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Toxicity to aquatic algae and cyanobacteria

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Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22/10/2013 - 25/10/2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study according to international guideline (OECD guideline 201) under GLP. Validity criteria met, deviations from protocol did not adversely affect study integrity.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)
Deviations:
no
Principles of method if other than guideline:
Not relevant
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not relevant
Analytical monitoring:
yes
Details on sampling:
- Concentrations: nominal WAFs of 10, 18, 32, 56 and 100 mg/l.
- Sampling method: 40 ml samples for analysis were taken from extra vessels containing test solutions incubated without algae after 0 and 72 hours. Additionally, reserve samples of 40 ml were taken from all test solutions for possible analysis. If not already used, these samples were stored in a refrigerator for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.
- Sample storage conditions before analysis: Samples were stored in a refrigerator until analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The standard test procedures required generation of test solutions, which contained completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that would disturb the test system was prevented as much as possible (e.g. film of the test substance on the water surface). No correction was made for the purity/composition of the test substance. The batch of Blown Linseed Oil tested was a hazy yellow to brown viscous liquid and an UVCB substance. The test substance was not completely soluble in test medium at the loading rates prepared.
Preparation started with individual loading rates ranging from 1.0 to 100 mg/l. A 2-day period of magnetic stirring was applied to accelerate the dissolving of the test substance in the test medium. The obtained mixtures were left for a period of 1 hour to stabilise. Subsequently, the aqueous Water Accommodated Fractions (WAFs) were siphoned off and used as test solutions. After preparation, volumes of 50 ml were added to each replicate of the respective test concentration.
- Controls: Yes, controls and blanks withut algae
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The highest WAF in the final test was slightly hazy, whereas the remaining test solutions were all clear and colourless.
Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: NIVA CHL 1
- Source (laboratory, culture collection): In-house laboratory culture
- Method of cultivation: Algae stock cultures were started by inoculating growth medium with algal cells from a pure culture on agar. The suspensions were continuously aerated and exposed to light in a climate room at a temperature of 21-24°C. 3 days before the start of the test, cells from the algal stock culture were inoculated in culture medium at a cell density of 1 x 10^4 cells/ml. The pre-culture was maintained under the same conditions as used in the test. The cell density was measured immediately before use.

ACCLIMATION
- Acclimation period: 3 days
- Culturing media and conditions (same as test or not): same
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
Not relevant
Hardness:
24 mg/L CaCO3
Test temperature:
22.5 - 23.3ºC
pH:
7.4 - 8.1
Dissolved oxygen:
No data
Salinity:
Not relevant
Nominal and measured concentrations:
Nominal: 10, 18, 32, 56 and 100 mg/L WAF
Measured (t=0h, TOC, corrected for control): 0.5508, 1.2109, 2.7069, 4.0589 and 6.3469 mg/L
Measured (t=72h, TOC, corrected for control): 0.5281, 1.1415, 2.4805, 2.4995 and 5.9645 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Vessel
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 100 mL filled with 50 mL of test medium
- Aeration: None
- Initial cells density: 1.0 x 10^4 cells/mL
- Control end cells density: 53.6 x 10^4 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per control (no algae): 2

GROWTH MEDIUM
- Standard medium used: yes, Elendt M1/M2 medium
- Detailed composition if non-standard medium was used:

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Milli-RO water
NH4Cl 15 mg/l
MgCl2.6H2O 12 mg/l
CaCl2.2H2O 18 mg/l
MgSO4.7H2O 15 mg/l
KH2PO4 1.6 mg/l
FeCl3.6H2O 64 μg/l
Na2EDTA.2H2O 100 μg/l
H3BO3 185 μg/l
MnCl2.4H2O 415 μg/l
ZnCl2 3 μg/l
CoCl2.6H2O 1.5 μg/l
CuCl2.2H2O 0.01 μg/l
Na2MoO4.2H2O 7 μg/l
NaHCO3 50 mg/l
Hardness (Ca+Mg): 0.24 mmol/l
- Culture medium different from test medium: No
- Intervals of water quality measurement: pH at the beginning and end of the test, temperature was recorded continuously in a temperature control vessel

