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EC number: - | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Biodegradation in water: screening tests
Administrative data
Link to relevant study record(s)
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- Test dates: Jun.18 to Jul. 22, 2013
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- Contradictory results between O2 uptake and analytical measurements.
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.4-E (Determination of the "Ready" Biodegradability - Closed Bottle Test)
- Deviations:
- yes
- Remarks:
- No effect on the study results
- Qualifier:
- according to guideline
- Guideline:
- other: NF EN ISO 9439
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Inspection date: 14 June 2012; Signature date: 10 Jan 2013
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- sewage, predominantly domestic, adapted
- Details on inoculum:
- - Source of inoculum/activated sludge: derived from surface waters collected in the near area of Pau, France and receiving predominantly domestic sewage
- Storage conditions: 20 ± 1°C.
- Storage length: 5 days
- Preparation of inoculum for exposure: The freshly collected sample of surface water was previously pre-conditioned to the experimental conditions under aerobic conditions for 5 days at the test temperature in the mineral medium used for testing. the resulting solution was coninuously stirred for 6 days at 20 +/- 1°C. The preconditioned inoculum was further used at a rate of 2ml/L - Duration of test (contact time):
- 32 d
- Initial conc.:
- 3.04 mg/L
- Based on:
- act. ingr.
- Initial conc.:
- 24.73 mg/L
- Based on:
- not specified
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium:
Mineral stock solutions were prepared as follows:
Stock solution A:
Monopotassium dihydrogen orthophosphate, KH2PO4: 8.50 g/L
Dipotassium hydrogen orthophosphate, K2HPO4: 21.75 g/L
Disodium hydrogen orthophosphate, dihydrate, Na2HPO4 2H2O: 33.40 g/L
Anunonium chloride, NH4Cl: 0.50 g/L
Stock solution B:
Calcium chloride anhydrous, CaCl2: 27.50 g/L
Or Calcium chloride dihydrate, CaCl2 2H2O: 36.40 g/L
Stock solution C:
Magnesium sulphate heptahydrate, MgSO4 7H2O: 22.50 g/L
Stock solution D:
Iron (III) chloride hexahydrate, FeCl3 6H2O: 2.5 g/L
The mineral medium was prepared using 1 mL/L of each the stock solutions A, B, C and D. The mineral medium was strongly aerated for at least 20 minutes, and then allowed to stand for approximately 20 h at the test temperature before use.
- Test temperature: 20 ± 1 °C
- Aeration of mineral medium: at least 20 minutes
- Continuous darkness: yes
TEST SYSTEM
- Culturing apparatus: 250 mL BOD bottles
- Number of culture flasks/concentration: 2
- Measuring equipment: electrode
SAMPLING
- Sampling frequency: at test initiation and at the end of test (day 32).
CONTROL AND BLANK SYSTEM
- Inoculum blank: 300 mL of mineral medium + 0.6 mL of pre-conditioned inoculum.
- Toxicity control: 300 mL of the test item solution + 100 µL of the Aniline stock solution + 0.6 mL of pre-conditioned inoculum.
- Reference group: A stock solution was prepared using 0.1217 g Aniline for 20 mL of water. 300 mL of the above test item solution were added with 100 µL of the aniline stock solution and 0.6 mL of pre-conditioned inoculum. - Reference substance:
- aniline
- Remarks:
- Sigma Reference A9880 (> 99.0%), batch No.034K0126
- Test performance:
- The test was terminated after 32 days of incubation.
The test was considered as valid:
- Mean oxygen uptake in the blank vessels < 1.5 mg/L on day 32,
- The residual concentration of oxygen in the test vessels did not fall below 0.5 mg/L at any time,
- Differences of extremes of replicate values of the removal of the test item was less than 20 % of mean value at the end of test,
- Percentage biodegradation of the reference item (aniline) had reached the pass-level by day 14,
- In the toxic control series more than 25 % biodegradation had occurred on day 14. - Parameter:
- % degradation (test mat. analysis)
- Value:
- 53.7
- Sampling time:
- 28 d
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- < 60
- Sampling time:
- 32 d
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 28
- Sampling time:
- 28 d
- Details on results:
- The test was performed using a 3.04 mg/L test solution (mean measured concentration). In the test item treatment, the beginning of the 10-day window was observed between day 4 and day 8 of the test period but the pass-level (60 %) was not reached within the 32-day observation period. The oxygen concentration stabilized at approximately 6.0 mg/L on day 18 and onwards.
