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Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

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Link to relevant study record(s)

Reference
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991-09-17 to 1991-09-27
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Guideline study with acceptable restrictions. The study was conducted according to an appropriate OECD test guideline, with acceptable restrictions. The restriction was that the presence of undissolved test material (turbidity) in the water-accommodated fractions may have contributed to effects observed in the test.
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: Water-accommodated fractions of the test substance were prepared individually for each treatment. Appropriate amounts of the substance were added to 1 litre bottles and 1 litre of algal growth medium added. The media were then stirred vigorously for 24 hours at room temperature and then allowed to stand for 20 hours. After standing the upper approximately 350 mL of medium in each bottle was siphoned off for use as the test water-accommodated fractions. A parallel set of test media were prepared by direct addition of the substance to algal growth medium with no further treatment.

- Controls: algal growth medium

- Evidence of undissolved material (e.g. precipitate, surface film, etc): there was evidence of turbidity in the water-accommodated fractions.
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM

- Strain: SAG 86.81

- Source (laboratory, culture collection): Collection of Algal Cultures, Institute for Plant Physiology, University of Göttingen, Germany.

- Method of cultivation: not described

ACCLIMATION

- Culturing media and conditions: The test cultures were grown in algal growth medium prepared in accordance with the OECD guideline except that it contained 150 mg/l NaHCO3 and not 50 mg/l.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Remarks on exposure duration:
Nominal
Hardness:
no data
Test temperature:
23+/-1ºC
pH:
8.0-8.1 at start of test

7.9-8.3 throughout test
Dissolved oxygen:
no data
Salinity:
not applicable
Nominal and measured concentrations:
Nominal loading rates in water-accommodated fraction test: 0(Control), 320, 560, 1000, 1800, 3200 and 10000 mg/L.

Nominal loading rates in direct addition test: 0(Control), 10, 32, 100, 320 and 1000 mg/L.

There was evidence of turbidity in the water-accommodated fractions.
Details on test conditions:
TEST SYSTEM

- Test vessel: Conical flask

- Type: covered with silicone sponge caps

- Material, size, headspace, fill volume: glass, 200 mL with 100 ml of test medium

- Aeration: none

- Initial cells density: 12000-13000 cells/mL

- Control end cells density: 2500000-2840000 cells/mL

- No. of vessels per concentration (replicates): 2

- No. of vessels per control (replicates): 4


GROWTH MEDIUM

- Standard medium used: no

- Detailed composition if non-standard medium was used: OECD medium except that 150 mg/L of NaHCO3 was used instead of 50 mg/L


TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: The dilution water (algal growth medium) was prepared by addition of concentrated stock solutions of salts and nutrients to Milli-Q filtered water. The final medium was filter-sterilized before use.

- Culture medium different from test medium: no

- Intervals of water quality measurement: start and end of test


OTHER TEST CONDITIONS

- Sterile test conditions: yes

- Adjustment of pH: no

- Photoperiod: continuous

- Light intensity and quality: 120 μmol/s/m2 +/-20% from fluorescent lights in a Gallenkamp orbital shaker


EFFECT PARAMETERS MEASURED (with observation intervals if applicable):

- Determination of cell concentrations: Coulter Counter TAII and Perking Elmer 3000 Fluorescence Spectrometer


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 1.8 (water-accommodated fraction test), 3.3 (direct addition test)

- Range finding study

- Test concentrations: 10, 100, 1000, 10000 mg/L

- Results used to determine the conditions for the definitive study: Results indicated that toxic effects could be expected at loading rates of >100 mg/L.
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
EL10
Effect conc.:
1 600 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: yield after logistic growth based on cell numbers
Duration:
96 h
Dose descriptor:
EL50
Effect conc.:
2 600 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: yield after logistic growth based on cell numbers
Remarks on result:
other: 2000-3400
Duration:
96 h
Dose descriptor:
EL10
Effect conc.:
< 320 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: area under the growth curve based on cell numbers
Duration:
96 h
Dose descriptor:
EL50
Effect conc.:
2 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
biomass
Remarks:
area under the growth curve based on cell numbers
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
60 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: yield after logistic growth based on fluorescence measurements
Duration:
96 h
Dose descriptor:
EL50
Effect conc.:
700 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: yield after logistic growth based on fluorescence measurements
Remarks on result:
other: 230-2300
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
> 1 000 - < 1 800 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: area under the growth curve based on fluorescence measurements
Duration:
96 h
Dose descriptor:
EL50
Effect conc.:
> 1 800 - < 32 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: area under the growth curve based on fluorescence measurements
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
water-accommodated fraction
Basis for effect:
other: yield and biomass
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
added directly to medium
Basis for effect:
other: yield and biomass
Details on results:
- Exponential growth in the control (for algal test): yes

- Observation of abnormalities (for algal test): no

- Any stimulation of growth found in any treatment: no

- Effect concentrations exceeding solubility of substance in test medium: yes, there was evidence of turbidity in the water-accommodated fractions. The results are above the expected water solubility of the substance (<1 mg/L).
Reported statistics and error estimates:
The area under the growth curve (biomass) and logistic growth parameters were calculated from the cell concentration data in accordance with the description of the test method.

