Registration Dossier
Registration Dossier
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EC number: 832-827-5 | CAS number: -
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:
- April 18, 2019 to May 16, 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)
- Version / remarks:
- Commission Regulation 440/2008/EC, Method C.4-D of May 30, 2008: Manometric Respirometry Test (EEC Publication No. L 142/496, May 2008)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
- Version / remarks:
- OECD Guideline for Testing of Chemicals No. 301 F: "Ready Biodegradability: Manometric Respirometry Test", adopted July 17, 1992
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, adapted
- Details on inoculum:
- Species/Origin: Aerobic activated sludge, microorganisms from a domestic waste water treatment plant was supplied by the sewage treatment plant Rossdorf, Germany.
Conditioning: The aerobic activated sludge used for this study was deposited for 15 min, washed by centrifugation and the supernatant liquid phase was decanted. The solid material was re-suspended in test water and centrifuged again. This procedure was done three times. An aliquot of the final sludge suspension was weighed, dried and the ratio of wet sludge to its dry weight was determined. Based on this ratio, calculated aliquots of washed sludge suspension, corresponding to 3.5 g dry material per litre were mixed with test water and aerated overnight. This suspension was used for the experiment. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 24.95 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Test Units
Type and Size: Manometric Test System with test flasks containing a volume of approximately 500 mL.
Apparatus: BSB/BOD-Sensor-System
Principle: The test flasks prepared were incubated at 22°C ± 1°C. The pressure decrease in the reaction vessels was measured over complete experimental phase of 28 days using the BSB/BODSensor-System. The test flasks were closed gas-tight by a measuring head. Potassium hydroxide solution (45%) was used for trapping the produced carbon dioxide. The amount of O2 consumed by the activated sludge was calculated from the decrease of pressure in the reaction vessel.
Identification: Each test unit was uniquely identified with the study number, treatment and replicate number.
Test Conditions
Surrounding Type: Climatised chamber
Temperature: 22°C ± 1°C
Light Conditions: Darkness
pH-Value of Test Solutions: 7.6 (measured at the start of the test) 7.5 to 7.6 (measured at the end of the test)
Recording: Test conditions (temperature) were recorded continuously with suitable instruments, documented in the raw data and reported in the final report.
(Short-term deviations (< 2 hours) from the recommended temperature range do normally not result in major disturbances of the test performance and were not reported.)
Course of the Test
Preparation of Test Flasks: The amounts of test item and reference item were directly weighed into the test flasks and the appropriate volume of test water was added to the test flasks.
Afterwards silicone oil AR 20 at a concentration of about 1% of the final flask volume was added to all flasks in order to improve the bioavailability of the mixture constituents. Due to constant stirring over the course of the test the silicone oil was mixed continuously with the test item and test water and provided the desired function of improving bioavailability of the test item.
Incubation: The closed test flasks were incubated in a climatised chamber under continuous stirring. The consumption of oxygen was determined by measuring the change of pressure in the flasks. Evolved carbon dioxide was absorbed in an aqueous solution (45%) of potassium hydroxide.
Test Duration: 28 days
Test Parameters
Measurement of Oxygen: The change of pressure in the test flasks was measured by means of a manometric method (BSB/BOD-Sensor-System).
Temperature: The temperature was recorded by means of the automated software AMR Wincontrol©.
pH-Value: pH-values were measured in procedure control, a separately prepared test flask with test item (to prevent loss of test item in the test flasks) and a separately prepared test flask without test item (control) at test start and in all flasks at the end of the test, except in the abiotic and toxicity control, using a pH-electrode WTW pH 340i. - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 34
- Sampling time:
- 28 d
- Details on results:
- Percentage Biodegradation: The criterion for ready biodegradability under the conditions of a manometric respirometry test is the degradation of the test item of at least 60%, reached within a 10-day window; the 10-day window starts when the degradation of the test item reaches at least 10% degradation.
The 10-day windows began on day 10 after application, the mean value was calculated to be 11% biodegradation (ThODNH4). Therefore, the end of the 10-day window was day 20. The degradation was 26% at day 20, the criterion of the 10 day window was not passed. The mean biodegradation percentage at the end of the 28-day exposure period was 34% (ThODNH4).
Biodegradation in the Toxicity Control
Percentage Biodegradation: In the toxicity control containing both, the test item and the reference item sodium benzoate, 33% (ThODNH4) biodegradation was noted within 14 days and 47% (ThODNH4) biodegradation after 28 days of incubation.
Abiotic Control
Oxygen Demand: The oxygen demand in the abiotic control was 0 mg/L during the test duration. There was no need to correct the degradation of the test item and toxicity control. - Results with reference substance:
- The reference item sodium benzoate was sufficiently degraded to 75% after 14 days and to 84% after 28 days of incubation.
Conclusion: The percentage biodegradation of the reference item confirms the suitability of the used aerobic activated sludge inoculum. - Validity criteria fulfilled:
- yes
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- The degradation rate of SynNova Base Oil did not reach 60% within the 10-day window or after 28 days.
Therefore, SynNova Base Oil is considered to be not readily biodegradable. - Executive summary:
Title: SynNova Base Oil: Ready Biodegradability in a Manometric Respirometry Test
Guidelines: This study was designed to comply with the following methods:
− Commission Regulation 440/2008/EC, Method C.4-D of May 30, 2008: Manometric Respirometry Test (EEC Publication No. L 142/496, May 2008)
− OECD Guideline for Testing of Chemicals No. 301 F: "Ready Biodegradability: Manometric Respirometry Test", adopted July 17, 1992
Material and Methods:
Test Item: SynNova Base Oil; Batch No.: TS20371
Test Species: Aerobic activated sludge (microorganisms from a domestic wastewater treatment plant) was supplied by the sewage treatment plant of Rossdorf, Germany.
Test Design: The test item SynNova Base Oil was investigated for its ready biodegradability in a manometric respirometry test over a period of 28 days. The biodegradation was followed by the oxygen uptake of the microorganisms during exposure. As a reference item sodium benzoate was tested simultaneously under the same conditions as the test item, and functioned as a procedure control.
Endpoints: Degradation rate of test item calculated by the oxygen consumption of the aerobic activated sludge microorganisms after 28 days of incubation.
Test Item Loading Rate (initial concentration in medium C0): 102.3 mg/L corresponding to an oxygen demand of about 349.3 mg/L (ThODNH4)
Reference Item: Sodium Benzoate
Reference Item Loading Rate: 103.7 mg/L corresponding to an oxygen demand of about 172.7 mg/L (ThODNH4)
Test Conditions: 22°C ± 1°C, darkness
Results:
Biodegradation of SynNova Base Oil: The criterion for ready biodegradability under the conditions of a manometric respirometry test is the degradation of the test item of at least 60%, reached within a 10-day window; the 10-day window starts when the degradation of the test item reaches at least 10% degradation.
The mean biodegradation of 10% of SynNova Base Oil was reached at day 10 (ThODNH4). At the end of the 10- day window at day 20, the mean degradation of SynNova Base Oil was 26% (ThODNH4) and therefore the 10 day window criterion was not passed. The mean biodegradation at test end after 28 days was 34% (ThODNH4).
Therefore, SynNova Base Oil is considered to be not readily biodegradable based on ThODNH4.
Biodegradation of Sodium Benzoate: The reference item sodium benzoate was sufficiently degraded to 75% after 14 days and to 84% after 28 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used.
Biodegradation of the Toxicity Control: In the toxicity control containing both, the test item and the reference item sodium benzoate, 33% (ThODNH4) biodegradation was noted within 14 days and 47% (ThODNH4) biodegradation after 28 days of incubation. According to the test guidelines, the test item can be assumed to be not inhibitory to the aerobic activated sludge microorganisms because degradation was >25% within 14 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- February 26, 2019 – March 26, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: Carbon dioxide evolution test, ISO 9439,
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Not adapted activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. 30 mg/l dry matter in the final mixture.
Sampling: February 21, 2019; 8:30 AM - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.7 mg/L
- Based on:
- test mat.
- Initial conc.:
- 20 mg/L
- Based on:
- TOC
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Test design and no. of replicates:
Blank control (medium + inoculum): 3*
Procedure control (medium + inoculum + reference item): 3*
Test suspension (medium + inoculum + test item): 3*
(Toxicity and Abiotic sterile controls were not conducted).
A background control (ultrapure water only) meant to determine the actual IC content of the air used to aerate the suspensions was also set up (one replicate).
*Due to the potential variability of the test system, three replicates were set up but only the two most similar replicates (i.e. showing the most similar curves of IC evolution vs. time, based on the smallest sum of squares of the differences) were considered for the evaluation of biodegradation.
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark
Reference value of the test item: Total Organic Carbon (TOC): 85.7%, determined by an external laboratory following guideline DIN 13137
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equals to 20 mg TOC /l; which corresponds to a nominal concentration of 23.7 mg/l of test item and 34.4 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted to glass fibre filters, the filters were dried and introduced in the test vessels. The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions is measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach).
Calculations: The degradation in unit i at time t (in % of the TOC) is calculated as follows:
Di = [(ICi – ICB)/TOCi] x 100
ICi IC concentration in unit i at time t, in mg/l
ICB IC concentration in the blank control at time t, in mg/l
TOCi initially applied TOC concentration in unit i, in mg/l - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 61
- Sampling time:
- 28 d
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 35
- Sampling time:
- 10 d
- Details on results:
- The biodegradability of TS20343, based on CO2 evolution and compared to the TOC, was calculated to be 61% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of about 7 days.
The biodegradation of TS20343 reached 35% at the end of the 10-day window. - Results with reference substance:
- The positive control, sodium benzoate, reached 83% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20343 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore, cannot be termed as readily biodegradable. However, the pass level of 60% was reached after 28 days.
- Executive summary:
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20343, based on CO2evolution and compared to the TOC, was calculated to be 61% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of about 7 days.
The biodegradation of TS20343 reached 35% at the end of the 10-day window.
The positive control, sodium benzoate, reached 83% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20343 did not reach the pass level of 60% for ready biodegradability in the CO2Evolution Test within the 10-d window, and therefore, cannot be termed as readily biodegradable. However, the pass level of 60% was reached after 28 days.
All validity criteria were fulfilled.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- May 28, 2019 – June 25, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: International Organization for Standardization: Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Not adapted activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. 30 mg/l dry matter in the final mixture.
Sampling: May 23, 2019; 7:30 AM - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.6 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Treatments and no. of replicates:
Blank control (medium + inoculum): 3 1
Procedure control (medium + inoculum + reference item): 3 1
Test suspension (medium + inoculum + test item): 3 1
(Toxicity and Abiotic sterile controls were not conducted).
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark.
Reference value of the test item: Total Organic Carbon (TOC): 85.7% (sponsor’s information)
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equaled approximately 20 mg TOC /l (exact values indicated under Table 1); which corresponds to a nominal concentration of 23.6 mg/l of test item and 34.3 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted onto glass fibre filters, the filters were dried and introduced into the test vessels.
The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions was measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach). - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 65
- Sampling time:
- 28 d
- Details on results:
- The biodegradability of TS20817, based on CO2 evolution and compared to the TOC, was calculated to be 65% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20817 reached 34% at the end of the 10-day window.
TS20817 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days. - Results with reference substance:
- The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20817 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Executive summary:
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20817, based on CO2evolution and compared to the TOC, was calculated to be 65% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20817 reached 34% at the end of the 10-day window.
The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20817 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- May 28, 2019 – June 25, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: International Organization for Standardization: Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Not adapted activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. 30 mg/l dry matter in the final mixture.
Sampling: May 23, 2019; 7:30 AM - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.6 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Treatments and no. of replicates:
Blank control (medium + inoculum): 3 1
Procedure control (medium + inoculum + reference item): 3 1
Test suspension (medium + inoculum + test item): 3 1
(Toxicity and Abiotic sterile controls were not conducted).
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark.
Reference value of the test item: Total Organic Carbon (TOC): 85.7% (sponsor’s information)
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equaled approximately 20 mg TOC /l (exact values indicated under Table 1); which corresponds to a nominal concentration of 23.6 mg/l of test item and 34.3 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted onto glass fibre filters, the filters were dried and introduced into the test vessels.
The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions was measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach). - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 66
- Sampling time:
- 28 d
- Details on results:
- The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 66% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20819 reached 41% at the end of the 10-day window.
TS20819 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days. - Results with reference substance:
- The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20819 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Executive summary:
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20819, based on CO2evolution and compared to the TOC, was calculated to be 66% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20819 reached 41% at the end of the 10-day window.
