Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 275-833-8 | CAS number: 71675-87-1
- 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
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 17 February 2015 - 24 February 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of preregistration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Name of test material (as cited in study report): Acilian (4-Amino-5-ethylsulfonyl-2-methoxybenzoic acid)
- batch No.of test material:GS00012
- Expiration date of the lot/batch:February 28, 2015
- Purity test date:March 13, 2014
- Purity: 98.0 – 101.5% (nominal)
100.9% (analytical), according to certificate of analysis
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:At room temperature (20 ± 5 °C), in the dark
- Stability under test conditions: yes - Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: Nominal concentrations of test item : Nominal:100, 32, 10, 3.2 and 1.0 mg test item/L and a 0 for control
Mean measured concentrations: 98.0, 28.8, 8.3, 2.59 and 0.87 mg test item/L and 0 (control).
- Sampling method:
Analysis of the Test Item Concentrations
Duplicate samples from the freshly prepared test media (containing algae) of all test concentrations and from the control were taken at the start of the test. For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of all test concentrations and the control (containing algae) were taken at the end of the test (after the 72 hours test period) by pouring together the contents of the test beakers of each treatment. All samples were diluted by a factor of 2 with acetonitrile.One additional sample of the blank control and the dilution solvent were taken at test start and test end without any sample treatment.
- Analyses:The concentrations of the test item were analysed in the duplicate test media samples from all test concentrations and in the duplicate control samples each from both sampling times (0 and 72 hours). All samples were centrifuged before analysis, in order to remove the algae.
- Fortified Samples:Two stock solutions were prepared. Approximately 50 mg of the test item were dissolved (1 minute ultrasonification, 30 minutes stirring) in 50 mL test water to obtain a stock solution of approximately 1 g test item/L. Appropriate amounts of these stock solutions were diluted with test water to obtain fortified samples at a level of 0.6, 15 and 140 mg test item/L.
- Sample storage conditions before analysis:All samples were stored in a freezer (≤ - 10 °C), protected from light until analysis was performed.
Standard Solutions used for the Quantification:
- Stock Solution:The test item was used to prepare a stock solution. 50.20 mg of the test item were dissolved in 50 mL acetonitrile to obtain a stock solution of approximately 1 g test item/L.
- Standard Solutions:Appropriate amounts of the stock solution were diluted with acetonitrile / test water (1/1, v/v) to obtain standard solutions in the range from 0.1 to 20 mg test item/L.
Analysis of Treatment Samples and Control Samples:
- Sample Preparation:The samples were shaken and where appropriate treated with ultrasound for 1 minute after thawing to obtain homogenous samples. The samples were diluted with acetonitrile by factor 2 (see chapter 6.10). The samples were further diluted with acetonitrile / test water (1/1, v/v) to match the calibration range, if necessary. The samples were centrifuged (13000 rpm, 10 minutes) before analysis.
- Sample storage conditions before analysis: All samples were stored in a freezer (≤ - 10 °C), protected from light until analysis was performed. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution of 100mg test item / L was prepared by dissolving 100.8 mg test item was dissolved into 1008 mL test water by intense stirring for 30 minutes and ultrasonic treatment for 2 minutes.The pH of the stock solution was adjusted from 6.3 to 8.1 with 1 M NaOH.Adequate volumes of this stock solution were diluted with test water to prepare the test media of the desired test concentrations. The test media were prepared just before introduction of the algae (= start of the test).
- Controls: test water was used without addition of the test item
- Evidence of undissolved material (e.g. precipitate, surface film, etc.):No - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata,
- Strain: 61.81 SAG
- Source (laboratory, culture collection):The algae were supplied by the Sammlung von Algenkulturen, Albrecht-von-Haller-Institut für Pflanzenwissenschaften, Universität Göttingen", 37073 Göttingen, Germany.
- Method of cultivation:The algae were cultivated in the laboratories of ibacon under standardised conditions according to the test guidelines.
ACCLIMATION
- Acclimation period:4 days
- Any deformed or abnormal cells observed: NO - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Hardness:
- 0.24 mmol/L (= 24 mg/L) as CaCO3.
