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EC number: 200-625-0 | CAS number: 66-27-3
- 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
Long-term toxicity to fish
Administrative data
Link to relevant study record(s)
- Endpoint:
- fish life cycle toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Justification for type of information:
- Data is from peer reviewed journal
- Qualifier:
- according to guideline
- Guideline:
- other: refer below principle
- Principles of method if other than guideline:
- Combine multigeneration reproductive and genotoxicity study of methyl methanesulfonate in zebrafish
- GLP compliance:
- no
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Methyl methanesulfonate
- Molecular formula (if other than submission substance): C2H6O3S
- Molecular weight (if other than submission substance): 110.1324 g/mol
- Smiles notation (if other than submission substance): COS(=O)(=O)C
- InChI: 1S/C2H6O3S/c1-5-6(2,3)4/h1-2H3
- Substance type: Organic
- Physical state: Liquid - Analytical monitoring:
- yes
- Details on sampling:
- Details on sampling
- Concentrations: 0, 0.2 and 0.4 mg/L
- Sampling method: 10 mL tap
water containing 0, 0.2 and 0.4 mg/L MMS in glass beakers at 26 ± 1 ˚C
- Sample storage conditions before analysis: 26 ± 1 ˚C and a photoperiod of 12 h:12 h - Vehicle:
- yes
- Details on test solutions:
- Details on test solutions
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 200 mL tap water containing 0, 0.2 and 0.4mg/L MMS in glass beakers at 26 ± 1˚C and a photoperiod of 12 h: 12 h, which was maintained throughout the whole experiment.
- Eluate: No data available
- Differential loading: Yes, flow of stock solutions (52, 104, 208 mg/L MMS) was adjusted by means of peristaltic pumps
- Controls: Tap water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Water
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 10 L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No data available - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- TEST ORGANISM
- Common name: Zebrafish
- Strain: Danio rerio, West Aquarium wild
- Source: Laboratory
- Age at study initiation (mean and range, SD): 220 days post fertilization
- Length at study initiation (length definition, mean, range and SD): No data available
- Weight at study initiation (mean and range, SD): No data available
- Method of breeding: Fertilization
- Feeding during test: Yes
- Food type: Artemia nauplii, powder food and flakes.
- Amount: No data available
- Frequency: Artemia nauplii fed once, powder food, three times and flakes two times a day
ACCLIMATION
- Acclimation period: 1–2 h
- Acclimation conditions (same as test or not): Same
- Type and amount of food: Artemia nauplii fed once, powder food, three times and flakes two times a day
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): No data available
QUARANTINE (wild caught)
- Duration: No data available
- Health/mortality: No data available - Test type:
- flow-through
- Water media type:
- freshwater
- Total exposure duration:
- 30 d
- Post exposure observation period:
- 370 days post fertilization
- Hardness:
- 12–18˚dH
- Test temperature:
- 25 ± 2 ˚C
- pH:
- 7.0–8.3
- Dissolved oxygen:
- No data available
- Salinity:
- No data available
- Nominal and measured concentrations:
- 0.2 and 0.4 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Tank
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 10 L
- Aeration: yes
- Type of flow-through (e.g. peristaltic or proportional diluter): peristaltic
- Renewal rate of test solution (frequency/flow rate): Renewed every 48 hours
- No. of organisms per vessel:
- No. of vessels per concentration (replicates): Triplicate
- No. of vessels per control (replicates): Triplicate
- No. of vessels per vehicle control (replicates): Triplicate
- Biomass loading rate: No data available
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: No data available
- Total organic carbon: No data available
- Particulate matter: No data available
- Metals: No data available
- Pesticides: No data available
- Chlorine: No data available
- Alkalinity: No data available
- Ca/mg ratio: No data available
- Conductivity: 522–828 lS/cm
- Culture medium different from test medium: No data available
- Intervals of water quality measurement: Weekly
OTHER TEST CONDITIONS
- Adjustment of pH: No data available
- Photoperiod: 12 h:12 h
- Light intensity: No data available
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, Growth, Body weight, length, Fertility and fertilization, Teratogenicity in embryos and early larval stages, Genotoxicity and histopathology were measured.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data available
- Justification for using less concentrations than requested by guideline: Since all MMS-exposed spawning groups failed to spawn, and fish exposed to 4 mg/L MMS also showed mortalities, MMS concentrations were reduced from 2 and 4 to 0.2 and 0.4 mg/L, respectively
- Range finding study: No data available
- Test concentrations: 0.2 and 0.4 mg/L
- Results used to determine the conditions for the definitive study: No data available - Reference substance (positive control):
- not specified
- Key result
- Duration:
- 30 d
- Dose descriptor:
- EC50
- Effect conc.:
- 0.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Effect on survival , growth, length of fish, Fertility and fertilization, Teratogenicity in embryos and early larval stages, Genotoxicity and Histopathology
- Remarks on result:
- other: At 300 dpf, all remaining individuals exposed to 0.4 mg/L MMS were sacrificed because of mortality, macroscopic pathology and consistent lack of spawn. F0 control fish as well as fish exposed to 0.2 mg/L MMS were sacrificed at 370 dpf.
- Details on results:
- - Mortality/survival at embryo and larval stages: Yes, 50 % mortality was observed.