The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% for ready biodegradability in the CO2Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- September 26, 2019 – October 24, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: ISO - Water quality: ISO 9439
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. Not adapted to the test item but aerated 7 days prior to testing.
Sampling: September 19, 2019; 8:30 AM
Concentration: 30 mg/l dry matter in the final mixture. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.4 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Treatments and no. of replicates:
Blank control (medium + inoculum): 3 1
Procedure control (medium + inoculum + reference item): 3 1
Test suspension (medium + inoculum + test item): 3 1
(Toxicity and Abiotic sterile controls were not conducted).
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark
Reference value of the test item: Total Organic Carbon (TOC): 85.7%, based on sponsor’s information.
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equaled approximately 20 mg TOC /l; which corresponds to a nominal concentration of 23.4 mg/l of test item and 34.0 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted onto glass fibre filters, the filters were dried and introduced into the test vessels.
The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). After application in the test vessels, the glass fiber filters were disintegrated/sheared at the beginning of the test so as to optimize the contact area of the inoculum with the test item. pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions was measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach). - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 71
- Sampling time:
- 28 d
- Details on results:
- The biodegradability of TS20817, based on CO2 evolution and compared to the TOC, was calculated to be 71% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 8 days.
The biodegradation of TS20817 reached 39% at the end of the 10-day window.
TS20817 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days. - Results with reference substance:
- The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20817 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Executive summary:
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20817, based on CO2 evolution and compared to the TOC, was calculated to be 71% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 8 days.
The biodegradation of TS20817 reached 39% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20817 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- September 26, 2019 – October 24, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: ISO - Water quality: ISO 9439
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. Not adapted to the test item but aerated 7 days prior to testing.
Sampling: September 19, 2019; 8:30 AM
Concentration: 30 mg/l dry matter in the final mixture. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.3 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Treatments and no. of replicates:
Blank control (medium + inoculum): 3
Procedure control (medium + inoculum + reference item): 3
Test suspension (medium + inoculum + test item): 3
(Toxicity and Abiotic sterile controls were not conducted).
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark
Reference value of the test item: Total Organic Carbon (TOC): 85.7%, based on sponsor’s information.
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equaled approximately 20 mg TOC /l; which corresponds to a nominal concentration of 23.4 mg/l of test item and 34.0 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted onto glass fibre filters, the filters were dried and introduced into the test vessels.
The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). After application in the test vessels, the glass fiber filters were disintegrated/sheared at the beginning of the test so as to optimize the contact area of the inoculum with the test item. pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions was measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach). - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 78
- Sampling time:
- 28 d
- Details on results:
- The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 45% at the end of the 10-day window.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days. - Results with reference substance:
- The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Executive summary:
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 45% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- September 26, 2019 – October 24, 2019
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Version / remarks:
- OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: ISO - Water quality: ISO 9439
- Version / remarks:
- International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- Activated sludge from the aeration tank of the ARA Werdhölzli (CH-8048 Zürich), a municipal biological waste water treatment plant. Not adapted to the test item but aerated 7 days prior to testing.
Sampling: September 19, 2019; 8:30 AM
Concentration: 30 mg/l dry matter in the final mixture. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 23.4 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- CO2 evolution
- Details on study design:
- Treatments and no. of replicates:
Blank control (medium + inoculum): 3
Procedure control (medium + inoculum + reference item): 3
Test suspension (medium + inoculum + test item): 3
(Toxicity and Abiotic sterile controls were not conducted).
Test units: 2.3 l glass bottles with 2.0 l test mixture, fitted to gas-absorption bottles containing 140 ml of 0.13 M potassium hydroxide (KOH); through aeration with CO2-free air, the CO2 produced by mineralization of the test item is trapped in the KOH solution.
Test medium: Mineral salts medium according to OECD guideline 301 B
Incubation: 28 days, 22±2 °C, continuous stirring and aeration, in the dark
Reference value of the test item: Total Organic Carbon (TOC): 85.7%, based on sponsor’s information.
Reference item and reference value: Sodium benzoate (C7H5NaO2), TOC = 58.3% (based on molecular formula)
Test concentrations: in each test vessel, the applied TOC content of the test and/or reference item equaled approximately 20 mg TOC /l; which corresponds to a nominal concentration of 23.4 mg/l of test item and 34.0 mg/l of reference item.
Application of test + reference items: The test item was applied with the glass fibre filter method: a test item’s stock solution in hexane was pipetted onto glass fibre filters, the filters were dried and introduced into the test vessels.
The reference item was applied by direct addition. Blank and procedure controls were also supplemented with glass fibre filters (treated with pure hexane). After application in the test vessels, the glass fiber filters were disintegrated/sheared at the beginning of the test so as to optimize the contact area of the inoculum with the test item. pH of test suspensions: 7.2–7.6
Measurements: The CO2 trapped in the KOH solutions was measured as Inorganic Carbon (IC) with a TOC-L CSH TOC-Analyzer (Shimadzu Schweiz GmbH, Römerstr. 3, CH-4153 Reinach). - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 78
- Sampling time:
- 28 d
- Details on results:
- The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 39% at the end of the 10-day window.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days. - Results with reference substance:
- The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable, but failing 10-day window
- Conclusions:
- TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Executive summary:
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 39% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- September 20, 2019 to October 18, 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
- Version / remarks:
- OECD.OECD Guidelines for Testing of Chemicals, 301F Manometric Respirometry Test. Paris: OECD. Adopted 17th July, 1992.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation, 301F Ready Biodegradability: Manometric Respirometry Test [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 56-61.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation, Introduction [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 6-8.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- Dangerous Chemicals Administration. GB/T 21801- 2008 Chemicals-Ready Biodegradability: Manometric Respirometry Test [S]. Beijing: Standards Press of China, 2008.
- Deviations:
- yes
- Remarks:
- See "Any other infromation" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S]. Beijing: China Environmental Science Press. 2004.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- Urban construction industry standard of the People's Republic of China. CJ/T 221-2005 Examination methods for sludge of Urban Sewage Treatment Plant [S]. Beijing: China Standards Press. 2006.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- State Environmental Protection Administration of the People's Republic of China. HJ 828-2017 Water quality-Determination of Chemical oxygen demand (CODcr) [S]. Ministry of Environmental Protection, 2017.
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- China National standardizing committee. GB/T 27849-2011 Chemicals-Degradation screening test - Chemical oxygen demand [S]. China Standards Press, 2011
- Deviations:
- yes
- Remarks:
- See "Any other information" for details
- GLP compliance:
- yes
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic (adaptation not specified)
- Details on inoculum:
- Name: Activated sludge
Source: Aeration tank of Liede Sewage Treatment Plant of Guangzhou
Batch No.: IAs20190917-1
Selection reason: The inoculum is recommended by "The Guidelines for the Testing of Chemicals".
Treatment: Collected a fresh sample of activated sludge from the aeration tank of a sewage treatment plant. After removing any coarse particles and impurities on the surface, the sludge was washed with the test medium once (1100 g, 10 min), the supernatant liquid phase was decanted and the solids were re-suspended in the test medium. The suspended solids of the sludge concentration was determined and adjusted to 3.7 g/L. Thereafter, the sludge was kept in aeration until required. When testing, the inoculum was added to each test vessel to give a final concentration of 30 mg suspended solids per liter.
Aerobic plate count: The aerobic plate count in the test system was 1.1e+7 CFU/L. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 20 mg/L
- Based on:
- test mat.
- Initial conc.:
- 54 mg/L
- Based on:
- COD
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Test medium
Test medium was prepared with deionized water.
Test methods
Test groups
Bottles 1, 2 & A (test suspension): Containing test substance, inoculum and test medium
Bottles 3, 4 (inoculum blank): Containing inoculum and test medium
Bottle 5 (procedure control): Containing reference substance, inoculum and test medium
Bottle 6 (toxicity control): Containing test substance, reference substance, inoculum and test medium
Note: A clean and dry glass cover was respectively added into bottles 3, 4, 5.
Additon of the test substance
7.2 mg of the test substance was respectively weighed on a clean and dry cover glass, and added directly into the bottles 1, 2, A. Then 362 mL of test medium was respectively joined into each bottle, and took them an ultrasound treatment for 10 min.
4.9 mg of the test substance was weighed on a clean and dry cover glass, and added directly into the bottle 6. Then 241 mL of test medium was joined into the bottle, and took it an ultrasound treatment for 10 min.
The purity of the test substance is 100% (UVCB).
Preparation of the reference substance stock solution
120 mg of the sodium benzoate was weighed into a 100 mL volumetric flask, then dissolved with test medium, and diluted with test medium to the mark to obtain the stock solution with the concentration of 1200 mg/L.
Preparation of the test systems
The test systems are prepared according to Table 1. The test substance concentration was
20 mg/L, giving 54 mg/L of COD. The reference substance concentration was 34 mg/L, giving 56 mg/L of ThOD. Total COD and ThOD of the toxicity control was 110 mg/L. The concentration of the inoculum was 30 mg/L in all vessels.
Test conditions
All the test vessels were stirred and incubated in the OxiTop® Control BOD Test System at 22°C±1°C in the dark and closed condition over 28 days.
Determination of Chemical Oxygen Demand (COD) for the test substance
Since the test substance is UVCB, the COD value was determined to use as a replacement of ThOD to calculate the percentage biodegradation. The COD values of test substance were measured as 2.798 mg/mg, 2.660 mg/mg, 2.742 mg/mg, with a mean of 2.73 mg/mg. - Reference substance:
- benzoic acid, sodium salt
- Test performance:
- (1) The oxygen uptake of the inoculum blank was 28.6 mg/L, which was less than 60 mg/L.
(2) At the end of the test, the pH values of each test bottle were 7.27 - 7.40, which were within the range of 6.0 - 8.5.
(3) The difference of extremes of replicate values of the removal of the test substance was less than 20% during the test.
(4) The percentage biodegradation of the procedure control and the toxicity control were 81.3% and 44.0%, which had reached the pass levels of 60% and 25% of ThOD within 14 days. - Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- > 34 - < 52.9
- Sampling time:
- 28 d
- Details on results:
- Under the conditions of the study, the percentage biodegradation of the test substance on 28 d were respectively 52.9% and 34.0%, with a mean of 43.4%.
- Results with reference substance:
- On day 14 of the test, the percentage biodegradation of the procedure control (reference substance) and the toxicity control were 81.3% and 44.0%, respectively, which had reached the pass levels of 60% and 25% of the theoretical oxygen demand (ThOD). It showed that the inoculum activity met the validity of the test, and the test substance was considered not to have a toxic effect on the sewage sludge micro-organisms used in the study.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- other: No conclusion is given as to whether the test substance can be considered as readily biodegradable.
- Conclusions:
- Under the conditions of the study, the percentage biodegradation of the test substance expressed as BOD was 43.4% at the end of the test.
Since the test substance is a mixture consisting of structurally similar chemicals (UVCB), it was not confirmed that whether a sequential biodegradation of the individual structures was taking place, the 10-day window for ready biodegradability was not applied to interpret the results of the test based on “The Guidelines for the Testing of Chemicals Degradation”. And the guideline doesn't also give the clear directions on how to evaluate the test results of such mixtures. Therefore, no conclusions is given as to whether the test substance can be considered as readily biodegradable. - Executive summary:
Purpose of the study
Determine the ready biodegradability of the test substance in the dark at 22°C±1°C over 28 days.
Guidelines
[1] Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation,301FReady Biodegradability: Manometric Respirometry Test[M]. Second Edition. Beijing: China Environmental Science Press. 2013: 56-61.
[2] Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation, Introduction [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 6-8.
[3] Dangerous Chemicals Administration. GB/T 21801- 2008 Chemicals-Ready
Biodegradability: Manometric Respirometry Test [S]. Beijing: Standards Press of China, 2008.
[4] OECD.OECD Guidelines for Testing of Chemicals, 301F Manometric Respirometry Test. Paris: OECD. Adopted 17th July, 1992.
[5] State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S]. Beijing: China Environmental Science Press. 2004.
[6] Urban construction industry standard of the People's Republic of China. CJ/T 221-2005 Examination methods for sludge of Urban Sewage Treatment Plant [S]. Beijing: China Standards Press. 2006.
[7] State Environmental Protection Administration of the People's Republic of China. HJ 828-2017 Water quality-Determination of Chemical oxygen demand (CODcr) [S].Ministry of Environmental Protection, 2017.
[8] China National standardizing committee. GB/T 27849-2011 Chemicals-Degradation screening test - Chemical oxygen demand [S]. China Standards Press, 2011.