ELx values with respect to logistic growth were calculated using a parametric model developed by Kooijman et al (1983).

NOELR values were determined by visual comparison of the growth curves for treated and control cultures.

Kooijman, S.A.L.M., A.O. Hanstveit and H. Oldersma. (1983). Parametric analysis of population growth in bioassays. Water Research, 17: 527-538.

Table 1. Results of test with test media prepared as water-accommodated fractions (mean cell concentrations in cells/ml in each treatment loading rate)

Time (hours)  Control  Control  0.32 g/L  0.56 g/L  1.0 g/L  1.8 g/L  3.2 g/L  10 g/L
 0  13000  12000  12000  12000  12000  12000  12000  12000
 23.5  56000  62000 58000   197000  142000  39000  -  94000
 48.5  278000  306000  304000  300000  170000  294000  248000  3000
 73.5  879000  1216000  851000  410000  836000  815000  245000  313000
 97.0  1961000  3070000  1665000  728000  1648000  1416000  385000  725000

Table 2. Results of test with test substance added directly (mean cell concentrations in cells/ml in each treatment loading rate)

 Time (hours)  Control  Control  10 mg/L  32 mg/L  100 mg/L  320 mg/L  1000 mg/L
 0  12000  12000  12000  12000  12000  12000  12000
 19.5  43000  39000  43000  44000  40000  50000  36000
 46.5  333000  329000  345000  327000  330000  337000  333000
 71.5  1250000  1126000  1132000  960000  1001000  1009000  1040000
 93.5  3005000  2675000  2803000  2228000  2536000  2479000  2209000
Validity criteria fulfilled:
yes
Conclusions:
A 96-h EL50 value of 700 mg/L and a 96-h EL10 of 60 mg/L have been determined for the effects of the test substance on yield of Scenedesmus subspicatus as determined by fluorescence measurement. A NOELR of 1000 mg/l has been estimated from the test results by the report authors but does not seem realistic given the EL50 and EL10 values. The results are above the expected water solubility of the test substance (<1 mg/L). There was evidence of undissolved test material (turbidity) in the water-accommodated fractions and this may have contributed to effects observed at the high test substance loading rates utilised in the test.

Description of key information

EL50 (96 h) >1000 mg/l (nominal) and EL10 (96 h): >1000 mg/l (nominal) based on: biomass and yield in Scenedesmus subspicatus (new name: Desmodesmus subspicatus), in media prepared by direct addition, and an EL50 of 700 mg/l (nominal) based on: yield in Scenedesmus subspicatus (new name: Desmodesmus subspicatus), in media prepared by water accommodated fraction. The EL50 value of 700 mg/l is selected as the key value for this endpoint as it is a definitive value, not a limit value (>).

Key value for chemical safety assessment

Additional information

A 96-hour EL50 value of 700 mg/l and a 96-hour EL10 of 60 mg/l have been determined for the effects of bis[3-triethoxysilylpropyl]polysulfides (CAS 211519-85-6, EC 915-673-4) on the yield of Scenedesmus subspicatus as determined by fluorescence measurement (Evonik Degussa, 1992a). A NOELR of 1000 mg/l has been estimated from the test results by the report authors but does not seem realistic given the EL50 and EL10 values. This test was conducted in media prepared by Water Accommodated Fractions (WAFs). The results are above the expected water solubility of the test substance (approximately 0.01 mg/l at 20°C). There was evidence of undissolved test material (turbidity) in the water-accommodated fractions and this may have contributed to effects observed at the high test substance loading rates utilised in the test. Based on the test media preparation methods and static exposure regime, it is likely that the test organisms would have been exposed to a mixture of the parent substance, the hydrolysis products and undissolved test substance and/or precipitated by-products. Insoluble oligomers may have precipitated out of solution to an extent during the media preparation and in the course of the test. The test results have been interpreted in this context. This is reflected in the observation of turbidity in test media, although no analytical verification was performed. In view of the media preparation method and duration of study, the effects seen are attributed to precipitated oligomeric or cross-linked by-products. Physical effects are likely to have dominated any intrinsic toxic effects.

In the same study, a 96-hour EL50 value of >1000 mg/l (nominal) and an EL10 of >1000 mg/l (nominal, highest concentration tested) have been determined for the effects of bis[3-triethoxysilylpropyl]polysulfides (CAS 211519-85-6) on the yield of Scenedesmus subspicatus as determined by fluorescence measurement (Evonik Degussa, 1992a). This test was conducted in media prepared by direct addition, intending to represent the properties of the parent substance. Based on the test media preparation methods and static exposure regime, it is likely that the test organisms would have been exposed to a mixture of the parent substance, the hydrolysis products and undissolved test substance and/or precipitated by-products. Insoluble oligomers may have precipitated out of solution to an extent in the course of the test. The test results have been interpreted in this context. No analytical verification was performed. In view of the media preparation method and duration of study, the effects seen are attributed to a mixture of parent, silanol hydrolysis product and oligomeric or cross-linked by-products. Physical effects such as decrease in light intensity could have contributed by affecting growth rate.

The 96-hour EL50 value of 700 mg/l is considered to be the appropriate value for the registration substance.