In the Toxic group, 25% biodegradation was achieved within 8 days. Test item was thus considered as not inhibitory.
Analytical check of the test item in the test solutions and in the toxic control proved that test item was decreased to approximately 45-55% of the initial concentration on day 15 and onwards. Test item was thus considered as not easily biodegradable. - Results with reference substance:
- Aniline: The beginning of the 10-day window was observed between day 4 and day 8 of the test period. The pass-level (60 %) was reached on day 8: mean value for biodegradation was 64.9 %. At the end of the test period the biodegradation was > 90 %.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- For the test item, the pass-level (60 %) was not reached within the 32-day observation period. Thus, test item should be regarded as not readily biodegradable according to this test.
- Executive summary:
The Ready Biodegradability of test item was determined by the Closed bottle test. For the test item, the pass-level (60 %) was not reached within the 32-day observation period.
Thus, test item should be regarded as not readily biodegradable according to this test.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- Test dates: Dec.16, 2013 to Jan. 14, 2014
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- unsuitable test system
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)
- Qualifier:
- according to guideline
- Guideline:
- other: NF EN ISO 9439
- Principles of method if other than guideline:
- Not applicable
- GLP compliance:
- no
- Details on inoculum:
- - Source of inoculum/activated sludge: Aeration tank of sewage treatment plant, (Pierre-Bénite) Rhône
- Pretreatment: pre-conditioning, 5 days
- Initial cell/biomass concentration: 1.62 x 10^6 micro -organisms/ml of inoculum - Duration of test (contact time):
- 28 d
- Initial conc.:
- 24.15 mg/L
- Based on:
- test mat.
- Initial conc.:
- 24.73 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- TOC removal
- Details on study design:
- TEST CONDITIONS
- Composition of medium: Mineral medium prepared according to OECD guideline 301B
- Test temperature: 21°C ±1°C
TEST SYSTEM
- Number of culture flasks/concentration: 2
- Measuring equipment: Ba(OH)2 titration with HCl (0,05 M) - Reference substance:
- benzoic acid, sodium salt
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Remarks:
- 10-day window from day 8 to day 18
- Value:
- 24
- Sampling time:
- 18 d
- Key result
- Parameter:
- % degradation (CO2 evolution)
- Value:
- 28
- Sampling time:
- 28 d
- Results with reference substance:
- Biodegradability of the reference substance (Na benzoate) was found to be 71.8 % in 14 days
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- For the test item, biodegradability was < 60 % after 28-day. Thus, test item should be regarded as not readily biodegradable according to this test.
- Executive summary:
The Ready Biodegradability of test item was determined by the CO2 Evolution Test according to OECD Guideline 301 B. For the test item biodegradability was < 60 % after 28-day.
Thus, test item should be regarded as not readily biodegradable according to this test.
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From 26 November to 28 January 2022
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
- Qualifier:
- according to guideline
- Guideline:
- ISO 10707 Water quality - Evaluation in an aqueous medium of the "ultimate" aerobic biodegradability of organic compounds - Method by analysis of biochemical oxygen demand (closed bottle test)
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.4-E (Determination of the "Ready" Biodegradability - Closed Bottle Test)
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- inspected on 12 - 13 November 2019 / signed on 09 december 2019
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, non-adapted
- Details on inoculum:
- Activated sludge was obtained from the wastewater treatment plant Nieuwgraaf in Duiven, The Netherlands. This plant is an activated sludge treatment plant treating predominantly domestic wastewater. The dry weight of the inoculum was determined by filtrating 50 mL of the activated sludge over a preweighed 12 μm cellulose nitrate filter. This filter was dried for 1.5 hour at 103.0 °C and weighed after cooling. Dry weight was calculated by subtracting the weight of the filters and dividing the difference by the filtered volume. The measured dry weight of the inoculum was 3.0 g/L.
The activated sludge was preconditioned to reduce the endogenous respiration rates. To this end the inoculum was diluted in aerated Closed Bottle test medium to 0.4 g Dry weight (DW)/L of activated sludge and aerated for one week. The preconditioned inoculum was diluted further to a dry weight concentration of 2 mg/L in the BOD bottles (van Ginkel and Stroo, 1992). The Colony forming units (CFU) of the preconditioned and diluted inoculum was determined by a colony count method based on the ISO 6222 (1999) guideline. The preconditioned and diluted inoculum as used in the closed bottles (2 mg/L dry weight) was diluted 10x and 100x in a sterile peptone solution (1 g/L). Subsequently 1 ml of the peptone dilutions was transferred on a sterile petri dish and yeast extract agar was added. The yeast extract agar contained per liter of water 6 g tryptone, 3 g yeast extract and 15 g agar.
