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EC number: 241-922-5 | CAS number: 18015-76-4
- 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
Link to relevant study record(s)
- Endpoint:
- short-term toxicity to aquatic invertebrates
- Type of information:
- other: read across from analogue substance
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
- Version / remarks:
- April 13, 2004
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: highest concentration 2.5 mg/l and 0.16 mg/l
- Sampling method: The samples for analysis (0 hours) were prepared at the beginning of the test and immediately delivered in transport box to analytical laboratory. The samples were analysed on the day of delivery. The samples for analysis at the end of the test (48 hours) were delivered to analytical laboratory immediately after the end of testing. The samples were analysed on the day of delivery. All samples were stored at laboratory temperature. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Preparation of solutions of the test item for preliminary and definite test
The two stock solutions of the test item with different pH value (4.5 and 7.8) were prepared in the dilution water for preliminary tests. The pH was adjusted with hydrochloric acid. The one stock solution of the test item with pH 7.8 was prepared in the dilution water for definitive test.
5 mg of the test item was weighed into 500 mL of the dilution water for the preliminary tests and for definitive test. The concentrations of solutions used in the preliminary and definitive test were obtained by dilution of the stock solution with dilution water.
The test item is ionised chemical and coloured dye that shows different forms at different pH, namely EC 255-288-2 (acidic pH) and its carbinol base EC 208-109-7 (basic pH) . For this reason (according OECD GD No.23, chapter 7, and its different subsections 7.6 Coloured test chemicals and 7.8 Ionised test chemicals and after consultation with sponsor) two preliminary tests were performed with different pH to determine the most toxic form.
The preliminary tests were performed at pH 4.5 and 7.8 in a range of the test item nominal concentrations 0.1 – 5 mg·L-1 in a static design. Samples for analytical determination were taken at the beginning and at the end of the test from the highest concentration (5.0 mg·L-1) and the lowest concentration (0.1 mg·L-1 ) The analytical results showed, that the test item was stable in the dilution water at conditions of the test in the highest concentration (5.0 mg·L-1). In the lowest concentration (0.1 mg·L-1) the test item was not stable in the dilution water at conditions of the test. The reason is probably due to its photodegration properties.
In the preliminary test with pH 7.8 slightly higher toxicity was observed than in the preliminary test with pH 4.5.
On the basis of the preliminary tests results the definitive test was performed at pH 7.8 and in the dark. The definitive test was performed in a range of nominal concentrations from 0.04 mg·L-1 to 2.5 mg·L-1. The geometric concentration series of the test item with factor 2.0 was used for the test.
Main test concentrations: 2.5, 1.25, 0.63, 0.32, 0.16, 0.08 and 0.04 mg·L-1. As reported in the section of the validation of the analytical method, due to the analytical conditions acidic pH (mobile phase has pH 3) the sampled test solutions were acidifed before the analyses and therefore it is possible to quantify the total amount of test substance but not the ratio of each single chemical species (cationic vs. carbinol). - Test organisms (species):
- Daphnia magna
- Details on test organisms:
- TEST ORGANISM
- Common name: Daphnia Magna Straus
- Strain/clone:
- Justification for species other than prescribed by test guideline: the prescribed organism was used
- Age at study initiation (mean and range, SD): young daphnia born in 24h
- Weight at study initiation (mean and range, SD): not relevant
- Length at study initiation (length definition, mean, range and SD): not reported
- Source: internal breeding, obtained by aperiodic parthenogenesis
- Feeding during test: no feeding
The grow-up gravid females are separated into dilution water. Young daphnia born
in 24 hours are segregated to the handling container from which they entered the experiment. At handling a care should be taken of not to penetrate the air under animal’s shell.
ACCLIMATION
Adaptation: Young daphnia used for preliminary test with pH 4.5 were born into dilution water with pH 6.0. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 48 h
- Post exposure observation period:
- none
- Test temperature:
- Range of temperature: 19.0– 19.9 °C
- pH:
- 7.7-7.8
- Dissolved oxygen:
- 8.6-9.2 mg/l
- Nominal and measured concentrations:
- nominal concentrations tested : 0.16 and 2.5 mg/l
measured concentrations tested: 0.17 (0% ) and 3.8 (+ 3.6%) mg/l - Details on test conditions:
- TEST SYSTEM
- Volume of test item solutions: 50 mL (least 2 mL per individual)
- Aeration: no aeration
- No. of organisms per vessel: 5 animals per replicate
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates):
TEST MEDIUM / WATER PARAMETERS
The holding and culturing of the Daphnia is performed at conditions (light, temperature, medium) similar to those which are used in the test.