Principle
20 mg/L of the test substance in the test system (giving 54 mg COD/L, within 50 - 100 mg COD/L) as the nominal sole source of organic carbon, was stirred in a closed bottle at 22°C±1°C for up to 28 days. The consumption of oxygen was determined by measuring the change of pressure in the apparatus. Evolved carbon dioxide was absorbed by sodium hydroxide. The amount of oxygen taken up by the microbial population during biodegradation of the test substance (corrected for uptake by inoculum blank, run in parallel) was expressed as a percentage of ThOD or COD.
Test conditions
Test method: Ready Biodegradability: Manometric Respirometry Test
Test substance: SynNova Base Oil
Test groups
Test suspension (bottles 1, 2, A)
Inoculum blank (bottles 3, 4)
Procedure control (bottle 5)
Toxicity control (bottle 6)
Test substance concentration: 20 mg/L (giving 54 mg/L of COD)
Reference substance concentration: 34 mg/L (giving 56 mg/L of ThOD)
Inoculum: Activated sludge
Inoculum concentration : 30 mg/L (aerobic plate count was 1.1´107 CFU/L)
Test temperature: 21.2°C - 23.3°C (within 22°C±1°C)
pH values of test solution: 7.34 - 7.39 (at the start of the test, within 7.4±0.2); 7.27 - 7.40 (at the end of the test)
Test period: 28 d (in the dark and seal)
Results
On day 14 of the test, the percentage biodegradation of the procedure control (reference substance) and the toxicity control were 81.3% and 44.0%, respectively, which had reached the pass levels of 60% and 25% of the theoretical oxygen demand (ThOD). The oxygen uptake of the inoculum blank was 28.6 mg/L, which was less than 60 mg/L. At the end of the test, the pH values of each test bottle were 7.27 - 7.40, which were within the range of 6.0 - 8.5. The difference of extremes of replicate values of the removal of the test substance was less than 20% during the test. The above results were all met the requirement of the test, which revealed that the test results were valid.
Under the conditions of the study, the percentage biodegradation of the test substance on 28 d were respectively 52.9% and 34.0%, with a mean of 43.4%.
Conclusion
Under the conditions of the study, the percentage biodegradation of the test substance expressed as BOD was 43.4% at the end of the test.
Since the test substance is a mixture consisting of structurally similar chemicals (UVCB), it was not confirmed that whether a sequential biodegradation of the individual structures was taking place, the 10-day window for ready biodegradability was not applied to interpret the results of the test based on “The Guidelines for the Testing of Chemicals Degradation”. And the guideline doesn't also give the clear directions on how to evaluate the test results of such mixtures. Therefore, no conclusions is given as to whether the test substance can be considered as readily biodegradable.
- Endpoint:
- biodegradation in water: inherent biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- September 27, 2019 to December 10, 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))
- Version / remarks:
- OECD. OECD Guidelines for Testing of Chemicals, 302C Inherent Biodegradability: Modified MITI Test (Ⅱ). Paris: OECD. Adopted: 12 th May, 1981.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation, 302C Inherent Biodegradability: Modified MITI Test (Ⅱ) [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 74-81.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- The national hazardous chemical management standardization technical committee. GB/T 21818-2008 Chemical Inherent Biodegradability: Modified MITI Test (Ⅱ) [S]. Beijing: China Standards Press.2008.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S]. Beijing: China Environmental Science Press. 2004.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- State Environmental Protection Administration of the People's Republic of China. HJ 828-2017 Water quality-Determination of Chemical oxygen demand (CODcr) [S]. Ministry of Environmental Protection, 2017.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: National Guideline
- Version / remarks:
- China National standardizing committee. GB/T 27849-2011 Chemicals-Degradation screening test - Chemical oxygen demand [S]. China Standards Press, 2011.
- Deviations:
- no
- GLP compliance:
- yes
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- mixture of sewage, soil and natural water
- Details on inoculum:
- Batch No.: I20190613-1
Names, sources and collection dates of the inoculum are reported in table form - see "Any other information" for details.
Fresh samples from the ten sites, mainly in areas where varieties of chemicals were used and discharged were collected. Sites samples included sewage treatment works, industrial waste-water treatment works, rivers, lakes, seas, 1L samples of sludge, surface soil, water, etc. were collected and mixed thoroughly together. After removing of floating matter and allowing to stand, the supernatant was adjusted to pH = 7.0 ± 1.0 with phosphoric acid. An appropriate volume of the filtered supernatant was filled in a vessel and the liquid was aerated for about 23.5 h. 30 min after ceasing the aeration of the solution obtained above, one third of the whole volume of the supernatant was discarded and an equal volume of a solution containing 0.1% each of glucose, peptone and monopotassium phosphate was added to the remaining portion of the supernatant and aeration re-commenced. This procedure was repeated once per day. The unit was maintained in a good operation and kept at 25°C ± 2°C with the pH = 7.0 ± 1.0; the final obtained sludge should settle well and be aerate continuously.
Before the study, the dry weight of the inoculum was determined and the initial concentration of the inoculum was adjusted to 4.0 g per liter, then kept it under aerobic at the test temperature until required. - Duration of test (contact time):
- 28 d
- Initial conc.:
- 30 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Test medium
The test medium was prepared with the deionized water.
Test methods
Test groups
Bottles 1 and A (abiotic control): ontaining test substance, sterilant and deionized water
Bottles 2, 3, 4 and B (test suspension): Containing test substance, inoculum and test medium
Bottle 5 (procedure control): Containing reference substance, inoculum and test medium
Bottle 6 (inoculum blank 1): Containing inoculum and test medium (inoculum concentration was 100 mg/L)
Bottle 7 (inoculum blank 2): Containing inoculum and test medium (inoculum concentration was 30 mg/L)
Bottles 8 and C (Toxicity control): Containing test substance, reference substance, inoculum and test medium (inoculum concentrations were 100 mg/L)
Note:
1. Bottles A, B and C for determination of pH values at the start of the test.
2. A clean and dry glass cover was added into bottle 6.
Additon of the test substance
15.0 mg of the test substance was respectively weighed on a clean and dry cover glass, and added them directly into the bottles 1, A, 2, 3, 4, B, 8 and C. Then deionized water or test medium were respectively added into each bottle, and took them an ultrasound treatment for 10 min.
The purity is 100% (UVCB).
Preparation of the reference substance stock solution
200 mg of the sodium benzoate was weighed into a 200 mL volumetric flask and dissolved with test medium, and filled up to the mark to obtain a stock solution with the concentration of 1000 mg/L.
Addition of sterilant
250 mg of mercuric chloride were weighed into bottles 1 and A.
Preparation of the test system
The final volume of each test system was 500 mL. The test substance final concentration was 30 mg/L, the reference substance final concentration was 100 mg/L, and the inoculum final concentrations were respectively 100 mg/L (test suspension, inoculum blank 1, toxicity control) and 30 mg/L (procedure control, inoculum blank 2).
Test conditions
All test bottles were stirred and incubated on C.E.S Respirometer Systems at 25°C ± 2°C in the dark and sealed condition over 28 days.
Determination of Chemical Oxygen Demand (COD) for the test substance
Since the test substance is UVCB, the COD value was determined to use as a replacement of ThOD to calculate the percentage biodegradation. The COD values of test substance were measured as 2.798 mg/mg, 2.660 mg/mg, 2.742 mg/mg, with a mean of 2.73 mg/mg. - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 44.5
- Sampling time:
- 28 d
- Key result
- Parameter:
- % degradation (O2 consumption)
- Remarks:
- primary rate
- Value:
- 47.2
- Sampling time:
- 28 d
- Details on results:
- Test temperature was within the range of 24.9°C - 26.5°C during the test. At the start of the test, pH values of each test bottle were within the range of 7.46 - 7.50 (pH value of abiotic control was 4.22). At the end of the test, pH values of each test bottle were within the range of 6.5 - 8.27 (pH value of abiotic control was 5.1).
Percentage biodegradation of two replicates of the test substance on 28 d calculated with BOD were respectively 47.8% and 41.2%, with a mean of 44.5%. Percentage biodegradation of the toxicity control on 7 d and 14 d were 53.0% and 63.2%, respectively.
On day 28, the residual test substance was extracted and measured. Results revealed that, the concentration of residual test substance in the abiotic control was 17.8 mg/L (abiotic degradation was 40.7%). The residual test substance concentrations in the test suspensions (three replicates) were 10.5 mg/L, 9.2 mg/L, 8.5 mg/L, and the primary biodegradation rates were respectively 41.0%, 48.3%, 52.2% , with a mean of 47.2%.
To verify whether the extraction and concentration analysis methods used in the study met the requirements, recovery tests were conducted. Prepared triplicate test solutions of the abiotic control and the test suspensions with the test substance concentrations of 30 mg/L, respectively, then extracted the test substance and determined the test substance concentrations. The average percent recovery of the abiotic control and the test suspension were 85.7% (RSD: 3.6%) and 85.0% (RSD: 4.5%), respectively. It revealed that the recovery methods for the test substance used in the study were valid. - Results with reference substance:
- Sodium benzoate was used as reference substance, the percentage biodegradation of the procedure control was 81.6% (>40%) on 7 d and was 90.0% (>65%) on 14 d of the test.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- inherently biodegradable
- Conclusions:
- Under the conditions of the study, percentage biodegradation rate of the test substance on day 28 which was calculated by BOD was 44.5%, and the primary biodegradation rates which was calculated by residual amount of the substance was 47.2%, therefore the test substance can be considered to be primary inherent biodegradable.
- Executive summary:
Purpose of the study
Determine the Inherent Biodegradability of the test substance in the dark at 25 °C ± 2 °C.
Guidelines
[1] Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals Degradation and Accumulation,Inherent Biodegradability: Modified MITI Test (Ⅱ) [M]. Second Edition. Beijing: China Environmental Science Press. 2013: 74-81.
[2] The national hazardous chemical management standardization technical committee. GB/T 21818-2008 Chemical Inherent Biodegradability: Modified MITI Test (Ⅱ) [S]. Beijing: China Standards Press.2008.
[3] OECD. OECD Guidelines for Testing of Chemicals,Inherent Biodegradability: Modified MITI Test (II). Paris: OECD. Adopted: 12th May, 1981.
[4] State Environmental Protection Administration of China. HJ/T 153-2004 The Guidelines for the test of chemical [S]. Beijing: China Environmental Science Press. 2004.
[5] State Environmental Protection Administration of the People's Republic of China. HJ 828-2017 Water quality-Determination of Chemical oxygen demand (CODcr) [S].Ministry of Environmental Protection, 2017.
[6] China National standardizing committee. GB/T 27849-2011 Chemicals-Degradation screening test - Chemical oxygen demand [S]. China Standards Press, 2011.
Principle
Test substance as sole source of organic carbon, no adaptation of micro-organisms to test substance, an automated closed-system oxygen consumption measuring apparatus (BOD-meter) is used. Test substance to be tested was inoculated in the testing vessels with micro-organisms. During the test period, the biological oxygen demand was measured continuously by means of a BOD-meter. Biodegradation was calculated on the basis of BOD and supplement chemical analysis, such as measurement of the concentration of the residual test substance.
Test conditions
Test substance: SynNova Base Oil
Test groups:
Abiotic control (bottles 1 and A)
Test suspension (bottles 2, 3, 4 and B)
Procedure control (bottle 5)
Inoculum blank 1 (bottle 6)
Inoculum blank 2 (bottle 7)
Toxicity control (bottles 8 and C)
Test substance concentration: 30 mg/L
Reference substance concentration: 100 mg/L
Sterilant final concentration: 500 mg/L
Inoculum concentration: 100 mg/L (test suspension, inoculum blank 1 and toxicity control); 30 mg/L (procedure control and inoculum blank 2)
Total volume: 500 mL
Test temperature: 24.9°C - 26.5°C (within 25°C±2°C)
Test period: 28 d (in the dark)
pH values: 7.46 - 7.50 (at the start of the test, 4.22 in abiotic control); 6.5 - 8.27 (at the end of the test, 5.1 in abiotic control)
Results
Sodium benzoate was used as reference substance, the percentage biodegradation of the procedure control was 81.6% (>40%) on 7 d and was 90.0% (>65%) on 14 d of the test.
Percentage biodegradation of two replicates of the test substance on 28 d calculated with BOD were respectively 47.8% and 41.2%, with a mean of 44.5%. Percentage biodegradation of the toxicity control on 7 d and 14 d were 53.0% and 63.2%, respectively.
On day 28, the residual test substance was extracted and measured. Results revealed that, the concentration of residual test substance in the abiotic control was 17.8 mg/L (abiotic degradation was 40.7%). The residual test substance concentrations in the test suspensions (three replicates) were 10.5 mg/L, 9.2 mg/L, 8.5 mg/L, and the primary biodegradation rates were respectively 41.0%, 48.3%, 52.2%, with a mean of 47.2%.
