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EC number: - | CAS number: -
- 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:
- July 2020
- 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)
- Version / remarks:
- 28.7.2011
- Deviations:
- yes
- Remarks:
- The stock solutions to be autoclaved were autoclaved for 20 minutes instead of the required 15 minutes. This can be considered uncritical, as the goal of sterility is achieved.
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Name: AM(pfa)4
Appearance: white powder
Homogeneity: homogeneous - Analytical monitoring:
- yes
- Details on sampling:
- The TOC content of the test item in the stock solution (c = 10 mg/L) was measured in extra replicates without algae cells at the beginning and at the end of the test.
Sample preparation: At the start of the test 5 mL of each test item solution and the control were acidified with 50 µL HNO3 (65 % in water) and measured via ICP-OES. At the end of the test 10 mL of each test item solution and the control were acidified with 100 µL HNO3 (65 % in water) and measured via ICP-OES.
On each sampling day, validity of calibration was controlled by measuring QC samples (30 µg/L). The mean QC recoveries per sequence (119.6% and 117.0%) lay outside the demanded range of 100 ±15%. But this is only a minor deviation and no back calculation from alkali metal to the test item is possible. Therefore, this deviation can be considered as uncritical. The QC recoveries were calculated using the following equations: QC recovery in % = measured concentration / nominal concentration * 100% - Vehicle:
- no
- Details on test solutions:
- Storage: The test item was stored in the test facility in a closed vessel at room temperature (17.3 – 23.0 °C), kept under inert gas.
A stock solution containing 10.2 mg/L test item in algal medium (demineralised water enriched with minerals but without algae) was prepared. The lower treatments (0.1 / 0.32 / 1 / 3.2 mg/L) were prepared by dilution of this stock solution with algal medium. - Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- Test System: Selection of the test system was made following the proposal of the guidelines with the following specification: Unicellular freshwater green alga, genus Desmodesmus, species subspicatus, SAG Strain Number 86.81, taxonomic position Chlorophyta – Chlorophyceae.
Origin and Culture: The culture of Desmodesmus subspicatus was obtained in May 2020 by MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen). The algae are kept as stock culture on solid agar at 2 - 8 °C. From the stock culture, a permanent culture was prepared. From an aliquot of the permanent culture, the pre-culture was prepared. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- Temperature: 21.2 – 22.2 °C
- pH:
- 7.2 - 8.2
- Nominal and measured concentrations:
- 0.1 / 0.32 / 1 / 3.2 / 10 mg/L nominal concentration - The tested concentrations were based on non GLP pre-tests.
- Details on test conditions:
- Date: 06. – 09. Jul. 2020
Vessels: glass flasks total volume 65 mL
Lighting: within the specified range (4440 – 8880 Lux)
Replicates for treatments: 3 vessels, filled with 45 ±1 mL test solution and algae, 1 vessel, filled with 45 ±1 mL test solution without algae for analytical determination
Blank Control: 6 vessels, each filled with 45 ±1 mL algal medium and algae, 1 vessel filled with 45 ±1 mL algal medium without algae for analytical determination - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.14 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.14 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.43 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.43 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 2.96 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.61 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.55 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.:
- 0.17 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Details on results:
- The study was performed using 5 concentrations ranging from 0.1 to 10 mg/L (nominal). Incubation time (test system Desmodesmus subspicatus) was 72 hours. The cell concentration of each replicate was determined by measuring the cell numbers every 24 hours with an electronic particle counter. Growth rate µ and the yield were determined from the cell number at the respective observation times.
Significant inhibition of algal growth was observed at the following concentrations: 0.32 / 1 / 3.2 / 10 mg/L nominal for growth rate and yield.
At the beginning and at the end of the test, the content of Alkali (A) as component of the test item in the test solutions was determined using ICP-OES-determination. Additional, at the start and at the end of the test, the content of the test item in the stock solution (c = 10 mg/L nominal) was estimated by determination of the total organic carbon (TOC) content in the test solutions using a carbon analyser.