For holding and culturing of the daphnia the M4 medium was used (according to Annex 2
of the EU method C.2 and according to Annex 3 of the OECD TG 202).
For test the dilution water was used (according to Annex 2 of the EU method C.2 and according to Annex 3 of the OECD TG 202). Dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D into 1 000 mL of deionized water.
Because culturing and test medium were different (M4 and dilution water), we included pre-test acclimation period: brood daphnids were maintained in dilution water for 48 hours prior to the start of the test.
Adaptation: Young daphnia used for preliminary test with pH 4.5 were born into dilution water with pH 6.0.
Dilution water
A. 117.6 g CaCl2·2H20 in 1L deionized water
(Batch No.: PP/2021/00741, exp.: 03/2023, Lach-Ner, s. r.o.)
B. 49.3 g MgSO4·7H2O in 1L deionized water
(Batch No.: PP/2018/08687, exp.: 04/2023, Lach-Ner, s. r.o.)
C. 25.9 g NaHCO3 in 1L deionized water
(Batch No.: PP/2019/11502, exp.: 09/2024, Lach-Ner, s. r.o.)
D. 2.3 g KCl in 1L deionized water
(Batch No.: PP/2021/08772, exp.: 06/2024, Lach-Ner, s. r.o.)
The dilution water should have a pH 7.8 0.2, values measured in this study were 7.9 in the preliminary tests and 7.8 in the definitive test.
The content of sum Ca + Mg ions should have the value of 2.50 0.10 mmol·L-1, values measured in this study were 2.52 and 2.53 mmol·L-1 in the preliminary tests and
2.56 mmol·L-1 in the definitive test.
The dilution water for preliminary test with pH 4.5 was adjusted at pH 4.5. The pH was adjusted with hydrochloric acid.
Measured values meet the requirements.
OTHER TEST CONDITIONS
- Adjustment of pH: with hdrochloridric acid
- Photoperiod: dark in the main test
- Light intensity: none
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
immobilisation
VEHICLE CONTROL PERFORMED: no
RANGE-FINDING STUDY
- Test concentrations: 5.0, 2.5, 1.0, 0.5 and 0.1 mg·L-1
- Results used to determine the conditions for the definitive study: see table below - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Duration:
- 48 h
- Dose descriptor:
- EC50
- Effect conc.:
- ca. 0.22 mg/L
- 95% CI:
- > 0.17 - < 0.27
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mobility
- Duration:
- 48 h
- Dose descriptor:
- EC100
- Effect conc.:
- ca. 1.25 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mobility
- Duration:
- 48 h
- Dose descriptor:
- EC0
- Effect conc.:
- ca. 0.04 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- mobility
- Results with reference substance (positive control):
- The sensitivity of the test species and correctness of test performance is periodically verified
in six-month period by testing with the reference item, potassium dichromate.
The results of the verification test with K2Cr2O7, carried out in period from 09.05. to 13.05.2022 are the following:
24 hour – EC50 = 1.13 mg·L-1 (95% confidence limit: 0.94 – 1.47 mg·L-1)
48 hour – EC50 = 0.67 mg·L-1 (95% confidence limit: 0.58 – 0.76 mg·L-1)
b) Interlaboratory test data
From the result of the last interlaboratory test arranged by CSlab spol. s r.o. (accredited provider of proficiency testing) in 2019 we have obtained data for potassium dichromate
48h – EC50 generated at 7 laboratories. The following reference range of values could be calculated from those data:
48 hour – EC50 = 0.50 – 1.16 mg·L-1
c) Method validity evaluation
The immobilisation of the test organism by the reference item (48 hour – EC50) obtained
from our last reference test meets the calculated range from the interlaboratory test. - Validity criteria fulfilled:
- yes
- Conclusions:
- The analogue substance was tested for short-term toxicity to daphnia according to OECD 202. Under the experimental conditions the EC50, 48 h is equal to 0.22 mg/l.
