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EC number: 211-063-0 | CAS number: 628-96-6
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- Ecotoxicological Summary
- Aquatic toxicity
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- Short-term toxicity to fish
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- Long-term toxicity to aquatic invertebrates
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Toxicity to microorganisms
Administrative data
- Endpoint:
- activated sludge respiration inhibition testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2010-07-27 to 2010-07-28
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 010
- Report date:
- 2010
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Ethylene dinitrate
- EC Number:
- 211-063-0
- EC Name:
- Ethylene dinitrate
- Cas Number:
- 628-96-6
- Molecular formula:
- C2H4N2O6
- IUPAC Name:
- ethane-1,2-diyl dinitrate
Constituent 1
- Specific details on test material used for the study:
- Test substance was produced on site in a micro-reactor and used on the same day it was produced. Purity was reported as >99% using HPLC; batch number was EGDN-LN-20100727. Stability of the test solutions was checked at the start and end of the study.
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- The measured concentrations at t = 0 h were in a range of 96 - 128 % of the nominal concentrations. The measured concentrations at t = 3 h were in a range of 85 - 105 % of the nominal concentrations and 77 - 101 % of the start concentrations. Therefore the determination of the results was based on the mean of measured concentrations.
Test solutions
- Vehicle:
- no
Test organisms
- Test organisms (species):
- activated sludge, domestic
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 3 h
Test conditions
- Test temperature:
- 20 ± 2 °C
- Details on test conditions:
- 16 mL nutrient solution was mixed with 225 mL water in the first control vessel, then 259 mL inoculum were added. The bottle was closed immediately and stirred on a mag-netic stirrer without aeration. Five minutes later, the second control vessel was prepared likewise.
Again five minutes later, the first positive control treatment was prepared by mixing 16 mL nutrient solution with the appropriate amount of the stock solution and adding water to give 241 mL. Then 259 mL inoculum was added. The following positive control, test item and control treatments were prepared likewise in five minute intervals.
The content of the test item in the test vessels was measured at the start after one minute stirring and at the end of the test before oxygen measurement.
After 2 hours and 59 minutes the test vessels were shaken for one minute to guarantee a sufficient oxygen concentration in the test solution. Then the narrow neck bottle was filled to the rim with an aliquot of the content of the first vessel, a magnetic stirrer was added and the oxygen electrode was tightly fit into the vessel. Care was taken that no air could reach the contents of the vessel. The oxygen concentration of the vessel was measured every 5 - 10 seconds over a period of approx. 3 - 5 minutes. The interface of the oxygen meter was connected to a personal computer, feeding the measured values with their corresponding times in a table. - Reference substance (positive control):
- yes
- Remarks:
- 3,5-Dichlorophenol (CAS 591-35-5) was used as positive control. A stock solution in deionised water containing 500 mg/L (nominal) was freshly prepared for each experiment.
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- > 5.6 - < 23 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 % Conf. interval
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 160 - < 530 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 % Conf. interval
- Results with reference substance (positive control):
- 3,5-Dichlorophenol (CAS 591-35-5) was used as positive control. A stock solution in deionised water containing 500 mg/L (nominal) was freshly prepared for each experiment.
Any other information on results incl. tables
Nominal Conc. in mg/L |
Measured Conc. in mg/L |
% Recovery |
% of Nominal Concentration |
Mean of Measured Conc. in mg/L |
||
after 3 h |
||||||
0 h |
3 h |
|
0 h |
3 h |
||
1 |
1.3 |
1.0 |
80 |
128 |
102 |
1.2 |
3.2 |
3.3 |
2.7 |
82 |
104 |
85 |
3.0 |
10 |
11.3 |
8.7 |
77 |
113 |
87 |
10.0 |
32 |
32.3 |
29.8 |
92 |
101 |
93 |
31.0 |
100 |
107.0 |
105.0 |
98 |
107 |
105 |
106.0 |
320 |
317.5 |
313.7 |
99 |
99 |
98 |
315.6 |
1000 |
968.8 |
975.5 |
101 |
97 |
98 |
972.1 |
1000 |
958.1 |
956.1 |
100 |
96 |
96 |
957.1 |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- The inhibitory effect of ethylene glycol dinitrate on the respiration rate of activated sludge was determined in a study according to OECD guideline 209.
The following results were determined
3h NOEC = 10 mg/L (95% confid. interval: not determinable)
3h EC10 = 13 mg/L (95% confid. interval: 5.6 - 23 mg/L)
3h EC20 = 37 mg/L (95% confid. interval: 20 - 62 mg/L)
3h EC50 = 260 mg/L (95% confid. interval: 160 - 530 mg/L) - Executive summary:
One valid experiment was performed.
The main study was performed using seven concentrations, ranging from 1000 to 1.0 mg/L (nominal concentration).
Due to the volatility of the test item, the study was performed in a closed system without aeration and with analytical determination.
The dry matter of the activated sludge was determined as 2.9 g suspended solids/L, giving a concentration of 1.5 g suspended solids/L in the test. 3,5-Dichlorophenole was used as positive control. Four concentrations were tested; an EC50 of 7.7 mg/L (95% confidence interval: 2.8 – 12 mg/L) was determined, which lies within the range of 5 – 30 mg/L.
The following results for the test item Ethylene Glycol Dinitrate were determined:
Biological Results Test Item
Parameter
Value
95% Confidence Interval
NOEC
10 mg/L
n. d.
3h-EC10
13 mg/L
5.6 – 23 mg/L
3h-EC20
37 mg/L
20 – 62 mg/L
3h-EC50
260 mg/L
160 – 530 mg/L
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