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EC number: 270-192-0 | CAS number: 68412-48-6
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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:
- October 26, 2017 to November 23, 2017
- 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 (adaptation not specified)
- 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 set to deposit for 30 minutes, then 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.:
- >= 24.9 - <= 25 other: mg/flask
- 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, Aqualytic Dortmund, Germany
Principle: The test flasks were prepared and 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, Aqualytic Dortmund, Germany. 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: 7.2 to 7.6 (measured at the start of the test) 7.2 to 7.6 (measured at the end of the test)
Recording: Test conditions (temperature) was 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 recorded.)
Test Water
Reconstituted Test Water: Analytical grade salts were added to pure water to prepare the following stock solutions:
a) 8.5 g KH2PO4, 21.75 g K2HPO4, 33.4 g Na2HPO4 x 2H2O, 0.5 g NH4Cl filled up with pure water to
1000 mL volume; The pH-value was 7.4.
b) 11.25 g MgSO4 x 7 H2O filled up with pure water to 500 mL volume
c) 18.2 g CaCl2 x 2 H2O filled up with pure water to 500 mL volume
d) 0.125 g FeCl3 x 6 H2O filled up with pure water to 500 mL volume
In order to avoid precipitation of iron hydroxide in the stock solution d), one drop of concentrated HCl per litre was added before storage.
50 mL of stock solution a) and 5 mL of the stock solutions b) to d) were combined and filled up to a final volume of 5000 mL with pure water.
Course of the Test
Preparation of Test Flasks: The amounts of test item and reference item were directly weighed into the test flasks. No emulsifiers or solvents were used, but the solutions were dispersed by stirring to achieve a homogeneous solution 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, Aqualytic Dortmund, Germany) each day.
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
- Test performance:
- 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.
2-Propanone, reaction products with diphenylamine never reached 10% biodegradation. The mean biodegradation at test end after 28 days was 0% (ThODNH4 and ThODNO3). - Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 0
- Sampling time:
- 28 d
- Details on results:
- Biodegradation of Test Item
Percentage Biodegradation: The mean biodegradation percentage at the end of the 28-day exposure period was 0% (based on ThODNH4 and ThODNO3).
Conclusion: 2-Propanone, reaction products with diphenylamine is considered to be not readily biodegradable.
Biodegradation in the Toxicity Control
Percentage Biodegradation: In the toxicity control containing both, the test item and the reference item sodium benzoate, 26% and 25% after 14 days and to 27% and 26% after 28 days of incubation (ThODNH4.and ThODNO3). 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.
Conclusion: 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.
Abiotic Control
Oxygen Demand: The oxygen demand in the abiotic control was 0 mg/L during the test duration. There was no use to correct the degradation of the test item and toxicity control. - Results with reference substance:
- Percentage Biodegradation: The reference item sodium benzoate was sufficiently degraded to 69% after 14 days and to 67% 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 mean biodegradation percentage at the end of the 28-day exposure period was 0% (based on ThODNH4 and ThODNO3).
2-Propanone, reaction products with diphenylamine is considered to be not readily biodegradable. - Executive summary:
Summary
Title: 2-Propanone, reaction products with diphenylamine: 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: 2-Propanone, reaction products with diphenylamine; Batch No.: EL5D06K347
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 2-Propanone, reaction products with diphenylamine 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 296.5 mg/L (ThODNH4) and 319.5 mg/L (ThODNO3)
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 2-Propanone, reaction products with diphenylamine: 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.
2-Propanone, reaction products with diphenylamine never reached 10% biodegradation. The mean biodegradation at test end after 28 days was 0% (ThODNH4and ThODNO3).
Therefore, 2-Propanone, reaction products with diphenylamine is considered to be not readily biodegradable based on ThODNH4.and ThODNO3
Biodegradation of Sodium Benzoate: The reference item sodium benzoate was sufficiently degraded to 69% after 14 days and to 67% 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, 26% and 25% after 14 days and to 27% and 26% after 28 days of incubation (ThODNH4.and ThODNO3). 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.
