colcemid solution (10 µg/ml) to block cells at the metaphase-stage of mitosis. Harvest times were 20 hours and 44 hours from the beginning of treatment, i.e. approximately 1.5 cell cycle times and 24 hours after.
After hypotonic treatment (KCl 0.075 M), the cells were fixed in a methanol/acetic acid mixture (3/1; v/v), spread on glass slides and stained with Giemsa. All the slides were coded for scoring.
The doses of DI-tert-DODECYL DISULFIDE were as follows:
First test with and without S9 mix: 50, 100, 200, 400, 600 and 800 µg/rnl for treatment and 400, 600 and 800 µg/ml for chromosomal aberrations scoring. 800 µg/ml being the lowest precipitating dose.
Second test with and without S9 mix: 200, 400, 600 and 800 µg/ml for treatment and 400, 600 and 800 µg/ml for chromosomal aberrations scoring. The doses of the positive controls were 3 µg/ml mitomycin C (MMC) without S9 mix (3 hours of treatment) or 0.2 µg/ml (continuous treatment) and 50 µg/ml of cyclophosphamide (CPA) with S9 mix.
The frequency of cells with structural chromosome aberrations in the negative, vehicle and positive controls was as specified in the acceptance criteria and within the range of the historical data. The test substance did not induce any significant increase in the frequency of cells with structural chromosome aberrations, with or without S9 mix, for both of the two tests and for the two harvest times. Under these experimental conditions, DI-tert-DODECYL DISULFIDE did not induce chromosome aberrations in cultured human lymphocytes.
Referenceopen allclose all
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
DI-TERT-DODECYL DISULPHIDEt is not classified for germ cell mutations according to CLP and GHS Regulations.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.