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EC number: - | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
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- Density
- Particle size distribution (Granulometry)
- Vapour pressure
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- Endpoint summary
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
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- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
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Biodegradation in water: screening tests
Administrative data
- Endpoint:
- biodegradation in water: ready biodegradability
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 21 December 2010 to 18 January 2011
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study conducted to OECD test guidance in compliance with GLP.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 011
- Report date:
- 2011
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
- Deviations:
- no
- GLP compliance:
- yes
Test material
- Details on test material:
- Name: Reactive Blue F08-0170
Constituent 1
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Study design
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge (adaptation not specified)
- Details on inoculum:
- Name: Activated sludge filtrate
Source: The secondary of Liede Sewage Treatment Plant of Guangzhou
Selection reason: The inoculum is recommended by "The Guidelines for the Testing of Chemicals"
Treatment: The sludge was removed any coarse particles and impurities on the surface, then was washed with the mineral medium (1100g, 10min, repeated for 5 times) , the supernatant liquid phase was decanted and the solid material resuspended in a mineral medium. The sludge was mixed by vortex for 5min and filtered by qualitative filter paper. The filtrate was kept aerobic until use. 1mL of filtrate per liter of medium was inoculated in the test system - Duration of test (contact time):
- 28 d
Initial test substance concentration
- Initial conc.:
- 5 mg/L
- Based on:
- test mat.
Parameter followed for biodegradation estimation
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- Test Medium
The test medium was prepared with the deionised water.
Instruments and equipments
Electronic balance: BSA223S Sartorius Scientific Instruments (Beijing) Co. Ltd.
Electronic balance: AG245 METTLER TOLEDO
High speed refrigerated: BECKMAN
centrifuge Avanti J-301
Magnetic stirrer :S21-1 Shanghai Sile Instrument Co. Ltd.
Aeration pump: IPX4 ACO-9610 Guangdong Haili Grpoup Co. Ltd.
Dissolved oxygen analyzer: WTW
Oxi 730
Test methods
Selection of the test method
The test substance is easily soluble in water, while the relevant physical and chemical properties of the test substance including the vapour pressure is unknown, so the ready biodegradability: closed bottle test was adopted.
Addition of the test substance
70.0mg test substance was dissolved in some test medium. The solution was transferred to 200ml volumetric flask. The stock solution was diluted with test medium to 200mL, and the stock solution with the concentration of 350mg/L was obtained.
Preparation of reference substance stock solution
50.0mg sodium benzoate was dissolved in some test medium, then transferred to 50mL volumetric flask. The stock solution was diluted with test medium to 50mL, and the stock solution with the concentration of 1000mg/L was obtained.
Test grouping
Test suspension: 20 BOD bottles containing test substance and inoculum
Inoculum blank: 20 BOD bottles containing only inoculum
Procedure control: 10 BOD bottles containing reference compound and inoculum
Toxicity control: 10 BOD bottles containing test substance, reference compound and inoculum
Preparation of test system
Addition of the test medium firstly: 6800mL, 6800mL, 4100mL and 4100mL of fully-aerated mineral medium was added to flask 1, flask 2, flask 3 and flask 4 respectively. The medium was ready for use after standing for 20h at 20°C
Addition of test substance stock solution: 100mL and 60mL test substance stock solution was added to flask 1 and flask 4 respectively
Addition of reference substance stock solution: 8.4mL reference substance stock solution was added to flask 3 and flask 4 respectively
Addition of activated sludge filtrate: 7.0mL, 7.0mL, 4.2mL and 4.2mL of activated sludge filtrate were added to flask1, flask 2, flask 3 and flask 4
Addition of the test medium lastly: The final volumes of suspensions in flask1, flask 2, flask 3 and flask 4 were made up to 7000mL, 7000mL, 4200mL and 4200mL respectively, by the addition of fully-aerated mineral medium. The suspensions were mixed uniformly.
Dispense reaction solutions: Reaction solutions were dispensed immediately into BOD bottles by hose from the 3/4 quarter under solution surface. All the BOD bottles were completely filled and were taped gently to remove any air bubbles.
