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- Life Cycle description
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Biodegradation in water and sediment: simulation tests
Administrative data
Link to relevant study record(s)
- Endpoint:
- biodegradation in water: sewage treatment simulation testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2017-03-21 to 2017-04-26
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 303 A (Simulation Test - Aerobic Sewage Treatment. A: Activated Sludge Units)
- Version / remarks:
- 2001
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- 2017-01-03
- Radiolabelling:
- no
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, non-adapted
- Details on inoculum:
- - Source of activated sludge (e.g. location, contamination history): Municipal sewage treatment plant, Hildesheim, Germany, receives predominantly municipal sewage and hardly industrial chemical waste
- Pretreatment: The activated sludge was washed once with tap water and maintained in an aerobic condition by aeration at room temperature until use
- Biomass concentration used in test: 2.8 g DW/L (starting concentration) - Duration of test (contact time):
- 39 d
- Initial conc.:
- 10 mg/L
- Based on:
- other: Cinfluent
- Remarks:
- Nominal test item concentration
- Parameter followed for biodegradation estimation:
- DOC removal
- test mat. analysis
- Details on study design:
- TEST CONDITIONS
- Volume of test solution/treatment: test volume in the aeration vessel was 372 mL
- Composition of medium: Domestic sewage with a DOC of approx. 50 - 100 mg/L was used as organic medium. Preparation of organic medium with tap water or synthetic sewage was not necessary.
- Test temperature: 21 °C - 22 °C (in the activated sludge vessel)
- pH: 7.65 - 8.07
- Aeration of dilution water: not relevant
- Suspended solids concentration: 2.8 g DW/L (activated sludge)
- Continuous darkness: no
- Any indication of the test material adsorbing to the walls of the test apparatus: The test item has a high potential for adsorption, therefore, the study was performed in a Husmann unit which consists completely of glass. The test item was continuously pumped through the system and the activated sludge settled in the settling tank was pumped back into the aeration vessel by an air lift pump.
- Other:
Incubation: Incubation took place in an activated sludge unit at 20 - 25 °C. The aeration vessel was aerated continuously. Airflow was adjusted to maintain an oxygen level of at least 2 mg/L
Sludge retention time (SRT): 10 d, 1/10 of the volume of the activated sludge in the aeration vessel was removed daily. The complete volume of about 37.2 mL was removed once per day directly from the aeration vessel.
Initial dry weight content: 2.8 g/L
Mean hydraulic retention time. 6h, the total influent flow of 62 mL/h was maintained by dosing the test item stock solution and organic medium in appropriate amounts. Both solutions were pumped directly into the aeration vessels.
Influent flow: Organic medium: 62 mL/h, test item solution: 124 µL/h
Sludge retention time: 10 d
Wastage sludge rate: ca. 37.2 mL/d
TEST SYSTEM
- Measuring equipment:
Test vessels: Hunsmann unit consisting of an aeration vessel and a separator
pH-value, O2, temperature: pH meter, HQ 40d multi, Hach Lange, Hach Lange pH meter, Multi 350 i, WTW Datalogger, 174T, Testo
Pumps: Precision syringe pump, Kd Scientific Peristaltic pump, ISMATEC
Aeration: Medo Kompressor, Rebie
DOC: Multi N/C 3100, Analytik Jena
Balances: Balance, Kern Analytical Balance, Sartorius
Dry weight: Drying oven T6, Thermo Scientific Drying oven, Binder
- Other:
Set up: The test system was started with activated sludge with a dry weight content of 2.8 g DW/L. To re-suspend activated sludge adhering to the walls of the aeration vessel, a magnetic bar was placed inside the aeration vessel permanently. For resuspension, the magnetic bar was swiped over the glass walls with a second magnetic bar from outside of the vessel. Due to the geometry of the air pump slight loss of sludge occurred until day 22, furthermore the sedimentation decreased. Therefore 50 mL fresh sludge (2.95 g DW/L) was added on day 22 (day 17 of test item application) to aeration vessel.
