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EC number: 222-492-8 | CAS number: 3495-36-1
- 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
Biodegradation in water: screening tests
Administrative data
Link to relevant study record(s)
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From April 23, 1992 to May 2, 1992
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- A) The study was conducted according to OECD Guideline 301D and EEC Directive 67/548 Annex V C.6 as published in 84/449/EEC. B) Well-defined information were provided on: 1. Test system (inoculum and composition) 2. Compositional information of the test and reference material (identity and purity) 3. Dosing procedure information (Controls and their composition) 4. Test results and observations (Kinetics of % biodegradation, oxygen concentration and depletion) 5. Definite conclusion
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.6 (Degradation: Chemical Oxygen Demand)
- Version / remarks:
- EEC Directive 67/548 Annex V C.6 as published in 84/449/EEC
- Deviations:
- no
- GLP compliance:
- yes
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- - Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Aeration stage of HRC Limited sewage treatment plant treating predominantly domestic sewage
- Laboratory culture: No
- Method of cultivation: Not applicable
- Storage conditions: Allowed to settle at room temperature
- Storage length: No data
- Preparation of inoculum for exposure: The supernatant of the settled sludge was filtered after the first 250 mL was discarded.
- Pretreatment: No
- Concentration of sludge: No data
- Initial cell/biomass concentration: No data
- Water filtered: Yes
- Type and size of filter used, if any: Whatman GF filter paper - Duration of test (contact time):
- ca. 28 d
- Initial conc.:
- ca. 45 mg/L
- Based on:
- act. ingr.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium: Analytical grade salts and buffers were dissolved in osmosis purified and deionised water to prepare the nutrient medium.
- Additional substrate: None
- Solubilising agent (type and concentration if used): None
- Test temperature: 20 ±1 °C
- pH: No data
- pH adjusted: Yes
- CEC (meq/100 g): No data
- Aeration of dilution water: No
- Suspended solids concentration: No data
- Continuous darkness: Yes, darkened bottles used
- Other: None
TEST SYSTEM
- Culturing apparatus: 280 mL darkened glass Biological Oxygen Demand (BOD) bottles
- Number of culture flasks/concentration: Two per sampling time
- Method used to create aerobic conditions: Test performed under anerobic conditions
- Method used to create anaerobic conditions: Bottles firmly stoppered to exclude all air bubbles
- Measuring equipment: Yellow Springs oxygen meter and BOD Probe
- Test performed in closed vessels due to significant volatility of test substance: Not applicable
- Test performed in open system: No
- Details of trap for CO2 and volatile organics if used: No
SAMPLING
- Sampling frequency: Dissolved oxygen concentrations for each test medium were determined, in duplicate at Days 0, 5, 15 and 28
- Sampling method: By means of a Yellow Springs BOD Probe (Model 50B)
- Sterility check if applicable: Not applicable
- Sample storage before analysis: No data
CONTROL AND BLANK SYSTEM
- Inoculum blank: Inoculated nutrient medium
- Abiotic sterile control: Non-inoculated nutrient medium
- Toxicity control: Containing both test item (45 mg/L) and reference material (3 mg/L) in test medium
- Procedure control: Containing reference material without test item
STATISTICAL METHODS: No data - Reference substance:
- benzoic acid, sodium salt
- Remarks:
- Batch No. 2923430L
- Preliminary study:
- Not applicable
- Test performance:
- No data
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- ca. 83
- Sampling time:
- 28 d
- Details on results:
- % Degradation after 28 d:
Test item - 83 %
Toxicity control - 72 % - Results with reference substance:
- % Degradation after 28 d:
Reference material - 92 %
ThOD value (mg O2/mg):
Reference material - 1.67
Estimated oxygen depletion for 3 mg/L is 5.01 mg O2/L. - Validity criteria fulfilled:
- yes
- Remarks:
- 1. Reference material yielded ≥ 60 % degradation by Day 14; 2. Oxygen depletion of the control did not exceed 30 % of the oxygen in the test bottle; 3. Residual oxygen concentration in the test bottles did not fall below 0.5 mg/L
- Interpretation of results:
- readily biodegradable
- Conclusions:
- Under the study conditions the test substance attained 83 % degradation within 28 days and the pass level of 60 % was reached within 10 days of exceeding the 10 % level, therefore the test substance can be considered to be readily degradable.