- Test temperature:
- 21 to 22 °C
- pH:
- 8.1 to 8.2 at test start and
9.3 to 9.5 at test end - Nominal and measured concentrations:
- Nominal:100, 32, 10, 3.2 and 1.0 mg test item/L and a control
Mean measured concentrations: 98.0, 28.8, 8.3, 2.59 and 0.87 mg test item/L and 0 (control). - Details on test conditions:
- TEST SYSTEM
- Test vessel:Erlenmeyer flasks of 50 mL volume with 50 mL of test medium.
- Type (delete if not applicable): closed.
- Aeration: continuously stirred with magnetic stirrers.
- Initial cells density: 5000 cells per mL test medium.
- Control end cells density: was determined by spectrophotometrical measurement : (See EFFECT PARAMETERS MEASURED)
- No. of vessels per concentration (replicates):3
- No. of vessels per control (replicates):6
GROWTH MEDIUM
- Standard medium used: yes - OECD medium
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted Water, according the OECD TG 201
- Culture medium different from test medium: NO
OTHER TEST CONDITIONS
- Adjustment of pH:The pH was adjusted with 1 M NaOH to pH 8.1.
- Photoperiod: Continuous illumination
- Light intensity and quality:Mean light intensity: 5655 lux (range: 5420 to 6180 lux). Measured once during the test at 6 posotions distributed over the experimental area at the surface of the test media.
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]:Spectrophotometer: Defined volumes of the algal suspensions from all replicates and from the blanks were sampled after 24, 48 and 72 hours of exposure, and were not replaced. The algal cell densities were calculated by subtracting the absorption of the blanks, from each of the measured absorption of the test media (with algae).
- Other : EC50,EC20,EC10, NOEC, LOEC for Yield and growth rate
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.13
- Range finding study: Pre-experiments were performed to determine a suitable concentration range and to establish suitable methods to prepare the test solutions. The pre-experiments were not performed in compliance with the GLP-Regulations and are excluded from the Statement of Compliance in the final report. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 98 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- > 98 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- > 98 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 28.8 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- ca. 98 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- - Exponential growth in the control (for algal test): no
- Observation of abnormalities (for algal test): NO
- Unusual cell shape:NO
- Colour differences: YES-After 24 hours until the end of the test, the test item caused colouration of the test medium in the concentrations of nominal 32 and 100 mg test item/L. Only after 48 hours the test item caused colouration of the test medium in the concentration of nominal 10 mg test item/L (see Table 11).
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: NO - Results with reference substance (positive control):
- - Results with reference substance valid : YES
Parameter: (0-72 h): EC50
Yield [mg test item/L]: 0.454
Growth rate [mg test item/L]: 1.05
Biomass [mg test item/L]: 0.492
95 % conf. Interval for : Yield= 0.439 - 0.471, Growth rate =1.02 - 1.08, Biomass = 0.475-0.510
Parameter: (0-72 h): EC20
Yield [mg test item/L]: 0.265
Growth rate [mg test item/L]: 0.520,
Biomass [mg test item/L]: 0.290
95 % conf. Interval for : Yield= 0.250 - 0.279, Growth rate =0.493 - 0.546, Biomass = 0.274 - 0.305
Parameter: (0-72 h): EC10
Yield [mg test item/L]: 0.200
Growth rate [mg test item/L]: 0.361,
Biomass [mg test item/L]: 0.220
95 % conf. Interval for : Yield= 0.185 - 0.214, Growth rate =0.335 - 0.385, Biomass = 0.204 - 0.235
Parameter: (0-72 h): NOEC
Yield [mg test item/L]: 0.094
Growth rate [mg test item/L]: 0.094,
Biomass [mg test item/L]: 0.094
Parameter: (0-72 h): LOEC
Yield [mg test item/L]: 0.210
Growth rate [mg test item/L]: 0.210
Biomass [mg test item/L]: 0.210
Values refer to nominal test concentrations - Reported statistics and error estimates:
- Calculations were made with the probit method, whereas analyses were conducted with Professional, Version 2.10.05, ToxRat Solutions GmbH., and the Williams t-test Procedure.
