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EC number: 209-533-5 | CAS number: 584-13-4
- 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
Toxicity to aquatic algae and cyanobacteria
Administrative data
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 09.09.2021 to 23.09.2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- other: Study plan
- Title:
- Unnamed
- Year:
- 2 021
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- Method C.3. - Freshwater Algae and Cyanobacteria, Growth Inhibition Test, Commission Regulation (EU) No. 2016/266.
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- Adopted March 23, 2006 (Annex 5 corrected: 28 July 2011)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Annex: Test Facility GLP Certificate
Test material
- Reference substance name:
- 4H-1,2,4-triazol-4-ylamine
- EC Number:
- 209-533-5
- EC Name:
- 4H-1,2,4-triazol-4-ylamine
- Cas Number:
- 584-13-4
- Molecular formula:
- C2H4N4
- IUPAC Name:
- 4H-1,2,4-triazol-4-ylamine
- Test material form:
- solid
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- Limit test
- Concentrations: The samples for analysis were taken from the stock solution (100 mg·L-1) with and without algae.
- 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 analyzed on the day of delivery. The samples for analysis at the end of the test (72 hours) were delivered to analytical laboratory immediately after the end of testing. The samples were analyzed on the day of delivery.
- Sample storage conditions before analysis: All samples were stored at laboratory temperature.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
The stock solutions of the test item were prepared in the test medium. 100 mg of the test item was weighed into 1000 mL of the test medium for the preliminary and limit test. The concentrations of test solutions used in the preliminary test were obtained by dilution of the stock solution with test medium.
The exact concentrations are given on tables in Chapter 4.1. of report.
The stock solution of the test item in the test medium was prepared for the preliminary/limit test. The test was performed in a range nominal concentrations from 1 mg·L-1 to 100 mg·L-1. Testing mixtures were prepared by dosing appropriate volumes of stock solution of the test item and inoculum into volumetric flasks. Testing mixtures were incubated in the Erlenmeyers’ flasks. The test contained three (preliminary) or six parallel series (limit test) of the test item concentrations and three (six) controls without the test item. The volume of the test solution was 50 mL in each testing flask. The initial density of 5 000 cells per mL was used in the test. The flasks were placed on a shaker under the lighting ramp and were incubated under continuous illumination and shaking for 72 hours. At the beginning and the end of the test the pH of the test mixtures was measured. The light intensity and temperature were measured every 24 hours. The cell density was measured by direct counting of living cells in Burker´s counting chamber.
Test organisms
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Strain: Brinkmann 1953/SAG 86.81
- Source (laboratory, culture collection): The collection of autotrophic organisms of The Botanic Institute of the Czech Academy of Science, Třeboň
- Age of inoculum (at test initiation): 3-4 days
- Method of cultivation: The strain culture was preinoculated from the stock solution and cultivated in flasks with the test medium on indirect daylight at laboratory temperature.
ACCLIMATION
- Acclimation period: 3-4 days
- Culturing media and conditions: same as test
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
Test conditions
- Test temperature:
- 21-24°C controlled at +/- 2°C; limit test 22.0 – 22.5 °C
- pH:
- 8
- Conductivity:
- 1.74 µS·cm-1
- Nominal and measured concentrations:
- Limit test:
nominal 100 mg.L-1, measured 98.1 mg.L-1 (0 hours) and 99.1 mg.L-1 (72 hours);
nominal 100 mg.L-1 w.a. (without algae), measured 98.9 mg.L-1 (0 hours) and 101.2 mg.L-1 (72 hours) - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyers flasks with stoppers venting air
- Aeration: no
- Initial cells density: 5000 cells in a 1 mL
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water with conductivity smaller than 5 μS.cm-1 was used for the preparation of solutions. All chemicals used were of analytical-grade.
- According to Method C.3 Algal Inhibition Test were prepared four basic stock solutions. The test medium was prepared by admixing of basic stock solutions. The solutions were kept in the dark at temperature of 4 +/- 1°C in fridge.
OTHER TEST CONDITIONS
- Lighting: continuous white light, 4 440 lux to 8 880 lux; limit test 7 600 – 7 620 lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- The density of algae culture was evaluated microscopically at 24, 48 and 72 hours.
(At the beginning and the end of the test the pH of the test mixtures was measured. The light intensity and temperature were measured every 24 hours. )
TEST CONCENTRATIONS
Range finding study
- Test concentrations: 100, 50, 10, 5 and 1 mg.L-1 (nominal concentration)
Based on low toxicity of the test item found in the preliminary test, the limit test was performed subsequently. - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
Results and discussion
Effect concentrationsopen allclose all
- Key result
- Duration:
- 72 h
- Dose descriptor:
- other: ErC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- other: EyC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 72 h
- Dose descriptor:
- other: NOECr
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- other: NOECy
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Results with reference substance (positive control):
- - Results with reference substance valid?
Yes. The inhibition of test organisms by the reference substance (72h – ErC50) obtained from last reference test meets the calculated range from the interlaboratory test.
Reference test: 72 hour – ErC50 = 1.01 mg.L-1 (95% confidence limit: 0.66 – 1.53 mg.L-1)
Interlaboratory test: 72 hour – ErC50 = 0.57 – 1.33 mg.L-1
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- The inhibition of growth rate was 1.0 % and inhibition of yield was 3.7 % in the limit test. Therefore exact values of ErC50 and EyC50 could not be calculated and the value of EC is given in the form of a range.
The determination of NOEC value was done by evaluation of the results from preliminary test using statistical software ToxRat Professional Version 3.3.0
72 hour – ErC50 > 100 mg·L-1 (nominal concentration)
72 hour – EyC50 ˃ 100 mg·L-1 (nominal concentration)
72 hour – NOECr ˃ 100 mg·L-1 (nominal concentration)
72 hour – NOECy ˃ 100 mg·L-1 (nominal concentration) - Executive summary:
The test item, 4H-1,2,4-triazol-4-amine, was tested for growth inhibition on algae Desmodesmus subspicatus.
The test was performed according to method C.3. - Freshwater Algae and Cyanobacteria, Growth Inhibition Test, Commission Regulation (EU) No.2016/266.
The preliminary test was performed in a range of the test item nominal concentrations 1 – 100 mg·L-1.
The inhibition of growth rate was 0.8 % in the highest concentration and the inhibition of yield was 3.0 % in the highest concentration in the preliminary test. Based on low toxicity of the test item found in the preliminary test, the limit test was performed subsequently.
Samples for analytical determination were taken at the beginning and at the end of the test. The analytical results showed, that the test item was sufficiently stable in test medium in the conditions of the test, therefore the limit test was performed as static one.
The concentration of 100 mg·L-1 was tested in the limit test. The inhibition of growth rate was 1.0 % and inhibition of yield was 3.7 %.
Samples for analytical determination were taken at the beginning and at the end of the test.
There is stated in the guideline that, if evidence is available to demonstrate that the concentration of the test item has been satisfactorily maintained within ± 20 percent of the nominal or measured initial concentration throughout the test, the results can be based on nominal or measured initial values.
In this study concentration of the test item has been satisfactorily maintained within ± 20 percent of the nominal concentration throughout the test.
The nominal concentrations were used for all evaluations and results.
Test results:
72 hour – ErC50 > 100 mg·L-1 (nominal concentration)
72 hour – EyC50 ˃ 100 mg·L-1 (nominal concentration)
72 hour – NOECr ˃ 100 mg·L-1 (nominal concentration)
72 hour – NOECy ˃ 100 mg·L-1 (nominal concentration)
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