Test substance concentration: 5.0mg/L
Reference substance concentration: 2.0mg/L
Concentration of inoculums: 1ml filtrate/L medium
Test conditions
All the BOD bottles were incubated at 20°C±2°C in the dark.
Analysis and determination
Duplicate bottles of all series were withdrawn for dissolved oxygen analysis firstly, and then for NO2--N and NO3--N analysis at time intervals (0d, 3d, 7d, 10d, 14d, 17d, 21d, 24d and 28d) over the 28 days incubation. Procedure and toxicity controls were analyzed until 14d.
Reference substance
- Reference substance:
- benzoic acid, sodium salt
Results and discussion
- Preliminary study:
- Not applicable.
- Test performance:
- The oxygen depletion of the inoculum blank (0.72mg/L) was lesser than 1.5mg/L on the28th day.
The residual concentration of oxygen in the test bottles did not fall below 0.5 mg/L at any time.
The difference of replicate values of the removal of the test chemical at the end of the test was less than 20%.
The percentage degradation of the reference compound (73.2%) had reached the pass levels (60%) by day 14.
The percentage degradation of the toxicity control was 28.9% by day 14, greater than 25%.
The test is considered to have fulfilled the validity criteria of the specified method.
% Degradation
- Parameter:
- % degradation (O2 consumption)
- Value:
- 0
- Sampling time:
- 28 d
- Details on results:
- At the start of the test, NO2¯-N and NO3¯-N concentrations of the test suspensions were 0.065mg/L, 0.042mg/L(2 replicates) and 0.040mg/L, 0.009mg/L(2 replicates) respectively. NO2¯-N and NO3¯-N concentrations of the inoculum blanks were 0.0.054mg/L, 0.054mg/L(2 replicates) and 0.012mg/L, 0.012mg/L(2 replicates) respectively. At the end of the test, NO2¯-N and NO3¯-N concentrations of the test suspensions were 0.025mg/L, 0.031mg/L (2 replicates) and 0.142mg/L, 0.142mg/L (2 replicates) NO2¯-N and NO3¯-N concentrations of the inoculum blanks were 0.026mg/L, 0.047mg/L (2 replicates) and 0.144mg/L, 0.143mg/L (2 replicates) respectively.
The results revealed that there was no nitrification occurred in bottle 1, while there was nitrification occurred in bottle 2, which resulted in 0.007mg/L of oxygen consumed.
During the test period, the measured minimum dissolved oxygen concentration was 6.16mg/L, which revealed that the residual concentration of oxygen in the test bottles were greater than 0.5mg/L. The oxygen depletion of the inoculum blank (0.72mg/L) was lesser than 1.5mg/L on the 28th day, which met the test requirement.
The percentage biodegradation of parallel of the test substance were 0%, so the test substance was not degradable during the test.
BOD5 / COD results
- Results with reference substance:
- The percentage biodegradation of reference substance and the toxicity control was 73.2% and 28.5% respectively on the 14th day, which showed that the activity of the inoculum met the requirement of the test. Statistical analysis was carried out using t-test on the oxygen depletion at the end of the test for the control and test substance vessels to determine any significant differences between the test and control groups. The analysis results revealed that there were no statistically significant differences (P≥0.05) between the oxygen depletion for the test and group groups. So the test substance was therefore considered not to have a toxic effect on the sewage sludge micro-organisms used in the study.
