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EC number: 701-301-6 | CAS number: -
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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:
- From 18 January 2016 to 11 March 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry 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: 1000100003
- Expiration date of the lot/batch: 13.10.2017
- Purity: 92% - Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, domestic, non-adapted
- Details on inoculum:
- - Source of inoculum/activated sludge: Sewage Treatment Plant ”Czajka” , Warsaw receiving predominantly domestic sewage.
- Storage conditions: No storage
- Preparation of inoculum for exposure: Preconditioned, the inoculum was aerated, at the test temperature of 22 °C, until application next day.
- Pretreatment: The coarse particles were removed by settling and the supernatant was discarded. The sludge was washed in the mineral medium. The concentrated sludge was suspended in mineral medium to yield a concentration of 3-5 g suspended solids/l.
- Water filtered: no - Duration of test (contact time):
- 28 d
- Initial conc.:
- 100 mg/L
- Based on:
- test mat.
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium: inoculum 30 mg/l suspended solids, 50-100mg ThOD/l, concentration of elements in mineral medium:
P =116 mg/l, N = 1.3 mg/l, Na=86 mg/l, K = 122 mg/l, Mg = 2.2 mg/l, Ca 9.9 mg/l, Fe = 0.05-0.1 mg/l.
- Solubilising agent (type and concentration if used): Mineral medium was performed mixing 10 ml of solution containing:
Monopotassium dihydrogen orthophosphate, KH2PO4 8.50 g/l
Dipotassium monohydrogen orthophosphate, K2HPO4 21.75 g/l
Disodium monohydrogen orthophosphate dihydrate Na2HPO4 · 2 H2O 33.40 g/l
Ammonium chloride, NH4Cl 0.50 g/l
with 800 ml of dilution water. Then it was added 1 ml of calcium chloride, anhydrous, CaCl2 27.50 g/l, 1ml of magnesium sulphate heptahydrate, MgSO4 · 7 H2O 22.50 g/l and 1 ml of Iron(III) chloride hexahydrate, FeC13 · 6H2O 0.25 g/l and it was made up to 11 with dilution water.
- Test temperature: 22 ± 2°C
- pH: 7.4 ± 0.2
- pH adjusted: yes
- Suspended solids concentration: 30 mg/l SS
- Organic carbon of dilution water: 3 mg/l of organic carbon
TEST SYSTEM
- Culturing apparatus: thermo-cabinet WTW TS 606 CZ-G/3-VAR
- Number of culture flasks/concentration:
Flasks# 1, 2, 3: Test suspension containing test item (100 mg/l) and inoculum 30 mg/l SS.
Flasks# 7, 8 and 9: Inoculum blank containing only inoculum 30 mg/l SS.
Flask# 10, 11 and 12: Procedure control containing reference item (sodium acetate 100 mg/l) and inoculum 30 mg/SS.
Flask# 4, 5 and 6: Toxicity control: containing test item, reference item at the same concentrations as in the individual solutions and inoculum 30 mg/l SS.
- Method used to create aerobic conditions: stirrers were used and it was checked for air-tightness.
- Measuring equipment: closed WTW OxiTop OC 110 respirometer for BOD determination and spectrophotometer Hach DR 3900 and 45600 reactor COD concentration measurements.
- Test performed in closed vessels due to significant volatility of test substance: yes
- Details of trap for CO2 and volatile organics if used: The carbon dioxide, evolved during test item degradation, was absorbed in a solution of potassium hydroxide.
SAMPLING
- Sampling frequency: The data were read out every 112 min
- Sampling method: a controller was used
CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, three inoculum blanks
- Abiotic sterile control: yes, only mineral medium was used.
- Toxicity control: yes, containing test item, reference item at the same concentrations as in the individual solutions and inoculum 30 mg/l SS
STATISTICAL METHODS: The calculations and the graphs were performed using SigmaP lot 9.0 software of SYSTAT Software, Inc., USA. - Reference substance:
- acetic acid, sodium salt
- Key result
- Parameter:
- % degradation (O2 consumption)
- Remarks:
- BOD/ThOD
- Value:
- 82.5
- St. dev.:
- 0.3
- Sampling time:
- 28 d
- Parameter:
- % degradation (O2 consumption)
- Remarks:
- BOD/ThOD
- Value:
- 60
- Sampling time:
- 7 d
- Details on results:
- At the 28th day of the test the aerobic biodegradation of the testing:
- In the toxicity test the biodegradation was equal to 67.4% in 14 days.As more than 25% of biodegradation (based on ThOD NH4) occurred in 14 days, the test item can not be assumed to be inhibitory.
