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EC number: 276-817-3 | CAS number: 72749-80-5
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
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- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- 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
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
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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:
- 2016-07-14 to 2016-08-11
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)
- Deviations:
- no
- GLP compliance:
- yes
Test material
- Reference substance name:
- Hydrogen 9-amino-7-phenyl-5-(phenylamino)-4,10-disulphonatobenzo[a]phenazinium, disodium salt
- EC Number:
- 276-817-3
- EC Name:
- Hydrogen 9-amino-7-phenyl-5-(phenylamino)-4,10-disulphonatobenzo[a]phenazinium, disodium salt
- Cas Number:
- 72749-80-5
- Molecular formula:
- C28H20N4O6S2.2Na
- IUPAC Name:
- Benzo[a]phenazinium, 9-amino-7-phenyl-5-(phenylamino)-4,10-disulfo-, inner salt, sodium salt (1:2)
- Test material form:
- solid: particulate/powder
Constituent 1
Study design
- Oxygen conditions:
- aerobic
- Inoculum or test system:
- activated sludge, non-adapted
- Details on inoculum:
- - Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Activated sludge from the municipal sewage treatment plant, 31137 Hildesheim, Germany
- Receipt: 2016-07-07
- Preparation of inoculum for exposure: The activated sludge was washed twice with chlorine free tap water. After the second washing the settled sludge was resuspeded in mineral salts medium and was maintained in an aerobic condition by aeration for 2 hours. Thereafter the sludge was homogenized with a blender. After sedimentation the supernatant was decanted and maintained in an aerobic condition by aeration with CO2-free air for 7 days. 10 mL/L were used to initiate inoculation.
- Initial cell/biomass concentration: approx. 3.5 x 10e7 CFU/L in the final test solution
- Water filtered: no - Duration of test (contact time):
- 28 d
Initial test substance concentration
- Initial conc.:
- 34 mg/L
- Based on:
- test mat.
Parameter followed for biodegradation estimation
- Parameter followed for biodegradation estimation:
- O2 consumption
- Details on study design:
- TEST CONDITIONS
- Composition of medium: Mineral salts medium acc. to OECD 301F
- Additional substrate: No
- Test temperature: Nominal: 20 - 24, ± 1 °C; Actual: 20.2 – 20.7 °C
- pH:
pH-Values at Test Start and Test End
pH-Value
Start End
Inoculum Control 7.72 1) 7.69
2) 7.66
Functional Control 7.72 7.87
Test Item 7.71 1) 7.66
2) 7.66
Toxicity Control 7.70 7.87
- pH adjusted: no
- Dispersion treatment: Continuous stirring
- Continuous darkness: Dark, in an incubator
TEST SYSTEM
- Culturing apparatus: Incubator
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: Closed bottles with a rubber sleeve with soda lime
- Measuring equipment: The bottles were closed with OxiTOP measuring heads.
The oxygen consumption was determined in the incubation vessels by the OxiTop® measuring system at 360 measuring points (every 112 minutes) during the 28 d incubation period.
SAMPLING
- Sampling frequency: 360 measuring points (every 112 minutes) during the 28 d incubation period.
- Sampling method: Pressure measurements directly from the test vessels
CONTROL AND BLANK SYSTEM
- Inoculum blank: Test medium without test and reference item
- Abiotic sterile control: No
- Toxicity control: Test item and reference item in test concentrations
STATISTICAL METHODS:
Calculation of the theoretical oxygen demand (ThOD) of the test item and functional control.
According to OECD-guideline 301, the index of the atoms of the item CcHhClclNnNanaOoPpSs was used for the calculation of the ThOD.
The oxygen depletion of the test and reference item was calculated out of the difference of the test and inoculum control replicates.
The biodegradation of the test and reference item was calculated out of the BOD in relation to the ThOD.
Reference substance
- Reference substance:
- benzoic acid, sodium salt
Results and discussion
- Preliminary study:
- No preliminary study
- Test performance:
- Based on the calculated oxygen demand, the test concentration the test vessels of 34 mg/L, corresponding to an oxygen demand of 53.4 mg O2/L (67.3 mg O2/L with nitrification) in the vessel, was selected. The test solutions were prepared in measuring flasks and were placed in brown glass bottles as incubation vessels (inoculum control, functional control and test item) or were prepared directly in brown glass bottles (toxicity control):
• two for the inoculum control (C1, C2)
• one for the functional control (R1)
• two for the test item (P1, P2)
• one for the toxicity control (T1)
Inoculum control
The inoculated test medium, consisting of the required volumes of mineral medium stock solutions, ultrapure water and inoculum, were prepared in a measuring flask. 250 mL of this solution were filled in the brown glass bottles of the inoculum control, using a 250 mL measuring flask.
