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EC number: 947-155-9 | CAS number: -
- Life Cycle description
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- Endpoint summary
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- Ecotoxicological Summary
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- Long-term toxicity to aquatic invertebrates
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Toxicity to microorganisms
Administrative data
Link to relevant study record(s)
- Endpoint:
- activated sludge respiration inhibition testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- June 20th, 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M FMRD, S352546
- Expiration date: 2020.05.01
- Purity: 99.7%
- Physical state: Clear and colorless liquid
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored at room temperature (15-25°C).
- Stability under test conditions: Due to analytical issus, stability during the test is unknown. - Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
-Methods: Test substance directly weighed into the test vessel and distilled water added.
-Controls: blank (water only), abiotic, procedural control - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Name and location of sewage treatment plant where inoculum was collected:
Activated sludge from aeration tank in the Shenyang North Sewage Treatment Plant, PRC
- Preparation of inoculum for exposure: Upon arrival at the laboratory, coarse particles and solids were removed by settling for 15 minutes. The decanted sludge was centrifuged for 5 minutes (2500 rpm) to obtain a sewage pellet and clear supernatant. The sludge pellet was washed by resuspension in dechlorinated tap water, and the above process was repeated. The washed, resuspended sludge was dried (10 mL suspended sludge, 5 replicates, at 105 C for 1 hour and 40 mins) to determine an initial suspended solids concentration and was further diluted to 3 g sludge solids/L. Sludge was fed with synthetic sewage feed (50 mL synthetic sewage feed/L activated sludge) and aerated one day prior to test.
- Preparation of inoculum for exposure: Sludge concentration was measured again on day of test and adjusted as needed to obtain 3 g sludge solids/L. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- 20.4-21.6 °C
- pH:
- 7.18-7.99
- Nominal and measured concentrations:
- Nominal: 0 (dilution water control), 10, 100, 300, and 1000 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Glass flasks, containing 500 mL volume
- Type: open during aeration
- Aeration: Yes, clean oil-free air, 0.5-1.0 mL/ min
- No. of vessels per concentration: The 10, 100, and 300 mg/L solutions were tested with 1 replicate and 1000 mg/L solution tested in triplicate and one abiotic control (1000 mg/L)
- No. of vessels per control (replicates): 4
- No. of vessels per abiotic control (replicates): 1
- No. of vessels per reference substance (replicates): 1 replicate at each of the five concentrations.
- Sludge concentration: 3 g/L
- Nutrients provided for bacteria: synthetic sewage per OECD 209 (peptone 16g, meat extract 11g, urea 3g, NaCl 0.7 g, CaCl2.2H2O 0.4 g, MgSO4.7H2O 0.2 g, K2HPO4 2.8g, (dissolved in 1 L of water and filtered).
- Test setup: For the exposure period, flasks received either pure test substance or reference substance stock solution. Distilled water was added to bring total volume to 234 mL. 16 mL synthetic sewage and 250 mL diluted sludge were then added and the flasks aerated until measurement. Solution pH was measured after aeration began.
- Nitrification inhibition used: None
- Biomass loading rate: 1.5 g sludge solids/L
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled tap-water
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Details on termination of incubation: After the 3-hour exposure, a sample was transferred to a measuring cell. Using a HACH HQ40d oxygen probe with a sleeve adaptor, the probe was inserted into the flask and magnetic stirrer started. Dissolved oxygen concentration was continuously measured for 10 minutes.
EFFECT PARAMETERS MEASURED : Dissolved oxygen concentration. For each exposure, dissolved oxygen depletion and time interval were correlated and the slope of the regression line was calculated as the respiration rate.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: variable
- Justification for using fewer concentrations than requested by guideline: test done as combined rangefinder/limit test - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks on result:
- other: Average percent inhibition at 1000 mg/L was 1.1%.
- Details on results:
- - Blank controls oxygen uptake rate: 20.7 mg/(g*hr), Details in Table 2
- Coefficient of variation of oxygen uptake rate in control replicates: 1.4% - Results with reference substance (positive control):
- - Results with reference substance valid: Yes
- EC50: 6.0 mg/L (95 CI: 3.9-9.2). The EC50 was within the 5 to 25 mg/L range considered acceptable for the test. - Reported statistics and error estimates:
- For the study summary, a t-test was run on the difference in respiration rate in the 1000 mg/L concentrations and blank control, following report submission. No difference in mean rate was found (p=0.63).
- Validity criteria fulfilled:
- yes
- Remarks:
- Control oxygen uptake rate > 20 mg O2/gram sludge/hour (20.7 mg O2/(h.g SS)); coeff. of variation <15% for control replicates (1.4%); ref. substance EC50 in range 5-25 mg/L (6.0 mg/L).