To verify whether the extraction and concentration analysis methods used in the study met the requirements, recovery tests were conducted. Prepared triplicate test solutions of the abiotic control and the test suspensions with the test substance concentrations of 30 mg/L, respectively, then extracted the test substance and determined the test substance concentrations. The average percent recovery of the abiotic control and the test suspension were 85.7% (RSD: 3.6%) and 85.0% (RSD: 4.5%), respectively. It revealed that the recovery methods for the test substance used in the study were valid.
Discussion
The Inherent Biodegradability: Modified MITI Test (II) of SynNova Base Oil had been performed for twice, with the C.E.S Respirometer Systems.
In one of the studies, the oxygen consumption of the inoculum blank was 123.3mg/L, which significantly deviated from the historical data in our laboratory (The range of oxygen consumption in 64 inoculum blanks of 2013 to 2019 was 31.0 – 103.8 mg/L, with a mean of 66.3mg/L), so the results were considered unreliable and no further statistics was made.
In the other study, the primary biodegradation rates were respectively 41.0%, 48.3%, 52.2%, the BOD biodegradation rates were 18.1%, 47.8% and 41.2%. It showed that BOD biodegradation rate of replicate 1 had great difference on the other two replicates, which failed to reflect the real biodegradation rate, so that BOD biodegradation rate of replicate 1 (18.1%) for was considered as outlier, and eventually the BOD biodegradation percentage of the test substance was decided to the average biodegradation rate with replicates 2 and 3 of 44.5%. In the meantime, the ready biodegradability test (301F) of the test substance showed a BOD biodegradation rate of 43.4% on 28 days, and it believed that the average biodegradation rate with replicates 2 and 3 of 44.5% was more able to reflect the reality of biodegradation rate for the test substance.
Conclusion
Under the conditions of the study, percentage biodegradation rate of the test substance on day 28 which was calculated by BOD was 44.5%, and the primary biodegradation rates which was calculated by residual amount of the substance was 47.2%, therefore the test substance can be considered to be primary inherent biodegradable.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- April 24, 2020 to August 12, 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
- Version / remarks:
- Biodegradability Test of Chemical Substances by Microorganisms (corresponding to 301F) (Japanese notification, Yakushokuhatsu 0331 No. 7, Heisei 23.03.29 Seikyoku No. 5, Kanpokihatsu No. 110331009, March 31, 2011; the latest revision, July 1, 2019)
- Deviations:
- not specified
- GLP compliance:
- yes
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge, domestic, adapted
- Details on inoculum:
- Inoculum: Activated sludge from domestic sewage treatment plant
Source: Sewage treatment plant mainly handling domestic wastewater in Shiga
Date of collection: April 23, 2020
Mixed liquor suspended solids (MLSS): 3300 mg/L (BOD measurement) - Duration of test (contact time):
- 28 d
- Initial conc.:
- 100 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Degradability study conditions
This study was conducted under the following conditions.
Apparatus
Closed system apparatus for measuring oxygen consumption, OM-3 lO0A (ID code: P), Ohkura Electric Co.
Data processing program: BOD_ CSCL301 F (Ver.1.0)
Conditions
Temperature: 22 ± 1°C
Exposure period: 28 days (BOD measurement)
Stirring: Continuous stirring with magnetic stirrer
Test vessel: Culture bottle, glass
CO2 absorbent: Soda lime, for carbon dioxide absorption, Kanto Chemical Co., Inc.
Test volume: 300mL
Concentration: Test substance: 100 mg/L
Reference substance: 100 mg/L
Inoculum: 30 mg/L
Test bottle contents and addition of the test substance
Outline of the test solution are shown in table format, see “Any other information” for details. The test substance or reference substance and inoculum were added to the media in test vessels.
Management and observation during BOD measurement period
In the apparatus, the temperature, stirring, recording meter and air-tightness of the closed system were checked once a day except holidays during BOD measurement period. Observation was made on color of the contents in test bottles using the [Inoculum blank] as a comparative reference.
After the end of BOD measurement, observations on color of the contents in test bottles and growth of the sludge using the [Inoculum blank] as a comparative reference and dissolution state of the test substance were also made.
pH measurement
At the start and the end of BOD measurement, pH values of the test solutions were measured.
1) Sample for analysis
At the start of BOD measurement: Test solutions of the [Inoculum blank]
At the end of BOD measurement: All test solutions
2) Apparatus
pH meter: F-52 , HORIBA, Ltd.
DOC measurement
The DOC in the processed samples (DOC phase) of the [Sludge + test substance] and the [Inoculum blank] were measured under the following apparatus and conditions.
Apparatus
TOC analyzer: multi N/C 2100S (No. 1), Analytik Jena Japan Co., Ltd.
Data-processing program: TOC_JENA (Ver.2.0)
Conditions
Furnace temperature: 800°C
Injection volume: 300 μL
Repeated number: n=3 (adopt mean value)
Calibration curve
The following standard solutions were injected into the TOC analyzer. The peak area (Y) was plotted against the concentration (X) to obtain the linear regression, Y = a + bX. Amount of DOC in each bottle was calculated from the linear regression.
Total carbon (TC): 0.500, 40.0 and 80.0 mgC/L standard solutions (aqueous solutions of potassium hydrogen phthalate)
Inorganic carbon (IC): 0.500, 5.00 and 10.0 mgC/L standard solutions (aqueous solution of sodium hydrogen carbonate and sodium carbonate) - Reference substance:
- benzoic acid, sodium salt
- Test performance:
- All of the validity criteria given in the guideline were fulfilled and hence the test was judged valid.
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 34
- Sampling time:
- 28 d
- Details on results:
- Factors which might have affected the reliability of the test results
There was no specific factor which might have affected the reliability of the test results.
Observation of the contents after the end of BOD measurement
The content in the [Sludge + test substance ]-1, 2 (bottle 1 and 2), and the [Sludge + reference substance] (bottle 3) were colorless.
Growth of the sludge was observed in the [Sludge+ test substance]-1, 2 and the [Sludge + reference substance].
The test substance was observed to be insolved in the [Sludge + test substance ]-1 and 2.
pH measurement
At the end of BOD measurement, the pH values were 7.5 and 7.5 for the [Sludge+ test substance]-! and 2, respectively.
Degradability based on the BOD
The BOD*1 in the [Sludge+ test substance]-! and 2 after 28 days were 32.3 and 37.0 mgO2 (as corrected with the average value in the [Inoculum blank]), respectively.
(* 1 Theoretical value= 103.3 mgO2)
The degradabilities based on the BOD after 28 days were calculated to be 31 % and 36% for the [Sludge+ test substance]-1 and 2, respectively.
Degradability based on the DOC
The DOC*2 in the [Sludge+ test substance]-1 and 2 after 28 days were 0.8 and 1.3 mgC (as corrected with the average value in the [Inoculum blank]), respectively.
(*2 Theoretical value= 25.5 mgC)
In this test, the degradability was not calculated because test substance was hardly soluble in water.
Degradability based on the residual test substance amount
The residual amounts of the test substance *3 in the [Sludge + test substance ]-1 and 2 after 28 days were 16.9 and 11.1 mg, respectively.
(*3 Initial amount= 30.0 mg)
The degradabilities based on the residual amount of the test substance after 28 days were calculated to be 44% and 63% for the [Sludge+ test substance ]-1, 2, respectively.
The GPC chromatogram of the [Sludge+ test substance]-1, 2 compared to the peak shape of the standard solution of the test substance, the peak height at retention time of about 11-12 min was slight increased.
Confirmation of presence or absence of transformation products with reversed phase
HPLC
The HPLC chromatograms of the [Sludge + test substance ]-1, 2 were compared to the one of the standard solution of the test substance. On the organic phase of the [Sludge + test substance]-1, 2, at retention time of about 4.6 - 5.3 min, new peak derived from transformation product was detected. - Results with reference substance:
- The degradabilities based on the BOD 14 days was calculated to be 88%, for the [Sludge + reference substance].
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- It can be concluded that the test substance is not readily biodegradable and transformation product was generated under the conditions of this test.
There is a trend that the BOD curve has been keeping elevation after 28 days. Whilst the test substance does not degrade within the specified 28-day time span of the test, it is anticipated that degradation will occur within a short time frame.
The detected transformation products were those in which the end of the structure of the test substance was a carboxyl group. - Executive summary:
Ready Biodegradability Test of SynNova Base Oil (Study No. A190954)
METHOD
This study was conducted in accordance with the Test Method Relating to New Chemical Substances <Biodegradability Test of Chemical Substances by Microorganisms (corresponding to 301F)> (Japanese notification, Yak.ushokuhatsu 0331 No. 7, Heisei 23.03.29 Seikyoku No. 5, Kanpokihatsu No. 110331009, March 31, 2011; the latest revision, July 1, 2019).
Test bottle contents
Bottle 1, 2: [Sludge+ test substance]-1, -2 (Test suspension)
Bottle 3: [Sludge + reference substance] (Activity control)
Bottle 4, 5: [Inoculum blank]-1, -2 (Control blank)
(test substance: 100 mg/L, reference substance: 100 mg/L, inoculum: 30 mg/L)
Measurements
Biochemical oxygen demand (BOD) for 28 days [measured with a closed system apparatus for measuring oxygen consumption]
Dissolved organic carbon (DOC) [measured with a TOC analyzer]
Residual amount of the test substance [measured with a gel permeation chromatograph (GPC)]
Confirmation of presence or absence of transformation products [ measured with a reversed phase high performance liquid chromatograph (RP-HPLC)]
RESULTS
Measured values after 28 days
Measurement
[Sludge + test substance]
Theoretical value
1
2
BOD*1, mgO2
32.3
37.0
103.3
DOC*1, mgC
0.8
1.3
25.5
Test substance
mg
16.9
11.1
30.0
%
56
37
-
*1 Values of [Sludge + test substance] were corrected with average BOC of DOC value of [Inoculum blank].
Degradability after 28 days
[Sludge + test substance]
1
2
Average
BOD, %
31
36
34
DOC*2, %
NA
NA
-
Test substance, %
44
63
54
NA: Not available
*2 Since the test substance is hardly soluble in water, degradability was not calculated.
On the RP-HPLC chromatogram, new peak derived from transformation product was detected.
CONCLUSION
It can be concluded that the test substance is not readily biodegradable and transformation product was generated under the conditions of this test.
There is a trend that the BOD curve has been keeping elevation after 28 days. Whilst the test substance does not degrade within the specified 28-day time span of the test, it is anticipated that degradation will occur within a short time frame.
The detected transformation products were those in which the end of the structure of the test substance was a carboxyl group.
- Endpoint:
- biodegradation in water: inherent biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- July-August 2020
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))
- Version / remarks:
- OECD Guideline for Testing of Chemicals No.302C (1981) "Inherent Biodegradability: Modified MITI Test (II)"
- Deviations:
- not specified
- GLP compliance:
- no
- Specific details on test material used for the study:
- No further details specified in the study report.
- Oxygen conditions:
- anaerobic
- Inoculum or test system:
- activated sludge (adaptation not specified)
- Details on inoculum:
- Not specified in the study report.
- Duration of test (contact time):
- 28 d
- Initial conc.:
- 30 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST BOTTLE CONTENTS
Test substance: 30 mg/L
Inoculum: 100 mg/L
· [Water + test substance] (Abiotic control)
· [Sludge + test substance] (Test suspension)
· [Inoculum blank] (Control blank)
MEASUREMENTS
Biochemical oxygen demand (BOD) for 28 days [measured with a closed system apparatus for measuring oxygen consumption]
Dissolved organic carbon (DOC) [measured with a TOC analyzer] - Reference substance:
- not specified
- Preliminary study:
- degradability results based on the BOD (av. 36%) and the residual amount of the test substance (av. 64%).
- Test performance:
- Not specified in the study report
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 36
- Sampling time:
- 28 d
- Details on results:
- No further details specified in the study report.
- Validity criteria fulfilled:
- not specified
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- From the degradability results based on the BOD (av. 36%) and the residual amount of the test substance (av. 64%), it can be concluded that the test substance is not readily biodegradable under the conditions of this test.
- Executive summary:
The study is conducted to evaluate the inherent biodegradability test of the test substance.
Study conducted in accordance with: OECD Guideline for Testing of Chemicals No.302C (1981) "Inherent Biodegradability: Modified MITI Test (II)"
From the degradability results based on the BOD (av. 36%) and the residual amount of the test substance (av. 64%), it can be concluded that the test substance is not readily biodegradable under the conditions of this test.
