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EC number: 205-741-5 | CAS number: 149-45-1
- 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
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 28 May - 16 Aug 2018
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
Data source
Referenceopen allclose all
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- 23. Mar. 2006, Annex 5 corrected: 28 July 2011
- Deviations:
- yes
- Remarks:
- See section "Any other information on material and methods incl tables"
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- adopted 07. December 2015
- Deviations:
- yes
- Remarks:
- See section "Any other information on material and methods incl tables"
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany
Test material
- Reference substance name:
- Disodium 4,5-dihydroxybenzene-1,3-disulphonate
- EC Number:
- 205-741-5
- EC Name:
- Disodium 4,5-dihydroxybenzene-1,3-disulphonate
- Cas Number:
- 149-45-1
- Molecular formula:
- C6H6O8S2.2Na
- IUPAC Name:
- disodium 4,5-dihydroxybenzene-1,3-disulphonate
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: Control and all test item concentrations;
- Sampling method: Sampling at test start and test end
Test solutions
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 500 mL of a stock solution containing 100.0 mg/L (experiment I) and 10.0 mg/L (experiment II) test item in algal medium was prepared. The resulting solution was clear. Lower test concentrations were prepared by dilution of the stock solution in algal medium.
- Controls: Yes, algae medium only
Test organisms
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Common name: Green alga
- Strain: 86.81
- Source: Originally obtained in January 2016 from MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen), Germany
- Age of inoculum : 4 day (pre-culture)
- Method of cultivation: 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.
ACCLIMATION
- Culturing media and conditions (same as test or not): Same for pre-culture
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
Test conditions
- Test temperature:
- Experiment I: 21.6 – 22.6 °C
Experiment II: 19.4 – 23.3 °C (see section "Any other information on material and methods incl tables" for deviation) - pH:
- Experiment I:
8.1 - 8.3 (Control)
7.4 - 8.2 (Test item concentrations)
Experiment II:
8.3 - 8.9 (Control)
7.8 - 8.9 (Test item concentrations) - Nominal and measured concentrations:
- Nominal: 0.1 / 0.32 / 1 / 3.2 / 10 / 32 / 100 mg/L (experiment I) For details on measured concentrations please refer to table 7 to 10 in section "Any other information on results incl tables".
Nominal: 0.0032 / 0.01 / 0.032 / 0.1 / 0.32 / 1 mg/L (experiment II) For details on measured concentrations please refer to table 7 to 10 in section "Any other information on results incl tables". - Details on test conditions:
- TEST SYSTEM
Test vessel:
- Type: open (covered with perforated plastic foil acting as a stopper)
- Material, size, headspace, fill volume: glass flasks total volume 65 mL, 45 ± 1 mL of the respective test solution
- Initial cells density: 2.1 *10E+03 cells/mL (experiment I and II)
- Control end cells density: 405477 (experiment I); 336973 cells/mL (expermient II)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- Others: Shaken on an orbital shaker to keep the algae in suspension
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: algal medium corresponds to the OECD TG 201 medium was producded with deionised sterilized water
- Culture medium different from test medium: stock culture are kept on solid agar; liquid algal medium (corresponds to the OECD TG 201 medium) was used for test and pre-culture
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod: continous illumination
- Light intensity and quality: 5000 lux
EFFECT PARAMETERS MEASURED :
- Determination of cell concentrations: cell number was determined with an electronic particle counter
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.2
- Range finding study: yes, non GLP pre-tests, no information on results given - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.49 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% confidence interval
- Remarks:
- 0.36 – 0.65 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.07 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% confidence interval
- Remarks:
- 0.06 – 0.09 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.02 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): Yes
- Observation of abnormalities (for algal test): No algae cells were observed at test item concentrations of 1 - 100 mg/L.
- Results with reference substance (positive control):
- ErC50 (72 h): 0.79 mg/L, test performed in January 2018
- Reported statistics and error estimates:
- Calculation of results was performed with the help of validated software (Microsoft Excel ®). The estimation of the biological data was accomplished using the software ToxRat® Professional, version 3.2.1. The details of calculation are stated in chapter 19 Annex 5: Statistical calculation using ToxRat® Professional 3.2.1.