OTHER TEST CONDITIONS
- Photoperiod: Continuous
- Light intensity and quality: TLD-lamps with a light intensity within the range of 91 to 97 uE/m2^/s.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: At the beginning of the test, cells were counted using a microscope and a counting chamber. Thereafter cell densities were determined by spectrophotometric measurement of samples at 720 nm using a spectrophotometer with immersion probe (pathlength =20 mm). Algal medium was used as blank and the extra replicates as background for the treated solutions.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study
- Test concentrations: 1, 10 amd 100 mg/L
- Results used to determine the conditions for the definitive study: Yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
56 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
61 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 95%CL: 56 - 64 mg/L
Key result
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
41 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95%CL: 37 - 44 mg/L
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
75 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95%CL: 72 - 78 mg/L
Details on results:
- Exponential growth in the control (for algal test): yes
- Observation of abnormalities: Microscopic observations at the end of the test revealed a normal and healthy appearance of the
exposed cells when compared to the control.
- Any stimulation of growth found in any treatment: Yes, in the 10, 18 and 32 mg/L treatments
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The highest WAF in the final test was slightly hazy, whereas the remaining test solutions were all clear and colourless.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50: 1.3 mg/L
Reported statistics and error estimates:
For determination of the NOEL and the EL50 the approaches recommended in the OECD guideline 201 were used. An effect was considered to be significant if statistical analysis of the data obtained for the test concentrations compared with those obtained in the negative control revealed significant inhibition of growth rate or inhibition of yield (Williams Multiple Sequential t-test Procedure, α=0.05, one-sided, smaller). Additionally, the EL10 and EL20 were determined to meet the recommendations as put down in "A Review of Statistical Data Analysis and Experimental Design in OECD Aquatic Toxicology Test Guidelines" by S. Pack, August 1993. Calculation of the ELX values was based on probit analysis using linear max. likelihood regression with the percentages of growth rate inhibition and the percentages of yield inhibition versus the logarithms of the corresponding loading rate initially applied. The calculations were performed with ToxRat Professional v. 2.10.05 (ToxRat Solutions® GmbH, Germany).

Individual cell densities

Individual cell densities (x 10^4 cells/mL)
WAF (mg/L)
Time Replicate Control 10 18 32 56 100
0h 1 1 1 1 1 1 1
2 1 1 1 1 1 1
3 1 1 1 1 1 1
4 1
5 1
6 1
n: 6 3 3 3 3 3
Mean: 1 1 1 1 1 1
St. Dev.: 0 0 0 0 0 0
CV: 0 0 0 0 0 0
24h 1 4.151 4.260 3.766 3.696 3.333 2.388
2 4.083 3.445 3.749 3.575 3.083 2.351
3 4.039 3.575 3.948 3.661 3.124 2.004
4 4.208
5 4.168
6 4.377
n: 6 3 3 3 3 3
Mean: 4.2 3.8 3.8 3.6 3.2 2.2
St. Dev.: 0.1 0.4 0.1 0.1 0.1 0.2
CV: 2.8 11.6 2.9 1.7 4.2 9.4
48h 1 13.541 17.622 13.761 16.622 9.688 5.601
2 12.700 14.183 14.275 15.189 10.544 5.754
3 12.632 15.508 17.171 17.360 9.019 5.361
4 14.506
5 13.915
6 14.616
n: 6 3 3 3 3 3
Mean: 13.7 15.8 15.1 16.4 9.8 5.6
St. Dev.: 0.9 1.7 1.8 1.1 0.8 0.2
CV: 6.3 11 12.2 6.7 7.8 3.6
72h 1 55.207 77.040 55.616 69.421 38.076 14.146
2 45.952 60.703 54.419 61.237 42.250 17.598
3 51.912 65.086 71.855 67.536 34.898 15.414
4 56.225
5 54.715
6 57.483
n: 6 3 3 3 3 3
Mean: 53.6 67.8 60.6 66.1 38.4 15.7
St. Dev.: 4.2 8.8 9.7 4.3 3.7 1.7
CV: 7.8 13 16.1 6.5 9.6 11.1

Individual growth rates (day^-1)

