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EC number: 221-070-0 | CAS number: 2997-92-4
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Density
- Particle size distribution (Granulometry)
- Vapour pressure
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- Auto flammability
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- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
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- Additional physico-chemical information
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- Endpoint summary
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- Environmental data
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- 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
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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:
- 07.02.12-15.02.12
- 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
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Laboratory culture: sludge was taken from ESN (Stadtentsorgung Neustadt) STP D-67435 NW-Lachen-Speyerdorf
- Method of cultivation:
- Preparation of inoculum for exposure:
- Pretreatment: sludge was filtrated, washed with tap water and afterwards resuspended in tap water. Sludge was aerated during the whole period from sampling until usage in test. Sludge was fed daily with 50 mL/L synthetic sewage feed.
- Initial biomass concentration: 1.25 g suspended solids/L, respectively 1.51 g suspended solids/L, dry matter in the test - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Hardness:
- 1.08 mmol
- Test temperature:
- 16.9 - 18.8 °C (pre-test)
18.9 - 19.7 °C (main study) - pH:
- 8.18
- Nominal and measured concentrations:
- pre-test: 1000, 100, 10, 1 mg/L (nominal)
main study: 3200, 1000, 320, 100, 32, 10 mg/L (nominal) - Details on test conditions:
- TEST SYSTEM
- Test vessel: beakers
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: glass, 500 mL
- Aeration: purified air, using Pasteur pipettes, flow approx. 0.75 L/min
- Type of flow-through (e.g. peristaltic or proportional diluter): -
- Renewal rate of test solution (frequency/flow rate): -
- No. of organisms per vessel: 250 mL inoculum
- No. of vessels per concentration (replicates): positive control: one replicate(/concentration; pre-test: one replicate (lower concentrations), five replicates (highest concentrations); main study: two replicates/treatment
- No. of vessels per control (replicates): two replicates before and two after measuring positive control and test item, respectively
- No. of vessels per vehicle control (replicates): -
- Biomass loading rate:
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Total organic carbon: 0.5 mg/l
- Particulate matter:
- Metals:< 0.007 mg/L
- Pesticides: < LOD
- Chlorine: 12 mg/L
- Alkalinity: not stated
- Ca/mg ratio: 7
- Conductivity: 223 µS/cm
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Adjustment of pH: -
- Photoperiod: -
- Light intensity: -
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : inhibition of respiration rate
TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:1000, 100, 10, 1 mg/L (nominal)
- Results used to determine the conditions for the definitive study: - Reference substance (positive control):
- yes
- Remarks:
- 3,5 DCP (CAS# 591-35-5)
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 7 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: 95% CI 3-13 mg/L
- Duration:
- 3 h
- Dose descriptor:
- other: EC20
- Effect conc.:
- 28 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: 95% CI 14-44 mg/L
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 360 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: 95% CI 280-510 mg/L
- Results with reference substance (positive control):
- pre-test: positve control (3,5-DCP) 7.8 mg/L
main test: positive control (3,5-DCP) 11.3 mg/L - Reported statistics and error estimates:
- linear regression
- Validity criteria fulfilled:
- yes
- Remarks:
- The results of the positive controls were within the stated range of tests with 3,5-DCP. Furthermore the CV of the oxygen control were less than 30 % and the O2 uptake of the blanks was more than 20 mg O2/ g activated sludge.
- Conclusions:
- Based on experimental observations the effect concentrations EC10(3h) = 7 mg/L and EC50(3h)= 360 mg/L were calculated. All validity criteria were fulfilled and hence the study is considered adequate for regulatory purposes.
- Executive summary:
Two valid experiments were performed. In the pre-test, the test item was tested using four concentrations (ranging from 1000 to 1 mg/L nominal concentration). Duration of the test was three hours. Activated sludge was used as inoculum. It was taken from a domestic sewage treatment plant and washed before usage. The dry matter was determined as 2.5 g suspended solids/, giving a concentration of 1.25 g suspended solids/L in the test.
3,5 -dichlorophenol was used as positive control. Four concentrations were tested; an EC50 of 7.8 mg/L (95% confidence interval: < 12 mg/L) was determined, which lies within the range of 2 -25 mg/L.
As inhibition > 20% was observed, a study plan amendment was issued and a main study was performed. The main study was performed likewise using six concentrations, ranging from 3200 to 10 mg/L nominal concentration. the dry matter of the activated sludge was determined as 3.02 g suspended solids/L, giving a concentration of 1.51 g suspended solids/L in the test. The EC50 of the positive control was determined as 11.3 mg/L (95% confidence interval: 8.6 - 14.0 mg/L), which lies within the range of 2 -25 mg/L.
The following results for 2,2'-azobis(2 -amidinopropane)-dihydrochloride (= 2,2'-azobis[2 -methylpropionamidine] dihydrochloride) were determined:
3h EC10 = 7 mg/L (95% confidence interval: 3 -13 mg/L)
3h EC20 = 28 mg/L (95% confidence interval: 14 -44 mg/L)
3h EC50 = 360 mg/L (95% confidence interval: 280 -510 mg/L)
Reference
Vessel No |
content |
concentration in mg/L |
O2consumption in mg//L*min) |
O2consumption in mg//L*h) |
Inhibition in % |
pH |
1 |
control |
0 |
0.5680 |
34.078 |
0 |
8.3 |
2 |
control |
0 |
0.5909 |
35.452 |
0 |
8.2 |
3 |
positive control |
5.0 |
0.4870 |
29.221 |
24.9 |
8.2 |
4 |
positive control |
10.1 |
0.3492 |
20.950 |
46.2 |
8.2 |
5 |
positive control |
20.2 |
0.1937 |
11.624 |
70.1 |
8.4 |
6 |
positive control |
40.4 |
0.1176 |
7.059 |
81.9 |
8.4 |
7 |
control |
0 |
0.6507 |
39.044 |
0 |
8.3 |
8 |
control |
0 |
0.6829 |
40.971 |
0 |
8.2 |
9 |
test item |
3201.2 |
0.1721 |
10.325 |
73.5 |
8.3 |
10 |
test item |
3200.2 |
0.1927 |
11.560 |
70.3 |
8.3 |
11 |
test item |
1000.2 |
0.2320 |
13.918 |
64.2 |
8.4 |
12 |
test item |
1000.4 |
0.2350 |
14.102 |
63.8 |
8.3 |
13 |
test item |
320.4 |
0.3145 |
18.872 |
51.5 |
8.3 |
14 |
test item |
321.0 |
0.3072 |
18.433 |
52.6 |
8.3 |
15 |
test item |
100.4 |
0.3919 |
23.517 |
39.6 |
8.3 |
16 |
test item |
98.8 |
0.4042 |
24.254 |
37.7 |
8.3 |
17 |
test item |
31.4 |
0.5397 |
32.380 |
16.8 |
8.2 |
18 |
test item |
33.0 |
0.5313 |
31.879 |
18.1 |
8.2 |
19 |
test item |
10.2 |
0.6529 |
39.174 |
-0.7 |
8.2 |
20 |
test item |
11.2 |
0.6462 |
38.774 |
0.4 |
8.2 |
21 |
control |
0 |
0.6979 |
41.875 |
0 |
8.2 |
22 |
control |
0 |
0.7010 |
42.061 |
0 |
8.2 |
Description of key information
Key value is based on experimental results observed in an OECD 209 guideline study.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 360 mg/L
- EC10 or NOEC for microorganisms:
- 7 mg/L
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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