Steady state phase: After a period in which the system had stabilized and the removal of DOC of the organic medium was efficient (> 80 %) the test item solution was dosed with a syringe pump into the aeration vessel fo the test item unit. Throughout the test, the sludge adhering to the walls of the activated sludge units was re-suspended at least daily. Regularly all tubes and tubing were checked and cleaned to prevent a growth of biofilm as far as possible. The rate of ultimate biodegradation of the organic medium was checked by determination of the DOC in the influents and effluents in regular time intervals during this phase of the test. The elimination of the test item was determined by UPLC MS/MS.
Replicates: One replicate
Water: Tap water for the preparation or organic medium. Demineralized water for the preparation of the synthetic sewage stock solutions.
Domestic sewage: Domestic sewage was collected from the primary sedimentation tank, free of coarse particles. The DOC and pH were measured from each batch. Each batch was stored at about 4 °C to reduce microbial activity. Domestic sewage of the same source as for activated sludge was used.
Synthetic sewage: DOC adjustment of the domestic sewage with synthetic sewage was not necessary.
Application: The test item was continuously applied to the test item unit by a syringe pump. As the test item stock solution was added directly to the activated sludge unit, no biodegradation may have occurred before it had reached the aeration vessel.
SAMPLING
- Sampling frequency: Influent: (Test item stock solution):
Adaption: every new batch,
Plateau phase: every new batch
Effluent: Adaption: At least 2 x per week, Plateau phase: At least weekdays
- Sampling method used per analysis type: Composite sampling of the effluent. An aliquot of the sample was stabilized with trifluoroacetic acid (1% v/v) and preconcentrated by SPE
- Sample storage before analysis: Samples taken on weekends were acidified with 1% TFA (v/v) and further sample preperation was done the following weekday. Samples taken on weekdays were processed on the same day. Prepared samples were stored at ambient temperature.
DESCRIPTION OF CONTROL AND/OR BLANK TREATMENT PREPARATION
CONTROL AND BLANK SYSTEM
- Inoculum blank: An activated sludge unit without test item treatment was used as control. This unit was used exclusively to generate matrix for spike controls. Due to decreasing settling capability of the activated sludge, the control unit was stopped on day 25 (day 20 of test application).
STATISTICAL METHODS:
- Software used for calculations: Excel, Microsoft Corporation and Sigma Plot, Spss Inc. - Reference substance:
- other: none
- Key result
- % Degr.:
- 99.87
- St. dev.:
- 0.054
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- based on the analysis of the seven main components of the test substance
- % Degr.:
- 99.36
- St. dev.:
- 0.34
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- N'-C14-diGly
- % Degr.:
- 99.9
- St. dev.:
- 0.061
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- N'-C14-Gly
- % Degr.:
- 99.35
- St. dev.:
- 0.238
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- C14-PDA
- % Degr.:
- 99.84
- St. dev.:
- 0.043
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- N'-C12-diGly
- % Degr.:
- 99.98
- St. dev.:
- 0.012
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- N'-C12-Gly
- % Degr.:
- 99.97
- St. dev.:
- 0.022
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- C12-PDA
- % Degr.:
- 99.97
- St. dev.:
- 0.033
- Parameter:
- test mat. analysis
- Sampling time:
- 24 d
- Remarks on result:
- other: elimination
- Remarks:
- N-C12-Gly x HCI
- Transformation products:
- not measured
- Evaporation of parent compound:
- not specified
- Volatile metabolites:
- not specified
- Residues:
- not specified
- Details on results:
- TEST CONDITIONS
- Aerobicity (or anaerobicity), moisture, temperature and other experimental conditions maintained throughout the study: At start of test item application, the pH was slightly aove 8.0, but decreased to the required range within 7 days. During the plateau phase, the pH-value was in the required range of 7.5 ± 0.5. The temperature in the aeration vessels was in the range of 21.0 - 22.7 °C.