Based on the molecular formula, a recalculation from the measured A concentrations to the whole test item was possible. The back calculated concentrations lay between 89 % and 136 % of the nominal concentrations at the beginning of the test and between 93 % and 144 % of the nominal concentrations at the end of the test. Therefore, the determination of the results was based on the mean of the measured concentrations (see OECD 201 (§ 39)).
The measured concentrations of the TOC analysis showed that the test item was present in the test solution of 10 mg/L (nominal) throughout the test.
The 72h-EC50 values of potassium dichromate were determined in a separate reference test. For the estimation of the 72h-EC50 values of the positive control, the fits showed sufficient statistical correspondence of the data with the dose-response-equation. The values were within the range of the laboratory.
The pH of the blank control should not fluctuate by more than 1.5 units. There was no change of the pH in the blank control. All validity criteria were met. No observations were made which might cause doubts concerning the validity of the study outcome. The result of the test can be considered valid. - Results with reference substance (positive control):
- Positive Control: Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a separate reference test.
The results of the last reference study (202007R301) with the positive control K2Cr2O7 (performed under GLP conditions in May 2020) are presented as follows: 72h ErC50 was 1.00 mg/L (95% confidence interval 0.97 – 1.03 mg/L) and 72h EyC50 was 0.35 mg/L (95% confidence interval 0.32 – 0.39 mg/L). - Reported statistics and error estimates:
- STATISTICS AND BIOLOGICAL RESULTS
Calculation of results was performed with the help of validated software (Microsoft Excel®). The estimation of the biological data was accomplished using the software Tox-Rat® Professional, version 3.3.0. Statistical calculation was performed using ToxRat® Professional 3.3.0. - Validity criteria fulfilled:
- yes
- Conclusions:
- Growth inhibition to Desmodesmus subspicatus through the test item was observed and the NOEC for growth rate was found 0.14 mg/L (72 h), whereas the ErC50 was determined as 2.96 mg/L. The LOEC for growth rate was 0.43 mg/L.
- Executive summary:
The study was performed using 5 concentrations ranging from 0.1 to 10 mg/L (nominal). Incubation time (test system Desmodesmus subspicatus) was 72 hours. The cell concentration of each replicate was determined by measuring the cell numbers every 24 hours with an electronic particle counter. Growth rate µ and the yield were determined from the cell number at the respective observation times.
Significant inhibition of algal growth was observed at the following concentrations: 0.32 / 1 / 3.2 / 10 mg/L nominal for growth rate and yield.
At the beginning and at the end of the test, the content of Alkali (A) as component of the test item in the test solutions was determined using ICP-OES-determination. Additional, at the start and at the end of the test, the content of the test item in the stock solution (c = 10 mg/L nominal) was estimated by determination of the total organic carbon (TOC) content in the test solution using a carbon analyser.
Considering the molecular formula, a recalculation from the measured Alkali concentrations to the whole test item was possible. The back calculated concentrations lay between 89 % and 136 % of the nominal concentrations at the beginning of the test and between 93 % and 144 % of the nominal concentrations at the end of the test. Therefore, the determination of the results was based on the mean of the measured concentrations (see OECD 201 (§ 39)).
The measured concentrations of the TOC analysis also showed that the test item was present in the test solution of 10 mg/L (nominal) throughout the test. A recalculation was not performed, because this additional analysis should only show that test item is present in the solutions.
The 72h-EC50 values of potassium dichromate (K2Cr2O7, CAS No. 7778-50-9) were determined in a separate reference test and the values were found within the range of the laboratory (growth rate 0.65 - 1.10 mg/L, yield 0.21 – 0.66 mg/L).