Yeast extract agar plates were incubated for 68 hours at a temperature ranging from 22.7 – 22.9 °C. Only CFU counts between 30 and 300 were regarded as accurate and accepted for calculation of the CFU content. The inoculum concentration in the BOD bottles determined by colony count was 9.105 CFU/L. - Duration of test (contact time):
- 60 d
- Initial conc.:
- 2 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Test bottles
The test was performed in 0.30 L BOD (biological oxygen demand) bottles with glass stoppers.
Nutrients and stocks
Deionized water used in the Closed Bottle test contained per liter of water 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.41 mg Na2HPO4.2H2O, 22.5 mg MgSO4·7H2O, 27.5 mg CaCl2 2H2O, 0.25 mg FeCl3·6H2O. Ammonium chloride was omitted from the medium to prevent nitrification that is not related to the biodegradation of the test substance.
Accurate administering of the test substance was accomplished by preparing a solid stock of 3.0 mg of test substance per g of silica gel in a 50-mL serum flask. Only part of the top layer of the silica gel was brought into contact with the test substance. The serum flask was closed with a screw cap and the content was mixed vigorously until there were no more lumps in the silica gel, indicating homogeneous distribution of the test substance. Subsequently, 0.20 g of silica gel with the test substance was added to the test bottles. The resulting concentration of test substance in the bottles was 2.0 mg/L. Next the bottles were filled with nutrient medium with inoculum and closed. Sodium acetate was added to the bottles using aqueous stock solution of 1.0 g/L.
Test procedures
The Closed Bottle test (OECD TG 301D) was performed according to the study plan. The study plan was developed from ISO Test Guidelines (1994). Use was made of 10 bottles containing only inoculum, 10 bottles containing inoculum and silica gel (0.2 g silica gel / bottle), 10 bottles containing inoculum and silica gel with test substance, 6 bottles containing inoculum and sodium acetate. The concentrations of the test substance, and sodium acetate in the bottles were 2.0 mg/L and 6.7 mg/L, respectively. Each of the prepared solutions was dispensed into the respective group of BOD bottles so that all bottles were completely filled without air bubbles. The zero-time bottles were immediately analyzed for dissolved oxygen using an oxygen electrode. The remaining bottles were closed and incubated in the dark. Two duplicate bottles of all series were withdrawn for analyses of the dissolved oxygen concentration at day 7, 14, 21, and 28. One extension from the protocol of the Closed Bottle test was introduced. The Closed Bottle test was prolonged by measuring the course of the oxygen decrease at day 42 and 60 using the bottles of day 28 and a special funnel. This funnel fitted exactly in the BOD bottle. Subsequently, the oxygen electrode was inserted in the BOD bottle to measure the oxygen concentration. The medium dissipated by the electrode was collected in the funnel. After withdrawal of the oxygen electrode the medium collected flowed back into the BOD bottle, followed by removal of the funnel and closing of the BOD bottle (van Ginkel and Stroo 1992).
Analyses
The dissolved oxygen concentrations were determined electrochemically using an oxygen electrode and meter (WTW). The pH was measured using an Eutech pH meter. The temperature was measured and recorded with a sensor connected to a data logger. - Reference substance:
- acetic acid, sodium salt
- Remarks:
- Purity: 99.8%; Batch n° BCCD4856
- Preliminary study:
- A non-GLP screening test was performed prior the GLP final test in order to determine the most suitable inoculum for the degradation of the test substance, in the Closed Bottle test.
Substance is classified not ready biodegradable based on the 16% biodegradation achieved at day 84 in the OECD 301D test using activated sludge as inoculum and 4% biodegradation achieved at day 84 with river water.
For the final GLP test it is recommended to use activated sludge as inoculum. - Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 3
- Sampling time:
- 28 d
- Parameter:
- % degradation (O2 consumption)
- Value:
- 8
- Sampling time:
- 60 d
- Details on results:
- The molecular formula of the known isomer constituents of the test substance was used as representative for all organics present in the test substance (assuming 100% purity of test substance). The theoretical oxygen demand (ThOD) of test substance calculated from this molecular formula and used to calculate the biodegradation percentages is 3.00 g oxygen/g test substance. The ThOD of sodium acetate is 0.78 g oxygen/g sodium acetate.