- Validity criteria fulfilled:
- yes
- Remarks:
- Control increased by a factor of 256 (> 16-fold growth), Coefficient of Variation of Sectional (Daily) Growth Rates in Control Cultures: 5.8% (<35%). Coefficient of Variation of Average Growth between Control Replicates:0.7% (<7%)
- Conclusions:
- The EC50 (based on growth inhibition rate and geometric mean measured concentrations) of the test item in green algae was > 98.0 mg test item /L after 72 hours of exposure. The EC10 (growth) > 98.0, EC10 (yield) egual to 79.6 mg test item/L, and NOEC 28.8 mg test item /L respectively. The yield inhibition and growth inhibition are observed in the concentration > 98 mg test item/L.
- Executive summary:
The green algae growth inhibition test was performed according to OECD Guideline 201 (GLP study). The purpose was to determine the inhibitory effect of the test item Acilian on the growth of the freshwater green algae Pseudokirchneriella subcapitata. The initial density(1 x 105 cells/mL) of Pseudokirchneriella subcapitata were exposed to geometric mean measured concentrations of 0 (control, 6 replicates), 0.87 mg test item /L, 2.59 mg test item /L, 8.30 mg test item /L, 28.8 mg test item /L, and 98.0 mg test item /L (3 replicates) for 72 hours under static conditions.The number of algal cells was determined by spectrophotometric measurement after 24, 48, and 72 hours of the exposure. Based on these measurements the growth rate and yield were calculated. After 24 hours until the end of the test, the test item caused colouration of the test medium in the nominal concentrations 32 and 100 mg test item/L. After 48 hours the test item caused colouration of the test medium in the nominal concentration 10 mg test item/L. The endpoints values based on the geometric mean measured test item concentrations : In 72 hours, measured EC50 value was > 98.0 mg test item /L, NOEC value 28.8 mg test item /L and LOEC egual to 98.0 mg test item /L for the growth and yield rate. EC10 (growth) > 98 mg test item/L and EC10 (yield) egual to 79.6 mg test item/L.
Reference
Table No.1: Algal Cell Densities during the Test Period of 72 Hours
Measured concentration [mg test item/L] |
Flask
|
Density of algal cells [10000/mL] after |
||
|
No. |
24 hours |
48 hours |
72 hours |
Control |
1 |
3.316 |
19.098 |
121.001 |
|
2 |
3.650 |
23.033 |
129.657 |
|
3 |
3.650 |
22.246 |
124.149 |
|
4 |
3.316 |
21.852 |
132.805 |
|
5 |
2.983 |
22.639 |
132.411 |
|
6 |
3.316 |
22.639 |
126.903 |
|
m |
3.372 |
21.918 |
127.821 |
|
s |
0.251 |
1.438 |
4.693 |
0.870 |
1 |
3.650 |
22.246 |
132.018 |
|
2 |
3.650 |
24.606 |
131.624 |
|
3 |
3.650 |
21.852 |
127.690 |
|
m |
3.650 |
22.902 |
130.444 |
|
s |
0.000 |
1.490 |
2.393 |
2.59 |
1 |
3.316 |
21.459 |
120.608 |
|
2 |
3.650 |
22.639 |
124.936 |
|
3 |
3.650 |
23.033 |
125.329 |
|
m |
3.539 |
22.377 |
123.624 |
|
s |
0.193 |
0.819 |
2.620 |
8.30 |
1 |
3.983 |
24.213 |
135.165 |
|
2 |
3.983 |
26.574 |
139.985 |
|
3 |
3.650 |
23.426 |
123.362 |
|
m |
3.872 |
24.738 |
131.755 |
|
s |
