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EC number: 250-774-0 | CAS number: 31714-55-3
- 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
Short-term toxicity to aquatic invertebrates
Administrative data
- Endpoint:
- short-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 08 December 2017 - 22 January 2018
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
- Version / remarks:
- April 13, 2004
- Deviations:
- yes
- Remarks:
- methodological limitations, e.g. test conc. spacing factor was 2.5 (instead of 2.2), discrepancy between results of solubility test and actually chosen test conc., inconsistent analytical procedures
- Principles of method if other than guideline:
- - The highest achievable test concentration (5 mg/L) did not result in 100% immobilisation.
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Hydrogen bis[1-[(5-chloro-2-hydroxyphenyl)azo]-2-naphtholato(2-)]chromate(1-)
- EC Number:
- 250-774-0
- EC Name:
- Hydrogen bis[1-[(5-chloro-2-hydroxyphenyl)azo]-2-naphtholato(2-)]chromate(1-)
- Cas Number:
- 31714-55-3
- Molecular formula:
- C32H19Cl2CrN4O4
- IUPAC Name:
- [1-{[5-chloro-2-(hydroxy-kappaO)phenyl]diazenyl}-2-naphtholato(2-)-kappaO]{1-[(5-chloro-2-hydroxyphenyl)diazenyl]-2-naphtholato-kappaO}chromium
- Test material form:
- solid: particulate/powder
- Details on test material:
- - Appearance: Black powder
- Storage conditions: At room temperature
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- - Sampling method: test samples from each replicate were drawn and pooled and mixed together for each group
- Volume: 10 mL (from each replicate)
- Frequency: at t=0 and t=48 h
- Sample storage conditions before analysis: not applicable, samples were analysed directly without storage
Test solutions
- Vehicle:
- yes
- Remarks:
- dimethyl formamide
- Details on test solutions:
- TEST FOR SOLUBILITY:
Before conducting the preliminary range finding study, the test item was checked for solubility and precipitation in algal medium, acetone and dimethyl formamide (DMF). Based on the results, DMF was chosen as the appropriate vehicle for the test item.
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a quantity of 13, 32 and 80 mg test item was dissolved in 10 mL DMF to obtain the nominal concentrations of 1.3 mg/mL (stock A), 3.2 mg/mL (stock B) and 8.0 mg/mL (stock C), and a quantity of 20 and 50 mg test item was dissolved in 1 mL DMF to obtain the nominal concentrations of 20.0 mg/mL (stock D) and 50.0 mg/mL (stock E) , respectively.
Volumes of 10 µL from the stock solutions A to E and 1000 µL blank medium with 5 daphnids were taken and diluted to 100 mL with reconstituted water in respective beakers to obtain the nominal test concentrations of 0.13, 0.32, 0.80, 2.00 and 5.00 mg/L.
- Controls: for the blank control group, 1000 µL blank medium with 5 daphnids was transferred using a micropipette and diluted to 100 mL with reconstituted water. For the vehicle control group, 10 µL DMF and 1000 µL blank medium with 5 daphnids were taken and diluted to 100 mL with reconstituted water.
Test organisms
- Test organisms (species):
- Daphnia magna
- Details on test organisms:
- TEST ORGANISM
- Common name: daphnia
- Details on organisms: healthy daphnia were used, showing no signs of stress such as presence of males and ephippia, a delay in the production of the first brood, discolouration, etc. and having normal behavior
- Age at study initiation: <24 hours (first instar nymph)
- Source (Master culture): MicroBio Test Inc, Kleimoer 15, 9030 Mariakerke (Gent), Belgium.
- Feeding during test: no
ACCLIMATION
- Acclimation period: >48 hours
- Acclimation conditions: same as test
- Type and amount of food: living algal cells
- Health during acclimation: no mortality observed
METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: first instars were collected using a micropipette
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 48 h
Test conditions
- Hardness:
- 196 mg CaCO3/L at t=0 in the main test
- Test temperature:
- 20.1-20.7 °C throughout the main test
- pH:
- 7.76-7.95 throughout the main test
- Dissolved oxygen:
- 8.74-8.93 mg/L throughout the main test
- Nominal and measured concentrations:
- Nominal concentrations: 0.13, 0.32, 0.8, 2.0 and 5.0 mg/L
Measured concentrations: 0.13, 0.32, 0.80, 1.93, 4.97 mg/L at t=0 and 0.12, 0.30, 0.81, 2.00 and 5.18 mg/L at t=48 h, respectively, corresponding to above stated nominal concentrations.
The measured concentrations at t=48 h were 90 - 104% of nominal concentrations. - Details on test conditions:
- TEST SYSTEM
- Test vessel: glass beaker with a fill volume of 600 mL
- Type: open
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per blank control (replicates): 4
- No. of vessels per vehicle control (replicates): 4
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dilution water was according to OECD 202. The water was aerated prior to use so that dissolved oxygen concentration had reached saturation.
- Intervals of water quality measurement: Temperature, pH, and dissolved oxygen content (using Multi portable meter) were recorded at 0 and 48 h. Total hardness was measured once during the test period using the titrimetric method.
- Other: Mobility of the daphnids was assessed by gently swirling the test container for 15 seconds and observing their swimming behaviour. Daphnids unable to swim during the agitation of the test container and remaining settled on the water surface or settled at the bottom of the test vessel were recorded as immobile.
OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/8 h dark
- Light intensity: 1320-1350 Lux
EFFECT PARAMETERS MEASURED: immobility was recorded at t=0, t=24 and t=48.