Toxicity
Inhibition of the degradation of a well-degradable compound, e.g. sodium acetate by the test substance in the Closed Bottle test is optional in the OECD guideline and was not determined because possible toxicity of the test substances to microorganisms degrading acetate is not relevant. Inhibition can be detected prior to the onset of the biodegradation through suppression of the oxygen consumption in the presence of the test substance (higher oxygen concentration in bottles with test substance compared to the concentration in the control bottles). Inhibition of the endogenous respiration of the inoculum by the test substance at day 7 was not detected. Therefore, no inhibition of the biodegradation due to the "high" initial test substance concentration is expected.
Test conditions
At the start of the test the pH of the media in the duplicate control, reference substance, control with silica gel, and test substance bottles was 7.0. The pH of the medium in the duplicate reference bottles measured at day 14 was 6.9. The pH of the medium in the duplicate bottles at day 60 was 7.1, 7.1 and 7.0 for the control, the control with silica gel and the test substance bottles, respectively. The temperature ranged from 22.4 to 22.9 °C which is within the prescribed temperature range of 20 to 24°C.
Validity of the test
The validity of the test is demonstrated by an endogenous respiration of 0.80 mg/L at day 28 . Furthermore, the differences of the replicate values at day 28 were less than 20%. The biodegradation percentage of the reference compound, sodium acetate, at day 14 was 79%. Finally, the validity of the test is shown by oxygen concentrations >0.5 mg/L in all bottles during the test period.
Biodegradability
TEst item was biodegraded by 3% at day 28 in the Closed Bottle test. The test substance should therefore not be classified as readily biodegradable. In the prolonged Closed Bottle test (enhanced biodegradability test) the test substance was biodegraded by 8% at day 60. Biodegradation percentages below 20% suggest that there is only minor partial degradation of the test substance achieved in the (prolonged) Closed Bottle test. Test item is therefore regarded not biodegradable in the OECD 301D ready biodegradation screening test. The lack of complete biodegradation in the Closed Bottle test does not mean that the test substance is recalcitrant in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test.
Test item is classified as not biodegradable in ready biodegradation screening test based on <20% biodegradation achieved in the OECD 301D test at day 60 - Results with reference substance:
- The biodegradation percentage of the reference compound, sodium acetate, at day 14 was 79%
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- not readily biodegradable
- Conclusions:
- The test item was biodegraded by 3% at day 28 in the Closed Bottle test. The test substance should therefore not be classified as readily biodegradable. In the prolonged Closed Bottle test (enhanced biodegradability test) the test substance was biodegraded by 8% at day 60. Biodegradation percentages below 20% suggest that there is only minor partial degradation of the test substance achieved in the (prolonged) Closed Bottle test. Test item is therefore regarded not biodegradable in the OECD 301D ready biodegradation screening test. The lack of complete biodegradation in the Closed Bottle test does not mean that the test substance is recalcitrant in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test.
The test item is classified as not biodegradable in ready biodegradation screening test based on <20% biodegradation achieved in the OECD 301D test at day 60 - Executive summary:
To assess the biotic degradation of test item a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the Closed Bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice.
The test substance (2 mg/L) was exposed to activated sludge, which was spiked to a mineral nutrient solution, dosed in closed bottles, and incubated in the dark at 22.4 to 22.9 °C for 60 days. The degradation of the test item was assessed by the measurement of oxygen consumption. According to the results of this study, the test item did not cause a reduction in the endogenous respiration at day 7. The test substance is therefore considered to be non-inhibitory to the inoculum. the test item was biodegraded by 3% at day 28 in the Closed Bottle test. The test substance should therefore not be classified as readily biodegradable. In the prolonged Closed Bottle test (enhanced biodegradability test) the test substance was biodegraded by 8% at day 60. Biodegradation percentages below 20% suggest that there is only minor partial degradation of the test substance achieved in the (prolonged) Closed Bottle test. the test item is therefore regarded not biodegradable in the OECD 301D ready biodegradation screening test. The lack of complete biodegradation in the Closed Bottle test does not mean that the test substance is recalcitrant in nature because the stringency of the test procedures could account for the recalcitrance in the Closed Bottle test.
the test item is classified as not biodegradable in ready biodegradation screening test based on <20% biodegradation achieved in the OECD 301D test at day 60. The test is valid as shown by an endogenous respiration of 0.80 mg/L and by the total mineralization of the reference compound, sodium acetate. Sodium acetate was degraded by 79% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.
Referenceopen allclose all
Active Ingredient Content
Measured initial concentrations ranged between 95.0 and 104.7 % of the nominal value for the Test group, and between 96.9 and 104.8 % for the Toxic group. Mean value was 3.04 mg/L. The treatment application was considered as valid. The initial concentration was thus taken as 3.04 mg a.i./L, corresponding to 9.12 ThOD/L.