- Executive summary:
A study was conducted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 301D and EEC Directive 67/548 Annex V C.6 as published in 84/449/EEC, in compliance with GLP. The test substance with the standard nutrient medium was kept in sealed Biological Oxygen Demand (BOD) bottles (darkened glass) at 20°C for 28 days. Non-inoculated nutrient medium (vehicle control), inoculated nutrient medium (inoculum control), toxicity control consisting of test substance (45 mg/L) and the reference material (3 mg/L) in nutrient medium (procedure control) were run in parallel for validation purposes. Dissolved oxygen concentrations for each test medium were determined, in duplicate on Days 0, 5, 15 and 28 by means of a Yellow Springs oxygen meter and BOD Probe. The reference material and the inhibition check attained 92 and 72% biodegradation, respectively, after 28 days. Under the study conditions, the test substance attained 83% degradation within 28 days and the pass level of 60% was reached within 10 days of exceeding the 10% level, therefore the test substance can be considered to be readily degradable (Douglas, 1992).
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From June 2, 2008 to August 11, 2008
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: SEPA, the Guidelines for the Testing of Chemical No. 301D “Closed Bottle Test” (May 2004)
- Deviations:
- no
- GLP compliance:
- yes
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- - Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Aeration tank of Shenyang Fairy River Sewage Treatment Center (treats predominantly domestic waste)
- Laboratory culture: No
- Method of cultivation: Not applicable
- Storage conditions: Kept under aeration till the test
- Storage length: 5 d
- Preparation of inoculum for exposure: After passing the sample through a fine 20-mesh sieve, 1 L of the sub-sample was removed and the remaining centrifuged at 3000 rpm for 1 min. The concentrated sludge, after removing the supernatant, was resuspended in mineral test medium.
- Pretreatment: No
- Concentration of sludge: No data
- Initial cell/biomass concentration: No data
- Water filtered: No data
- Type and size of filter used, if any: No data - Duration of test (contact time):
- ca. 28 d
- Initial conc.:
- ca. 47.12 mg/L
- Based on:
- act. ingr.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium:
Preparation of mineral nutrient medium
The nutrient medium was composed of four stock solutions the composition of which are as follows:
Solution A (pH 7.4):
Ingredient weight (g/L)
KH2PO4 8.5
K2HPO4 21.75
Na2HPO4.2H2O 33.4
NH4Cl 0.5
Solution B:
27.50 g anhydrous CaCl2 in 1 L water
Solution C:
22.50 g MgSO4.7H2O in 1 L water
Solution D:
0.25 g FeCl3.6H2O in 1 L water
For each liter of mineral medium, 1 mL stock solution A was added to approximately 800 mL deionized water and the mixture stirred before adding, sequentially, 1 mL each of stock solutions B, C and D. After mixing, the mixture was made to 1 L with deionised water. The pH was maintained in the range of 7.4±0.2.
- Additional substrate: None
- Solubilising agent (type and concentration if used): None
- Test temperature: 20 ± 2 °C
- pH: 7.4± 0.2
- pH adjusted: Yes
- CEC (meq/100 g): No data
- Aeration of dilution water: Strongly aerated for 4 h and then allowed to stand for 20 h
- Suspended solids concentration: 3-5 g/L
- Continuous darkness: Yes, test performed in dark and nutrient medium prepared in brown glass bottle
TEST SYSTEM
- Culturing apparatus: BOD flasks (100~125mL) with glass stopper
- Number of culture flasks/concentration:
35 replicates for each concentration in test suspension, inoculum control and process control
21 replicates for the toxicity control
- Method used to create aerobic conditions: Test performed under anerobic condition
- Method used to create anaerobic conditions:
1. The BOD flasks were firmly stoppered with glass stoppers.
2. All transfer and filling operations of the air-saturated medium were done by siphons and the outlet maintained below the surface of the medium.
- Measuring equipment: O2-electrode and detecting instrument
- Test performed in closed vessels due to significant volatility of test substance: Not applicable
- Test performed in open system: No
- Details of trap for CO2 and volatile organics if used: Not applicable
SAMPLING
- Sampling frequency: 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 24, and 28 d after treatment
- Sampling method: Dissolved oxygen concentration for each test medium were determined in triplicate by the Oxygen Electrode method at each sampling frequency
- Sterility check if applicable: Not applicable
- Sample storage before analysis: No data
CONTROL AND BLANK SYSTEM
- Inoculum blank: Test system with the inoculum (added at 0.05 mL/L to all test flasks) and without the test and reference substances
- Abiotic sterile control: No data
- Toxicity control: 283.3 mg/L of the test substance and 5 mL of the stock solution of reference substance in the nutrient medium with inoculum
- Other:
Procedure/Process control - Reference substance added at 2.024 mg/L in the inoculated medium from a stock solution of 2024 mg/L.