- Conclusions:
- 3 hour EC50 >1000 mg/L in activated sludge for respiration inhibition (OECD 209)
- Executive summary:
The toxicity of MTDID 44428 to activated sludge was assessed according to the OECD 209 guideline in a definitive test conducted at 10, 100, 300, and 1000 mg/L. The validity criteria were met based on an acceptable level of oxygen uptake in control samples, acceptable variation among control replicates and a typical level of inhibition with the positive control. The 3-hour EC50 (respiration) of MTDID 44428 to activated sludge is >1000 mg/L, as only 1.1% inhibition was observed at the highest concentration. There was no significant difference between the respiration rates of the blank control and the 1000 mg/L concentration. The study followed an international standard method and is GLP compliant. Therefore the study is deemed reliable without restrictions and can be utilized to calculate a PNEC for STP.
Reference
Table 1. Dissolved oxygen concentration for total respiration during 10-minute measurement period after 3 hour contact time
Oxygen |
Test |
Nominal |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
Blank |
FBT1 |
/ |
8.61 |
8.10 |
7.59 |
7.08 |
6.57 |
6.06 |
5.55 |
5.04 |
4.53 |
4.02 |
|
FBT2 |
/ |
7.76 |
7.24 |
6.72 |
6.20 |
5.68 |
5.16 |
4.65 |
4.14 |
3.63 |
3.12 |
|
FBT3 |
/ |
7.24 |
6.72 |
6.20 |
5.68 |
5.16 |
4.64 |
4.12 |
3.61 |
3.10 |
2.59 |
|
FBT4 |
/ |
6.20 |
5.67 |
5.14 |
4.61 |
4.08 |
3.55 |
3.03 |
2.51 |
1.99 |
1.47 |
Reference |
FRT1 |
0.5 |
7.77 |
7.27 |
6.77 |
6.27 |
5.77 |
5.28 |
4.79 |
4.30 |
3.82 |
3.34 |
|
FRT2 |
1.3 |
6.85 |
6.40 |
5.98 |
5.55 |
5.11 |
4.68 |
4.24 |
3.82 |
3.37 |
2.93 |
|
FRT3 |
3.5 |
7.63 |
7.19 |
6.78 |
6.37 |
5.98 |
5.60 |
5.22 |
4.85 |
4.49 |
4.13 |
|
FRT4 |
9.4 |
7.52 |
7.30 |
7.08 |
6.86 |
6.64 |
6.42 |
6.20 |
5.97 |
5.74 |
5.51 |
|
FRT5 |
25 |
9.42 |
9.39 |
9.33 |
9.27 |
9.21 |
9.14 |
9.08 |
9.02 |
8.96 |
8.90 |
Test |
FTT1 |
10 |
7.77 |
7.26 |
6.77 |
6.28 |
5.77 |
5.28 |
4.77 |
4.26 |
3.75 |
3.24 |
|
FTT2 |
100 |
7.85 |
7.32 |
6.80 |
6.31 |
5.79 |
5.24 |
4.72 |
4.15 |
3.60 |
3.05 |
|
FTT3 |
300 |
7.70 |
7.16 |
6.64 |
6.14 |
5.64 |
5.14 |
4.64 |
4.15 |
3.65 |
3.13 |
|
FTT4-1 |
1000 |
6.88 |
6.36 |
5.84 |
5.32 |
4.81 |
4.29 |
3.77 |
3.24 |
2.72 |
2.18 |
|
FTT4-2 |
1000 |
6.96 |
6.42 |
5.91 |
5.39 |
4.86 |
4.34 |
3.82 |
3.29 |
2.77 |
2.24 |
|
FTT4-3 |
1000 |
7.20 |
6.71 |
6.19 |
5.67 |
5.16 |
4.67 |
4.19 |
3.72 |
3.25 |
2.79 |
|
FA |
1000 |
8.53 |
8.53 |
8.52 |
8.50 |
8.49 |
8.47 |
8.47 |
8.46 |
8.44 |
8.44 |
Table 2. Oxygen uptake rate and percent inhibition of total respiration after 3 hour contact time
Oxygen uptake |
Test vessel No. |
Nominal Concentration (mg/L) |
1-2 |
2-3 |
3-4 |
4-5 |
5-6 |
6-7 |
7-8 |
8-9 |
9-10 |
Average |
R:mg/(L·h) |
Rs: mg/(g·h) |
% Inhibition |
|
Blank control (Total) |
FBT1 |
/ |
0.51 |
0.51 |