- The oxygen uptake of the inoculum blank was equal to 54.8 mg/l in 28 days.
- The pH values of all flasks were inside the range 7.19-8.60. - Results with reference substance:
- The reference item reached 78.4% of biodegradation and the level for ready biodegradability by 5 days.
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- readily biodegradable
- Conclusions:
- The test substance can be considered readily biodegradable within 28 days based on the oxygen demand (BOD and ThOD).
- Executive summary:
A 28-day ready aerobic biodegradability test through the manometric respirometry method was performed with the test substance according to OECD 301F / EC C.4 – D under GLP conditions. A 100 mg/L of test item was inoculated with non-adapted activated sludge (30 mg/L SS) and incubated under aerobic conditions in a closed respirometer flask at constant temperature 22ºC(± 2) for 28 days. A blank test, a procedure test with reference substance (sodium acetate) and a toxicity test were run in parallel. The validity criteria was considered fulfilled. The substance was mostly degraded (82.5%) within the 28-day period based on BOD (based on measurements of oxygen uptake) and ThOD. Thus the test substance can be deemed to be readily biodegradable.
Reference
Sample oxygen uptake: biodegradability
|
time, days |
||||||||||||
3 |
5 |
7 |
9 |
12 |
14 |
16 |
18 |
21 |
23 |
25 |
28 |
||
Test item O2uptake, mg/l |
a1 |
52.4 |
83.6 |
102.0 |
115.0 |
128.9 |
138.9 |
164.0 |
165.0 |
168.0 |
169.9 |
172.0 |
173.6 |
a2 |
53.9 |
86.2 |
105.5 |
117.5 |
129.4 |
141.2 |
160.9 |
165.8 |
169.2 |
169.7 |
172.2 |
174.5 |
|
a3 |
48.4 |
78.3 |
98.2 |
112.2 |
125.3 |
137.9 |
160.7 |
162.6 |
167.0 |
169.3 |
172.3 |
174.0 |
|
am. avg |
51.5 |
82.7 |
101.9 |
114.9 |
127.9 |
139.4 |
161.9 |
164.5 |
168.1 |
169.6 |
172.2 |
174.0 |
|
Blank test O2uptake. mg/l |
b1 |
6.7 |
13.8 |
17.7 |
22.9 |
25.6 |
33.3 |
51.6 |
53.5 |
56.2 |
56.3 |
56.3 |
59.7 |
b2 |
3.3 |
8.8 |
13.3 |
17.4 |
19.7 |
27.8 |
45.0 |
46.2 |
48.1 |
48.2 |
50.6 |
50.6 |
|
b3 |
9.0 |
13.6 |
17.5 |
19.7 |
25.6 |
30.3 |
46.8 |
47.8 |
50.8 |
50.6 |
51.0 |
54.1 |
|
bm. avg |
6.3 |
12.1 |
16.2 |
20.0 |
23.6 |
30.5 |
47.8 |
49.2 |
51.7 |
51.7 |
52.6 |
54.8 |
|
Reference item O2uptake. mg/l |
w1 |
52.0 |
64.0 |
73.6 |
79.3 |
87.9 |
95.3 |
113.2 |
113.5 |
115.0 |
115.7 |
118.2 |
118.8 |
w2 |
44.6 |
57.2 |
67.9 |
73.5 |
79.9 |
90.7 |
107.2 |
107.0 |
110.8 |
113.0 |
113.0 |
115.4 |
|
w3 |
48.6 |
61.7 |
71.1 |
76.7 |
82.1 |
90.3 |
107.2 |
107.5 |
109.8 |
110.0 |
110.0 |
113.6 |
|
wm. avg |
48.4 |
61.0 |
70.9 |
76.5 |
83.3 |
92.1 |
109.2 |
109.3 |
111.9 |
112.9 |
113.7 |
115.9 |
|
Toxicity control O2uptake. mg/l |
tox1 |
63.2 |
91.5 |
118.7 |
141.5 |
163.7 |