On day 15. Mean recovery of the test item was 56.2 % for the test group and 48.9 % for the Toxic group.
On day 28, mean recovery was 46.3 % for the test group and 44.4 % for the Toxic group.
On day 32. The primary degradation of the test item was considered as stabilized: on this day, mean recovery was 47.6 % for the test group and 43.2 % for the Toxic group.
Table 5.2.1.1: Mean Measured concentrations of test item throughout the test period
Day |
|
Test item (mg/L) |
% of nominal value |
Day 0 |
Test group |
3.03 |
98.2 |
Toxic control |
3.07 |
99.7 |
|
Day 15 |
Test group |
1.73 |
56.2 |
Toxic control |
1.51 |
48.9 |
|
Day 28 |
Test group |
1.43 |
46.3 |
Toxic control |
1.37 |
44.4 |
|
Day 32 |
Test group |
1.47 |
47.6 |
Toxic control |
1.33 |
43.2 |
Oxygen Uptake and Biodegradation
The initial value for dissolved oxygen concentration was 8.57 mg/L for the control, 8.61 mg/L for the test item treatment, 8.59 mg/L for the toxic control, and 8.62 mg/L for the reference item treatment.
The consumption of oxygen in the controls was less than 1.5 mg/L over the test period, as required. For the test item treatment, the oxygen concentration stabilised at approximately 6.0 mg/L on day 18 and onwards. The test was continued until day 32 without changes.
The calculated values for biodegradation, %, are presented below.
Table 5.2.1.2: Calculated mean biodegradation for test item based on oxygen uptake, %
Day |
Test item |
Toxic control |
Anilline (reference item) |
0 |
0 |
0 |
0 |
4 |
9.59 |
6.50 |
5.12 |
8 |
17.27 |
35.50 |
64.87 |
11 |
20.78 |
37.65 |
74.10 |
15 |
24.84 |
44.58 |
74.51 |
18 |
29.88 |
47.15 |
85.47 |
21 |
29.99 |
44.51 |
91.93 |
28 |
28.46 |
46.15 |
93.37 |
32 |
29.44 |
50.22 |
93.16 |
Test item: The beginning of the 10-day window was observed between day 4 and day 8 of the test period but the pass-level (60 %) was not reached within the 32-day testing period: the biodegradation was 29% as a maximum.
Toxic control: Based on total ThOD, the biodegradation was more than 25 % up from day 8 of the test period.
Table 5.2.1.1: CO2release and biodegradability of test item and reference substance
Time (days) |
Test item |
Reference substance (Na benzoate) |
||
Average Cumulated CO2released (mg) |
Biodegradability (%) |
CO2released * (mg) |
Biodegradability (%) |
|
2 |
0.57 |
0.49 |
18.94 |
17.22 |
3 |
2.20 |
1.91 |
32.90 |
29.93 |
4 |
3.38 |
2.94 |
36.70 |
33.39 |
7 |
8.69 |
7.58 |
58.04 |
52.80 |
8 |
11.70 |
10.18 |
67.96 |
61.82 |
11 |
14.36 |
12.44 |
74.40 |
67.68 |
14 |
19.87 |
17.24 |
78.88 |
71.75 |
18 |
27.25 |
23.72 |
85.36 |
77.65 |
22 |
29.00 |
25.22 |
92.35 |
84.00 |
25 |
29.76 |
25.90 |
95.02 |
86.43 |
28 |
32.52 |
28.34 |
97.01 |
88.25 |
*CO2released = total CO2production –average CO2production in blanks
Validity of the Test
Test was considered to be valid as
- Biodegradability of the reference substance (Na benzoate) was 71.8 % in 14 days
- Total CO2 released in blanks was 48.57 et 39.47 mg CO2/l in 28 days
- Difference of extremes of replicate value (removal of test chemical) was less than 20%
Table 5.2.1/1: Dissolved oxygen concentrations (mg/L) in the closed bottles
Time (days) | Oxygen concentration (mg/L) | |||
| Mc | Ma | Mcs | Mt |
0 | 9.00 | 9.00 | 9.00 | 9.00 |
| 9.00 | 9.00 | 9.00 | 9.00 |
Mean (M) | 9.00 | 9.00 | 9.00 | 9.00 |
7 | 8.6 | 4.7 | 8.3 | 8.3 |
| 8.5 | 4.6 | 8.4 | 8.4 |
Mean (M) | 8.55 | 4.65 | 8.35 | 8.35 |
14 | 8.3 | 4.3 | 8.2 | 8.1 |
| 8.4 | 4.1 | 8.3 | 8.1 |
Mean (M) | 8.35 | 4.20 | 8.25 | 8.10 |
21 | 8.4 |
| 8.2 | 7.9 |
| 8.4 |
| 8.2 | 7.9 |
Mean (M) | 8.4 |
| 8.2 | 7.9 |
28 | 8.2 |
| 8.0 | 7.9 |
| 8.2 |
| 8.0 | 7.8 |
Mean (M) | 8.2 |
| 8.00 | 7.85 |
42 | 8.1 |
| 7.9 | 7.7 |
| 8.0 |
| 7.9 | 7.5 |
mean | 8.05 |
| 7.90 | 7.60 |
60 | 8.0 |
| 7.8 | 7.4 |
| 7.9 |
| 7.9 | 7.3 |
Mean | 7.95 |
| 7.85 | 7.35 |
Mc = Mineral nutrient solution with only inoculum.