There is a trend that the BOD curve has been keeping elevation after 28 days. This would indicate that the study has a lag time, and post 28-days further degradation would be anticipated. Therefore, whilst the substance does not degrade within the specified 28-day time span of the test, it is anticipated that degradation will occur within a short time frame.
It is suggested that the test substance was transformed since a new peak was observed on the chromatogram of HPLC.
Referenceopen allclose all
Cumulative Biochemical Oxygen Demand (mg O2/L) in Test Flasks during the Test Period of 28 Days
Time (days) |
Flask No. |
||||||
1 |
2 |
3 |
4 |
5 |
6 |
7 |
|
1 |
0 |
0 |
5 |
5 |
45 |
0 |
30 |
2 |
5 |
5 |
5 |
5 |
85 |
0 |
85 |
3 |
5 |
5 |
5 |
5 |
100 |
0 |
105 |
4 |
10 |
10 |
5 |
10 |
110 |
0 |
125 |
5 |
10 |
10 |
10 |
10 |
125 |
0 |
135 |
6 |
15 |
15 |
10 |
10 |
130 |
0 |
140 |
7 |
20 |
20 |
10 |
15 |
135 |
0 |
145 |
8 |
30 |
35 |
15 |
15 |
140 |
0 |
145 |
9 |
40 |
50 |
15 |
15 |
145 |
0 |
155 |
10 |
50 |
60 |
15 |
20 |
145 |
0 |
165 |
11 |
55 |
70 |
20 |
20 |
145 |
0 |
170 |
12 |
65 |
75 |
20 |
20 |
150 |
0 |
175 |
13 |
70 |
80 |
20 |
20 |
150 |
0 |
185 |
14 |
80 |
85 |
20 |
20 |
150 |
0 |
190 |
15 |
85 |
90 |
20 |
20 |
155 |
0 |
195 |
16 |
95 |
95 |
20 |
20 |
155 |
0 |
200 |
17 |
100 |
95 |
20 |
20 |
160 |
0 |
205 |
18 |
110 |
100 |
20 |
20 |
160 |
0 |
210 |
19 |
115 |
100 |
20 |
20 |
160 |
0 |
215 |
20 |
120 |
100 |
20 |
20 |
160 |
0 |
220 |
21 |
125 |
100 |
20 |
20 |
160 |
0 |
225 |
22 |
130 |
105 |
20 |
20 |
160 |
0 |
230 |
23 |
135 |
105 |
20 |
20 |
165 |
0 |
235 |
24 |
145 |
105 |
20 |
20 |
165 |
0 |
240 |
25 |
145 |
110 |
20 |
20 |
165 |
0 |
240 |
26 |
150 |
110 |
20 |
20 |
165 |
0 |
245 |
27 |
155 |
115 |
20 |
20 |
165 |
0 |
255 |
28 |
160 |
115 |
20 |
20 |
165 |
0 |
260 |
Flasks 1 and 2: SynNova Base Oil; Flasks 3 and 4: inoculum control; Flask 5: reference (procedure control); Flask 6: abiotic control; Flaks 7: toxicity control
Percentage Biodegradation of Test Item, of Sodium Benzoate and of the Toxicity Control based on ThODNH4
Time (days) |
Percentage Biodegradation |
|||
SynNova Base Oil1 |
Sodium Benzoate2 |
Toxicity control1,2 |
||
Flask 1 [%] |
Flask 2 [%] |
Flask 5 [%] |
Flask 7 [%] |
|
1 |
0 |
0 |
23 |
5 |
2 |
0 |
0 |
46 |
16 |
3 |
0 |
0 |
55 |
19 |
4 |
1 |
1 |
59 |
23 |
5 |
0 |
0 |
67 |
24 |
6 |
1 |
1 |
69 |
25 |
7 |
2 |
2 |
71 |
26 |
8 |
4 |
6 |
72 |
25 |
9 |
7 |
10 |
75 |
27 |
10 |
9 |
12 |
74 |
29 |
11 |
10 |
14 |
72 |
29 |
12 |
13 |
16 |
75 |
30 |
13 |
14 |
17 |
75 |
32 |
14 |
17 |
18 |
75 |
33 |
15 |
19 |
20 |
78 |
34 |
16 |
22 |
21 |
78 |
35 |
17 |
23 |
21 |
81 |
36 |
18 |
26 |
23 |
81 |
37 |
19 |
27 |
23 |
81 |
38 |
20 |
29 |
23 |
81 |
39 |
21 |
30 |
23 |
81 |
40 |
22 |
32 |
24 |
81 |
41 |
23 |
33 |
24 |
84 |
42 |
24 |
36 |
24 |
84 |
43 |
25 |
36 |
26 |
84 |
43 |
26 |
38 |
26 |
84 |
44 |
27 |
39 |
27 |
84 |
46 |
28 |
40 |
27 |
84 |
47 |
1ThODNH4of SynNova Base Oil: 3.416 mg O2/mg test item
2ThODNH4of sodium benzoate: 1.66 mg O2/mg reference item
pH-Values at the End of the Test
Flask No. |
Treatment |
pH-value |
1 |
SynNova Base Oil |
7.5 |
2 |
SynNova Base Oil |
7.6 |
3 |
Inoculum Control |
7.5 |
4 |
Inoculum Control |
7.5 |
5 |
Reference item (procedure control) |
7.6 |
Inorganic carbon produced by the test and control units, and corresponding degradation data
Time [days] |
Blank control |
Procedure control |
Test Suspension |
Toxicity control |
Abiotic sterile control |
|||||||
Replicate 1 |
Replicate 2 |
Mean |
||||||||||
CO2prod. [mg IC/l] |
CO2prod. [mg IC/l] |
Degrad. [%] |
CO2prod. [mg IC/l] |
Degrad. [%] |
CO2prod. [mg IC/l] |
Degrad. [%] |
Degrad. [%] |
CO2prod. [mg IC/l] |
Degard. [%] |
CO2prod. [mg IC/l] |
Degrad. [%] |
|
0 |
0.0 |
0.0 |
0 |
0.0 |
0.0 |
0.0 |
0.0 |
0 |
n.d. |
-- |
n.d. |
-- |
1 |
0.0 |
0.8 |
4 |
0.3 |
1.5 |
0.1 |
0.9 |
1 |
n.d. |
-- |
n.d |
-- |
3 |
0.6 |
9.6 |
45 |
1.3 |
3.6 |
1.1 |
2.6 |
3 |
n.d. |
-- |
n.d |
-- |
7 |
1.8 |
15.5 |
69 |
4.3 |
12.3 |
3.3 |
7.4 |
10 |
n.d. |
-- |
n.d |
-- |
10 |
2.5 |
17.1 |
73 |
5.5 |
14.7 |
5.3 |
13.9 |
14 |
n.d. |
-- |
n.d |
-- |
14 |
3.1 |
19.7 |
83 |
7.5 |
21.9 |
9.2 |
30.3 |
26 |
n.d. |
-- |
n.d |
-- |
17 |
4.1 |
21.5 |
87 |
10.3 |
30.8 |
12.0 |
39.4 |
35 |
n.d. |
-- |
n.d |
-- |
21 |
5.4 |
22.7 |
87 |
14.1 |
43.0 |
14.6 |
45.4 |
44 |
n.d. |
-- |
n.d |
-- |
24 |
5.9 |
23.4 |
87 |
15.6 |
48.0 |
16.2 |
51.1 |
50 |
n.d. |
-- |
n.d |
-- |
27 |
5.9 |
24.7 |
94 |
18.4 |
61.8 |
17.5 |
57.2 |
60 |
n.d. |
-- |
n.d |
-- |
28 |
6.0 |
24.8 |
94 |
18.6 |
62.2 |
18.3 |
60.3 |
61 |
n.d. |
-- |
n.d |
-- |
CO2prod. [mg IC/l]: mg/l of inorganic carbon produced in the vessel; n.d.: not determined.
Degrad. [%]: Biodegradation in % of the TOC; the total TOC concentrations taken into account for the calculations are 20.3 and 20.3 mg/l for replicate 1 and 2 of the test suspension, resp. and 20.0 mg/l for the procedure control.
Validity criteria
Validity criterion |
Required |
Achieved |
Inorganic carbon produced in the blank control after 28 days [mg/l] |
≤ 19.1 |
6.0 |
Degradation of the mixture in the toxicity control after 14 days [%] |
≥ 25 |
n.d. |
Difference of extremes of replicate values (after 28 days) [%] |
≤ 20 |
3% |
Degradation of the reference item after 14 days [%] |
≥ 60 |
83 |
n.d.: not determined
Inorganic carbon produced by the test and control units, and corresponding degradation data
Time [days] |
Blank control |
Procedure control |
Test Suspension |
Toxicity control |
Abiotic sterile control |
|||||
(Replicates 1 / 2) |
Mean |
|||||||||
CO2prod. [mg IC/l]* |
CO2prod. [mg IC/l]** |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
|
0 |
0.0 |
0.0 |
0 |
0.0 / 0.0 |
0.0 / 0.0 |
0 |
n.d. |
- |
n.d. |
- |
1 |
1.1 |
3.4 |
11 |
2.7 / 1.2 |
7.9 / 0.3 |
4 |
n.d. |
- |
n.d. |
- |
6 |
3.6 |
18.0 |
72 |
5.7 / 3.5 |
10.5 / -0.2 |
5 |
n.d. |
- |
n.d. |
- |
7 |
4.0 |
18.7 |
74 |
6.7 / 4.3 |
13.5 / 1.6 |
8 |
n.d. |
- |
n.d. |
- |
10 |
5.3 |
21.0 |
79 |
8.8 / 5.9 |
17.2 / 3.1 |
10 |
n.d. |
- |
n.d. |
- |
14 |
6.8 |
22.9 |
80 |
11.6 / 8.5 |
23.7 / 8.0 |
16 |
n.d. |
- |
n.d. |
- |
17 |
7.9 |
24.2 |
81 |
13.7 / 10.2 |
28.6 / 11.4 |
20 |
n.d. |
- |
n.d. |
- |
21 |
9.1 |
25.8 |
84 |
18.8 / 15.3 |
48.3 / 30.9 |
40 |
n.d. |
- |
n.d. |
- |
24 |
10.0 |
27.3 |
87 |
20.8 / 18.5 |
53.5 / 42.2 |
48 |
n.d. |
- |
n.d. |
- |
28 |
11.5 |
29.4 |
89 |
26.5 / 22.9 |
73.8 / 56.2 |
65 |
n.d. |
- |
n.d. |
- |
CO2prod. [mg IC/l]: mg/l of inorganic carbon produced per liter of medium.
Deg [%]: Biodegradation in % of the TOC; the total TOC concentrations taken into account for the calculations are 20.3 and 20.3 mg/l for replicates 1 and 2 of the test suspension, resp., and 20.0 mg/l for the procedure control.
*3 replicates; **2 replicates; n.d.: not determined
Inorganic carbon produced by the test and control units, and corresponding degradation data
Time [days] |
Blank control |
Procedure control |
Test Suspension |
Toxicity control |
Abiotic sterile control |
|||||
(Replicates 1 / 2) |
Mean |
|||||||||
CO2prod. [mg IC/l]* |
CO2prod. [mg IC/l]** |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
|
0 |
0.0 |
0.0 |
0 |
0.0 / 0.0 |
0.0 / 0.0 |
0 |
n.d. |
- |
n.d. |
- |
1 |
0.5 |
0.9 |
2 |
0.3 / 0.4 |
-0.7 / -0.4 |
-1 |
n.d. |
- |
n.d. |
- |
4 |
1.4 |
11.8 |
53 |
2.4 / 1.6 |
5.3 / 1.2 |
3 |
n.d. |
- |
n.d. |
- |
7 |
2.3 |
15.6 |
67 |
3.3 / 3.6 |
5.3 / 6.4 |
6 |
n.d. |
- |
n.d. |
- |
11 |
3.3 |
17.7 |
73 |
6.6 / 7.0 |
16.7 / 18.5 |
18 |
n.d. |
- |
n.d. |
- |
14 |
3.9 |
18.9 |
76 |
8.6 / 9.2 |
23.4 / 26.5 |
25 |
n.d. |
- |
n.d. |
- |
18 |
4.9 |
20.6 |
79 |
12.3 / 12.6 |
37.0 / 38.4 |
38 |
n.d. |
- |
n.d. |
- |
21 |
5.7 |
21.7 |
81 |
15.3 / 15.1 |
47.8 / 46.9 |
47 |
n.d. |
- |
n.d. |
- |
25 |
6.5 |
22.8 |
82 |
18.6 / 15.8 |
60.1 / 46.1 |
53 |
n.d. |
- |
n.d. |
- |
27 |
7.1 |
23.3 |
82 |
19.8 / 19.0 |
63.2 / 59.1 |
61 |
n.d. |
- |
n.d. |
- |
28 |
8.3 |
25.6 |
87 |
23.6 / 21.4 |
76.3 / 65.4 |
71 |
n.d. |
- |
n.d. |
- |
CO2prod. [mg IC/l]: mg/l of inorganic carbon produced per liter of medium.