0.51 |
0.51 |
0.51 |
0.51 |
0.51 |
0.51 |
0.51 |
0.510 |
30.6 |
20.4 |
/ |
|
|
FBT2 |
/ |
0.52 |
0.52 |
0.52 |
0.52 |
0.52 |
0.51 |
0.51 |
0.51 |
0.51 |
0.516 |
31.0 |
20.7 |
/ |
|
|
FBT3 |
/ |
0.52 |
0.52 |
0.52 |
0.52 |
0.52 |
0.52 |
0.51 |
0.51 |
0.51 |
0.517 |
31.0 |
20.7 |
/ |
|
|
FBT4 |
/ |
0.53 |
0.53 |
0.53 |
0.53 |
0.53 |
0.52 |
0.52 |
0.52 |
0.52 |
0.526 |
31.6 |
21.1 |
/ |
|
Reference item (Total) |
FRT1 |
0.5 |
0.50 |
0.50 |
0.50 |
0.50 |
0.49 |
0.49 |
0.49 |
0.48 |
0.48 |
0.492 |
29.5 |
19.7 |
4.8% |
|
|
FRT2 |
1.3 |
0.45 |
0.42 |
0.43 |
0.44 |
0.43 |
0.44 |
0.42 |
0.45 |
0.44 |
0.436 |
26.2 |
17.5 |
15.5% |
|
|
FRT3 |
3.5 |
0.44 |
0.41 |
0.41 |
0.39 |
0.38 |
0.38 |
0.37 |
0.36 |
0.36 |
0.389 |
23.3 |
15.5 |
25.1% |
|
|
FRT4 |
9.4 |
0.22 |
0.22 |
0.22 |
0.22 |
0.22 |
0.22 |
0.23 |
0.23 |
0.23 |
0.223 |
13.4 |
8.9 |
57.0% |
|
|
FRT5 |
25 |
0.03 |
0.06 |
0.06 |
0.06 |
0.07 |
0.06 |
0.06 |
0.06 |
0.06 |
0.058 |
3.5 |
2.3 |
88.9% |
|
Test item |
FTT1 |
10 |
0.51 |
0.49 |
0.49 |
0.51 |
0.49 |
0.51 |
0.51 |
0.51 |
0.51 |
0.503 |
30.2 |
20.1 |
2.9% |
|
|
FTT2 |
100 |
0.53 |
0.52 |
0.49 |
0.52 |
0.55 |
0.52 |
0.57 |
0.55 |
0.55 |
0.533 |
32.0 |
21.3 |
-2.9% |
|
|
FTT3 |
300 |
0.54 |
0.52 |
0.50 |
0.50 |
0.50 |
0.50 |
0.49 |
0.50 |
0.52 |
0.508 |
30.5 |
20.3 |
1.9% |
|
|
FTT4-1 |
1000 |
0.52 |
0.52 |
0.52 |
0.51 |
0.52 |
0.52 |
0.53 |
0.52 |
0.54 |
0.522 |
31.3 |
20.9 |
-1.0% |
|
|
FTT4-2 |
1000 |
0.54 |
0.51 |
0.52 |
0.53 |
0.52 |
0.52 |
0.53 |
0.52 |
0.53 |
0.524 |
31.4 |
20.9 |
-1.0% |
|
|
FTT4-3 |
1000 |
0.49 |
0.52 |
0.52 |
0.51 |
0.49 |
0.48 |
0.47 |
0.47 |
0.46 |
0.490 |
29.4 |
19.6 |
5.3% |
|
|
FA |
1000 |
0.00 |
0.01 |
0.02 |
0.01 |
0.02 |
0.00 |
0.01 |
0.02 |
0.00 |
0.010 |
/ |
/ |
/ |
Note: R: mg/(L·h), oxygen uptake rate; Rs: mg/(g·h), specific respiration rate
Average of I% was calculated based on average of Rs.
Description of key information
The toxicity of MTDID 44428 to activated sludge was assessed according to the OECD 209 guideline in a definitive test conducted at 10, 100, 300, and 1000 mg/L. The validity criteria were met based on an acceptable level of oxygen uptake in control samples, acceptable variation among control replicates and a typical level of inhibition with the positive control. The 3-hour EC50 (respiration) of MTDID 44428 to activated sludge is >1000 mg/L, as only 1.1% inhibition was observed at the highest concentration.There was no significant difference between the respiration rates of the blank control and the 1000 mg/L concentration.The study followed an international standard method and is GLP compliant.Therefore the study is deemed reliable without restrictions and can be utilized to calculate a PNEC for STP.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 1 000 mg/L
Additional information
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