Mcs = Mineral nutrient solution with inoculum, and silica gel
Mt = Mineral nutrient solution with inoculum, test substance (2.0 mg/L test substance) and silica gel
Ma = Mineral nutrient solution with inoculum and sodium acetate (6.7 mg/L).
Table 5.2.1/2: Oxygen consumption (mg/L) and the percentages biodegradation of the test substance (BOD/ThOD) and sodium acetate (BOD/ThOD) in the Closed Bottle test
Time (days) | Oxygen consumption (mg/L) | Biodegradation (%) | ||
| Test substance | Acetate | Test substance | Acetate |
0 | 0.00 | 0.00 | 0 | 0 |
7 | 0.00 | 3.90 | 0 | 75 |
14 | 0.15 | 4.15 | 3 | 79 |
21 | 0.30 |
| 5 |
|
28 | 0.15 |
| 3 |
|
42 | 0.30 |
| 5 |
|
60 | 0.50 |
| 8 |
|
Description of key information
OECD Guideline 301D, GLP, Key study, validity 1:
3% biodegradation after 28 days and 8% biodegradation after 60 days, with activated sludge
Not readily biodegradable
Key value for chemical safety assessment
- Biodegradation in water:
- not biodegradable
- Type of water:
- freshwater
Additional information
To assess the biodegradation potential of the registered substance, three experimental studies are available.
The most recent study (Nouryon, 2022), assessed as the key study, was performed on the registered substance to assess the ready biodegradability of the test item in an aerobic aqueous medium, according to OECD Guideline 301D, , performed under GLP (with statement included in the report). The silica gel with test substance (2 mg/L) was exposed to activated sludge, which was spiked to a mineral nutrient solution, dosed in closed bottles, and incubated in the dark at 22.4 to 22.9 °C for 60 days. The degradation of the test item was assessed by the measurement of oxygen consumption. According to the results of this study, the test item did not cause a reduction in the endogenous respiration at day 7. The test substance is therefore considered to be non-inhibitory to the inoculum. The test item was biodegraded by 3% at day 28 in the Closed Bottle test. In the prolonged Closed Bottle test (enhanced biodegradability test) the test substance was biodegraded by 8% at day 60. the test substance is classified as not biodegradable in ready biodegradation screening test based on <20% biodegradation achieved in the OECD 301D test at day 60. The test is valid.
The second study (Phytosafe, 2013) was also performed on the registered substance according to the closed-bottle test method. The solution of the test substance (3.04 mg/L) in mineral medium was inoculated with a relatively small number of micro-organisms from a mixed population and kept in completely filled, closed bottles in the dark at constant temperature (20 +/- 1 °C). Degradation was followed by analysis of dissolved oxygen over a 28-day period. Additional analysis assessments were performed at test initiation and at test completion in the test vessels for quantification of the test item, so as to judge upon the primary degradation over the test period. The calculated values for biodegradation of the test substance were 28% at day 28 base on oxygen up-take but measured concentration of test item at day 28 was 46.3%. Therefore, the test substance can be considered as not readily biodegradable. This result was slightly different that of the key study because of purity of the sample.
A third study was also available according to OECD guideline 301 B and NF EN ISO 9439, but not performed under GLP. The test item was biodegraded by 28% at day 28. Therefore, the test substance can be considered as not readily biodegradable. This study supports result of the key study. This result was slightly different that of the key study because of purity of the sample.
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