Any other information on results incl. tables
Table 1 – Composition of test systems in flasks
Test groups |
Test suspension
Flask 1 |
Inoculum blank
Flask 2 |
Reference control Flask 3 |
Toxicity control
Flask 4 |
Test medium added firstly (mL) |
6800 |
6800 |
4100 |
4100 |
Test substance stock solution (mL) |
100 |
- |
- |
60 |
Reference substance stock solution (mL) |
- |
- |
8.4 |
8.4 |
Activated sludge filter (mL) |
7.0 |
7.0 |
4.2 |
4.2 |
Test medium added lastly (mL) |
93.0 |
193.0 |
87.4 |
27.4 |
Final volume (mL) |
7000 |
7000 |
4200 |
4200 |
Test substance concentration (mg/L) |
5.0 |
- |
- |
5.0 |
Reference substance concentration (mg/L) |
- |
- |
2.0 |
2.0 |
Table 2 – NO2+-N, NO3+-N contents of the test suspensions during the test
Time (d) |
Test suspension (T) |
Inoculum blank (b) |
||||||
T1 |
T2 |
B1 |
B2 |
|||||
NO3¯-N |
NO2¯-N |
NO3¯-N |
NO2¯-N |
NO3¯-N |
NO2¯-N |
NO3¯-N |
NO2¯-N |
|
0 |
0.040 |
0.065 |
0.009 |
0.042 |
0.012 |
0.054 |
0.012 |
0.054 |
3 |
0.012 |
0.084 |
0.011 |
0.077 |
0.012 |
0.055 |
0.013 |
0.074 |
7 |
0.012 |
0.106 |
0.010 |
0.075 |
0.012 |
0.091 |
0.012 |
0.090 |
28 |
0.142 |
0.025 |
0.142 |
0.031 |
0.144 |
0.026 |
0.143 |
0.047 |
Consumed oxygen (mg/L) |
0.466 |
0.608 |
0.601 |
|||||
Consumed oxygen of nitrification (mg/L) |
0 |
0.007 |
- |
Table 3 – DO of the test suspensions during the test
Time (d) |
Test suspension (T) |
Inoculum blank (B) |
Reference control (R) |
Toxicity control (C) |
||||
T1 |
T2 |
B1 |
B2 |
R1 |
R2 |
C1 |
C2 |
|
0 |
8.64 |
8.58 |
8.65 |
8.62 |
8.57 |
8.65 |
8.55 |
8.57 |
3 |
8.93 |
8.85 |
9.09 |
9.08 |
7.09 |
7.13 |
7.16 |
7.14 |
7 |
8.93 |
8.87 |
8.97 |
8.96 |
6.84 |
6.86 |
6.88 |
6.86 |
10 |
8.73 |
8.86 |
8.78 |
8.73 |
6.46 |
6.41 |
6.62 |
6.67 |
14 |
8.79 |
8.85 |
8.65 |
8.68 |
6.16 |
6.17 |
6.68 |
6.71 |
21 |
9.03 |
9.08 |
8.89 |
8.66 |
- |
- |
- |
- |
28 |
8.37 |
8.34 |
8.31 |
8.25 |
- |
- |
- |
- |
Table 4 – Percentage biodegradation during the test
Test groups |
Time (d) |
||||
7 |
14 |
21 |
28 |
||
Test substance |
Percentage biodegradation (%) |
-9.0 -9.0 |
-4.7 -8.4 |
-2.6 -3.6 |
-2.2 -5.2 |
Reference substance |
Percentage biodegradation (%) |
51.8 53.6 52.7 |
72.2 74.3 73.2 |
- - - |
- - - |
Average (%) |
|||||
Toxicity control |
Percentage biodegradation (%) |
25.5 26.1 25.8 |
28.5 28.4 28.5 |
- - - |
- - - |
Average (%) |
Note: The percentage was regarded as 0% when calculated value was negative.
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- under test conditions no biodegradation observed
- Conclusions:
- The test substance cannot be considered to be readily biodegradable under the conditions of the study presently performed.
- Executive summary:
Ready Biodegradability: Closed Bottle Test of Reactive Blue F08-0170 was performed in compliance with OECD Guidelines for the Testing of Chemicals, 301D Closed Bottle Test. Paris: OECD, Adopted 17 July 1992 and the Principles of Good Laboratory Practice (GLP), the test was conducted according to the study plan and SOP, and the results and this report accurately and completely reflect the raw data.
Analysis and determination
The dissolved oxygen, NO2¯-N and NO3¯-N concentrations of the test solution was measured at intervals.
Results
Consumed oxygen of inoculum blank (28d): 0.72mg/L
Percentage biodegradation
Reference substance (14d): 73.2%
Toxicity control (14d): 28.5%
Test substance (28d): 0%
Conclusion
The test substance cannot be considered to be readily biodegradable under the conditions of the study presently performed.
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Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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