0.193 |
1.638 |
7.531 |
28.8 |
1 |
3.650 |
25.787 |
137.919 |
|
2 |
4.317 |
23.820 |
133.985 |
|
3 |
3.316 |
22.246 |
123.362 |
|
m |
3.761 |
23.951 |
131.755 |
|
s |
0.509 |
1.774 |
7.531 |
98.0 |
1 |
2.983 |
19.492 |
109.951 |
|
2 |
3.316 |
17.131 |
103.296 |
|
3 |
3.316 |
19.098 |
117.067 |
|
m |
3.205 |
18.574 |
109.985 |
|
s |
0.193 |
1.265 |
6.894 |
m: mean value; s: standard deviation
Table No.2: Yield during the Test Period of 72 Hours
Measured concentration [mg test item/L] |
Flask
|
Yield y [10000 cells/mL] after |
||
|
No. |
24 hours |
48 hours |
72 hours |
Control |
1 |
2.816 |
18.598 |
120.501 |
|
2 |
3.150 |
22.533 |
129.157 |
|
3 |
3.150 |
21.746 |
123.649 |
|
4 |
2.816 |
21.352 |
132.305 |
|
5 |
2.483 |
22.139 |
131.911 |
|
6 |
2.816 |
22.139 |
126.403 |
|
m |
2.872 |
21.418 |
127.321 |
|
s |
0.251 |
1.438 |
4.693 |
0.870 |
1 |
3.150 |
21.746 |
131.518 |
|
2 |
3.150 |
24.106 |
131.124 |
|
3 |
3.150 |
21.352 |
127.190 |
|
m |
3.150 |
22.402 |
129.944 |
|
s |
0.000 |
1.490 |
2.393 |
2.59 |
1 |
2.816 |
20.959 |
120.108 |
|
2 |
3.150 |
22.139 |
124.436 |
|
3 |
3.150 |
22.533 |
124.829 |
|
m |
3.039 |
21.877 |
123.124 |
|
s |
0.193 |
0.819 |
2.620 |
8.30 |
1 |
3.483 |
23.713 |
134.665 |
|
2 |
3.483 |
26.074 |
138.993 |
|
3 |
3.150 |
22.926 |
129.944 |
|
m |
3.372 |
24.238 |
134.534 |
|
s |
0.193 |
1.638 |
4.526 |
28.8 |
1 |
3.150 |
25.287 |
137.419 |
|
2 |
3.817 |
23.320 |
133.485 |
|
3 |
2.816 |
21.746 |
122.862 |
|
m |
3.261 |
23.451 |
131.255 |
|
s |
0.509 |
1.774 |
7.531 |
98.0 |
1 |
2.483 |
18.992 |
109.091 |
|
2 |
2.816 |
16.631 |
102.796 |
|
3 |
2.816 |
18.598 |
116.567 |
|
m |
2.705 |
18.074 |
109.485 |
|
s |
0.193 |
1.265 |
6.894 |
m: mean value; s: standard deviation
Cell density of 5000 algal cells/mL at test start are subtracted from the yield values
Table NO.3: Growth rate during the Test Period of 72 Hours
Measured concentration [mg test item/L] |
Flask
|
Growth ratesμ[1/day] after |
||
|
No. |
0 - 24 hours |
0 - 48 hours |
0 - 72 hours |
Control |
1 |
1.892 |
1.821 |
1.830 |
|
2 |
1.988 |
1.915 |
1.853 |
|
3 |
1.988 |
1.898 |
1.838 |
|
4 |
1.892 |
1.889 |
1.861 |
|
5 |
1.786 |
1.906 |
1.860 |
|
6 |
1.892 |
1.906 |
1.846 |
|
m |
1.906 |
1.889 |
1.848 |
|
s |
0.075 |
0.034 |
0.012 |
0.870 |
1 |
1.988 |
1.898 |
1.859 |
|
2 |
1.988 |
1.948 |
1.858 |
|
3 |
1.988 |
1.889 |
1.848 |
|
m |
1.988 |
1.911 |
1.855 |
|
s |
0.00 |
0.032 |
0.006 |
2.59 |
1 |
1.892 |
1.880 |
1.829 |
|
2 |
1.988 |
1.906 |
1.840 |
|
3 |
1.988 |
1.915 |
1.841 |
|
m |
1.956 |
1.900 |
1.837 |
|
s |
0.055 |
0.018 |
0.007 |
8.30 |
1 |
2.075 |
1.940 |
1.867 |
|
2 |
2.075 |
1.987 |
1.877 |
|
3 |
1.988 |
1.923 |
1.855 |
|
m |
2.046 |
1.950 |
1.866 |
|
s |
0.050 |
0.033 |
0.011 |
28.8 |
1 |
1.988 |
1.972 |
1.873 |
|
2 |
2.156 |
1.932 |
1.864 |
|
3 |
1.892 |
1.898 |
1.836 |
|
m |
2.012 |
1.934 |
1.858 |
|
s |
0.133 |
0.037 |
0.019 |
98.0 |
1 |
1.786 |
1.832 |
1.797 |
|
2 |
1.892 |
1767 |
1.777 |
|
3 |
1.892 |
1.821 |
1.819 |
|
m |
1.857 |
1.807 |
1.797 |
|
s |
0.061 |
0.035 |
0.021 |