- Executive summary:
The analogue substance was tested for short-term toxicity to daphnia according to OECD 202. The substance is an ionic dye that shows two different chemical structures depending on the pH of the solution, i.e. the target substance and its carbinol base. Following the OECD 23 , preliminary tests at different pH were performed for assessing the most toxic design for the main test and the relavant test substance concentrations. After the dose range finding the pH equal to 7.8 was selected with a range of concentrations from 0.04 to 2.5 mg/l (nominal concentrations) spacing with a factor of 2. Moreover, due to potential photodegradation occuring at the lowest test concentration, the main test was performed in the dark. The analytical monitoring of the test concentrations was performed by HPLC-MS with measured concentrations within the 20% of the nominal values. Five animals of the species Daphnia Magna per replicate and four replicate per concentrations were tested and after 48h the mobility of the organisms was recorded. Under the experimental conditions the EC50,48 h resulted equal to 0.22 mg/l.
Reference
Preliminary test at pH 4.5 | ||||||||||
Nominal concentration | mg·L-1 | 5.0 | 2.5 | 1.0 | 0.5 | 0.1 | Control | |||
Number of mobile animals | 24 hours/1st series | 0 | 0 | 1 | 3 | 5 | 5 | |||
Number of mobile animals | 24 hours/2nd series | 0 | 0 | 0 | 3 | 5 | 5 | |||
Number of mobile animals | 24 hours/3rd series | 0 | 0 | 1 | 3 | 4 | 5 | |||
Number of mobile animals | 24 hours/4th series | 0 | 0 | 2 | 4 | 4 | 5 | |||
Number of mobile animals | 24 hours/total | 0 | 0 | 4 | 13 | 18 | 20 | |||
Immobilisation % | 24 hours | 100 | 100 | 80 | 35 | 10 | 0 | |||
Number of mobile animals | 48 hours/1st series | 0 | 0 | 0 | 3 | 3 | 5 | |||
Number of mobile animals | 48 hours/2nd series | 0 | 0 | 0 | 1 | 3 | 5 | |||
Number of mobile animals | 48 hours/3rd series | 0 | 0 | 0 | 1 | 3 | 5 | |||
Number of mobile animals | 48 hours/4th series | 0 | 0 | 0 | 1 | 2 | 5 | |||
Number of mobile animals | 48 hours/total | 0 | 0 | 0 | 6 | 11 | 20 | |||
Immobilisation % | 48 hours | 100 | 100 | 100 | 70 | 45 | 0 | |||
Preliminary test at pH 7.8 | ||||||||||
Nominal concentration | mg·L-1 | 5.0 | 2.5 | 1.0 | 0.5 | 0.1 | Control | |||
Number of mobile animals | 24 hours/1st series | 0 | 0 | 1 | 2 | 5 | 5 | |||
Number of mobile animals | 24 hours/2nd series | 0 | 0 | 0 | 3 | 3 | 5 | |||
Number of mobile animals | 24 hours/3rd series | 0 | 0 | 0 | 3 | 4 | 5 | |||
Number of mobile animals | 24 hours/4th series | 0 | 0 | 2 | 3 | 3 | 5 | |||
Number of mobile animals | 24 hours/total | 0 | 0 | 3 | 11 | 15 | 20 | |||
Immobilisation % | 24 hours | 100 | 100 | 85 | 45 | 25 | 0 | |||
Number of mobile animals | 48 hours/1st series | 0 | 0 | 0 | 0 | 3 | 5 | |||
Number of mobile animals | 48 hours/2nd series | 0 | 0 | 0 | 1 | 2 | 5 | |||
Number of mobile animals | 48 hours/3rd series | 0 | 0 | 0 | 1 | 3 | 5 | |||
Number of mobile animals | 48 hours/4th series | 0 | 0 | 0 | 0 | 2 | 5 | |||
Number of mobile animals | 48 hours/total | 0 | 0 | 0 | 2 | 10 | 20 | |||
Immobilisation % | 48 hours | 100 | 100 | 100 | 90 | 50 | 0 | |||
Main test at pH 7.8 | ||||||||||
Nominal concentration | mg·L-1 | 2.5 | 1.25 | 0.63 | 0.32 | 0.16 | 0.08 | 0.04 | Control | |
Number of mobile animals | 24 hours | 0 | 0 | 0 | 4 | 4 | 5 | 5 | 5 | |
(1st series) | ||||||||||
Number of mobile animals | 24 hours | 0 | 0 | 0 | 3 | 5 | 5 | 5 | 5 | |