Functional control
The reference item was weighed out and transferred into a measuring flask, the required volumes of mineral medium stock solutions, ultrapure water and inoculum, were added. 250 mL of this solution were filled in the brown glass bottle of the functional control, using a 250 mL measuring flask.
Test item
The test item was weighed out and transferred into a measuring flask with ultrapure water and stirred overnight. The required volumes of mineral medium stock solutions, ultrapure water and inoculum, were added. 250 mL of this solution were filled in the brown glass bottle of the test item replicate, using a 250 mL measuring flask.
Toxicity control
The reference item was weighed out and directly transferred into the test vessel with 250 mL of the test item solution (see above), using a 250 mL measuring flask.
A rubber sleeve with soda lime was hung into the opening of the bot-tles to absorb evolved CO2. The bottles were closed with
OxiTop measuring heads and the measuring system was activated.
% Degradation
- Key result
- Parameter:
- % degradation (O2 consumption)
- Value:
- 3
- Sampling time:
- 28 d
- Details on results:
- The biodegradation in % was calculated based on the ThOD of 1.57 mg O2/mg test item.
Colony Forming Units of the Inoculum:
Colony forming units (CFU) of the inoculum were determined prior to test start by standard dilution plate count: 3.5 109 CFU/L. The CFU concentration of the inoculum corresponds to approx. 3.5 x 107 CFU/L in the final test solution.
Results of the Functional Control:
The pass level for ready biodegradation (> 60% degradation) was reached on day 3. The validity criterion that the degradation should be > 60% after 14 d was fulfilled. Stages of Biodegradation: The adaptation phase changed to degradation phase within 1 day (degradation > 10%) and the biodegradation came to a maximum of 100% on day 21.
Results of the Toxicity Control:
In the toxicity control the biodegradation achieved 59% after 14 days. After 28 days the biodegradation came to 61%. The validity criterion that the test item should not inhibit the biodegradation of the reference item was fulfilled.
Course and Stages of Biodegradation of Functional Control and Test Item:
The biodegradation of the test item is shown in comparison to the readily degradable functional control and the toxicity control. Both test item replicates did not reach the 10% level (beginning of biodegradation) until test end. The mean biodegradation on day 28 was 3%.
After a test period of 28 days, the test item is classified as not readily biodegradable within the 28 day period of the study.
BOD5 / COD results
- Results with reference substance:
- In the toxicity control the biodegradation achieved 59% after 14 days. After 28 days the biodegradation remained at 61%. The validity criterion that the test item should not inhibit the biodegradation of the reference item was fulfilled.
Any other information on results incl. tables
Biological Oxygen Demand (BOD) of Inoculum Control, Functional Control, Test Item and Toxicity Control
|
|
BOD |
||||||||
Date |
[d] |
Inoculum |
Functional |
Test Item |
Toxicity Control |
|||||
|
|
|
R1 |
P1 |
P2 |
T1 |
||||
|
|
mean |
gross |
net |
gross |
net |
gross |
net |
gross |
net |
2016-07-15 |
1 |
0.6 |
1.6 |
1.0 |
1.3 |
0.6 |
0.0 |
-0.6 |
2.4 |
1.7 |
2016-07-16 |
2 |
1.6 |
32.0 |
30.4 |
2.5 |
0.9 |
1.2 |
-0.4 |
37.5 |
35.9 |
2016-07-17 |
3 |
3.4 |
51.9 |
48.5 |
4.4 |
1.0 |
4.0 |
0.6 |
51.4 |
48.0 |
2016-07-18 |
4 |
4.9 |
62.9 |
58.0 |
6.9 |
2.0 |
5.6 |
0.7 |
62.5 |
57.6 |
2016-07-19 |
5 |
5.6 |
68.5 |
62.9 |
8.4 |
2.8 |
6.6 |
1.0 |
69.7 |
64.1 |
2016-07-20 |
6 |
5.9 |
71.5 |
65.5 |
8.6 |
2.7 |
7.0 |
1.1 |
72.9 |
66.9 |
2016-07-21 |
7 |
7.1 |
73.6 |
66.6 |
9.9 |
2.9 |
8.2 |
1.1 |
75.0 |
68.0 |
2016-07-22 |
8 |
7.1 |
74.7 |
67.6 |
9.9 |
2.8 |
8.5 |
1.4 |
77.1 |