Deg [%]: Biodegradation in % of the TOC; the total TOC concentrations taken into account for the calculations are 20.0 and 20.0 mg/l for replicates 1 and 2 of the test suspension, resp., and 19.8 mg/l for the procedure control.
*3 replicates; **2 replicates; n.d.: not determined
Inorganic carbon produced by the test and control units, and corresponding degradation data
Time [days] |
Blank control |
Procedure control |
Test Suspension |
Toxicity control |
Abiotic sterile control |
|||||
(Replicates 1 / 2) |
Mean |
|||||||||
CO2prod. [mg IC/l]* |
CO2prod. [mg IC/l]** |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
|
0 |
0.0 |
0.0 |
0 |
0.0 / 0.0 |
0.0 / 0.0 |
0 |
n.d. |
- |
n.d. |
- |
1 |
0.5 |
0.9 |
2 |
0.6 / 0.7 |
0.4 / 1.1 |
1 |
n.d. |
- |
n.d. |
- |
4 |
1.4 |
11.8 |
53 |
1.7 / 2.3 |
1.8 / 4.5 |
3 |
n.d. |
- |
n.d. |
- |
7 |
2.3 |
15.6 |
67 |
3.7 / 4.3 |
7.2 / 10.1 |
9 |
n.d. |
- |
n.d. |
- |
11 |
3.3 |
17.7 |
73 |
8.9 / 7.4 |
28.3 / 20.6 |
24 |
n.d. |
- |
n.d. |
- |
14 |
3.9 |
18.9 |
76 |
11.2 / 10.4 |
36.3 / 32.5 |
34 |
n.d. |
- |
n.d. |
- |
18 |
4.9 |
20.6 |
79 |
15.4 / 13.2 |
52.3 / 41.5 |
47 |
n.d. |
- |
n.d. |
- |
21 |
5.7 |
21.7 |
81 |
17.6 / 15.3 |
59.3 / 47.8 |
54 |
n.d. |
- |
n.d. |
- |
25 |
6.5 |
22.8 |
82 |
20.9 / 18.5 |
72.0 / 59.8 |
66 |
n.d. |
- |
n.d. |
- |
27 |
7.1 |
23.3 |
82 |
22.6 / 20.0 |
77.3 / 64.2 |
71 |
n.d. |
- |
n.d. |
- |
28 |
8.3 |
25.6 |
87 |
24.6 / 23.2 |
81.3 / 74.3 |
78 |
n.d. |
- |
n.d. |
- |
CO2prod. [mg IC/l]: mg/l of inorganic carbon produced per liter of medium.
Deg [%]: Biodegradation in % of the TOC; the total TOC concentrations taken into account for the calculations are 20.0 and 20.0 mg/l for replicates 1 and 2 of the test suspension, resp., and 19.8 mg/l for the procedure control.
*3 replicates; **2 replicates; n.d.: not determined
Inorganic carbon produced by the test and control units, and corresponding degradation data
Time [days] |
Blank control |
Procedure control |
Test Suspension |
Toxicity control |
Abiotic sterile control |
|||||
(Replicates 1 / 2) |
Mean |
|||||||||
CO2prod. [mg IC/l]* |
CO2prod. [mg IC/l]** |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
CO2prod. [mg IC/l] |
Deg. [%] |
|
0 |
0.0 |
0.0 |
0 |
0.0 / 0.0 |
0.0 / 0.0 |
0 |
n.d. |
- |
n.d. |
- |
1 |
0.5 |
0.9 |
2 |
0.9 / 0.8 |
1.9 / 1.4 |
2 |
n.d. |
- |
n.d. |
- |
4 |
1.4 |
11.8 |
53 |
2.4 / 1.7 |
5.3 / 1.5 |
3 |
n.d. |
- |
n.d. |
- |
7 |
2.3 |
15.6 |
67 |
5.0 / 3.5 |
13.8 / 6.2 |
10 |
n.d. |
- |
n.d. |
- |
11 |
3.3 |
17.7 |
73 |
8.0 / 6.6 |
23.5 / 16.3 |
20 |
n.d. |
- |
n.d. |
- |
14 |
3.9 |
18.9 |
76 |
10.6 / 8.5 |
33.4 / 23.1 |
28 |
n.d. |
- |
n.d. |
- |
18 |
4.9 |
20.6 |
79 |
14.7 / 12.4 |
48.9 / 37.4 |
43 |
n.d. |
- |
n.d. |
- |
21 |
5.7 |
21.7 |
81 |
17.0 / 15.8 |
56.4 / 50.3 |
53 |
n.d. |
- |
n.d. |
- |
25 |
6.5 |
22.8 |
82 |
19.9 / 19.4 |
66.9 / 64.3 |
66 |
n.d. |
- |
n.d. |
- |
27 |
7.1 |
23.3 |
82 |
21.2 / 20.7 |
70.3 / 67.9 |
69 |
n.d. |
- |
n.d. |
- |
28 |
8.3 |
25.6 |
87 |
25.0 / 23.0 |
83.4 / 73.3 |
78 |
n.d. |
- |
n.d. |
- |
CO2prod. [mg IC/l]: mg/l of inorganic carbon produced per liter of medium.
Deg [%]: Biodegradation in % of the TOC; the total TOC concentrations taken into account for the calculations are 20.0 and 20.0 mg/l for replicates 1 and 2 of the test suspension, resp., and 19.8 mg/l for the procedure control.
*3 replicates; **2 replicates; n.d.: not determined
BOD of each test bottle during the test (mg/L)
Time (d) |
Test suspension |
inoculum blank |
Procedure control |
Toxicity control |
|||
Bottle 1 |
Bottle 2 |
Bottle 3 |
Bottle 4 |
Mean |
Bottle 5 |
Bottle 6 |
|
1 |
8.5 |
7.3 |
9.6 |
9.6 |
9.6 |
31.0 |
32.4 |
2 |
10.7 |
9.6 |
11.3 |
11.3 |
11.3 |
43.7 |
40.9 |
3 |
12.4 |
11.9 |
13.6 |
13.0 |
13.3 |
49.36 |
50.7 |
4 |
16.4 |
14.1 |
15.3 |
14.7 |
15.0 |
55.0 |
56.4 |
5 |
21.5 |
18.1 |
17.0 |
15.8 |
16.4 |
60.6 |
59.2 |
6 |
24.3 |
20.3 |
17.5 |
15.8 |
16.7 |
60.6 |
60.6 |
7 |
27.3 |
23.7 |
18.7 |
17.5 |
18.1 |
63.4 |
64.8 |
8 |
29.4 |
24.9 |
19.2 |
17.5 |
18.4 |
64.8 |
66.2 |
9 |
31.7 |
26.0 |
19.8 |
18.1 |
19.0 |
64.8 |
67.6 |
10 |
34.5 |
28.8 |
21.5 |
19.8 |
20.7 |
70.5 |
71.9 |
11 |
35.6 |
28.8 |
20.9 |
18.7 |
19.8 |
66.2 |
67.6 |
12 |
36.2 |
29.4 |
19.8 |
18.1 |
19.0 |
62.0 |
66.2 |
13 |
38.4 |
31.7 |
21.5 |
19.8 |
20.7 |
67.6 |
70.5 |
14 |
39.6 |
32.8 |
21.5 |
19.8 |
20.7 |
66.2 |
69.0 |
15 |
40.7 |
33.3 |
22.0 |
19.8 |
20.9 |
66.2 |
69.0 |
16 |
42.4 |
34.5 |
22.0 |
20.3 |
21.2 |
66.2 |
70.5 |
17 |
44.1 |
35.6 |
23.2 |
20.9 |
22.1 |
67.6 |
71.9 |
18 |
45.8 |
37.3 |
24.3 |
22.0 |
23.2 |
70.5 |
74.7 |
19 |
46.9 |
38.4 |
24.9 |
23.2 |
24.1 |
71.9 |
74.7 |
20 |
49.2 |
40.1 |
26.6 |
23.7 |
25.2 |
73.3 |
77.5 |
21 |
50.3 |
41.8 |
27.1 |
24.9 |
26.0 |
74.7 |
78.9 |
22 |
50.9 |
41.3 |
26.6 |
23.7 |
25.2 |
73.3 |
77.5 |
23 |
52.0 |
43.0 |
27.7 |
24.9 |
26.3 |
76.1 |
80.3 |
24 |
53.7 |
44.1 |
28.8 |
25.4 |
27.1 |
77.5 |
81.7 |
25 |
54.8 |
45.2 |
29.4 |
26.0 |
27.7 |
78.9 |
84.5 |
26 |
55.4 |
45.8 |
30.0 |
26.0 |
28.0 |
77.5 |
83.1 |
27 |
56.0 |
46.3 |
30.0 |
26.0 |
28.0 |
78.9 |
83.1 |
28 |
57.1 |
46.9 |
30.5 |
26.6 |
28.6 |
80.3 |
83.1 |
Percentage biodegradation (%)
Time (d) |
Test suspension |
Procedure control |
Toxicity control |
|||
Bottle 1 |
Bottle 2 |
Average |
Difference |
Bottle 5 |
Bottle 6 |
|
1 |
-2.0 |
-4.3 |
-3.2 |
2.2 |
38.2 |
20.7 |
2 |
-1.1 |
-3.2 |
-2.1 |
2.0 |
57.9 |
26.9 |
3 |
-1.7 |
-2.6 |
-2.1 |
0.9 |
64.3 |
34.0 |
4 |
2.6 |
-1.7 |
1.3 |
4.3 |
71.4 |
37.6 |
5 |
9.4 |
3.2 |
6.3 |
6.3 |
78.9 |
38.9 |
6 |
14.2 |
6.8 |
10.5 |
7.4 |
78.5 |
40.0 |
7 |
17.0 |
10.4 |
13.7 |
6.7 |
80.9 |
42.4 |
8 |
20.5 |
12.1 |
16.3 |
8.3 |
83.0 |
43.5 |
9 |
23.6 |
13.1 |
18.3 |
10.6 |
81.9 |
44.2 |
10 |
25.6 |
15.1 |
20.4 |
10.6 |
89.0 |
46.6 |
11 |
29.3 |
16.7 |
23.0 |
12.6 |
82.9 |
43.4 |
12 |
31.9 |
19.4 |
25.6 |
12.6 |
76.9 |
43.0 |
13 |
32.9 |
20.5 |
26.7 |
12.4 |
83.8 |
45.3 |
14 |
35.1 |
22.5 |
28.8 |
12.6 |
81.3 |
44.0 |
15 |
36.7 |
23.0 |
29.8 |
13.7 |
80.9 |
43.7 |
16 |
39.4 |
24.7 |
32.0 |
14.6 |
80.4 |
44.9 |
17 |
40.8 |
25.1 |
33.0 |
15.7 |
81.3 |
45.3 |
18 |
41.9 |
26.2 |
34.1 |
15.7 |
84.6 |
46.9 |
19 |
42.3 |
26.6 |
34.4 |
15.7 |
85.4 |
46.0 |
20 |
44.5 |
27.7 |
36.1 |
16.9 |
86.0 |
47.6 |
21 |
45.0 |
29.3 |
37.1 |
15.7 |
87.0 |
48.1 |
22 |
47.7 |
29.9 |
38.8 |
17.8 |
86.0 |
47.6 |
23 |
47.6 |
30.9 |
39.3 |
16.7 |
88.9 |
49.1 |
24 |
49.3 |
31.5 |
40.4 |
17.8 |
90.0 |
49.6 |
25 |
50.2 |
32.4 |
41.3 |
17.8 |
91.4 |
51.6 |
26 |
50.7 |
33.0 |
41.8 |
17.8 |
88.4 |
50.1 |
27 |
51.8 |
33.9 |
42.9 |
18.0 |
90.9 |
50.1 |
28 |
52.9 |
34.0 |
43.4 |
18.9 |
92.4 |
49.6 |
Note:
①When there was a positive and negative in the percentage biodegradation of the test suspension, the negative was regarded as zero in the calculation of the mean.