STATISTICAL METHODS: Not applicable - Reference substance:
- benzoic acid, sodium salt
- Remarks:
- used at a concentration of 2.024 mg/L
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- ca. 79
- Sampling time:
- 28 d
- Details on results:
- The accumulated percentage biodegradation in activated sludge on 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 24, and 28 d after treatment were 0, 13.0, 26.7, 44.7, 60.8, 63.2, 66.8, 75.8, 76.4, 77.9, 78.9 and 79.0 respectively.
- Results with reference substance:
- The % biodegradation values of the reference material were as follows:
Day 2 - 32.0
Day 9 - 74.3
Day 28 - 87.7 - Validity criteria fulfilled:
- yes
- Remarks:
- 1. Dissolved oxygen did not exceed 1.5 mg/L after 28 d 2. Residual concentration of oxygen did not fall below 0.5 mg/L after 28 d 3. Percentage degradation in the toxicity control was 76.7 % after 14 d
- Interpretation of results:
- readily biodegradable
- Conclusions:
- Under the study conditions the test substance attained 79 % degradation within 28 days and the pass level of 60 % was reached within 10 days of exceeding the 10 % level, therefore the test substance can be considered to be readily degradable.
- Executive summary:
A study was conducted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 301D and SEPA Guideline 301D (closed bottle test), in compliance with GLP. The test substance with the standard nutrient medium was kept in sealed Biological Oxygen Demand (BOD) bottles (darkened glass) at 20°C for 28 days. Non-inoculated nutrient medium (vehicle control), inoculated nutrient medium (inoculum control), toxicity control consisting of test substance (45 mg/L) and the reference material (3 mg/L) in nutrient medium (procedure control) were run in parallel for validation purposes. Dissolved oxygen concentrations for each test medium were determined, in duplicate on Days 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 24 and 28 by means of a Yellow Springs oxygen meter and BOD Probe. The reference material and the inhibition check attained 87.7 and 76.7% biodegradation, respectively, after 28 days. Under the study conditions, the test substance attained 79% degradation within 28 days and the pass level of 60% was reached within 10 days of exceeding the 10% level, therefore the test substance can be considered to be readily degradable (Zhao, 2008).
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2000
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guidelines for Testing of Chemicals (1992) No. 306
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: US EPA Fate, Transport and Transformation Test Guidelines OPPTS 835.3160
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- natural water
- Details on inoculum:
- Test Species:
Mixed population of marine micro-organisms in sea water was used as test species (obtained from Huttoft on the East Coast of the United Kingdom).
Treatment: The sea water after filtration was maintained under continuous aeration in a temperature-controlled room at 21 °C for 8 d prior to the start of the study in order to reduce the basal BOD.
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Sea water obtained from Huttoft on the East Coast of the United Kingdom
- Laboratory culture: Yes
- Method of cultivation: The sea water after collection, was filtered and was aerated continuously under constant temperature.
- Storage conditions: Maintained under continuous aeration in a temperature controlled room at 21 °C
- Storage length: 8 d
- Preparation of inoculum for exposure: No data
- Pretreatment: 1 mL of buffered solutions and salt solutions were added to 1 L of aerated sea water
- Concentration of sludge: No data
- Initial cell/biomass concentration: No data
- Water filtered: Yes
- Type and size of filter used, if any: Coarse filter paper - Duration of test (contact time):
- >= 28 d
- Initial conc.:
- ca. 35 mg/L
- Based on:
- act. ingr.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium:
The composition of different solutions are given below:
Solution a
pH = 7.4 KH2PO4 8.50 g/L
K2HPO4 21.75 g/L
Na2HPO4.2H20 33.40 g/L
NH4Cl 0.50 g/L
Solution b CaCl2 27.50 g/L
Solution c MgSO4.7H2O 22.50 g/L
Solution d FeC13.6H2O 0.25 g/L
1 ml aliquot of each the above solutions was added to each liter of aerated sea water to prepare the test medium.
Test item concentration:
1000 mg of test item was dissolved in test medium and the volume adjusted to 1 L to give a 1000 mg/L stock solution. An aliquot (210 mL) stock solution was dispersed in a final volume of 6 L of test medium to give a concentration of 35 mg/L. The Chemical Oxygen Demand (COD) of the test material was determined to be 0.098 mg 02/mg. Hence if complete degradation of the test material occurred the oxygen depletion in the test vessels would be 3.43 mg 02/mg and as such deoxygenation of the test media would not occur. Therefore, a test concentration of 35 mg/L was employed in the study.