Any other information on results incl. tables
Table 1: Microscopical Observations
Experiment I |
|
Nominal Concentration in mg/L |
Normal and Healthy Appearance of the Algae |
Blank control |
Yes |
0.1 |
Yes |
0.32 |
Yes |
1 |
No cells visible |
3.2 |
No cells visible |
10 |
No cells visible |
32 |
No cells visible |
100 |
No cells visible |
Experiment II |
|
Nominal Concentration in mg/L |
Normal and Healthy Appearance of the Algae |
Blank control |
Yes |
0.0032 |
Yes |
0.01 |
Yes |
0.032 |
Yes |
0.1 |
Yes |
0.32 |
Yes |
1 |
No cells visible |
Treatment 0.0032 mg/L not used for evaluation, as some replicates were turbid
Table 2: Cell Number/mL (experiment I)
Nominal Concentration |
Parameter |
Cell Number/mL |
|||
in mg/L |
|
0 h |
24 h |
48 h |
72 h |
Blank control |
Mean |
2100 |
11550 |
74097 |
405477 |
Blank control |
SD |
0 |
723 |
3861 |
29378 |
0.1 |
Mean |
2100 |
8187 |
44100 |
183800 |
0.1 |
SD |
0 |
1110 |
4568 |
22795 |
0.32 |
Mean |
2100 |
7840 |
19440 |
84813 |
0.32 |
SD |
0 |
658 |
658 |
11680 |
1 |
Mean |
2100 |
6673 |
13133 |
16167 |
1 |
SD |
0 |
873 |
2487 |
892 |
3.2 |
Mean |
2100 |
7373 |
13527 |
13553 |
3.2 |
SD |
0 |
670 |
3606 |
1497 |
10 |
Mean |
2100 |
7680 |
13140 |
14487 |
10 |
SD |
0 |
500 |
302 |
1808 |
32 |
Mean |
2100 |
7527 |
12953 |
16793 |
32 |
SD |
0 |
687 |
424 |
884 |
100 |
Mean |
2100 |
6080 |
13893 |
16320 |
100 |
SD |
0 |
677 |
1230 |
1311 |
SD = Standard Deviation
Table 3: Cell Number/mL (experiment II)
Nominal Concentration in mg/L |
Parameter |
Cell Number/mL |
|
|
|
|
|
0 h |
24 h |
48 h |
72 h |
Blank control |
Mean |
2100 |
12493 |
57847 |
336973 |
Blank control |
SD |
0 |
1230 |
4134 |
24070 |
0.0032 |
Mean |
2100 |
10220 |
43587 |
250673 |
0.0032 |
SD |
0 |
1849 |
3507 |
16372 |
0.01 |
Mean |
2100 |
12513 |
62007 |
360980 |
0.01 |
SD |
0 |
232 |
3156 |
8152 |
0.032 |
Mean |
2100 |
12207 |
56987 |
349200 |
0.032 |
SD |
0 |
515 |
4268 |
25824 |
0.1 |
Mean |
2100 |
10133 |
41907 |
251713 |
0.1 |
SD |
0 |
1544 |
4818 |
26433 |
0.32 |
Mean |
2100 |
10033 |
26393 |
70013 |
0.32 |
SD |
0 |
674 |
1187 |
2807 |
1 |
Mean |
2100 |
9447 |
15433 |
20313 |
1 |
SD |
0 |
1467 |
1305 |
2066 |
SD = Standard Deviation; Treatment 0.0032 mg/L not used for evaluation, as some replicates were turbid
Table 4: Inhibition Values (experiment I), calculated by Microsoft Excel
Nominal concentration in mg/L |
% Inhibition |
|
|
Growth Rate (0-72h) |
Yield (0-72h) |
Blank control |
0 |
0 |
0.1 |
15.10 |
54.96 |
0.32 |
29.83 |
79.49 |
1 |
61.22 |
96.51 |
3.2 |
64.64 |
97.16 |
10 |
63.39 |
96.93 |
32 |
60.50 |
96.36 |
100 |
61.07 |
96.47 |
Table 5: Inhibition Values (experiment II), calculated by ToxRat
Nominal concentration in mg/L |
% Inhibition |
|
|
Growth Rate (0-72h) |
Yield (0-72h) |
Blank control |
0 |
0 |
0.0032 |
5.81 |
25.77 |
0.01 |
-1.4 |
-7.2 |
0.032 |
-0.7 |
-3.7 |
0.1 |
5.8 |
25.5 |
0.32 |
30.9 |
79.7 |
1 |
55.4 |
94.6 |
Negative inhibition values indicate a stimulation of algal growth compared to the blank control; Treatment 0.0032 mg/L not used for evaluation, as some replicates were turbid
Table 6: Biological Results of the Test Item
Parameter |
Value |
95% confidence interval |
NOEC (Growth Rate) 72 h |
0.02 mg/L |
-- |
NOEC (Yield) 72 h |
0.02 mg/L |
-- |
LOEC (Growth Rate) 72 h |
0.06 mg/L |
-- |
LOEC (Yield) 72 h |
0.06 mg/L |
-- |
72h ErC10 |
0.07 mg/L |
0.06 – 0.09 mg/L |
72h EyC10 |
0.04 mg/L |
0.03 – 0.05 mg/L |
72h ErC50 |
0.49 mg/L |
0.36 – 0.65 mg/L |
72h EyC50 |
0.11 mg/L |
0.07 – 0.15 mg/L |
Analytical Determination:
Experiment I
The measured concentrations in treatment 0.1 mg/L were below the LOQ (0.05 mg/L) at the start of the test and after 72 hours. The measured concentrations in treatment 0.32 mg/L were below the LOQ after 72h. Half the LOQ (0.025 mg/L) was used to calculate the geometric mean of the treatment 0.32 mg/L for the 72h value. This is permitted according to Guidance Document No. 23. The measurable concentrations lay between 41 % and 92 % of the nominal concentrations at the beginning of the test and between 50 % and 88 % of the nominal concentrations at the end of the test. The decrease in the recovery of the test item with decreasing concentration can be explained by the adsorption of dissolved test item onto the algae cells.