The following results for the test item AM(pfa)4 were determined:
Endpoint
NOEC
LOEC
EC10
EC50
Growth Rate
0.14 mg/L
0.43 mg/L
0.55 mg/L
2.96 mg/L
Yield
0.14 mg/L
0.43 mg/L
0.17 mg/L
0.61 mg/L
Reference
The cell numbers were determined with an electronic particle counter. The means and standard deviations of the cell numbers of the blank control and the treatments are presented in the following table:
Table 1: Cell Number/mL
Nominal conc. in mg/L | Parameter | Cell Number/mL | |||
0 h | 24 h | 48 h | 72 h | ||
Blank control | Mean | 2620 | 11137 | 95980 | 576437 |
SD | 0 | 679 | 13975 | 24683 | |
0.1 | Mean | 2620 | 9780 | 70907 | 621687 |
SD | 0 | 1693 | 16847 | 56361 | |
0.32 | Mean | 2620 | 9340 | 81907 | 351633 |
SD | 0 | 1413 | 24625 | 41412 | |
1 | Mean | 2620 | 7600 | 28293 | 132213 |
SD | 0 | 908 | 3668 | 10922 | |
3.2 | Mean | 2620 | 7513 | 12380 | 23873 |
SD | 0 | 2162 | 1044 | 5300 | |
10 | Mean | 2620 | 5300 | 4813 | 6693 |
SD | 0 | 710 | 613 | 656 |
SD = Standard Deviation
The light intensity is checked monthly and recorded in a control chart. The lighting corresponded to the specified range of 4440 - 8880 lux.
In the following table, the pH values measured at the start and the end of the test are stated.
Table 2: pH values
Nominal Concentration in mg/L | 0 h | 72 h |
Blank control | 7.7 | 7.7 |
0.1 | 7.9 | 8.2 |
0.32 | 7.9 | 7.6 |
1 | 7.3 | 7.5 |
3.2 | 7.3 | 7.4 |
1 | 7.2 | 7.4 |
Microscopical Observations: In the following table, the appearance of the algae at the end of the test is stated.
Table 3: Microscopical Observations
Nominal Concentration in mg/L | Normal and Healthy Appearance of the Algae |
Blank control | Yes |
0.1 | Yes |
0.32 | Yes |
1 | Yes |
3.2 | No cells visible |
10 | No cells visible |
Analytical Determination: At the beginning and at the end of the test, the content of Alkali (A) as component of the test item in the test solutions was determined using ICP-OES-determination. Additional, at the start and at the end of the test, the content of the test item in the stock solution (c = 10 mg/L nominal) was estimated by determination of the total organic carbon (TOC) content in the test solution using a carbon analyser.
Based on the molecular formula, a recalculation from the measured A concentrations to the whole test item was possible. The back calculated concentrations were between 89 % and 136 % of the nominal concentrations at the beginning of the test and between 93 % and 144 % of the nominal concentrations at the end of the test. Therefore, the determination of the results was based on the mean of the measured concentrations (see OECD 201 (§ 39)) and details are given in the following tables:
Table 4: Measured Concentrations of A at 0 h
Nominal conc. | Intensity | Measured Concentration | QC recovery | Dilution factor | Calculated Concentration |
mg/L | -- | µg/L | % | -- | µg/L |
QC sample (30 µg/L) | 413242.0 | 35.92 | 119.7 | -- | -- |
Blank control | 6446.7 | -0.03 | 119.6 | 1.01 | < LOQ |
0.1 | 17499.3 | 1.15 | 119.6 | 1.01 | 0.97 |
0.32 | 41324.3 | 3.63 | 119.6 | 1.01 | 3.07 |