- Test temperature: 20 °C
- pH: 7.4
- pH adjusted: Yes
- CEC (meq/100 g): No data
- Aeration of dilution water: Yes
- Suspended solids concentration: No data
- Continuous darkness: Yes
TEST SYSTEM
- Culturing apparatus: 250-300 mL Biological Oxygen Demand (BOD) bottles (darkened glass)
- Number of culture flasks/concentration: Two per concentration
- Method used to create aerobic conditions: Carried under anerobic condition
- Method used to create anaerobic conditions: BOD bottles were stoppered tightly to exclude all air bubbles
- Measuring equipment: Yellow Springs oxygen meter and BOD Probe.
- Test performed in closed vessels due to significant volatility of test substance: Not applicable
- Test performed in open system: Not applicable
- Details of trap for CO2 and volatile organics if used: None
SAMPLING
- Sampling frequency: Dissolved oxygen concentrations for each test medium were determined, in duplicate on Days 0, 3, 6, 9, 12, 15, 18, 21, 24 and 28
- Sampling method: Samples were analysed at said intervals by means of a Yellow Springs oxygen meter and BOD Probe.
CONTROL AND BLANK SYSTEM
- Inoculum blank: Test medium alone
- Toxicity control: The test material (35 mg/L) and the reference material (1mg/L) in test medium
STATISTICAL METHODS: No data - Reference substance:
- benzoic acid, sodium salt
- Remarks:
- Sodium Benzoate (Lot no. 77H05005) at a concentration of 2 mg/L
- Preliminary study:
- No
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- ca. 79
- Sampling time:
- 28 d
- Details on results:
- % Degradation after 28 d:
Dissolved Oxygen measurement (mg O2/L) after 28 d
1. Dissolved oxygen of test medium (control) was 7.20 and 7.10 at Day 0 in replicate 1 and 2 respectively and was reduced to 6.20 at Day 28 in both the replicates.
2. Dissolved oxygen of reference material in test medium was 7.15 and 7.10 at Day 0 in replicate 1 and 2 respectively and was reduced to 3.15 and 3.10 in replicate 1 and 2 respectively at Day 28.
3. Dissolved oxygen of test material in test medium was 7.15 at Day 0 in both replicates and was reduced to 3.55 and 3.45 at Day 28 in replicate 1 and 2 respectively.
4. Dissolved oxygen of reference material and test item in test medium was 7.10 at Day 0 in both replicates and was reduced to 1.80 and 1.75 at Day 28 in replicate 1 and 2 respectively.
- Results with reference substance:
- % Degradation after 28 d:
91 % degradation of reference material
Dissolved oxygen of reference material in the test medium was 7.15 and 7.10 at Day 0 in replicate 1 and 2 respectively and was reduced to 3.15 and 3.10 in replicate 1 and 2 respectively at Day 28. - Validity criteria fulfilled:
- yes
- Remarks:
- 1. Standard material yields ≥ 60% degradation by day 14 2. Oxygen depletion of the control did not exceed 30% of the oxygen in the test bottle after 28 days
- Interpretation of results:
- other: potential for biodegradation in the marine environment
- Conclusions:
- Under the test conditions the test substance attained 79 % degradation after 28 days and therefore can be considered to have the potential for biodegradation in marine environment.
- Executive summary:
A study was conduted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 306 and US EPA Fate, Transport and Transformation Test Guidelines OPPTS 835.3160, in compliance with GLP. The test substance, at a concentration of 35 mg/L, was exposed to marine microorganisms with test medium in sealed culture vessels in the dark at 20°C for 28 days. The degradation of the test substance was assessed by the determination of the amount of oxygen consumed. Control solutions containing the standard material, sodium benzoate, together with a toxicity control were used for validation purposes. Under the test conditions the test substance attained 79% degradation after 28 days and therefore can be considered to have the potential for biodegradation in marine environment (Mead, 2000).
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From September 4, 2000 to October 2, 2000
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guidelines for Testing of Chemicals (1992) No. 306, "ReadyBiodegradability in Sea Water: Closed Bottle Test"
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: US EPA Fate, Transport, and Transformation Test Guidelines OPPTS 835.3160
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- natural water
- Details on inoculum:
- Treatment: The sea water was filtered and maintained under continuous aeration in temperature controlled room at 21 °C for 10 d in order to reduce the basal BOD.
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Mixed population of marine micro-organisms in sea water was used as test species (obtained from Huttoft on the East Coast of the United Kingdom).