- Anomalies or problems encountered (if yes): Component 2 (N,N'-C12-diGly) was removed from the evaluation, because NMR analysis of the standard did not confirm the supposed identity. The standard of component 2 was identical with component 5 (N'-C12-diGly). Consequently, all analytical data obtained for component 5 (N'-C12-diGly) during the study were multiplied by factor 2.
These deviations were considered to have no impact on the integrity and validity of the study.
RESULTS OF SUPPLEMENTARY EXPERIMENT (if any):
Prior to study initiation, the loss of the test item to the application equipment (syringe, tubing) was assessed over a period of up to 6 days. No significant loss of the test item (based on TOC measurements) was observed. - Validity criteria fulfilled:
- yes
- Remarks:
- The degree of DOC elimination/degradation in the control unit was > 80 % after 6 days and no unusual observations were made.
- Conclusions:
- Based on the analysis of 7 components of the test substance in the effluent, the mean elimination rate at the plateau phase in relation to the total influent concentration was calculated. The elimination of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was 99.87% in a continuously operating activated sludge unit. The elimination of the 7 individual components was in the range 99.35% - 99.98%.
- Executive summary:
The elimination of the test item Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid in an activated sludge unit was determined over a test period of 34 days according to OECD guideline 303 A (2001).
The elimination of main 7 components of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was determined in a continuously operating laboratory activated sludge unit simulating the activated sludge process. The activated sludge unit was started with an activated sludge content of 2.8 g/L DW. After a short acclimatization period of the activated sludge (DOC elimination > 80%), the application of the test substance 2000 was initiated. Samples of the effluent were taken at regular intervals and LC-MS/MS analysis was carried out to determine the elimination of the test substance.
The nominal concentration of the test item stock solution was 5 g/L. The stock solution was dosed at a constant rate of 124 µg/h into the activated sludge unit, resulting in an influent concentration of 10 mg test substance/L.
Based on the analysis of 7 main constituents of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid in the effluent, the mean elimination rate at the plateau phase in relation to the total influent concentration was calculated. The elimination of test substance was 99.87% in a continuously operating activated sludge unit. The elimination of the 7 individual components was in the range 99.35% - 99.98%.
- Endpoint:
- biodegradation in water: sewage treatment simulation testing
- Type of information:
- experimental study
- Adequacy of study:
- other information
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- documentation insufficient for assessment
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- - OECD confirmatory test (according to: Bundesgesetzblatt (1977), part 1, p. 248 - 266) substance was tested with activated sludge. Therefore, the test substance and a mineral nutrient solution are continuosly added to an aeration tank. Activated sludge, taken from a municipal sewage treatment plant, migrates to a sedimentation tank and is afterwards pumped back to the aeration tank. Inflow and discharge are tested analytically for surfactants. For determination of a total degradation, an analysis of DOC/COD can be carried out. Confirmatory test is taking into account anionic and non-anionic surfactants (not cationic). Inflow surfactant concentrations are defined as: A-surfactant: 5-20 mg active ingredient/L, Nio-surfactant: 5-10 mg active ingredient/L, K-surfactant: 1-5 mg active ingredient/L
- GLP compliance:
- no
- Radiolabelling:
- no
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge (adaptation not specified)
- Details on source and properties of surface water:
- - synthetic waste water
- Duration of test (contact time):
- 2 wk
- Initial conc.:
- 1 mg/L
- Based on:
- act. ingr.
- Initial conc.:
- 2 mg/L
- Based on:
- act. ingr.
- Initial conc.:
- 5 mg/L
- Based on:
- act. ingr.
- Initial conc.:
- 10 mg/L
- Based on:
- act. ingr.