Experiment II
In treatment 0.0032 mg/L to 0.032 mg/L the measured concentrations at the beginning of the test were below the LOQ. At the end of the test, the concentrations in the treatments 0.0032 - 0.1 mg/L were below the LOQ. The measurable concentrations lay between 58 % and 72 % of the nominal concentrations at the beginning of the test and between 48 % and 58 % of the nominal concentrations at the end of the test and therefore not within ± 20% of the nominal values. Since the lower concentrations were prepared by dilution of the stock solution, the real concentrations were calculated on the basis of the geometric mean of the measured concentration in the highest concentrated treatment and the respective dilution factor. Geometric mean is calculated by multiplication of the n participating concentrations and taking the nth root. At the lowest concentration (0.0032 mg/L), the solution became partly turbid after 48 hours. This treatment is therefore excluded from the evaluation. Turbidity was not caused by the test item because in the 2 next higher concentrations the solutions were not turbid and no inhibition of algal growth was observed.
Table 7: Measured Concentrations and % of Nominal (Experiment I)
Nominal Concentration Test Item |
Measured concentration t = 0 h |
Measured concentration t = 72 h |
% of Nominal t = 0 h |
% of Nominal t = 72 h |
mg/L |
mg/L |
mg/L |
mg/L |
mg/L |
Blank control |
< LOQ |
< LOQ |
-- |
-- |
0.1 |
< LOQ |
< LOQ |
-- |
-- |
0.32 |
0.13 |
< LOQ |
41 |
-- |
1 |
0.67 |
0.50 |
67 |
50 |
3.2 |
2.57 |
2.25 |
80 |
70 |
10 |
8.83 |
7.78 |
88 |
78 |
32 |
28.58 |
25.70 |
89 |
80 |
100 |
92.39 |
87.55 |
92 |
88 |
LOQ (Limit of quantification) = 0.05 mg/L
Table 8: Measured Concentrations and % of Nominal (Experiment II)
Nominal Concentration Test Item |
Measured concentration t = 0 h |
Measured concentration t = 72 h |
% of Nominal t = 0 h |
% of Nominal t = 72 h |
mg/L |
mg/L |
mg/L |
mg/L |
mg/L |
Blank control |
< LOQ |
< LOQ |
-- |
-- |
0.0032 |
< LOQ |
< LOQ |
-- |
-- |
0.01 |
< LOQ |
< LOQ |
-- |
-- |
0.032 |
< LOQ |
< LOQ |
-- |
-- |
0.1 |
0.06 |
< |
LOQ |
58 |
0.32 |
0.21 |
0.15 |
65 |
48 |
1 |
0.72 |
0.58 |
72 |
58 |
LOQ (Limit of quantification) = 0.05 mg/L
Treatment 0.0032 mg/L not used for evaluation, as some replicates were turbid
Table 9: Geometric Mean (Experiment I)
Nominal Concentration Test Item |
Geometric Mean |
mg/L |
mg/L |
Blank control |
-- |
0.1 |
-- |
0.32 |
0.06 |
1 |
0.58 |
3.2 |
2.40 |
10 |
8.29 |
32 |
27.10 |
100 |
89.93 |
Table 10: Geometric Mean and calculated concentrations (Experiment II)
Nominal Concentration Test Item |
Geometric Mean |
Calculated concentrations |
mg/L |
mg/L |
mg/L |
Blank control |
-- |
-- |
0.0032 |
-- |
0.002 |
0.01 |
-- |
0.006 |
0.032 |
-- |
0.021 |
0.1 |
0.058 |
0.065 |
0.32 |
0.179 |
0.207 |
1 |
0.648 |
0.648 |
Treatment 0.0032 mg/L not used for evaluation, as some replicates were turbid
Table 10: Validity criteria for OECD 201.
Criterion from the guideline |
Outcome |
Validity criterion fulfilled |
The biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72-hour test period. |
193 (experiment I) 160 (experiment II) |
Yes Yes |
The mean coefficient of variation for section-by-section specific growth rates (days 0-1, 1-2 and 2-3, for 72-hour tests) in the control cultures must not exceed 35% |
6%(experiment I) 9%(experiment II)
|
Yes Yes |
The coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% in tests with Pseudokirchneriella subcapitata and Desmodesmus subspicatus. For other less frequently tested species, the value should not exceed 10%. |
1%(experiment I) 1%(experiment II)
|
Yes Yes |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Remarks:
- For further details please refer to “Any other information on results incl. tables”.
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