- Laboratory culture: No
- Method of cultivation: Not applicable
- Storage conditions: Under continuous aeration at a constant temperature of 21 °C
- Storage length: 10 d
- Preparation of inoculum for exposure: The sea water after collection was filtered.
- Pretreatment: No data
- Concentration of sludge: No data
- Initial cell/biomass concentration: No data
- Water filtered: Yes
- Type and size of filter used, if any: Coarse filter paper - Duration of test (contact time):
- ca. 28 d
- Initial conc.:
- ca. 40 mg/L
- Based on:
- COD
- Parameter followed for biodegradation estimation:
- DOC removal
- Details on study design:
- TEST CONDITIONS
- Composition of medium:
Solution a
pH = 7.4 KH2PO4 8.50 g/L
K2HPO4 21.75 g/L
Na2HPO4.2H20 33.40 g/L
NH4Cl 0.50 g/L
Solution b CaCl2 27.50 g/L
Solution c MgSO4.7H2O 22.50 g/L
Solution d FeC13.6H2O 0.25 g/L
1 mL aliquot of each the above solutions was added to each liter of aerated sea water to prepare the test medium.
- Concentration of test chemical:
500 mg of test item was dissolved in test medium and the volume adjusted to 1 L to give a 500 mg/l stock solution. An aliquot (480 Ll)of stock solution was dispersed in a final volume of 6 L of test medium to give a concentration of 40 mg/L.
The Chemical Oxygen Demand (COD) of the test material was determined to be 0.081 mg 02/mg. Hence if complete degradation of the test material occurred the oxygen depletion in the test vessels would be 3.20 mg 02/mg and as such deoxygenation of the test media would not occur. Therefore a test concentration of 40 mg/L was employed in the study.
Reference material:Sodium Benzoate (Sigma Lot No. 77H05005) at a concentration of 2 mg/L
- Dosing procedure: Following test solutions were inoculated in 250-300 mL Biological Oxygen Demand (BOD) bottles (darkened glass) in duplicate:
1.Test medium acting as control
2. Sodium Benzoate in test medium to give a concentration of 2 mg/L
3. The test item in test medium to give a concentration of 40 mg/L
4. The test item (40 mg/L) plus reference material (1mg/L) in test medium to act as toxicity control
The BOD bottles were firmly stoppered to exclude all air bubbles after transferring the test solutions and incubated in a temperature controlled water bath at 20 °C.
- Test temperature: 20 °C
- Test performed in open system:
- Details of trap for CO2 and volatile organics if used: No data
SAMPLING
- Sampling frequency: In duplicate, on Days 0, 3, 6, 9, 12, 15, 18, 21, 24 and 28
- Sampling method: Yellow Springs oxygen meter and BOD Probe
CONTROL AND BLANK SYSTEM
- Toxicity control: The test item (40 mg/L) and the reference material (1 mg/L) in test medium
- Vehicle control: Test medium alone
- Procedure control: With the reference material without test item in test medium
STATISTICAL METHODS: No data - Reference substance:
- benzoic acid, sodium salt
- Remarks:
- (Sigma Lot No. 77H05005) at a concentration of 2 mg/L
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- ca. 66
- Sampling time:
- 28 d
- Details on results:
- % Degradation after 28 d:
Test substance - 66 % degradation
Toxicity control - 66 % degradation
The oxygen depletion in the control vessels was 22 % of the initial dissolved oxygen concentration.
Oxygen concentration remained at 2.25 mg O2/L or greater in all test vessels and thus validation criteria for oxygen depletion was satisfied. - Key result
- Parameter:
- COD
- Value:
- ca. 0.1 other:
- Results with reference substance:
- % Degradation after 28 d:
Reference material - 75 % degradation
ThOD value (mg O2/mg):
Reference material: 1.67
Estimated oxygen depletion for 2 mg/L is 3.34 mg O2/L.
COD values (mg O2/mg):
Reference material: 1.69
Reference material (Sodium Benzoate) attained 75 % degradation after 28 d. This confirms the suitability of the test method and culture conditions. - Validity criteria fulfilled:
- yes
- Remarks:
- 1. Reference material yielded ≥ 60 % degradation by Day 14; 2. Oxygen depletion of the control did not exceed 30 % of the oxygen in the test bottle; 3. Residual oxygen concentration in the test bottles did not fall below 0.5 mg/L
- Interpretation of results:
- other: potential for biodegradation in the marine environment
- Conclusions:
- Under the study conditions the test substance attained 66% degradation after 28 days and therefore can be considered to have the potential for biodegradation in the marine environment.