m: mean value; s: standard deviation
Table No.4: Yield y and Percentage Inhibition of y during the Test Period
Measured concentration [mg test item/L] |
Yields y [10000 cells/mL] and % inhibition of y |
|||||
|
24 hours |
48 hours |
72 hours |
|||
|
y |
% |
y |
% |
y |
% |
Control |
2.872 |
- |
21.418 |
- |
127.321 |
- |
0.870 |
3.150 |
-9.7 |
22.402 |
-4.6 |
129.944 |
-2.1 |
2.59 |
3.039 |
-5.8 |
21.877 |
-2.1 |
123.124 |
3.3 |
8.30 |
3.372 |
-17.4 |
24.238 |
-13.2 |
134.534 |
-5.7 |
28.8 |
3.261 |
-13.5 |
23.451 |
-9.5 |
131.255 |
-3.1 |
98.0 |
2.705 |
5.8 |
18.074 |
15.6* |
109.485- |
14.0* |
negative values in ‘% inhibition’ indicate an increase in growth relative to that of the control
* mean value significantly different from the control (tested with Williams t-test, α = 0.05, one-sided)
Table No.5: Growth Rates μ and Percentage Inhibition of μ during the Test Period
Measured concentration [mg test item/L] |
Growth ratesμ[1/day] and % inhibition ofμ |
|||||
|
24 hours |
48 hours |
72 hours |
|||
|
μ |
% |
μ |
% |
μ |
% |
Control |
1.906 |
- |
1.889 |
- |
1.848 |
- |
0.870 |
1.988 |
-4.3 |
1.911 |
-1.2 |
1.855 |
-0.4 |
2.59 |
1.956 |
-2.6 |
1.900 |
-0.6 |
1.837 |
0.6 |
8.30 |
2.046 |
-7.3 |
1.950 |
-3.2 |
1.866 |
-1.0 |
28.8 |
2.012 |
-5.5 |
1.934 |
-2.3 |
1.858 |
-0.5 |
98.0 |
1.857 |
2.6 |
1.807 |
4.4* |
1.797 |
2.7* |
negative values in ‘% inhibition’ indicate an increase in growth relative to that of the control
* mean value significantly different from the control (tested with Williams t-test, α = 0.05, one-sided)
Temperature in the test media (water) during the test period:
0hours= 21ºC
24 hours =22ºC
48 hours =21ºC
72 hours=22ºC
Table No. 6 pH-Values in the Test Media at the Start and End of the Test
Measured concentration[mg test item/L] |
pH at 0hours |
pH at 72Hours |
Control |
8.2 |
9.3 |
0.870 |
8.1 |
9.5 |
2.59 |
8.1 |
9.5 |
8.30 |
8.1 |
9.5 |
28.8 |
8.1 |
9.4 |
98.0 |
8.1 |
9.3 |
Table No.7: Appearance of the Test Item in the Test Media
Measured concentration[mg test item/L] |
Appearance at 0hours |
Appearance at 24 Hours |
Appearance at 48 hours |
Appearance at 72 hours |
0.870 |
0 |
0 |
0 |
0 |
2.59 |
0 |
0 |
0 |
0 |
8.30 |
0 |
0 |
1 |
0 |
28.8 |
0 |
1 |
1 |
1 |
98.0 |
0 |
1+ |
1+ |
1+ |
Abbreviations:
0: No remarkable observations, clear test medium
1: Colouration caused by the test item
2: Turbidity caused by the test item
3: Inhomogeneous dispersion of the test item
4: Precipitation of the test item
5: Test item at the surface
6: Test item lying at the bottom of the test vessel
+: Effects were strongly pronounced
Effects in parenthesis were only slightly pronounced
Table No.8 Results for the Determination of the Test Item in the Test Samples
mple Description [mg test item/L] |
AGE |
Concentration found [mg test item/L]1 |
D.F. |
Concentration calculated [mg test item/L]1 |
Concentration nominal [mg test item/L]2 |
% of Nominal3 |
Geometric mean concentration [mg test item/L]5 |
|
|
day |
h |
|
|||||
Control |
0 |
0 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
