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EC number: 272-559-0 | CAS number: 68877-33-8
- 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
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- 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
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:
- 2003
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: Real Decreto 363/1995 (BOE núm. 133 de Junio de 1995).
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
- Version / remarks:
- 1984
- GLP compliance:
- no
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- Test concentrations: 10 mg/L, 32 mg/L, 100 mg/L, 320 mg/L and 1000 mg/L, plus two controls.
Preparation of stock and test solutions: A stock solution has been prepared with a concentration of 1 g/L of active matter of the test item. Therefore, 1.025 g of the test item has been dissolved with 100 mL of deionized water in a 100 mL Class A volumetric flask by shaking, so we have a 10250 mg/L stock solution. Because the study material presents an average of 97.54% of active matter, the stock solution contains 1000 mg/L of active matter. The different solutions used in the test have been prepared from this 1000 mg/L stock solution. - Test organisms (species):
- activated sludge, domestic
- Details on inoculum:
- - Preparation of inoculum for exposure: Based on the known concentration of the of dried activated sludge (7.2 g/L) from a domestic waste water treatment plant, a suspestion is prepared by adding water until the concentration os suspended matter is 4 g/L. Then, 200 ml of this suspended sluge solution, is added to the vessels with the rest of components up to a final volume of 500 mL. Therefore, the final concentration of activated sludge (supended solids) is 1.6 g/L
- Pretreatment: no
- Initial biomass concentration: 4g/L - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- 20±2 °C
- pH:
- pH values at initial times:
control (7.23)
10 mg/L (7,25)
32 mg/L (7,22)
100 mg/L (7,16)
320 mg/L (7,12)
1000 mg/L (7,06)
control (7,22) - Nominal and measured concentrations:
- Concentration of the test substance: 10 mg/L; 32 mg/L; 100 mg/L; 320 mg/L; 1000 mg/L and two controls.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: beaker
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: glass
- Aeration: yes, continous flux of 1L/min
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- No. of vessels per abiotic control (replicates): n.a.
- Sludge concentration (weight of dry solids per volume): 7.2 g/L
- Nutrients provided for bacteria: Residual synthetic water prepared according to OECD 209
—Peptone 16 g
—Meat extract 11 g
—Urea 3 g
—NaCl 0,7 g
—CaCl2.2H2O 0,4 g
—MgSO4.7H2O 0,2 g
—K2HPO4 2,8 g
Diluted to 1000 mL with deionized water
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated water
- Particulate matter: not specified
OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: not specified
- Light intensity: not specified
- Details on termination of incubation: The incubation period begins when 200 mL of activated sludge is added at 15 minute intervals to each of the test vessels. All containers are aerated for a period of 3 hours.
EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Oxygen measurements (mg/L) during the respiration inhibition test. Values were collected in 1 minute intervals for a period of 10 minutes (measured with an oxigen electrode cell). - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 262 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- act. ingr.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Details on results:
- Concentration of dissolved oxygen during the inhibition respiratory test where analyzed at 1 minute intervals.
Calculation of oxygen uptake rates are measured for each concentration and control.
Calculation of percentage of inhibition is also reported.
As a conclusion: the substance does not produce any respiratory inhibition in the activated sludges at the maximum concentration of tested substance corresponding to 100 mg/l.
CE50 (3h)>100mg/L - Results with reference substance (positive control):
- Respiration rate of the test control are: 23.7 mgO2/L.h and 24.36 mgO2/L.h. The difference between both values is 2,7% which is inside the acceptation criteria (15%).
CE50-3hr of 3,5 Dichlorophenol is 12 mg/L, which is between the acceptation range of the OECD 209 guide (5mg/L - 30mg/L). - Validity criteria fulfilled:
- yes
- Conclusions:
- The test item Direct Black 155 RBB (batch: 12336-d) has a 3h EC50 value equal to 262 mg/L of based on active matter.
- Executive summary:
The study of bacterial inhibition in activated sludge of the Direct Black 155 RBB dye 97.54% active matter (batch: 12336-d) has been carried out. For the determination of bacterial inhibition, OECD 209 (1984) and Royal Decree 363/1995 (BOE no. 133 of June 1995) have been followed. This method is used to evaluate the effect that the test substance has on microorganisms, measuring the respiration rate under certain conditions and at different concentrations. Activated sludge from an urban sewage treatment plant has been exposed to the dye Direct Black 155 RBB at test concentrations: 10, 32, 100, 320 and 1000 mg/L based on active material. The total duration of the test has been 3 hours. The result obtained in this test indicates that the dye Direct Black 155 RBB has an EC50 value (3h) equal to 262 mg/L based on active material after 3 hours of testing.
Reference
Table 1. Oxygen measurements (mg/L) during the respiration inhibition test:
conc (mg/L) | mg O2/L in the time interval (minutes | ||||||||||
0 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |
control | 7.35 | 6.90 | 6.46 | 5.99 | 5.56 | 5.08 | 4.65 | 4.19 | 3.73 | 3.28 | 2.84 |
10 | 7.28 | 6.82 | 6.36 | 5.94 | 5.45 | 5.01 | 4.57 | 4.11 | 3.64 | 3.20 | 2.75 |
32 | 7.26 | 6.86 | 6.42 | 6.02 | 5.60 | 5.19 | 4.77 | 4.38 | 3.95 | 3.54 | 3.12 |
100 | 7.00 | 6.67 | 6.36 | 6.06 | 5.73 | 5.40 | 5.09 | 4.77 | 4.46 | 4.14 | 3.84 |
320 | 7.22 | 7.04 | 6.85 | 6.65 | 6.47 | 6.27 | 6.08 | 5.88 | 5.79 | 5.51 | 5.31 |
1000 | 7.40 | 7.28 | 7.17 | 7.02 | 6.93 | 6.81 | 6.70 | 6.57 | 6.44 | 6.32 | 6.21 |
control | 7.32 | 6.89 | 6.43 | 5.97 | 5.55 | 5.09 | 4.63 | 4.15 | 3.74 | 2.30 | 2.85 |
Table 2. Respiration rate (mg L-1 h-1) obtained for each of the concentrations and controls:
control | 10 | 32 | 100 | 320 | 1000 | control | |
respiration rate (mg L-1 h-1) | 27.06 | 27.18 | 24.84 | 18.96 | 11.43 | 7.14 | 26.82 |
Table 3. % Inhibition of respiration at 3h:
conc (mg/L) | % Inhibition of respiration |
control | 0 |
10 | 0 |
32 | 7.8 |
100 | 29.6 |
320 | 57.5 |
1000 | 73.5 |
Description of key information
One study is available to determine the inhibition growth of micro-organisms of the substance. The study gives a value of EC50 of 262 mg/L, obtained by calculations based on experimental results.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 262 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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