- Executive summary:
A study was conducted to determine the biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 306 and US EPA Fate, Transport and Transformation Test Guideline OPPTS 835.3160, in compliance with GLP. The test substance, at a concentration of 40 mg/L, was exposed to marine microorganisms with test medium in sealed culture vessels in the dark at 20°C for 28 days. The degradation of the test substance was assessed by the determination of the amount of oxygen consumed. Control solutions containing the reference substance, sodium benzoate, together with a toxicity control were used for validation purposes. Under the study conditions, the test substance attained 66% degradation after 28 days and therefore can be considered to have the potential for biodegradation in the marine environment (Clark, 2000).
Referenceopen allclose all
Dissolved oxygen measurement (mg O2/L) after 28 d:
Test series |
Dissolved oxygen (mg O2/L) Days 0 5 15 28 |
||||
Dilution water without inoculum |
R1 |
8.90 |
8.78 |
8.74 |
8.80 |
R2 |
8.90 |
8.81 |
8.80 |
8.78 |
|
Dilution water with inoculum |
R1 |
8.90 |
8.80 |
8.63 |
8.54 |
R2 |
8.90 |
8.84 |
8.67 |
8.78 |
|
Test substance |
R1 |
8.90 |
7.81 |
5.62 |
5.38 |
R2 |
8.90 |
8.15 |
5.73 |
5.25 |
|
Sodium Benzoate in test medium (procedure control) |
R1 |
8.90 |
5.03 |
4.36 |
4.16 |
R2 |
8.90 |
4.93 |
4.10 |
3.92 |
|
Test substance and reference material (toxicity control) |
R1 |
8.90 |
NA |
NA |
2.87 |
R2 |
8.90 |
NA |
NA |
2.91 |
NA - Not applicable % degradation after 28 d:
Test series |
% degradation value Days 7 15 28 |
||
Test substance |
21 |
73 |
81 |
Sodium Benzoate in test medium (procedure control) |
77 |
88 |
92 |
Dissolved Oxygen Concentration in the test media:
Day |
IC-1 |
IC-2 |
IC-3 |
TS-1 |
TS-2 |
TS-3 |
BOD |
% Degradation |
0 |
8.63 |
8.63 |
8.62 |
8.49 |
8.53 |
8.48 |
- |
0.0 |
2 |
8.40 |
8.42 |
8.45 |
7.89 |
7.85 |
7.85 |
0.433 |
13.0 |
4 |
8.21 |
8.25 |
8.23 |
7.20 |
7.22 |
7.21 |
0.893 |
26.7 |
7 |
8.05 |
7.86 |
7.91 |
6.31 |
6.32 |
6.32 |
1.497 |
44.7 |
9 |
7.86 |
7.88 |
7.80 |
5.69 |
5.72 |
5.65 |
2.033 |
60.8 |
11 |
7.68 |
7.70 |
7.68 |
5.45 |
5.42 |
5.47 |
2.113 |
63.2 |
14 |
7.61 |
7.62 |
7.62 |
5.27 |
5.24 |
5.26 |
2.233 |
66.8 |
16 |
7.54 |
7.58 |
7.56 |
4.98 |
4.89 |
4.82 |
2.537 |
75.8 |
18 |
7.50 |
7.52 |
7.50 |
4.88 |
4.76 |
4.83 |
2.557 |
76.4 |
21 |
7.46 |
4.47 |
7.45 |
4.71 |
4.71 |
4.76 |
2.607 |
77.9 |
24 |
7.42 |
7.45 |
7.46 |
4.68 |
4.65 |
4.70 |
2.640 |
78.9 |
28 |
7.41 |
7.40 |
7.38 |
4.65 |
4.62 |
4.61 |
2.643 |
79.0 |
Note: IC means Inoculum Control; TS means Test Substance.