Reference
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 |
0 |
5 |
45 |
0 |
35 |
2 |
0 |
5 |
5 |
5 |
70 |
0 |
60 |
3 |
5 |
5 |
5 |
5 |
80 |
0 |
70 |
4 |
5 |
5 |
5 |
5 |
90 |
0 |
80 |
5 |
10 |
5 |
5 |
5 |
105 |
0 |
110 |
6 |
10 |
10 |
10 |
10 |
110 |
0 |
115 |
7 |
10 |
10 |
10 |
10 |
120 |
0 |
120 |
8 |
10 |
15 |
10 |
10 |
125 |
0 |
130 |
9 |
10 |
15 |
10 |
10 |
125 |
0 |
130 |
10 |
10 |
15 |
10 |
15 |
130 |
0 |
135 |
11 |
10 |
15 |
15 |
15 |
130 |
0 |
135 |
12 |
15 |
15 |
15 |
15 |
130 |
0 |
135 |
13 |
15 |
15 |
15 |
15 |
130 |
0 |
135 |
14 |
15 |
20 |
15 |
15 |
135 |
0 |
135 |
15 |
15 |
20 |
15 |
15 |
135 |
0 |
140 |
16 |
20 |
20 |
15 |
20 |
135 |
0 |
140 |
17 |
20 |
20 |
20 |
20 |
135 |
0 |
140 |
18 |
20 |
20 |
20 |
20 |
140 |
0 |
145 |
19 |
20 |
20 |
20 |
20 |
140 |
0 |
150 |
20 |
20 |
20 |
20 |
20 |
140 |
0 |
150 |
21 |
20 |
20 |
20 |
20 |
140 |
0 |
150 |
22 |
25 |
25 |
25 |
25 |
140 |
0 |
150 |
23 |
25 |
25 |
25 |
25 |
140 |
0 |
150 |
24 |
25 |
25 |
25 |
25 |
140 |
0 |
150 |
25 |
25 |
25 |
25 |
25 |
140 |
0 |
150 |
26 |
25 |
25 |
25 |
25 |
140 |
0 |
150 |
27 |
25 |
25 |
30 |
30 |
145 |
0 |
155 |
28 |
25 |
25 |
30 |
30 |
145 |
0 |
155 |
Flasks 1 and 2: 2-Propanone, reaction products with diphenylamine
Flasks 3 and 4: inoculum control
Flask 5: reference (procedure control)
Flask 6: abiotic control
Flask 7: toxicity control
Percentage Biodegradation of Test Item, of Sodium Benzoate and of the Toxicity Control based on ThODNH4
Time (days) |
Percentage Biodegradation1 |
|||
2-Propane, reaction products with diphenylamine1 |
Sodium Benzoate2 |
Toxicity control1,2 |
||
Flask 1 [%] |
Flask 2 [%] |
Flask 5 [%] |
Flask 7 [%] |
|
1 |
0 |
0 |
25 |
7 |
2 |
0 |
0 |
38 |
12 |
3 |
0 |
0 |
43 |
14 |
4 |
0 |
0 |
49 |
16 |
5 |
2 |
0 |
58 |
23 |
6 |
0 |
0 |
58 |
23 |
7 |
0 |
0 |
64 |
24 |
8 |
0 |
2 |
67 |
26 |
9 |
0 |
2 |
67 |
26 |
10 |
0 |
1 |
68 |
26 |
11 |
0 |
0 |
67 |
26 |
12 |
0 |
0 |
67 |
26 |
13 |
0 |
0 |
67 |
26 |
14 |
0 |
2 |
69 |
26 |
15 |
0 |
2 |
69 |
27 |
16 |
1 |
1 |
68 |
26 |
17 |
0 |
0 |
67 |
26 |
18 |
0 |
0 |
69 |
27 |
19 |
0 |
0 |
69 |
28 |
20 |
0 |
0 |
69 |
28 |
21 |
0 |
0 |
69 |
28 |
22 |
0 |
0 |
67 |
27 |
23 |
0 |
0 |
67 |
27 |
24 |
0 |
0 |
67 |
27 |
25 |
0 |
0 |
67 |
27 |
26 |
0 |
0 |
67 |
27 |
27 |
0 |
0 |
67 |
27 |
28 |
0 |
0 |
67 |
27 |
1 ThODNH4of 2-Propanone, reaction products with diphenylamine: 2.900 mg O2/mg test item
2 ThODNH4of sodium benzoate: 1.666 O2/mg reference item
Percentage Biodegradation of Test Item, of Sodium Benzoate and of the Toxicity Control based on ThODNO3
Time (days) |
Percentage Biodegradation1 |
|||