②When they were both negative in the percentage biodegradation of the test suspension, the mean was directly calculated.
pH values
Test groups |
Test suspension |
Inoculum blank |
Procedure control |
Toxicity control |
||
Bottle 1 |
Bottle 2 |
Bottle 3 |
Bottle 4 |
Bottle 5 |
Bottle 6 |
|
At the start of the test |
7.38 |
7.38 |
7.36 |
7.39 |
7.34 |
7.38 |
At the end of the test |
7.27 |
7.27 |
7.27 |
7.29 |
7.40 |
7.27 |
pH values of the test solutions
Groups |
Abiotic control |
Test suspension |
Procedure control |
Inoculum blank 1 |
Inoculum blank 2 |
Toxicity control |
||
Test bottle No. |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
At the start of the test |
4.22 |
7.47 |
7.47 |
7.47 |
7.50 |
7.47 |
7.50 |
7.46 |
At the end of the test |
5.1 |
6.5 |
6.5 |
6.5 |
8.27 |
7.04 |
7.15 |
7.91 |
Oxygen uptake of each test bottle during the test (mg O2)
Time (d) |
Abiotic control |
Test suspension |
Procedure control |
Inoculum blank 1 |
Inoculum blank 2 |
Toxicity control |
||
Bottle 1 |
Bottle 2 |
Bottle 3 |
Bottle 4 |
Bottle 5 |
Bottle 6 |
Bottle 7 |
Bottle 8 |
|
1 |
1.02 |
2.94 |
3.56 |
3.31 |
7.58 |
3.12 |
1.50 |
7.04 |
2 |
1.54 |
4.83 |
5.85 |
5.71 |
32.22 |
5.00 |
2.39 |
38.15 |
3 |
1.87 |
6.71 |
8.48 |
8.58 |
40.44 |
6.91 |
3.14 |
52.73 |
4 |
2.21 |
8.79 |
11.00 |
11.89 |
52.83 |
9.02 |
4.00 |
66.35 |
5 |
2.44 |
11.08 |
13.72 |
15.49 |
63.72 |
11.14 |
4.85 |
72.51 |
6 |
2.62 |
13.74 |
16.89 |
19.26 |
71.33 |
13.20 |
5.69 |
77.05 |
7 |
2.87 |
16.29 |
20.66 |
22.62 |
74.76 |
15.26 |
6.60 |
81.24 |
8 |
3.19 |
18.72 |
24.14 |
25.47 |
77.72 |
16.99 |
7.39 |
84.92 |
9 |
3.44 |
20.99 |
26.97 |
28.07 |
79.74 |
18.51 |
8.08 |
88.65 |
10 |
3.67 |
23.14 |
29.59 |
30.57 |
81.32 |
19.89 |
8.70 |
92.09 |
11 |
3.92 |
25.18 |
32.03 |
33.01 |
82.68 |
21.20 |
9.33 |
95.50 |
12 |
4.23 |
27.28 |
34.49 |
35.32 |
83.99 |
22.45 |
9.93 |
98.56 |
13 |
4.52 |
29.18 |
36.94 |
37.46 |
85.22 |
23.74 |
10.60 |
101.23 |
14 |
4.79 |
30.84 |
39.13 |
39.42 |
86.44 |
24.97 |
11.25 |
103.56 |
15 |
5.02 |
32.34 |
41.09 |
41.15 |
87.74 |
26.09 |
11.87 |
105.65 |
16 |
5.27 |
33.84 |
43.13 |
42.82 |
89.19 |
27.26 |
12.54 |
107.85 |
17 |
5.54 |
35.44 |
45.38 |
44.44 |
90.69 |
28.45 |
13.27 |
110.29 |
18 |
5.79 |
37.19 |
47.36 |
46.02 |
92.07 |
29.78 |
14.06 |
113.10 |
19 |
6.04 |
38.96 |
49.21 |
47.46 |
93.25 |
31.28 |
14.87 |
116.02 |
20 |
6.73 |
40.98 |
51.33 |
49.15 |
94.67 |
33.32 |
15.89 |
119.06 |
21 |
6.73 |
42.25 |
52.48 |
50.25 |
95.27 |
34.80 |
16.29 |
120.93 |
22 |
6.73 |
43.55 |
53.75 |
51.50 |
96.02 |
36.32 |
16.70 |
122.76 |
23 |
6.81 |
44.67 |
54.98 |
52.92 |
96.71 |
37.69 |
17.06 |
124.30 |
24 |
6.96 |
45.77 |
56.25 |
54.31 |
97.34 |
38.92 |
17.41 |
125.78 |
25 |
7.16 |
46.86 |
57.75 |
55.71 |
97.98 |
40.00 |
17.85 |
127.26 |
26 |
7.41 |
47.96 |
59.37 |
57.08 |
98.54 |
40.98 |
18.24 |
128.70 |
27 |
7.58 |
49.02 |
61.00 |
58.37 |
98.96 |
41.80 |
18.58 |
129.99 |
28 |
7.64 |
49.63 |
61.81 |
59.12 |
99.17 |
42.23 |
18.72 |
130.70 |
Percentage biodegradation (%)
Time (d) |
Test suspension |
Procedure control |
Toxicity control |
|||
Bottle 2 |
Bottle 3 |
Bottle 4 |
Mean |
Bottle 5 |
Bottle 8 |
|
1 |
-0.4 |
1.1 |
0.5 |
0.8 |
7.3 |
3.2 |
2 |
-0.4 |
2.1 |
1.7 |
1.9 |
35.7 |
26.6 |
3 |
-0.5 |
3.8 |
4.1 |
4.0 |
44.7 |
36.8 |
4 |
-0.6 |
4.8 |
7.0 |
5.9 |
58.5 |
46.1 |
5 |
-0.2 |
6.3 |
10.6 |
8.5 |
70.5 |
49.3 |
6 |
1.3 |
9.0 |
14.8 |
11.9 |
78.6 |
51.3 |
7 |
2.5 |
13.2 |
18.0 |
15.6 |
81.6 |
53.0 |
8 |
4.2 |
17.5 |
20.7 |
19.1 |
84.2 |
54.6 |
9 |
6.1 |
20.7 |
23.4 |
22.0 |
85.8 |
56.4 |
10 |
7.9 |
23.7 |
26.1 |
24.9 |
87.0 |
58.0 |
11 |
9.7 |
26.4 |
28.8 |
27.6 |
87.8 |
59.7 |
12 |
11.8 |
29.4 |
31.4 |
30.4 |
88.7 |
61.2 |
13 |
13.3 |
32.2 |
33.5 |
32.9 |
89.4 |
62.3 |
14 |
14.3 |
34.6 |
35.3 |
34.9 |
90.0 |
63.2 |
15 |
15.3 |
36.6 |
36.8 |
36.7 |
90.9 |
63.9 |
16 |
16.1 |
38.8 |
38.0 |
38.4 |
91.8 |
64.8 |
17 |
17.1 |
41.3 |
39.0 |
40.2 |
92.7 |
65.8 |
18 |
18.1 |
42.9 |
39.7 |
41.3 |
93.4 |
67.0 |
19 |
18.8 |
43.8 |
39.5 |
41.6 |
93.9 |
68.1 |
20 |
18.7 |
44.0 |
38.7 |
41.3 |
94.4 |
68.9 |
21 |
18.2 |
43.2 |
37.7 |
41.3 |
94.4 |
68.9 |
22 |
17.7 |
42.6 |
37.1 |
39.8 |
95.0 |
69.5 |
23 |
17.0 |
42.2 |
37.2 |
39.7 |
95.4 |
69.6 |
24 |
16.7 |
42.3 |
37.6 |
40.0 |
95.7 |
69.8 |
25 |
16.8 |
43.4 |
38.4 |
40.8 |
96.0 |
70.1 |
26 |
17.0 |
44.9 |
39.3 |
42.1 |
96.2 |
70.5 |
27 |
17.6 |
46.9 |
40.5 |
43.7 |
96.3 |
70.9 |
28 |
18.1 |
47.8 |
41.2 |
44.5 |
96.4 |
71.1 |
Note: Replicate 1 of the test suspension was omitted, and the mean biodegradation rate was calculated by replicates 2, 3.
Residual amount of the test substance
Test Groups |
Measured residual concentration (mg/L) |
Actual residual concentration of each group (mg/L) |
Abiotic degradation (%) |
Primary biodegradation (%) |
Average (%) |
Abiotic control |
35.6 |
17.8 |
40.7 |
-- |
-- |
Test suspension 1 |
21.0 |
10.5 |
-- |
41.0 |
47.2 |
Test suspension 2 |
18.4 |
9.2 |
-- |
48.3 |
|
Test suspension 3 |
17.0 |
8.5 |
-- |
52.2 |
|
Inoculum blank 1 |
<LOD |
0 |
-- |
-- |
-- |
Note: The LOD of the analytical method of the test substance is 0.644 mg/L.
Summary of the test results
BOD measurement, pH
Bottle No. |
Sample description |
BOD (mgO2) |
pH |
||||
Day 7 |
Day 14 |
Day 21 |
Day 28 |
Day 0 |
Day 28 |
||
1 |
Sludge + test substance-1 (Test suspension-1) |
8.6 |
26.1 |
32.4 |
36.1 |
- |
7.5 |
2 |
Sludge + test substance-2 (Test suspension-2) |
6.1 |
23.0 |
32.4 |
40.8 |
- |
7.5 |
3 |
Sludge + reference substance (Activity control) |
42.8 |
47.6 |
48.6 |
49.5 |
- |
8.1 |
4 |
Inoculum blank-1 (Control blank-1) |
2.8 |
3.5 |
3.5 |
3.9 |
7.6 |
7.5 |
5 |
Inoculum blank-2 (Control blank-2) |
2.8 |
3.4 |
3.4 |
3.6 |
7.7 |
7.5 |
Measured values (day 28)
|
Sludge + test substance |
Theoretical value |
||
No. 1 |
No. 2 |
|||
Bottle 1 |
Bottle 2 |
|||
BOD*1 |
mgO2 |
32.3 |
37.0 |
103.3 |
DOC*1 |
mgC |
0.8 |
1.3 |
25.5 |
Test substance (GPC) |
mg |
16.9 |
11.1 |
30.0 |
% |
56 |
37 |
- |
|
*1: Values of [Sludge + test substance] were corrected with the average BOD or DOC value of [Inoculum blank]
Degradability (%)
|
Sludge + test substance |
|||
No. 1 |
No. 2 |
Average |
||
Bottle 1 |
Bottle 2 |
|||
BOD |
% |
31 |
36 |
34 |
DOC*2 |
% |
NA |
NA |
- |
Test substance |
% |
44 |
63 |
54 |
NA: Not available
*2: Degradability was not calculated because the test substance was hardly soluble in water.