- Overall mortality/survival: 53 % were observed as compared to control.
- Days to hatch and numbers hatched: 96 days
- Data for length and weight of surviving fish:
Body weight: Significant increase in body weight was observed in 0.2 mg/L treated fish as compared to control.
Length: No effect on total length of F1 fish were observed in 0.2 mg/L dose as compared to control
- Type of and number with morphological abnormalities: Earlier spawn were observed in 0.2 mg/L treated fish as compared to control.
- Type of and number with behavioural abnormalities: No data available
- Other biological observations:
Fertilization: In 0.2 mg/L dose group, increase egg productions relative to the controls were observed.
Teratogenicity in embryos and early larval stages: Significantly more sublethal teratogenic effects in embryos and larvae were observed in 0.2 mg/L treated fish as compared to control.
Genotoxicity:
Percentage of DNA significantly elevated in tail and liver cells from sexes as well as female gills and male gonads of 0.2 mg/L treated fish as compared to control.
Histopathology:
Micronucleus frequencies were significantly increased in liver, gills and gonads of both male and female fish in 0.2 mg/L treated fish as compared to control.
- Effect concentrations exceeding solubility of substance in test medium: No data available
- Incidents in the course of the test which might have influenced the results: No data available - Results with reference substance (positive control):
- No data available
- Reported statistics and error estimates:
- No data available
- Validity criteria fulfilled:
- yes
- Conclusions:
- In a long term Combine multigeneration reproductive and genotoxicity study, Danio rerio, treated with Methyl methanesulfonate (MMS) in the concentration of 0, 0.2 and 0.4 mg/L for 30 days. In the study EC50 was considered to be 0.2 mg/L when Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS).
- Executive summary:
In a long term Combine multigeneration reproductive and genotoxicity study, Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) in the concentration of 0, 0.2 and 0.4 mg/L for30 days. Overall mortality, embryo and larval stages mortality were observed in 0.2 doses. Similarly, significant increase in body weight of fish, Earlier spawn and increase egg productions were observed in 0.2 mg/L dose. Significantly more sublethal teratogenic effects in embryos and larvae were observed in 0.2 mg/L treated fish as compared to control. In addition, significantly elevated percentage of DNA in tail and liver cells from sexes as well as female gills and male gonads and significantly increased micronucleus frequencies in liver, gills and gonads of both male and female fish in 0.2 mg/L treated fish were observed as compared to control. Therefore, EC 50 was considered to be 0.2 mg/L when Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) for 96 hours.
Reference
For F0 generation 0, 2, 4 and 8 mg/L dose were used and for F1 generation 0, 0.2 and 0.4 mg/L dose were used.
Description of key information
In a long term Combine multigeneration reproductive and genotoxicity study, Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) in the concentration of 0, 0.2 and 0.4 mg/L for30 days. Overall mortality, embryo and larval stages mortality were observed in 0.2 doses. Similarly, significant increase in body weight of fish, Earlier spawn and increase egg productions were observed in 0.2 mg/L dose. Significantly more sublethal teratogenic effects in embryos and larvae were observed in 0.2 mg/L treated fish as compared to control. In addition, significantly elevated percentage of DNA in tail and liver cells from sexes as well as female gills and male gonads and significantly increased micronucleus frequencies in liver, gills and gonads of both male and female fish in 0.2 mg/L treated fish were observed as compared to control. Therefore, EC 50 was considered to be 0.2 mg/L when Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) for 96 hours.
Key value for chemical safety assessment
Fresh water fish
Fresh water fish
- Effect concentration:
- 0.2 mg/L
Additional information
Two studies including experimental results of long term fish toxicity endpoint for test chemical methyl methanesulfonate (Cas no. 66-27-3) were summarized as followed:
First experimental study (Ecotoxicology (2013) 22:825–837)suggest a long term Combine multigeneration reproductive and genotoxicity study, Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) in the concentration of 0, 0.2 and 0.4 mg/L for 30 days. Overall mortality, embryo and larval stages mortality were observed in 0.2 doses. Similarly, significant increase in body weight of fish, Earlier spawn and increase egg productions were observed in 0.2 mg/L dose. Significantly more sublethal teratogenic effects in embryos and larvae were observed in 0.2 mg/L treated fish as compared to control. In addition, significantly elevated percentage of DNA in tail and liver cells from sexes as well as female gills and male gonads and significantly increased micronucleus frequencies in liver, gills and gonads of both male and female fish in 0.2 mg/L treated fish were observed as compared to control. Therefore, EC 50 was considered to be 0.2 mg/L when Danio rerio, West Aquarium wild Zebrafish treated with Methyl methanesulfonate (MMS) for 96 hours.
Also supporting study (ECOTOX database by US EPA; 2017) indicate In long term fish toxicity study the Lowest observable effect concentration (LOEC) on the basis of Genetics damage effect was observed to be 0.11 mg/l for exposure period of 21 days.This effect concentration indicate that the test substance methyl methanesulfonate was toxic to fish in long exposure period and thus consider for the aquatic classification.
Thus based on the above available studies it is concluded that the test substance methyl methanesulfonate (Cas no. 66-27-3) consider to be toxic to fish in long exposure period and thus consider for the aquatic classification in chronic category 2 as per the CLP criteria based on the EC50 value of 0.2 mg/L.
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