- Parameter followed for biodegradation estimation:
- DOC removal
- O2 consumption
- % Degr.:
- >= 92 - <= 96
- Parameter:
- DOC removal
- % Degr.:
- >= 93 - <= 97
- Parameter:
- O2 consumption
- Evaporation of parent compound:
- not measured
- Volatile metabolites:
- not measured
- Residues:
- not measured
- Validity criteria fulfilled:
- not applicable
- Conclusions:
- In the modified OECD confirmatory test, primary biodegradation of > 99% was observed for all tested inflow concentrations of the test item (1 - 10 mg a.i./L). DOC, COD, nitrogen compounds, Orange II analytics revealed that test item concentrations up to 50 mg/L (corresponding to 10 mg a.i./L) do not reduce the degradation capacity of the test plant.
- Executive summary:
In an OECD confirmatory test (according to Budesgesetzblatt (1977), part 1, p. 248 - 266): the removal of the test substance was tested at concentrations of 5, 10, 25 and 50 mg/L (corresponding to 1, 2, 5 and 10 mg a.i./L). Therefore, the test substance and a mineral nutrient solution are continuosly added to an aeration tank. Bacterial sludge then migrates to a sedimentation tank and is afterwards pumped back to the aeration tank. Inflow and discharge are tested analytically for surfactants. For determination of a total degradation, an analysis of DOC and COD was carried out. Confirmatory test is taking into account anionic and non-anionic surfactants (not cationic). Additionally nitrogen compunds were measured. Primary biodegradation of > 99% was observed for all tested inflow concentrations of the test item. DOC, COD, nitrogen compounds and photometric analytics revealed that test item concentrations up to 50 mg/L (corresponding to 10 mg a.i./L) do not reduce the degradation capacity of the test plant.
- Endpoint:
- biodegradation in water: sewage treatment simulation testing
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1991-11-28 to 1991-12-27
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 303 A (Simulation Test - Aerobic Sewage Treatment. A: Activated Sludge Units)
- Version / remarks:
- 1981
- Deviations:
- no
- GLP compliance:
- yes
- Radiolabelling:
- no
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic (adaptation not specified)
- Details on inoculum:
- - Source of inoculum/activated sludge: Oxidation ditch for domestic sewage treatment, Berkel, The Netherlands
- Laboratory culture: No
- Preparation of inoculum for exposure: Sieving (2 mm) and homogenising - Duration of test (contact time):
- 25 d
- Initial conc.:
- 120 mg/L
- Based on:
- DOC
- Parameter followed for biodegradation estimation:
- DOC removal
- Details on study design:
- TEST CONDITIONS
- Composition of nutrient feed solution:
Peptone 6400 mg/L, Meat extract 4400 mg/L, Urea 1200 mg/L, NaCl 280 mg/L, CaCl2 • 2 H2O 160 mg/L, MgSO4 • 7 H2O 80 mg/L,
K2HPO4 • 3 H2O 1120 mg/L
- Additional substrate: None; however, the dilution water (Milli-Q) contained 15–21 mg/l DOC between Dec 09 and Dec 18, 1991; this was corrected for in the calculations
- Test temperature: 18 ± 2°C
- pH: 6.9–7.4 (test unit), 6.6–7.3 (blank unit)
- pH adjusted: no
- Aeration of dilution water: Continuous aeration of the test solutions
- Suspended solids concentration: 2.0–2.5 g/L
- Continuous darkness: yes
- Other: Working-in time: 8 days
TEST SYSTEM
- Culturing apparatus: OECD confirmatory test units, continuous flow activated sludge system, Husmann apparatus, aeration vessel (contents: 3 L) and a separator (secondary clarifier, about 2 L)
- Number of culture flasks/concentration: 1
- Aeration device: Not specified
- Measuring equipment: OI Model 700 TOC analyser
- Test performed in closed vessels due to significant volatility of test substance: Not relevant
SAMPLING
- Sampling frequency: daily
CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
STATISTICAL METHODS:
Degradation rate according to test guideline. - % Degr.:
- 81
- St. dev.:
- 5
- Parameter:
- DOC removal
- Sampling time:
- 25 d
- Transformation products:
- not measured
- Validity criteria fulfilled:
- yes
- Conclusions:
- By means of the coupled units test (OECD 303A), Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was established to be ultimately biodegradable in sewage treatment plants.