177.2 |
200.0 |
205.0 |
211.3 |
217.8 |
221.4 |
227.6 |
tox2 |
70.1 |
101.7 |
128.2 |
149.3 |
171.9 |
184.6 |
208.1 |
213.0 |
217.1 |
219.0 |
222.0 |
225.8 |
|
tox3 |
74.8 |
102.9 |
128.1 |
146.5 |
167.4 |
179.5 |
203.3 |
208.3 |
214.4 |
217.7 |
220.6 |
225.3 |
|
toxm. avg |
69.4 |
98.7 |
125.0 |
145.8 |
167.6 |
180.4 |
203.8 |
208.8 |
214.3 |
218.2 |
221.3 |
226.2 |
|
Corrected test item O2uptake, mg/l |
(a1-bm) |
46.1 |
71.5 |
85.8 |
95.0 |
105.3 |
108.5 |
116.2 |
115.9 |
116.3 |
118.2 |
119.4 |
118.8 |
(a2-bm) |
47.6 |
74.1 |
89.4 |
97.5 |
105.8 |
110.7 |
113.1 |
116.6 |
117.5 |
118.0 |
119.6 |
119.7 |
|
(a3-bm) |
42.0 |
66.2 |
82.0 |
92.2 |
101.7 |
107.5 |
112.9 |
113.4 |
115.3 |
117.6 |
119.7 |
119.2 |
|
Reference item % degradation ThOD = 0.78 mgO2/mg C = 100 mg/l |
R1(w1) |
58.5 |
66.5 |
73.7 |
76.1 |
82.3 |
83.1 |
83.9 |
82.5 |
81.2 |
82.0 |
84.1 |
82.0 |
R1(w2) |
49.1 |
57.8 |
66.3 |
68.6 |
72.2 |
77.3 |
76.2 |
74.1 |
75.8 |
78.6 |
77.4 |
77.7 |
|
R3(w3) |
54.2 |
63.7 |
70.4 |
72.7 |
75.0 |
76.8 |
76.2 |
74.8 |
74.5 |
74.7 |
73.5 |
75.4 |
|
Rwavg |
54.0 |
62.7 |
70.2 |
72.4 |
76.5 |
79.1 |
78.8 |
77.2 |
77.2 |
78.5 |
78.3 |
78.4 |
|
Test item % degradation ThOD = 1.57 mgO2/mg C = 92 mg/l |
R1(a1) |
31.9 |
49.5 |
59.4 |
65.8 |
72.9 |
75.1 |
80.4 |
80.2 |
80.5 |
81.8 |
82.6 |
82.2 |
R1(a2) |
32.9 |
51.3 |
61.9 |
67.5 |
73.2 |
76.7 |
78.3 |
80.7 |
81.3 |
81.7 |
82.8 |
82.8 |
|
R3(a3) |
29.1 |
45.8 |
56.8 |
63.8 |
70.4 |
74.4 |
78.2 |
78.5 |
79.8 |
81.4 |
82.8 |
82.5 |
|
Raavg |
31.3 |
48.9 |
59.4 |
65.7 |
72.2 |
75.4 |
79.0 |
79.8 |
80.6 |
81.6 |
82.8 |
82.5 |
|
Toxicity control % degradation
|
R1(tox1) |
25.6 |
35.7 |
46.1 |
54.6 |
63.0 |
66.0 |
68.4 |
70.0 |
71.8 |
74.7 |
75.9 |
77.7 |
R2(tox2) |
28.7 |
40.3 |
50.4 |
58.1 |
66.6 |
69.3 |
72.1 |
73.6 |
74.3 |
75.2 |
76.1 |
76.9 |
|
R3(tox3) |
30.8 |
40.8 |
50.3 |
56.9 |
64.6 |
67.0 |
69.9 |
71.5 |
73.1 |
74.6 |
75.5 |
76.6 |
|
Rtoxavg |
28.3 |
38.9 |
48.9 |
56.5 |
64.7 |
67.4 |
70.1 |
71.7 |
73.1 |
74.8 |
75.8 |
77.1 |
The pH values of test flasks
flask # |
1 |
2 |
3 |
7 |
8 |
9 |
10 |
11 |
12 |
4 |
5 |
6 |
Test item |
Control |
Reference item |
Toxicity test |
|||||||||
initial |
7.12 |
7.22 |
7.42 |
7.48 |
7.50 |
7.54 |
7.57 |
7.47 |
7.46 |
7.45 |
7.45 |
7.46 |
final |
7.22 |
7.23 |
7.26 |
7.27 |
7.19 |
7.20 |
8.37 |
8.43 |
8.60 |
8.13 |
8.53 |
8.35 |
Description of key information
Key study: Test method according to OECD 301F. GLP study. The substance was mostly degraded (82.5%) within a 28-day period. The test substance is regarded as ready biodegradable.
Key value for chemical safety assessment
- Biodegradation in water:
- readily biodegradable
Additional information
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