VEHICLE CONTROL PERFORMED: yes
RANGE-FINDING STUDY
- Test concentrations: 0.0, 0.001, 0.01, 0.1, 1.0 and 5.0 mg/L
- Results used to determine the conditions for the definitive study: yes, the observed immobility was 30% at a test concentration of 1 mg/L and 60% at a concentration of 5.0 mg/L. The expected EC50 was between these two test concentrations and based on these result, the test concentrations for the main study were selected. - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate (January 2018)
Results and discussion
Effect concentrations
- Key result
- Duration:
- 48 h
- Dose descriptor:
- EC50
- Effect conc.:
- 2.73 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mobility
- Remarks on result:
- other: 95% CI: 1.62-4.61 mg/L
- Details on results:
- - Behavioural abnormalities: no
- Other biological observations: at t=48 h, lethargy was observed at test concentrations of 0.80, 2.00 and 5.00 mg/L
- Mortality of control: no mortality occured either in the blank control or in the vehicle control
- Measured concentrations at t=48 h were 90 - 104% of nominal concentrations. Based on these results, the effect parameters were based on nominal values.
- Test conditions were within the limits prescribed by OECD guideline 202 during the exposure period.
- For details on incidence of immobilisation, see table 2 in 'any other information on results' section. - Results with reference substance (positive control):
- - Results with reference substance valid?
Yes
- Tested concentrations: 0.09, 0.20, 0.44, 0.96 and 2.11 mg/L
- 48h-EC50: 0.24 mg/L - Reported statistics and error estimates:
- The 48 h EC50 and the associated 95% confidence limits were calculated following the Probit analysis method (Finney, 1971) using in-house developed, validated software.
Any other information on results incl. tables
Table 1 Measured concentrations of T-95/T-99 (main study)
Groups |
Nominal Conc. based on Purity of Test Item (mg/L) |
t=0 h |
t=48 h |
||||
Peak Area of Sample (Y) |
Analysed Conc. (mg/L) |
Recovery# (%) |
Peak Area of Sample (Y) |
Analysed Conc. (mg/L) |
Recovery# (%) |
||
G1 (0.00) |
0.00000 |
ND |
0.00000 |
0.00 |
ND |
0.00000 |
0.00 |
G2 (0.00) |
0.00000 |
ND |
0.00000 |
0.00 |
ND |
0.00000 |
0.00 |
G3 (0.13) |
0.12922 |
11518 |
0.13228 |
102.37 |
8875 |
0.11674 |
90.34 |
G4 (0.32) |
0.31808 |
27943 |
0.31881 |
100.23 |
24993 |
0.30040 |
94.44 |
G5 (0.80) |
0.79520 |
70654 |
0.80385 |
101.09 |
69301 |
0.80527 |
101.27 |
G6 (2.00) |
1.98800 |
169559 |
1.92704 |
96.93 |
174237 |
2.00097 |
100.65 |
G7 (5.00) |
4.97000 |
87373 |
4.96858 |
99.97 |
89605 |
5.18313 |
104.29 |
Intercept of Y-axis (a) |
-130.58 |
-1370.45 |
|||||
Slope of the line (b) |
88056.95 |
87761.04 |
ND = Not Detected, Conc. = Concentration.
# relative to nominal
Dilution factors for the samples of groups G1, G2, G3, G4, G5, G6 and G7 were 1.0, 1.0, 1.0, 1.0, 1.0, 1.0 and 5.0, respectively.
Table 2 Incidence of immobility (main study)
Group |
Nominal Test Concentration (mg/L) |
N° of Replicates |
N° of Daphnids/ group |
Immobility N° and % at |
|||||
0 h |
24 h |
48 h |
|||||||
N° |
% |
N° |
% |
N° |
% |
||||
G1 |
Control (0.0) |
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
G2 |
Vehicle control (0.0) |
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
G3 |
0.13 |
4 |
20 |
0 |
0 |
0 |
0 |
0 |
0 |
G4 |
0.32 |
4 |
20 |
0 |
0 |
0 |
0 |
1 |
5 |
G5 |
0.80 |
4 |
20 |
0 |
0 |
0 |
0 |
5 |
25 |
G6 |
2.00 |
4 |
20 |
0 |
0 |
0 |
0 |
8 |
40 |
G7 |
5.00 |
4 |
20 |
0 |
0 |
0 |
0 |
13 |
65 |
h = Hour
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Remarks:
- See 'overall remarks' section
- Conclusions:
- The 48h-EC50 of T95/T-99 towards Daphnia magna was determined to be 2.73 mg/L (95% CI: 1.62-4.61 mg/L).
- Executive summary:
In an short-term toxicity test performed according to OECD 202 and GLP principles, T-95/T-99 was assessed for its effect on Daphnia magna.
The test item was initially tested for solubility and dimethyl formamide was chosen as the most appropriate vehicle.
Based on a preliminary test, nominal concentrations of 0.13, 0.32, 0.8, 2.0 and 5.0 mg/L were chosen for the main test and both a blank control and a vehicle control were included. The test period was 48 hours, under static conditions, in which immobility was recorded at t=24 and t=48 h. Test samples for analysis of test concentrations were taken at t=0 and t=48 h. Results showed that the test item concnetrations were in the range of 90.3 -104.3% of nominal concentrations throughout the test period, and 88.2 -104.3% of initial measured concentrations at the end of the test. Based on these results, effect parameters were based on nominal concentrations.
The 48h-EC50 of T-95/T-99 towards Daphnia magna was determined to be 2.73 mg/L.
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