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
24h 1 1.423 1.449 1.326 1.307 1.204 0.87
2 1.407 1.237 1.321 1.274 1.126 0.855
3 1.396 1.274 1.373 1.298 1.139 0.695
4 1.437
5 1.427
6 1.476
n: 6 3 3 3 3 3
Mean: 1.428 1.320 1.340 1.293 1.156 0.807
St. Dev.: 0.028 0.113 0.029 0.017 0.042 0.097
CV: 2.000 8.600 2.100 1.200 3.600 12.000
48h 1 1.303 1.435 1.311 1.405 1.135 0.861
2 1.271 1.326 1.329 1.360 1.178 0.875
3 1.268 1.371 1.422 1.427 1.100 0.84
4 1.337
5 1.316
6 1.341
n: 6 3 3 3 3 3
Mean: 1.306 1.377 1.354 1.398 1.138 0.859
St. Dev.: 0.032 0.055 0.059 0.034 0.039 0.018
CV: 2.400 4.000 4.400 2.400 3.400 2.100
72h 1 1.337 1.451 1.339 1.413 1.213 0.833
2 1.276 1.369 1.332 1.372 1.248 0.956
3 1.317 1.392 1.425 1.404 1.184 0.912
4 1.343
5 1.334
6 1.350
n: 6 3 3 3 3 3
Mean: 1.326 1.404 1.366 1.396 1.215 0.917
St. Dev.: 0.027 0.043 0.052 0.022 0.032 0.037
CV: 2.000 3.000 3.800 1.600 2.600 4.000

Section-by-section growth rate

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
0 - 24h 1 1.400 1.400 1.300 1.300 1.200 0.9
2 1.400 1.200 1.300 1.300 1.100 0.9
3 1.400 1.300 1.400 1.300 1.100 0.7
4 1.400
5 1.400
6 1.500
n: 6 3 3 3 3 3
Mean: 1.400 1.300 1.300 1.300 1.200 0.800
St. Dev.: 0.030 0.110 0.030 0.020 0.040 0.100
CV: 2.000 8.600 2.100 1.300 3.600 12.000
24 - 48h 1 1.200 1.400 1.300 1.500 1.100 0.900
2 1.100 1.400 1.300 1.400 1.200 0.900
3 1.100 1.500 1.500 1.600 1.100 1.000
4 1.200
5 1.200
6 1.200
n: 6 3 3 3 3 3
Mean: 1.200 1.400 1.400 1.500 1.100 0.900
St. Dev.: 0.040 0.030 0.090 0.050 0.100 0.070
CV: 3.400 2.000 6.700 3.700 8.600 7.400
48 - 72h 1 1.400 1.500 1.400 1.400 1.400 0.900
2 1.300 1.500 1.300 1.400 1.400 1.100
3 1.400 1.400 1.400 1.400 1.400 1.100
4 1.400
5 1.400
6 1.400
n: 6 3 3 3 3 3
Mean: 1.400 1.500 1.400 1.400 1.400 1.000
St. Dev.: 0.050 0.030 0.050 0.040 0.020 0.100
CV: 3.300 1.700 3.400 2.500 1.300 9.500

Individual yields

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
24h 1 3.151 3.260 2.766 2.696 2.333 1.388
2 3.083 2.445 2.749 2.575 2.083 1.351
3 3.039 2.575 2.948 2.661 2.124 1.004
4 3.208
5 3.168
6 3.377
n: 6 3 3 3 3 3
Mean: 3.171 2.760 2.821 2.644 2.180 1.248
St. Dev.: 0.118 0.438 0.11 0.062 0.134 0.212
CV: 3.700 15.900 3.900 2.400 6.200 17.000
48h 1 12.541 16.622 12.761 15.622 8.688 4.601
2 11.700 13.183 13.275 14.189 9.544 4.754
3 11.632 14.508 16.171 16.360 8.019 4.361
4 13.506
5 12.915
6 13.616
n: 6 3 3 3 3 3
Mean: 12.652 14.771 14.069 15.390 8.750 4.572
St. Dev.: 0.859 1.735 1.838 1.104 0.764 0.198
CV: 6.800 11.700 13.100 7.200 8.700 4.300
72h 1 54.207 76.740 54.616 68.421 37.076 13.146
2 44.952 59.703 53.419 60.237 41.250 16.598
3 50.912 64.086 70.855 66.536 33.898 14.414
4 55.225
5 53.715
6 56.483
n: 6 3 3 3 3 3
Mean: 52.582 66.843 59.630 65.065 37.408 14.719
St. Dev.: 4.175 8.847 9.740 4.286 3.687 1.746
CV: 7.900 13.200 16.300 6.600 9.900 11.900
Validity criteria fulfilled:
yes
Remarks:
> 16-fold increase in controls, coefficients of variation for section-by-section specific growth rates and average specific growth rates were 9.8 and 2.0% respectively
Conclusions:
The acute toxicity (72h-ELr50) of Blown linseed oil towards Pseudokirchneriella subcapitata is > 100 mg/L.
Executive summary:

The acute toxicity of Blown linseed oil towards Pseudokirchnerialla subcapitata was investigated according to OECD guideline 201 under GLP. Algal cells were exposed to nominal WAFs of 10, 18, 32, 56 and 100 mg/L and were observed for 72 hours. Based on nominal loading rates the 72h-ErL10, 72h-EbL10, 72h-ErL50 and 72h-EbL50 were found to be 61, 41, >100 and 75 mg/L respectively.