1 | 113618.0 | 10.77 | 119.6 | 1.01 | 9.09 |
3.2 | 416320.0 | 36.15 | 119.6 | 1.01 | 30.54 |
10 | 1024140.0 | 75.39 | 119.6 | 1.01 | 63.68 |
QC sample (30 µg/L) | 411977.0 | 35.82 | 119.4 | -- | -- |
LOQ = 1 µg/L
Table 5: Measured Concentrations of A at 72 h
Nominal conc. | Intensity | Measured Concentration | QC recovery | Dilution factor | Calculated Concentration |
mg/L | -- | µg/L | % | -- | µg/L |
QC sample (30 µg/L) | 400896.0 | 34.99 | 116.6 | -- | -- |
Blank control | 6208.6 | -0.05 | 117.0 | 1.01 | < LOQ |
0.1 | 17869.0 | 1.19 | 117.0 | 1.01 | 1.03 |
0.32 | 40744.0 | 3.57 | 117.0 | 1.01 | 3.08 |
1 | 125174.0 | 11.86 | 117.0 | 1.01 | 10.24 |
3.2 | 421333.0 | 36.53 | 117.0 | 1.01 | 31.54 |
10 | 1048690.0 | 76.77 | 117.0 | 1.01 | 66.29 |
QC sample (30 µg/L) | 403645.0 | 35.19 | 117.3 | -- | -- |
LOQ = 1 µg/L
Table 6: Content of Chemical Elements in the Test Item
Based on the known molecular formula of the test item, the individual amounts of element weight, relative to total weight could be calculated and used for analysis, as outlined in a table in the original study report.
Table 7: Calculated test item concentrations
Nominal conc. mg/L | Calculated Concentration in mg/L | % of Nominal Concentration | Geometric Mean | ||
0 h | 72 h | 0 h | 72 h | mg/L | |
Blank control | -- | -- | -- | -- | -- |
0.1 | 0.14 | 0.14 | 136 | 144 | 0.14 |
0.32 | 0.43 | 0.43 | 134 | 135 | 0.43 |
1 | 1.28 | 1.44 | 128 | 144 | 1.35 |
3.2 | 4.29 | 4.43 | 134 | 138 | 4.35 |
10 | 8.94 | 9.30 | 89 | 93 | 9.12 |
The measured concentrations of the TOC analysis show that the test item was present in the test solution of 10 mg/L (nominal) throughout the test. A recalculation was not performed, because this additional analysis should only show that test item is present in the solutions.
Table 8: Measured Concentrations IC and TC
Nominal conc. Test Item | Measured TC | Measured TC | Measured IC | Measured IC |
mg/L | mg/L | mg/L | mg/L | mg/L |
Blank control | 8.11 | 8.02 | 7.37 | 7.45 |
10 | 9.86 | 9.62 | 7.12 | 7.16 |
LOQ (Limit of quantification) TC = 254.7 µg/L
LOQ (Limit of quantification) IC = 99.94 µg/L
Table 9: Calculated Concentrations TOC
Nominal conc. Test Item mg/L | Calculated TOC (TC-IC) t = 0 h | Calculated TOC (TC-IC)t = 72 h |
mg/L | mg/L | |
Blank control | 0.75 | 0.57 |
10 | 2.74 | 2.46 |
Growth Rate, Yield
From the cell numbers, the Growth Rate µ and the Yield were calculated. The means and standard deviations at the end of the test are given in the following table.
Table 10: Growth Rate µ, Yield
Nominal conc. in mg/L | Parameter | Growth Rate (0-72 h) [day-1] | Yield (0-72 h) [Cell Concentration/mL] |
Blank control | Mean | 1.80 | 573817 |
SD | 0.01 | 24683 | |
0.1 | Mean | 1.82 | 619067 |
SD | 0.03 | 56361 | |
0.32 | Mean | 1.63 | 349013 |
SD | 0.04 | 41412 | |
1 | Mean | 1.31 | 129593 |
SD | 0.03 | 10922 | |
3.2 | Mean | 0.73 | 21253 |
SD | 0.07 | 5300 | |
10 | Mean | 0.31 | 4073 |
SD | 0.03 | 656 |
SD = Standard Deviation
Inhibition: The following mean inhibition values were calculated for the Growth Rate µ and the Yield.