0.870 |
Control |
0 |
0 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
|
Control |
3 |
72 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
|
Control |
3 |
72 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
|
1 |
0 |
0 |
0.554 |
2 |
1.109 |
1.000 |
111 |
|
1 |
0 |
0 |
0.551 |
2 |
1.102 |
1.000 |
110 |
|
1 |
3 |
72 |
0.339 |
2 |
0.678 |
1.000 |
68 |
|
1 |
3 |
72 |
0.337 |
2 |
0.675 |
1.000 |
67 |
|
|
4geometric mean value (n=2): |
87 |
||||||
3.2 |
0 |
0 |
1.684 |
2 |
3.368 |
3.200 |
105 |
2.59 |
3.2 |
0 |
0 |
1.713 |
2 |
3.427 |
3.200 |
107 |
|
3.2 |
3 |
72 |
0.989 |
2 |
1.979 |
3.200 |
62 |
|
3.2 |
3 |
72 |
0.986 |
2 |
1.971 |
3.200 |
62 |
|
|
4geometric mean value (n=2): |
81 |
||||||
10 |
0 |
0 |
5.173 |
2 |
10.347 |
10.000 |
103 |
8.30 |
10 |
0 |
0 |
5.216 |
2 |
10.431 |
10.000 |
104 |
|
10 |
3 |
72 |
3.294 |
2 |
6.588 |
10.000 |
66 |
|
10 |
3 |
72 |
3.308 |
2 |
6.616 |
10.000 |
66 |
|
|
4geometric mean value (n=2): |
83 |
||||||
32 |
0 |
0 |
16.453 |
2 |
32.905 |
32.000 |
103 |
28.8 |
32 |
0 |
0 |
16.492 |
2 |
32.985 |
32.000 |
103 |
|
32 |
3 |
72 |
12.287 |
2 |
24.573 |
32.000 |
77 |
|
32 |
3 |
72 |
12.416 |
2 |
24.833 |
32.000 |
78 |
|
|
4geometric mean value (n=2): |
90 |
||||||
100 |
0 |
0 |
5.267 |
20 |
105.346 |
100.000 |
105 |
98.0 |
100 |
0 |
0 |
5.216 |
20 |
104.327 |
100.000 |
104 |
|
100 |
3 |
72 |
4.5969 |
20 |
91.382 |
100.000 |
91 |
|
100 |
3 |
72 |
4.550 |
20 |
90.991 |
100.000 |
91 |
|
|
4geometric mean value (n=2): |
98 |
1 The tabulated results represent rounded results calculated on the exact raw data
2 Nominal values given by the study director
3 The results represent rounded values
4 geometric mean values are calculated with the mean values of the fresh and aged samples
5 geometric mean concentrations are calculated with the geometric mean values of the recovery rates in percent
and the nominal test item concentrations
LOD: Limit of Detection = 0.09 mg test item/L
n.a.: not applicable
RSD: Relative Standard Deviation
D.F.: Dilution factor
Table No.9. Results for the Determination of the Test Item in the Fortified Samples
ple Description [mg test item/L] |
|
Concentration found [mg test item/L]1 |
D.F. |
Concentration calculated [mg test item/L]1 |
Concentration nominal [mg test item/L]2 |
% of Nominal3 |
|
|||
Analytical Blank Control |
0 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
||||
Analytical Blank Control |
0 |
<LOD |
2 |
n.a. |
0.000 |
n.a. |
||||
Fortified Sample |
0.6 |
0.337 |
2 |
0.674 |
0.599 |
112 |
||||
Fortified Sample |
0.6 |
0.335 |
2 |
0.670 |
0.599 |
112 |
||||
Fortified Sample |
0.6 |
0.334 |
2 |
0.668 |
0.596 |
112 |
||||
Fortified Sample |
0.6 |
0.336 |
2 |
0.672 |
0.596 |
113 |
||||
Fortified Sample |
0.6 |
0.336 |
2 |
0.672 |
0.596 |
113 |
||||
|
mean value (n=5): |
112 |
||||||||
RSD (n=5): |
0 |
|||||||||
Fortified Sample |
15 |
7.449 |
2 |
17.898 |
14.982 |
99 |
||||
Fortified Sample |
15 |
7.463 |
2 |
14.925 |
14.982 |
100 |
||||
Fortified Sample |
15 |
7.435 |
2 |