Dissolved Oxygen Concentration in the process control:
Day |
IC-1 |
IC-2 |
IC-3 |
PC-1 |
PC-2 |
PC-3 |
BOD |
% Degradation |
0 |
8.63 |
8.63 |
8.62 |
8.43 |
8.38 |
8.36 |
- |
0.0 |
2 |
8.40 |
8.42 |
8.45 |
7.00 |
7.15 |
7.15 |
1.087 |
32.2 |
4 |
8.21 |
8.25 |
8.23 |
6.47 |
6.47 |
6.48 |
1.520 |
45.1 |
7 |
8.05 |
7.86 |
7.91 |
5.45 |
5.40 |
5.51 |
2.250 |
66.7 |
9 |
7.86 |
7.88 |
7.80 |
5.12 |
5.10 |
5.10 |
2.503 |
74.3 |
11 |
7.68 |
7.70 |
7.68 |
4.82 |
4.78 |
4.87 |
2.627 |
77.9 |
14 |
7.61 |
7.62 |
7.62 |
4.69 |
4.65 |
4.64 |
2.720 |
80.7 |
16 |
7.54 |
7.58 |
7.56 |
4.52 |
4.54 |
4.51 |
2.800 |
83.1 |
18 |
7.50 |
7.52 |
7.50 |
4.48 |
4.42 |
4.40 |
2.837 |
84.1 |
21 |
7.46 |
4.47 |
7.45 |
4.35 |
4.36 |
4.38 |
2.860 |
84.8 |
24 |
7.42 |
7.45 |
7.46 |
4.25 |
4.31 |
4.32 |
2.913 |
86.4 |
28 |
7.41 |
7.40 |
7.38 |
4.22 |
4.21 |
4.18 |
2.957 |
87.7 |
Note: IC means Inoculum Control; PC means Process Control.
Dissolved Oxygen Determination in Toxicity Control:
Day |
IC-1 |
IC-2 |
IC-3 |
TC-1 |
TC-2 |
TC-3 |
BOD |
% Degradation |
0 |
8.63 |
8.63 |
8.62 |
8.87 |
8.88 |
8.88 |
- |
0.0 |
2 |
8.40 |
8.42 |
8.45 |
6.64 |
6.65 |
6.62 |
2.037 |
27.5 |
4 |
8.21 |
8.25 |
8.23 |
5.69 |
5.76 |
5.69 |
2.767 |
37.4 |
7 |
8.05 |
7.86 |
7.91 |
4.66 |
4.66 |
4.67 |
3.527 |
47.7 |
9 |
7.86 |
7.88 |
7.80 |
3.52 |
3.51 |
3.51 |
4.583 |
62.0 |
11 |
7.68 |
7.70 |
7.68 |
2.37 |
2.33 |
2.28 |
5.610 |
75.9 |
14 |
7.61 |
7.62 |
7.62 |
2.17 |
2.20 |
2.22 |
5.670 |
76.7 |
Note: IC means Inoculum Control; TC means Toxicity Control.
Calculated ThOD values:
For test substance - 0.071 mg O2/mg
For reference substance - 1.67 mg O2/mg
% Degradation after 28 d
Test substance - 79 % degradation
Reference material - 91% degradation
Toxicity control - 86 % degradation
The oxygen depletion in the control vessels was 13 % of the initial dissolved oxygen concentration.
Oxygen concentration remained at 1.75 mg O2/L or greater in all test vessels and thus the validation criteria for oxygen depletion was satisfied.
ThOD value (mg O2/mg)
Reference material: 1.67
Estimated oxygen depletion for 2 mg/L is 3.34 mg O2/L
COD values (mg O2/mg)
Test material : 0.098
Reference material: 1.69
Reference material (Sodium Benzoate) attained 91% degradation after 28 d. This confirms the suitability of the test method and culture conditions.
Dissolved Oxygen measurement (mg O2/L) after 28 d:
Test series |
Dissolved oxygen (mg O2/L) Days 0 9 18 28 |
||||
Test medium |
R1 |
7.20 |
6.80 |
6,40 |
6.20 |
R2 |
7.10 |
6.60 |
6.45 |
6.20 |
|
Sodium Benzoate in test medium (procedure control) |
R1 |
7.15 |
4.00 |
3.75 |
3.15 |
R2 |
7.10 |
4.00 |
3.80 |
3.10 |
|
Test material in test medium |
R1 |
7.15 |
5.80 |
5.20 |
3.55 |
R2 |
7.15 |
5.75 |
5.00 |
3.54 |
|
Test material and Sodium Benzoate in test medium (toxicity control) |
R1 |
7.10 |
3.70 |
2.60 |
1.80 |
R2 |
7.10 |
3.40 |
2.60 |
1.75 |
Mean % degradation value after 28 d:
Test series |
Mean % degradation value Days 3 9 18 28 |
|||
Sodium Benzoate in test medium (procedure control) |
43 |
30 |
79 |
91 |
Test material in test medium |
21 |
27 |
39 |
79 |
Test material and Sodium Benzoate in test medium (toxicity control) |
41 |
61 |
74 |
86 |
Dissolved oxygen measurement (mg O2/L) during the study period
Test series |
Dissolved oxygen (mg O2/L) Days 0 9 18 28 |
||||
Test medium |
R1 |
8.0 |
8.0 |
8.0 |
7.8 |
R2 |
8.0 |
8.0 |
8.0 |
7.9 |
|
Sodium Benzoate in test medium (procedure control) |
R1 |
8.0 |
7.9 |
8.0 |
7.8 |
R2 |
8.0 |
7.9 |
7.9 |
7.8 |
|
Test material in test medium |
R1 |
8.1 |
8.0 |
8.0 |
7.9 |
R2 |
8.1 |
8.0 |
7.9 |
7.8 |
|
Test material and Sodium Benzoate in test medium (toxicity control) |
R1 |
8.1 |
8.0 |
7.9 |
7.8 |
R2 |
8.1 |
7.9 |
8.0 |
7.8 |
Mean % degradation value after during the study period
Test series |
Mean % degradation value Days 3 9 18 28 |
|||
Sodium Benzoate in test medium (procedure control) |
63 |
72 |
74 |
75 |
Test material in test medium |
14 |
20 |
58 |
66 |
Test material and Sodium Benzoate in test medium (toxicity control) |
37 |
56 |
63 |
66 |
Description of key information
Key value for chemical safety assessment