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
See attached justification.
Reason / purpose for cross-reference:
read-across source
Key result
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
56 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Key result
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
61 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 95%CL: 56 - 64 mg/L
Key result
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
41 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95%CL: 37 - 44 mg/L
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
75 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 95%CL: 72 - 78 mg/L
Details on results:
- Exponential growth in the control (for algal test): yes
- Observation of abnormalities: Microscopic observations at the end of the test revealed a normal and healthy appearance of the
exposed cells when compared to the control.
- Any stimulation of growth found in any treatment: Yes, in the 10, 18 and 32 mg/L treatments
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The highest WAF in the final test was slightly hazy, whereas the remaining test solutions were all clear and colourless.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50: 1.3 mg/L
Reported statistics and error estimates:
For determination of the NOEL and the EL50 the approaches recommended in the OECD guideline 201 were used. An effect was considered to be significant if statistical analysis of the data obtained for the test concentrations compared with those obtained in the negative control revealed significant inhibition of growth rate or inhibition of yield (Williams Multiple Sequential t-test Procedure, α=0.05, one-sided, smaller). Additionally, the EL10 and EL20 were determined to meet the recommendations as put down in "A Review of Statistical Data Analysis and Experimental Design in OECD Aquatic Toxicology Test Guidelines" by S. Pack, August 1993. Calculation of the ELX values was based on probit analysis using linear max. likelihood regression with the percentages of growth rate inhibition and the percentages of yield inhibition versus the logarithms of the corresponding loading rate initially applied. The calculations were performed with ToxRat Professional v. 2.10.05 (ToxRat Solutions® GmbH, Germany).

Individual cell densities

Individual cell densities (x 10^4 cells/mL)
WAF (mg/L)
Time Replicate Control 10 18 32 56 100
0h 1 1 1 1 1 1 1
2 1 1 1 1 1 1
3 1 1 1 1 1 1
4 1
5 1
6 1
n: 6 3 3 3 3 3
Mean: 1 1 1 1 1 1
St. Dev.: 0 0 0 0 0 0
CV: 0 0 0 0 0 0
24h 1 4.151 4.260 3.766 3.696 3.333 2.388
2 4.083 3.445 3.749 3.575 3.083 2.351
3 4.039 3.575 3.948 3.661 3.124 2.004
4 4.208
5 4.168
6 4.377
n: 6 3 3 3 3 3
Mean: 4.2 3.8 3.8 3.6 3.2 2.2
St. Dev.: 0.1 0.4 0.1 0.1 0.1 0.2
CV: 2.8 11.6 2.9 1.7 4.2 9.4
48h 1 13.541 17.622 13.761 16.622 9.688 5.601
2 12.700 14.183 14.275 15.189 10.544 5.754
3 12.632 15.508 17.171 17.360 9.019 5.361
4 14.506
5 13.915
6 14.616
n: 6 3 3 3 3 3
Mean: 13.7 15.8 15.1 16.4 9.8 5.6
St. Dev.: 0.9 1.7 1.8 1.1 0.8 0.2
CV: 6.3 11 12.2 6.7 7.8 3.6
72h 1 55.207 77.040 55.616 69.421 38.076 14.146
2 45.952 60.703 54.419 61.237 42.250 17.598
3 51.912 65.086 71.855 67.536 34.898 15.414
4 56.225
5 54.715
6 57.483
n: 6 3 3 3 3 3
Mean: 53.6 67.8 60.6 66.1 38.4 15.7
St. Dev.: 4.2 8.8 9.7 4.3 3.7 1.7
CV: 7.8 13 16.1 6.5 9.6 11.1

Individual growth rates (day^-1)