14.870 |
14.907 |
100 |
||||
Fortified Sample |
15 |
7.246 |
2 |
14.493 |
14.907 |
97 |
||||
Fortified Sample |
15 |
7.361 |
2 |
14.721 |
14.907 |
99 |
||||
|
mean value (n=5): |
99 |
||||||||
RSD (n=5): |
1 |
|||||||||
Fortified Sample |
140 |
7.213 |
20 |
144.258 |
139.832 |
103 |
||||
Fortified Sample |
140 |
7.179 |
20 |
143.577 |
139.832 |
103 |
||||
Fortified Sample |
140 |
7.078 |
20 |
141.568 |
139.132 |
102 |
||||
Fortified Sample |
140 |
7.091 |
20 |
141.824 |
139.132 |
102 |
||||
Fortified Sample |
140 |
7.007 |
20 |
140.134 |
139.132 |
101 |
||||
|
mean value (n=5): |
102 |
||||||||
RSD (n=5): |
101 |
mean value (n=15): 105
RSD (n=15) : 6
1 The tabulated results represent rounded results calculated on the exact raw data
2 The results represent rounded values
LOD: Limit of Detection = 0.09 mg test item/L
n.a.: not applicable
RSD: Relative Standard Deviation
D.F.: Dilution factor
Results:
The EuC50, EuC20 and EuC10 (72 h) value were > 98.0 mg test item/L.
The EyC50 and EyC20 (72 h) value was > 98.0 mg test item/L.
The EyC10 (72 h) value was 79.6 mg test item/L.
The LOEC (72 h) for growth rate was 98.0 mg test item/L.
The NOEC (72 h) for growth rate was 28.8 mg test item/L.
The LOEC (72 h) for yield was 98.0 mg test item/L.
The NOEC (72 h) for yield was 28.8 mg test item/L.
Description of key information
Key study: OECD guideline 201, GLP study. In 72 hours, measured EC50 value was > 98.0 mg test item /L, NOEC value 28.8 mg test item /L and LOEC egual to 98.0 mg test item /L for the growth and yield rate. EC10 (growth) > 98 mg test item/L and EC10 (yield) egual to 79.6 mg test item/L. The yield inhibition and growth inhibition are observed in the concentration > 98 mg test item/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 98 mg/L
- EC10 or NOEC for freshwater algae:
- 28.8 mg/L
Additional information
Key study: OECD GUIDELINE 201, GLP study: The green algae growth inhibition test was performed according to OECD Guideline 201 (GLP study). The initial density of 1 x 105 cells/mL Pseudokirchneriella subcapitata were exposed to geometric mean measured concentrations for 72 hours under static conditions. The algal cells was determinated by spectrophotometric measurement after 24, 48, and 72 hours of the exposure. Based on these measurements the growth rate and yield were calculated. After 24 hours until finish, the test item caused colouration of the test medium in the nominal concentrations 32 and 100 mg test item/L, after 48 hours in the nominal concentration 10 mg test item/L. The endpoints values based on the geometric mean measured test item concentrations: During 72 hours, measured EC50 value was > 98.0 mg test item /L for the growth and yield rate, EC10 (growth) > 98 mg test item/L and EC10 (yield) egual to 79.6 mg test item/L. The NOEC value was 28.8 mg test item /L and LOEC egual to 98.0 mg test item /L for both of growth and yield rate.
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.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.