- Biodegradation in water:
- readily biodegradable
Additional information
Biodegradation in seawater:
A study was conducted to determine the biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 306 and US EPA Fate, Transport and Transformation Test Guideline OPPTS 835.3160, in compliance with GLP. The test substance, at a concentration of 40 mg/L, was exposed to marine microorganisms with test medium in sealed culture vessels in the dark at 20°C for 28 days. The degradation of the test substance was assessed by the determination of the amount of oxygen consumed. Control solutions containing the reference substance, sodium benzoate, together with a toxicity control were used for validation purposes. Under the study conditions, the test substance attained 66% degradation after 28 days and therefore can be considered to have the potential for biodegradation in the marine environment (Clark, 2000).
A study was conducted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 306 and US EPA Fate, Transport and Transformation Test Guidelines OPPTS 835.3160, in compliance with GLP. The test substance, at a concentration of 35 mg/L, was exposed to marine microorganisms with test medium in sealed culture vessels in the dark at 20 °C for 28 days. The degradation of the test substance was assessed by the determination of the amount of oxygen consumed. Control solutions containing the standard material, sodium benzoate, together with a toxicity control were used for validation purposes. Under the test conditions, the test substance attained 79% degradation after 28 days and therefore can be considered to have the potential for biodegradation in marine environment (Mead, 2000).
Biodegradation in freshwater:
A study was conducted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 301D and EEC Directive 67/548 Annex V C.6 as published in 84/449/EEC, in compliance with GLP. The test substance with the standard nutrient medium was kept in sealed Biological Oxygen Demand (BOD) bottles (darkened glass) at 20°C for 28 days. Non-inoculated nutrient medium (vehicle control), inoculated nutrient medium (inoculum control), toxicity control consisting of test substance (45 mg/L) and the reference material (3 mg/L) in nutrient medium (procedure control) were run in parallel for validation purposes. Dissolved oxygen concentrations for each test medium were determined, in duplicate on Days 0, 5, 15 and 28 by means of a Yellow Springs oxygen meter and BOD Probe. The reference material and the inhibition check attained 92 and 72% biodegradation, respectively, after 28 days.Under the study conditions, the test substance attained 83% degradation within 28 days and the pass level of 60% was reached within 10 days of exceeding the 10% level, therefore the test substance can be considered to be readily degradable (Douglas, 1992).
A study was conducted to determine the ready biodegradability of the test substance in an aerobic aqueous medium according to OECD Guideline 301D and SEPA Guideline 301D (closed bottle test), in compliance with GLP.The test substance with the standard nutrient medium was kept in sealed Biological Oxygen Demand (BOD) bottles (darkened glass) at 20°C for 28 days. Non-inoculated nutrient medium (vehicle control), inoculated nutrient medium (inoculum control), toxicity control consisting of test substance (45 mg/L) and the reference material (3 mg/L) in nutrient medium (procedure control) were run in parallel for validation purposes. Dissolved oxygen concentrations for each test medium were determined, in duplicate on Days 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 24 and 28 by means of a Yellow Springs oxygen meter and BOD Probe.The reference material and the inhibition check attained 87.7 and 76.7% biodegradation, respectively, after 28 days. Under the study conditions, the test substance attained 79% degradation within 28 days and the pass level of 60% was reached within 10 days of exceeding the 10% level, therefore the test substance can be considered to be readily degradable (Zhao, 2008).
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