Percentage biodegradation values after 7, 14, 21 and 28 days
Bottle No. |
Sample description |
|
Day 7 |
Day 14 |
Day 21 |
Day 28 |
ThOD mgO2 |
1 |
Sludge + test substance-1 (Test suspension-1) |
Measured value (mgO2) |
8.6 |
26.1 |
32.4 |
36.1 |
103.3 |
BODs – BODb (mgO2) |
5.8 |
22.6 |
28.9 |
32.3 |
|||
Degradability (%) |
6 |
22 |
28 |
31 |
|||
2 |
Sludge + test substance-2 (Test suspension-2) |
Measured value (mgO2) |
6.1 |
23.0 |
32.4 |
40.8 |
103.3 |
BODs – BODb (mgO2) |
3.3 |
19.5 |
28.9 |
37.0 |
|||
Degradability (%) |
3 |
19 |
28 |
36 |
|||
|
Average (Bottle 1,2) |
Degradability |
|
|
|
34 |
% |
3 |
Sludge + reference substance (activity control) |
Measured value (mgO2) |
42.8 |
47.6 |
48.6 |
49.5 |
50.0 |
BODs – BODb (mgO2) |
40.0 |
44.1 |
45.1 |
45.7 |
|||
Degradability (%) |
80 |
88 |
90 |
91 |
|||
4 |
Inoculum blank-1 (Control blank-1) |
Measured value (mgO2) |
2.8 |
3.5 |
3.5 |
3.9 |
- |
5 |
Inoculum blank-2 (Control blank-2) |
Measured value (mgO2) |
2.8 |
3.4 |
3.4 |
3.6 |
- |
|
Average (Bottle 4,5) |
BODb (mgO2) |
2.8 |
3.5 |
3.5 |
3.8 |
|
Equations:
Degradability (%) = (BODs-BODb)/ThODx100
BODs: Oxygen consumption (mgO2) in [Sludge + test substance] or [Sludge + reference substance]
BODb: Average of oxygen consumption (mgO2) in [Inoculum blank]
ThOD: Theoretical oxygen demand (mgO2) of the test substance or reference substance
Result of DOC measurement
Bottle No. |
Sample description |
Theoretical value |
Measured concentration |
DOC in bottle |
DOCs – DOCb |
Ratio to theoretical value |
Degradability |
mgC |
mgC/L |
mgC |
mgC |
% |
% |
||
1 |
Sludge + test substance-1 (Test suspension-1) |
25.5 |
3.669 |
1.1 |
0.8 |
3 |
NA |
2 |
Sludge + test substance-2 (Test suspension-2) |
25.5 |
51.77 |
1.6 |
1.3 |
5 |
NA |
Average (Bottle 1,2) |
- |
- |
- |
- |
4 |
- |
|
4 |
Inoculum blank-1 (Control blank-1) |
- |
1.136 |
0.3 |
- |
- |
- |
5 |
Inoculum blank-2 (Control blank-2) |
- |
1.145 |
0.3 |
- |
- |
- |
Average (Bottle 4,5) |
- |
- |
0.3 |
- |
- |
- |
|
NA: Not available
Equations:
Theoretical value = Added amount (mg) x Carbon ratio
Added amount: 30.0 mg
Carbon ration in the test substance: 0.851
DOC in Bottle (mgC) = Measured concentration x IV
IV: Volume of test solution in bottle 0.300L
Ratio to theoretical value = (DOCs-DOCb)/Theoretical value x 100
DOCs: DOC (mgC) in [Sludge + test substance]
DOCb: The average value of DOC (mgC) in [Inoculum blank]-1 and 2
Recovery, blank and detection limit
|
A |
B |
C |
D |
E |
Added amount |
Peak area |
Measured concentration |
Test substance in bottle |
Recovery |
|
|
mg |
mV∙sec |
mg/L |
mg |
% |
Recovery-1 |
30.0 |
3506.926 |
600 |
30.0 |
100.0 |
Recovery-2 |
30.0 |
3489.488 |
597 |
29.9 |
99.7 |
Blank |
0.0 |
ND |
<4 |
<0.2 |
- |
Detection limit |
|
20 |
4 |
0.2 |
- |
Average recovery |
|
|
|
|
99.9 |
ND: Not detected
Peak area of standard solution AS: 3504.907 mV∙sec
Concentration of standard solution CS: 600 mg/L
Final volume FV: 0.050 L
Sample volume SV: 0.300 L
Volume of test solution in bottle IV: 0.300 L
Equations:
C=B/ASxCS
D=CxFV/SVxIV
E=D/Ax100
Residual test substance amount in test bottles
|
A |
B |
C |
D |
E |
|
|
Bottle No. |
Sample description |
Initial amount |
Peak area |
Measured concentration |
Residual amount |
Residual test substance rate |
Degradability |
mg |
mV∙sec |
mg/L |
mg |
% |
% |
||
1 |
Sludge + test substance-1 (Test suspension-1) |
30.0 |
1978.856 |
337 |
16.9 |
56 |
44 |
2 |
Sludge + test substance-2 (Test suspension-2) |
30.0 |
1301.750 |
222 |
11.1 |
37 |
63 |
4 |
Inoculum blank-1 (Control blank-1) |
0.0 |
ND |
<4 |
<0.2 |
- |
- |
5 |
Inoculum blank-2 (Control blank-2) |
0.0 |
ND |
<4 |
<0.2 |
- |
- |
Detection limit |
- |
20 |
4 |
0.2 |
- |
- |
|
Average (Bottle 1,2) |
|
|
|
|
47 |
54 |
|
ND: Not detected
Peak area of standard solution AS: 3525.887 mV∙sec
Concentration of standard solution CS: 600 mg/L
Final volume FV: 0.050 L
Sample volume SV: 0.300 L
Volume of test solution in bottle IV: 0.300 L
Average recovery R: 99.9%
Equations:
C=B/ASxCS
D=CxFV/SVxIV/Rx100
E=D/Ax100
Degradability(%) = (Initial amount – Residual amount) / Initial amount x 100
RESULTS
|
Measurement |
[Sludge + test substance] |
[Water + test substance] |
Theoretical value |
|
BOD |
Measured value*1 |
mgO2 |
11.1 |
0.0 |
31.0 |
Degradability |
% |
36 |
- |
- |
|
DOC |
Measured value*1 |
mgC |
0.8 |
0.0 |
7.7 |
Detection rate |
% |
10 |
<1 |
- |
|
Degradability |
% |
NA |
- |
- |
|
Test substance |
Measured value |
mg |
3.2 |
9.0 |
9.0 |
Residual rate |
% |
36 |
100 |
- |
|
Degradability |
% |
64 |
- |
- |
|
*1 Values if [Sludge + test substance] were corrected with BOD or DOC value of [Inoculum blank].
Description of key information
Key value for chemical safety assessment
- Biodegradation in water:
- readily biodegradable but failing 10-day window
- Type of water:
- freshwater
Additional information
Arcadis - A19 -00070
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20343, based on CO2evolution and compared to the TOC, was calculated to be 61% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of about 7 days.
The biodegradation of TS20343 reached 35% at the end of the 10-day window.
The positive control, sodium benzoate, reached 83% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20343 did not reach the pass level of 60% for ready biodegradability in the CO2Evolution Test within the 10-d window, and therefore, cannot be termed as readily biodegradable. However, the pass level of 60% was reached after 28 days.
All validity criteria were fulfilled.
iBacon - 19/045 -914AN
Title: SynNova Base Oil: Ready Biodegradability in a Manometric Respirometry Test
Endpoints: Degradation rate of test item calculated by the oxygen consumption of the aerobic activated sludge microorganisms after 28 days of incubation.
Test Item Loading Rate (initial concentration in medium C0): 102.3 mg/L corresponding to an oxygen demand of about 349.3 mg/L (ThODNH4)
Reference Item: Sodium Benzoate
Reference Item Loading Rate: 103.7 mg/L corresponding to an oxygen demand of about 172.7 mg/L (ThODNH4)
Results:
Biodegradation of SynNova Base Oil: The mean biodegradation of 10% of SynNova Base Oil was reached at day 10 (ThODNH4). At the end of the 10- day window at day 20, the mean degradation of SynNova Base Oil was 26% (ThODNH4) and therefore the 10 day window criterion was not passed. The mean biodegradation at test end after 28 days was 34% (ThODNH4).
Therefore, SynNova Base Oil is considered to be not readily biodegradable based on ThODNH4.
Biodegradation of Sodium Benzoate: The reference item sodium benzoate was sufficiently degraded to 75% after 14 days and to 84% after 28 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used.
Biodegradation of the Toxicity Control: In the toxicity control containing both, the test item and the reference item sodium benzoate, 33% (ThODNH4) biodegradation was noted within 14 days and 47% (ThODNH4) biodegradation after 28 days of incubation. According to the test guidelines, the test item can be assumed to be not inhibitory to the aerobic activated sludge microorganisms because degradation was >25% within 14 days.
Arcadis - A19 -00601/e
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20817, based on CO2evolution and compared to the TOC, was calculated to be 65% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20817 reached 34% at the end of the 10-day window.
The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20817 did not reach the pass level of 60% for ready biodegradability in the CO2 Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
Arcadis - A19 -0061/f
Ready biodegradability in an aerobic aqueous medium CO2Evolution Test – OECD 301 B / ISO 9439
The biodegradability of TS20819, based on CO2evolution and compared to the TOC, was calculated to be 66% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 9 days.
The biodegradation of TS20819 reached 41% at the end of the 10-day window.
The positive control, sodium benzoate, reached 80% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% for ready biodegradability in the CO2Evolution Test within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
Arcadis - A19 -01461/a
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20817, based on CO2 evolution and compared to the TOC, was calculated to be 71% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 8 days.
The biodegradation of TS20817 reached 39% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20817 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
Arcadis-A19 -01461/b
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 45% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
Arcadis-A19 -01461/c
The applied test method is based on the guideline “OECD Guidelines for the Testing of Chemicals. Guideline 301: Ready Biodegradability, Method 301 B: CO2 Evolution Test, adopted July 1992” as well as on the norm “International Organization for Standardization – Water quality: Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium – Carbon dioxide evolution test, ISO 9439, 2nd edition 1999-03-01”.
The biodegradability of TS20819, based on CO2 evolution and compared to the TOC, was calculated to be 78% after an incubation time of 28 days.
Significant degradation of the test item was observed after a lag-phase of approximately 7 days.
The biodegradation of TS20819 reached 39% at the end of the 10-day window.
The positive control, sodium benzoate, reached 76% biodegradation after 14 days, thus confirming suitability of inoculum and test conditions.
TS20819 did not reach the pass level of 60% within the 10-d window, and therefore cannot be termed readily biodegradable. However, the pass level of 60% was reached within 28 days.
Guangdong Detection Center of Microbiology-2019ESG0177
OECD Guidelines for Testing of Chemicals, 301F Manometric Respirometry Test. Paris: OECD. Adopted 17th July, 1992.
Under the conditions of the study, the percentage biodegradation of the test substance on 28 d were respectively 52.9% and 34.0%, with a mean of 43.4%.
Since the test substance is a mixture consisting of structurally similar chemicals (UVCB), it was not confirmed that whether a sequential biodegradation of the individual structures was taking place, the 10-day window for ready biodegradability was not applied to interpret the results of the test based on “The Guidelines for the Testing of Chemicals Degradation”. And the guideline doesn't also give the clear directions on how to evaluate the test results of such mixtures. Therefore, no conclusions is given as to whether the test substance can be considered as readily biodegradable.
Guangdong Detection Center of Microbiology-2019ESG0178
OECD Guidelines for Testing of Chemicals,Inherent Biodegradability: Modified MITI Test (II). Paris: OECD. Adopted: 12th May, 1981.
On day 28, the residual test substance was extracted and measured. Results revealed that, the concentration of residual test substance in the abiotic control was 17.8 mg/L (abiotic degradation was 40.7%). The residual test substance concentrations in the test suspensions (three replicates) were 10.5 mg/L, 9.2 mg/L, 8.5 mg/L, and the primary biodegradation rates were respectively 41.0%, 48.3%, 52.2%, with a mean of 47.2%.
Discussion
The Inherent Biodegradability: Modified MITI Test (II) of SynNova Base Oil had been performed for twice, with the C.E.S Respirometer Systems.
In one of the studies, the oxygen consumption of the inoculum blank was 123.3mg/L, which significantly deviated from the historical data in our laboratory (The range of oxygen consumption in 64 inoculum blanks of 2013 to 2019 was 31.0 – 103.8 mg/L, with a mean of 66.3mg/L), so the results were considered unreliable and no further statistics was made.
In the other study, the primary biodegradation rates were respectively 41.0%, 48.3%, 52.2%, the BOD biodegradation rates were 18.1%, 47.8% and 41.2%. It showed that BOD biodegradation rate of replicate 1 had great difference on the other two replicates, which failed to reflect the real biodegradation rate, so that BOD biodegradation rate of replicate 1 (18.1%) for was considered as outlier, and eventually the BOD biodegradation percentage of the test substance was decided to the average biodegradation rate with replicates 2 and 3 of 44.5%. In the meantime, the ready biodegradability test (301F) of the test substance showed a BOD biodegradation rate of 43.4% on 28 days, and it believed that the average biodegradation rate with replicates 2 and 3 of 44.5% was more able to reflect the reality of biodegradation rate for the test substance.
Conclusion
Under the conditions of the study, percentage biodegradation rate of the test substance on day 28 which was calculated by BOD was 44.5%, and the primary biodegradation rates which was calculated by residual amount of the substance was 47.2%, therefore the test substance can be considered to be primary inherent biodegradable.
OECD 301C &F – LSI Mediscience
Ready biodegradability
It can be concluded that the test substance is not readily biodegradable and transformation product was generated under the conditions of this test.
There is a trend that the BOD curve has been keeping elevation after 28 days. Whilst the test substance does not degrade within the specified 28-day time span of the test, it is anticipated that degradation will occur within a short time frame.
The detected transformation products were those in which the end of the structure of the test substance was a carboxyl group.
Inherent biodegradability
From the degradability results based on the BOD (av. 36%) and the residual amount of the test substance (av. 64%), it can be concluded that the test substance is not readily biodegradable under the conditions of this test.
There is a trend that the BOD curve has been keeping elevation after 28 days. This would indicate that the study has a lag time, and post 28-days further degradation would be anticipated. Therefore, whilst the substance does not degrade within the specified 28-day time span of the test, it is anticipated that degradation will occur within a short time frame.
It is suggested that the test substance was transformed since a new peak was observed on the chromatogram of HPLC.
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