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EC number: 255-965-2 | CAS number: 42844-93-9
- 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
- Endpoint:
- activated sludge respiration inhibition testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2017-05-31 to 2017-06-01
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- According to OECD and GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 017
- Report date:
- 2017
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
- Deviations:
- no
- GLP compliance:
- yes
Test material
- Reference substance name:
- [1,3-dihydro-5,6-bis[[(2-hydroxy-1-naphthyl)methylene]amino]-2H-benzimidazol-2-onato(2-)-N5,N6,O5,O6]nickel
- EC Number:
- 255-965-2
- EC Name:
- [1,3-dihydro-5,6-bis[[(2-hydroxy-1-naphthyl)methylene]amino]-2H-benzimidazol-2-onato(2-)-N5,N6,O5,O6]nickel
- Cas Number:
- 42844-93-9
- Molecular formula:
- C29H18N4NiO3
- IUPAC Name:
- Nickel, [1,3-dihydro-5,6-bis[[[2-(hydroxy-kO)-1-naphthalenyl]methylene]amino-kN]-2H-benzimidazol-2-onato(2-)]-, (SP-4-2)-
- Test material form:
- solid: nanoform
Constituent 1
Sampling and analysis
- Analytical monitoring:
- no
Test solutions
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1000 mg/L were weighed out directly in Erlenmeyer flasks.
- Eluate: Synthetic waste water acc. to OECD 209
- Differential loadings: 1000 mg/L, 3 replicates
- Controls: Test medium without test or reference item. Triplicates at the beginning of the test + triplicates at the end of the test
-Reference item: three concentrations within the concentration range 58 - 180 mg/L and a dilution factor of 1.8., Triplicates
Test organisms
- Test organisms (species):
- activated sludge
- Details on inoculum:
- - Preparation of inoculum for exposure: Non-adapted activated sludge from the sewage plant at Hildesheim (Municipal sewage treatment plant of 31137 Hildesheim, Germany)
- Receipt: 2017-05-31
- Pretreatment: The sludge was washed twice with chlorine free tap water and adjusted to a dry sludge concentration of 3.0 g/L ± 10 %. The sludge was used within 24 h after sampling.
- Dry sludge concentration: 2.71 g/L, corresponding to 1.36 g/L in the test vessel
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 3 h
Test conditions
- Hardness:
- Not determined
- Test temperature:
- 20.8 °C
- pH:
- pH-value of the activated sludge 7.45
pH-value of the synthetic waste water 7.34 - Salinity:
- Not determined
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 3
- 500 mL Erlenmeyer flasks, DIN 12380
- Aeration: Permanent, to keep the dissolved oxygen concentration above 60 - 70 % saturation and to maintain the sludge flakes in suspension.
- Pretreatment: stirring overnight
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6 (Triplicates at the beginning of the test + triplicates at the end of the test.)
-No. of vessels per reference item (replicates): 9 (three concentrations within the range 58 - 180 mg/L and a dilution factor 1.8,at the end of the test)
- Composition of test medium:
Test Item Test concentration [mg/L]
1000
Test Item [mg] 500
Synthetic waste water [mL] 16
Demineralised water [mL] Filled up to 250 mL
Inoculum [mL] 250
Dilution Table for the Reference Item
Reference item concentration [mg/L] 58 100 180
Stock solution [µL]* 96.7 166.7 300
Synthetic waste water [mL] 16
Demineralised water [mL] Filled up to 250 mL
Inoculum [mL] 250
* Stock solution with 300g/L
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
After 3 h incubation, the oxygen concentration of each control, reference item and test item replicate was determined by means of oximeter every 5 seconds after transferring the solution to the measuring chamber and the oxygen depletion was recorded for at least 5 minutes with the software PreSens Measurement Studio 2.
TEST CONCENTRATIONS
Inhibitions in the Preliminary Tests
A preliminary test (non GLP) was carried out with the following test item concentrations of PV-Echtorange 6RL:
10 – 100 - 1000 mg/L with two replicates each. The pH was not adjusted.
In the preliminary test, additional replicates were prepared to determine the inhibition of nitrification. Two replicates 1000 mg/L were measured with addition of Allylthiourea ATU.
No indication for the requirement to measure the inhibition of nitrification in the definitive study was observed, even the inhibitions of the heterotrophic respiration or nitrification show no appreciable value.
Inhibitions in the Preliminary Tests
Nominal Test Item Concentration
[mg/L] pH
(test item in medium) Inhibition
[%]
1st preliminary test - without pH adjustment, direct aeration
10 - 1
- -3
100 - 0
- 3
1000 7.44 -4
- 5
Inhibitions in the Preliminary Test (with/ without ATU)
Nominal Test Item Concentration
[mg/L] Inhibition of total respiration
[%] Inhibition of heterotrophic respiration
[%] Inhibition of nitrification
[%]
1000 -4 -16 3
5 -13 15
Based on the results of the preliminary test it could concluded that the heterotrophic respiration as well as the nitrification are not inhibited. Therefore, the definitive study was carried out with the concentration of 1000 mg/L in 3 replicates (limit test to determine the NOEC). The concentration was weighed out directly in the Erlenmeyer flasks.