Table 11: Inhibition Values
Nominal conc. in mg/L | % Inhibition Growth Rate (0-72 h) | % Inhibition Yield (0-72 h) |
Blank control | 0 | 0 |
0.1 | -1.4 | -7.9 |
0.32 | 9.2 | 39.2 |
1 | 27.3 | 77.4 |
3.2 | 59.3 | 96.3 |
10 | 82.7 | 99.3 |
Negative inhibition values indicate a stimulation of algal growth compared to the blank control.
STATISTICS AND BIOLOGICAL RESULTS
Calculation of results was performed with the help of validated software (Microsoft Excel®). The estimation of the biological data was accomplished using the software Tox-Rat® Professional, version 3.3.0. Statistical calculation was performed using ToxRat® Professional 3.3.0.
The following values were determined.
Table 12: Biological Results of the Test Item
Parameter | Value | 95 % confidence interval |
NOEC (Growth Rate) 72 h | 0.14 mg/L | -- |
NOEC (Yield) 72 h | 0.14 mg/L | -- |
LOEC (Growth Rate) 72 h | 0.43 mg/L | -- |
LOEC (Yield) 72 h | 0.43 mg/L | -- |
72h ErC10 | 0.55 mg/L | 0.49 mg/L – 0.61 mg/L |
72h EyC10 | 0.17 mg/L | 0.11 mg/L – 0.26 mg/L |
72h ErC50 | 2.96 mg/L | 2.82 mg/L – 3.11 mg/L |
72h EyC50 | 0.61 mg/L | 0.49 mg/L – 0.75 mg/L |
VALIDITY
Increase Factor: The cell concentration in the blank control should increase by a factor of at least 16 within 72h.
Daily Growth Rates: Mean coefficient of variation of daily growth rates in the blank controls should be 35 % at the most. Coefficient of variation of average growth rate during the whole test period should be 7 % at the most.
Values and assessment
The daily growth rates of the blank controls were calculated. Means, standard deviations and coefficients of variation were determined. Values and assessment can be found in the following tables.
Table 13: Daily Growth Rates of the blank controls
Growth rates | days 0 – 1 | days 1 – 2 | days 2 – 3 | CV of sectional daily growth rates | days 0 – 3 |
Replicate 1 | 1.438 | 2.087 | 1.833 | 18 % | 1.786 |
Replicate 2 | 1.384 | 2.083 | 1.940 | 20 % | 1.803 |
Replicate 3 | 1.444 | 2.033 | 1.959 | 18 % | 1.812 |
Replicate 4 | 1.377 | 2.198 | 1.867 | 23 % | 1.814 |
Replicate 5 | 1.503 | 2.185 | 1.693 | 20 % | 1.794 |
Replicate 6 | 1.527 | 2.297 | 1.509 | 25 % | 1.778 |
Mean | 1.446 | 2.147 | 1.800 | 21 % | 1.798 |
SD | 0.061 | 0.097 | 0.171 | -- | 0.014 |
CV | 4 % | 5 % | 10 % | -- | 1 % |
CV = Coefficient of Variation
Table 14: Assessment
Parameter | Validity criteria | Observed value | Assessment |
Increase factor biomass | factor 16 in 72 h | 220 | Valid |
Mean coefficient of variation of daily growth rates | max. 35 % | 21 % | Valid |
Coefficient of variation of aver-age growth rate during the whole test period | max. 7 % | 1 % | Valid |
The growth curve shows that during the test exponential growth in the blank controls was given.
Description of key information
Growth inhibition to Desmodesmus subspicatus through the test item was observed and the NOEC for growth rate was found 0.14 mg/L (72 h), whereas the ErC50 was determined as 2.96 mg/L. The LOEC for growth rate was 0.43 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 2.96 mg/L
- EC10 or NOEC for freshwater algae:
- 0.14 mg/L
Additional information
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