- Executive summary:
The degradation of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was simulated in a coupled units test (OECD 303A). The performance of the test deviated from the guideline in several aspects. However, the fact that the inoculum originated from only a single source is considered to be without negative consequences since the microbial activity proved to be adequate. Deviations in the concentration of the synthetic sewage were compensated during dosing. Centrifugation may be considered a sufficiently effective method for removal of particulate matter from the effluent. In conclusion, the deviations from the guideline are considered to be insignificant for the validity of the study.
The DOC fed to the test system by means of the test substance was effectively removed from the test solution. The degradation rate was 81 ± 5% (mean ± SD).
The test substance is known to exhibit microbicidal properties. However, this had apparently no negative effects on the test system, presumably due to the high activity and concentration of the activated sludge.
Since the deviations from the test guideline as discussed above had no significant impact on the outcome of the study, the test is considered to be valid.
By means of the coupled units test (OECD 303A), Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was established to be ultimately biodegradable in sewage treatment plants.
Referenceopen allclose all
Table 1: Statistical evaluation of the elimination at plateau phase (n=18)
Elimination in % |
||||
|
N-C12-Gly x HCL |
C 12 PDA |
N'-C12-Gly |
N'-C12-diGly |
Mean
95% CI |
99.97
99.95 - 99.98 |
99.97
99.96 - 99.98 |
99.98
99.98 - 99.99 |
99.84
99.80 - 99.88 |
|
C14-PDA
|
N'-C14-GLY |
N'-C14-diGly |
Test substance |
Mean
95% CI |
99.35
99.23 - 99.46 |
99.90
99.87 - 99.93 |
99.36
99.19 - 99.53 |
99.87
99.84 - 99.89 |
CI= Confidence Interval
Ammonium content: < 0.1 mg/L, nitrite: not detectable in > 90% of all outflow measurements, total load of nitrogen: > 90% consisting of nitrate
Table 1: Surfactant specific degradation
Test substance concentration (mg/L) |
Exctinction |
|
Inflow |
Discharge |
|
0 |
0.064 |
0.001 |
25 |
0.981 |
0.001 |
50 |
1.907 |
0.001 |
The results indicate a nearly complete removal of the test substance in the coupled unit test. The plateau level was reached in 8 days and the mean degradation percentage was calculated from 10 plateau values to be 81 ± 5%.
Description of key information
The elimination of the 7 individual components of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was in the range 99.35% - 99.98% after 24 days.
Key value for chemical safety assessment
Additional information
The elimination of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid in an activated sludge unit was determined in two studies performed according to and similar to OECD guideline 303 A, respectively.
In the key study, the elimination of main 7 components of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acidwas determined in a continuously operating laboratory activated sludge unit simulating the activated sludge process. The activated sludge unit was started with an activated sludge content of 2.8 g/L DW. After a short acclimatization period of the activated sludge (DOC elimination > 80%), the application of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was initiated. Samples of the effluent were taken at regular intervals and LC-MS/MS analysis was carried out to determine the elimination of the test item.
The nominal concentration of the test item stock solution was 5 g/L. The stock solution was dosed at a constant rate of 124 µg/h into the activated sludge unit, resulting in an influent concentration of 10 mg test item/L.
Based on the analysis of 7 main constituents of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid in the effluent, the mean elimination rate at the plateau phase in relation to the total influent concentration was calculated. The elimination of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was 99.87% in a continuously operating activated sludge unit. The elimination of the 7 individual components was in the range of 99.35% - 99.98%.
This result is supported by another study in which the degradation of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was simulated in a coupled units test (OECD 303A). The performance of the test deviated from the guideline in several aspects. However, the deviations from the guideline are considered to be insignificant for the validity of the study. The DOC fed to the test system by means of the test substance was effectively removed from the test solution. The degradation rate was 81 ± 5% (mean ± SD).
By means of the coupled units test (OECD 303A), Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid was established to be ultimately biodegradable in sewage treatment plants.
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