Triplicates of the control without test item were included at the beginning and at the end of the test.
A fresh sample of activated sludge taken from the sewage plant Hildesheim was used for the test procedure. It was washed twice and adjusted with chlorine free tap water if necessary to a dry sludge concentration of 3.0 g/L ± 10 %.
The reference item was tested with three concentrations within the concentration range 58 - 180 mg/L and a dilution factor of 1.8. - Reference substance (positive control):
- yes
- Remarks:
- Copper (II) sulphate pentahydrate p.a.
Results and discussion
Effect concentrations
- Key result
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- >= 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- Reported statistics and error estimates:
- The NOEC was determined by calculation of statistical significance of the inhibition of respiration in comparison to the control. One Way Analysis of Variance (ANOVA) was used for NOEC calculation. When running a One Way Analysis of Variance a Normality test and an Equal Variance test were done first. P-value for both tests was 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) is =0.05.
The EC-value of the reference item was calculated by linear regression using software GraphPad Prism. Calculation of the confidence intervals for the EC50-value was carried out using standard procedures. The coefficient of variation of the oxygen uptake rates of the control replicates was calculated with standard methods using Excel.
Any other information on results incl. tables
From the oxygen depletion the oxygen uptake rates were calculated in mg O2/L×h, additionally the specific respiration rates based on the dry sludge concentration were calculated in mg O2/g×h. The inhibition of the oxygen uptake rates of the test and reference item concentrations in % was calculated in comparison to the oxygen uptake rates of the control.
Oxygen Uptake Rates and Specific Respiration Rates of the Control Replicates
|
Repl. |
Oxygen |
Oxygen Uptake Rate R |
Specific Respiration Rate Rs |
|
|
[mg O2/L] |
[mg O2/(L×h)] |
[mg O2/g×h] |
Control |
1 |
7.88 |
32.8 |
24.2 |
2 |
7.93 |
33.4 |
24.6 |
|
3 |
7.24 |
32.6 |
24.0 |
|
4 |
8.06 |
34.4 |
25.4 |
|
5 |
8.67 |
33.8 |
24.9 |
|
6 |
8.41 |
32.8 |
24.2 |
|
|
Mean value |
33.3 |
24.6 |
|
Standard deviation |
0.640 |
|
||
CV [%] |
1.92 |
*) determined at start of the measurement
Repl. = Replicate
CV = Coefficient of variation
Oxygen Uptake Rates and Inhibition of the Respiration of the Test Item Replicates
Test Item Concentration |
Repl. |
Oxygen |
Oxygen Uptake Rate R |
Inhibition |
Mean Inhibition |
[mg/L] |
|
[mg O2/L] |
[mg O2/(L×h)] |
[%] |
[%] |
1000 |
1 |
8.11 |
35.8 |
-8 |
-6 |
2 |
6.74 |
35.4 |
-6 |
||
3 |
8.01 |
35.1 |
-5 |
*) determined at start of the measurement
Repl. = Replicate
Oxygen Uptake Rates and Inhibition of the Respiration of the Reference Item Replicates
Reference Item Concentration |
Repl. |
Oxygen |
Oxygen Uptake Rate R |
Inhibition |
Mean Inhibition |
[mg/L] |
|
[mg O2/L] |
[mg O2/(L×h)] |
[%] |
[%] |
58 |
1 |
7.20 |
24.6 |
26 |
28 |
2 |
7.80 |
24.1 |
28 |
||
3 |
7.13 |
23.3 |
30 |
||
100 |
1 |
8.29 |
15.9 |
52 |
52 |
2 |
7.56 |
16.8 |
50 |
||
3 |
6.94 |
15.1 |
55 |
||
180 |
1 |
9.17 |
6.0 |
82 |
82 |
2 |
9.31 |
6.3 |
81 |
||
3 |
8.91 |
5.7 |
83 |
*) determined at start of the measurement
Repl. = Replicate
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- The NOEC of the test item is ≥ 1000 mg/L.
The test item caused no inhibiting effects on activated sludge of a municipal sewage treatment plant at the limit test item concentration of 1000 mg/L. - Executive summary:
A Respiration Inhibition Test with activated sludge according to OECD Guideline No. 209 (2010) was carried out with the test item (batch number: DEB2 188394) from 2017 -05 -31 to 2017 -06 -01, with the definitive exposure phase on 2017 -06 -01 at the test facility. The test system was activated sludge of the municipal treatment plant of 31137 Hildesheim, Germany. The test was carried out under static conditions with the limit test item concentration 1000 mg/L. The respiration rates of the control, reference and test item replicates were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. The mean inhibition of respiration for the test item replicates was -6% .
In order to check the activity of the test system and the test conditions a reference test was carried out with copper (II) sulphate pentahydrate as reference item and the reference toxicity was determined. The EC50-value for the reference item was 93.1 mg/L. The validity criteria of the test guideline were fulfilled.
NOEC of the test item
Test item Concentration
[mg/L]NOEC*
≥ 1000
*) No statistically significant inhibiting effect (P = 0.002), ANOVA
The NOEC of the test item is
≥1000 mg/L.the test item caused no inhibiting effects on activated sludge of a
municipal sewage treatment plant at the limit test item concentration of
1000 mg/L.
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