(2nd series) | ||||||||||
Number of mobile animals | 24 hours | 0 | 0 | 1 | 4 | 4 | 5 | 5 | 5 | |
(3rd series) | ||||||||||
Number of mobile animals | 24 hours | 0 | 1 | 1 | 4 | 5 | 5 | 5 | 5 | |
(4th series) | ||||||||||
Number of mobile animals | 24 hours/total | 0 | 1 | 2 | 15 | 18 | 20 | 20 | 20 | |
Immobilisation % | 24 hours | 100 | 95 | 90 | 25 | 10 | 0 | 0 | 0 | |
Number of mobile animals | 48 hours | 0 | 0 | 0 | 1 | 3 | 5 | 5 | 5 | |
(1st series) | ||||||||||
Number of mobile animals | 48 hours | 0 | 0 | 0 | 2 | 4 | 5 | 5 | 5 | |
(2nd series) | ||||||||||
Number of mobile animals | 48 hours | 0 | 0 | 1 | 2 | 3 | 4 | 5 | 5 | |
(3rd series) | ||||||||||
Number of mobile animals | 48 hours | 0 | 0 | 0 | 1 | 4 | 4 | 5 | 5 | |
(4th series) | ||||||||||
Number of mobile animals | 48 hours/total | 0 | 0 | 1 | 6 | 14 | 18 | 20 | 20 | |
Immobilisation % | 48 hours | 100 | 100 | 95 | 70 | 30 | 10 | 0 | 0 | |
Determined concentrations at the definitive test – pH 7.8 | ||
Nominal concentration mg/l | Measured concentration mg/l | Measured concentration mg/l |
0 hours | 48 hours | |
2.5 | 2.6 | 2.7 |
0.16 | 0.17 | 0.17 |
Description of key information
test item | year of the study | target/RA | purity | guidelines | results | notes | Data owner |
Basic Green 4 acetate | 2022 | RA | 89% without solvent | OECD 202/Short-term invertebrates | Daphnia Magna, static, pH 7.7-7.8 EC50, 48h ca 0.22 mg/l (nominal, mobility) | analytical monitoring performed via LC-MS | |
Basic Green oxalate | 2007 | no details | similar to OECD 202 | Daphnia Magna, static LC50, 48h ca 4.65 mg/l (nominal, mobility) | Aquatic ecotoxicity of two dyes and three antiparasitics of importance in aquaculture on employing Daphnia magna. | ||
Basic Green 4 oxalate | 1998 | no details | no details | Daphnia Magna LC50, 24h, 0.81 mg/l | only final results available | Dystar, report date 1998-05-26 | |
Basic Green 4 oxalate (2:3) | 1998 | no details | no details | Palaemonetes Kadiakensis LC50, 96h, 1.9 mg/l | |||
Basic Green 4 Chloride | 1998 | no details | no details | Palaemonetes Kadiakensis LC50, 96h, 0.81 mg/l | |||
Basic Green 4 X | 1988 | no details | similar to OECD 202 | Daphnia Magna, semistatic, pH 8.1-8.4 EC50, 48h, 0.81 mg/l EC100, 48h, 1.1 mg/l (nominal, geometric mean) | Dystar | Dystar, report# 41 A/88, Life cycle-Test mit Wasserfloehen Daphnia magna EC50 Immobilisierung und EC50 Reproduktion. | |
Basic Green X | 1993 | not reported-Handbook information | Daphnia Magna EC50, 48h, 0.13-0.29 mg/l | only final results available | Burchmore and Wilinson, pubblication | ||
Basic Green 4 Chloride | 1977 | no details | no details | ostracod (Cypridopsis sp.) and freshwater prawn (Palaemonetes kadiakensis). LC50, 96h 1.9-3.45 mg/l | only final results available | Bills T. D., Marking L.L, Chandler J.H.J., pubblication |
Key value for chemical safety assessment
Fresh water invertebrates
Fresh water invertebrates
- Dose descriptor:
- LC50
- Effect concentration:
- 0.22 mg/L
Additional information
The substance is an ionic dye that shows two different chemical structure based on the pH of the ecotox medium. After preliminary testing, the short term toxicity to aquatic invertebrates was tested at pH 7.8 in the dark, with HPLC-MS as analytical monitoring.
The OECD202 was performed on analogue substance 1, whose purity is within the boundary composition of the target substance.
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