2-Propane, reaction products with diphenylamine1 |
Sodium Benzoate2 |
Toxicity control1,2 |
||
Flask 1 [%] |
Flask 2 [%] |
Flask 5 [%] |
Flask 7 [%] |
|
1 |
0 |
0 |
25 |
7 |
2 |
0 |
0 |
38 |
11 |
3 |
0 |
0 |
43 |
13 |
4 |
0 |
0 |
49 |
15 |
5 |
2 |
0 |
58 |
22 |
6 |
0 |
0 |
58 |
22 |
7 |
0 |
0 |
64 |
23 |
8 |
0 |
2 |
67 |
25 |
9 |
0 |
2 |
67 |
25 |
10 |
0 |
1 |
68 |
25 |
11 |
0 |
0 |
67 |
25 |
12 |
0 |
0 |
67 |
25 |
13 |
0 |
0 |
67 |
25 |
14 |
0 |
2 |
69 |
26 |
15 |
0 |
2 |
69 |
26 |
16 |
1 |
1 |
68 |
25 |
17 |
0 |
0 |
67 |
25 |
18 |
0 |
0 |
69 |
26 |
19 |
0 |
0 |
69 |
27 |
20 |
0 |
0 |
69 |
27 |
21 |
0 |
0 |
69 |
27 |
22 |
0 |
0 |
67 |
26 |
23 |
0 |
0 |
67 |
26 |
24 |
0 |
0 |
67 |
26 |
25 |
0 |
0 |
67 |
26 |
26 |
0 |
0 |
67 |
26 |
27 |
0 |
0 |
67 |
26 |
28 |
0 |
0 |
67 |
26 |
1 ThODNO3of 2-Propanone, reaction products with diphenylamine: 3.125 mg O2/mg test item
2 ThODNO3of sodium benzoate: 1.666 O2/mg reference item
pH-Values at the End of the Test
Flask No. |
Treatment |
pH-value |
1 |
2-Propanone, reaction products with diphenylamine |
7.6 |
2 |
2-Propanone, reaction products with diphenylamine |
7.6 |
3 |
Inoculum control |
7.2 |
4 |
Inoculum control |
7.2 |
5 |
Reference item (procedure control) |
7.2 |
Description of key information
Ready Biodegradability (Manometric Respirometry Test)
2-Propanone, reaction products with diphenylamine is considered to be not readily biodegradable based on ThODNH4.and ThODNO3
Key value for chemical safety assessment
- Biodegradation in water:
- under test conditions no biodegradation observed
Additional information
Ready Biodegradability (Manometric Respirometry Test)
The test item 2-Propanone, reaction products with diphenylamine 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.
Results:
Biodegradation of 2-Propanone, reaction products with diphenylamine: 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.
2-Propanone, reaction products with diphenylamine never reached 10% biodegradation. The mean biodegradation at test end after 28 days was 0% (ThODNH4and ThODNO3).
Therefore, 2-Propanone, reaction products with diphenylamine is considered to be not readily biodegradable based on ThODNH4.and ThODNO3
Biodegradation of Sodium Benzoate: The reference item sodium benzoate was sufficiently degraded to 69% after 14 days and to 67% 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, 26% and 25% after 14 days and to 27% and 26% after 28 days of incubation (ThODNH4.and ThODNO3). 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.
[Type of water: freshwater]
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