70.0 |
2016-07-23 |
9 |
7.9 |
76.0 |
68.1 |
10.4 |
2.5 |
9.4 |
1.5 |
78.0 |
70.1 |
2016-07-24 |
10 |
8.0 |
77.2 |
69.2 |
11.0 |
3.0 |
9.9 |
1.9 |
79.4 |
71.4 |
2016-07-25 |
11 |
8.2 |
77.9 |
69.6 |
11.3 |
3.1 |
10.4 |
2.1 |
80.5 |
72.3 |
2016-07-26 |
12 |
9.0 |
78.9 |
69.9 |
11.3 |
2.3 |
11.3 |
2.3 |
82.5 |
73.4 |
2016-07-27 |
13 |
9.2 |
80.0 |
70.8 |
11.5 |
2.3 |
11.4 |
2.2 |
83.4 |
74.2 |
2016-07-28 |
14 |
8.6 |
80.3 |
71.7 |
12.7 |
4.1 |
11.4 |
2.9 |
84.5 |
75.9 |
2016-07-29 |
15 |
8.8 |
80.9 |
72.2 |
12.7 |
3.9 |
11.9 |
3.2 |
84.8 |
76.1 |
2016-07-30 |
16 |
9.6 |
81.9 |
72.3 |
12.7 |
3.1 |
12.6 |
3.0 |
86.0 |
76.4 |
2016-07-31 |
17 |
9.9 |
83.1 |
73.2 |
12.8 |
2.9 |
12.7 |
2.8 |
86.4 |
76.5 |
2016-08-01 |
18 |
10.2 |
83.1 |
72.9 |
12.9 |
2.7 |
12.7 |
2.5 |
87.4 |
77.2 |
2016-08-02 |
19 |
10.3 |
84.1 |
73.7 |
12.9 |
2.6 |
12.8 |
2.5 |
87.4 |
77.1 |
2016-08-03 |
20 |
10.6 |
84.6 |
74.0 |
13.2 |
2.6 |
13.1 |
2.5 |
88.0 |
77.4 |
2016-08-04 |
21 |
11.0 |
85.9 |
74.9 |
14.1 |
3.1 |
13.6 |
2.5 |
88.7 |
77.7 |
2016-08-05 |
22 |
11.2 |
86.0 |
74.8 |
13.9 |
2.6 |
14.0 |
2.7 |
88.8 |
77.6 |
2016-08-06 |
23 |
11.3 |
86.0 |
74.7 |
12.8 |
1.5 |
13.9 |
2.6 |
88.8 |
77.5 |
2016-08-07 |
24 |
11.8 |
86.5 |
74.7 |
12.9 |
1.1 |
14.1 |
2.3 |
89.3 |
77.5 |
2016-08-08 |
25 |
12.0 |
87.3 |
75.3 |
13.4 |
1.4 |
14.1 |
2.1 |
90.2 |
78.2 |
2016-08-09 |
26 |
12.2 |
87.4 |
75.2 |
13.6 |
1.3 |
14.1 |
1.9 |
90.2 |
78.0 |
2016-08-10 |
27 |
12.6 |
87.4 |
74.8 |
13.9 |
1.3 |
14.2 |
1.6 |
90.3 |
77.7 |
2016-08-11 |
28 |
13.3 |
88.6 |
75.2 |
14.4 |
1.1 |
15.5 |
2.2 |
91.5 |
78.1 |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Interpretation of results:
- other: under test conditions the test item is classified as not readily biodegradable
- Conclusions:
- The test item is classified as not readily biodegradable within the 28 day period of the study.
- Executive summary:
The ready biodegradability of the test item was determined with non-adapted activated sludge in the Manometric Respirometry Test for a period of 28 days. The study was conducted from 2016-07-14 to 2016-08-11 according to OECD guideline 301 F at the test facility. The test item concentration selected as appropriate was 34 mg/L, corresponding to a ThOD of 53.4 mg O2/L per test vessel
(83.6 mg O2/L per test vessel with nitrification). The oxygen was depleted by the respiration of bacteria and the degradation was followed by measuring the oxygen concentration. The biodegradation
rate is therefore expressed as the percentage BOD (biological oxygen demand) and was calculated for each study day.The mean oxygen depletion in the inoculum control was 13.3 mg O2/L on day 28.
In order to check the activity of the test system sodium benzoate was used as functional control. The pass level > 60% was reached on day 3. The biodegradation reached a maximum of 100% degradation on day 21.
In the toxicity control containing both test and reference item 59% degradation occurred within 14 days. After 28 days the biodegradation came to 61%. The degradation of the reference item was not inhibited by the test item.
The biodegradation of the test item is shown in comparison to the readily degradable functional control and the toxicity control. Both test item replicates did not reach the 10% level (beginning of biodegradation) until test end. The mean biodegradation on day 28 was 3%.
The validity criteria of the guideline are fulfilled.
The test item is classified as not readily biodegradable within the 28 day period of the study.
Biodegradation [%] of the Test Item in Comparison to the Functional Control and Toxicity Control
Biodegradation [%]
Study Day [d]
Replicate
7
14
21
28
Test Item
34mg/L1
5
8
6
2
2
2
5
5
4
Functional Control
45 mg/L89
95
100
100
Toxicity Control
34mg/L Test Item +
45 mg/L Reference Item53
59
60
61
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