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
24h 1 1.423 1.449 1.326 1.307 1.204 0.87
2 1.407 1.237 1.321 1.274 1.126 0.855
3 1.396 1.274 1.373 1.298 1.139 0.695
4 1.437
5 1.427
6 1.476
n: 6 3 3 3 3 3
Mean: 1.428 1.320 1.340 1.293 1.156 0.807
St. Dev.: 0.028 0.113 0.029 0.017 0.042 0.097
CV: 2.000 8.600 2.100 1.200 3.600 12.000
48h 1 1.303 1.435 1.311 1.405 1.135 0.861
2 1.271 1.326 1.329 1.360 1.178 0.875
3 1.268 1.371 1.422 1.427 1.100 0.84
4 1.337
5 1.316
6 1.341
n: 6 3 3 3 3 3
Mean: 1.306 1.377 1.354 1.398 1.138 0.859
St. Dev.: 0.032 0.055 0.059 0.034 0.039 0.018
CV: 2.400 4.000 4.400 2.400 3.400 2.100
72h 1 1.337 1.451 1.339 1.413 1.213 0.833
2 1.276 1.369 1.332 1.372 1.248 0.956
3 1.317 1.392 1.425 1.404 1.184 0.912
4 1.343
5 1.334
6 1.350
n: 6 3 3 3 3 3
Mean: 1.326 1.404 1.366 1.396 1.215 0.917
St. Dev.: 0.027 0.043 0.052 0.022 0.032 0.037
CV: 2.000 3.000 3.800 1.600 2.600 4.000

Section-by-section growth rate

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
0 - 24h 1 1.400 1.400 1.300 1.300 1.200 0.9
2 1.400 1.200 1.300 1.300 1.100 0.9
3 1.400 1.300 1.400 1.300 1.100 0.7
4 1.400
5 1.400
6 1.500
n: 6 3 3 3 3 3
Mean: 1.400 1.300 1.300 1.300 1.200 0.800
St. Dev.: 0.030 0.110 0.030 0.020 0.040 0.100
CV: 2.000 8.600 2.100 1.300 3.600 12.000
24 - 48h 1 1.200 1.400 1.300 1.500 1.100 0.900
2 1.100 1.400 1.300 1.400 1.200 0.900
3 1.100 1.500 1.500 1.600 1.100 1.000
4 1.200
5 1.200
6 1.200
n: 6 3 3 3 3 3
Mean: 1.200 1.400 1.400 1.500 1.100 0.900
St. Dev.: 0.040 0.030 0.090 0.050 0.100 0.070
CV: 3.400 2.000 6.700 3.700 8.600 7.400
48 - 72h 1 1.400 1.500 1.400 1.400 1.400 0.900
2 1.300 1.500 1.300 1.400 1.400 1.100
3 1.400 1.400 1.400 1.400 1.400 1.100
4 1.400
5 1.400
6 1.400
n: 6 3 3 3 3 3
Mean: 1.400 1.500 1.400 1.400 1.400 1.000
St. Dev.: 0.050 0.030 0.050 0.040 0.020 0.100
CV: 3.300 1.700 3.400 2.500 1.300 9.500

Individual yields

WAF (mg/L)
Time Replicate Control 10 18 32 56 100
24h 1 3.151 3.260 2.766 2.696 2.333 1.388
2 3.083 2.445 2.749 2.575 2.083 1.351
3 3.039 2.575 2.948 2.661 2.124 1.004
4 3.208
5 3.168
6 3.377
n: 6 3 3 3 3 3
Mean: 3.171 2.760 2.821 2.644 2.180 1.248
St. Dev.: 0.118 0.438 0.11 0.062 0.134 0.212
CV: 3.700 15.900 3.900 2.400 6.200 17.000
48h 1 12.541 16.622 12.761 15.622 8.688 4.601
2 11.700 13.183 13.275 14.189 9.544 4.754
3 11.632 14.508 16.171 16.360 8.019 4.361
4 13.506
5 12.915
6 13.616
n: 6 3 3 3 3 3
Mean: 12.652 14.771 14.069 15.390 8.750 4.572
St. Dev.: 0.859 1.735 1.838 1.104 0.764 0.198
CV: 6.800 11.700 13.100 7.200 8.700 4.300
72h 1 54.207 76.740 54.616 68.421 37.076 13.146
2 44.952 59.703 53.419 60.237 41.250 16.598
3 50.912 64.086 70.855 66.536 33.898 14.414
4 55.225
5 53.715
6 56.483
n: 6 3 3 3 3 3
Mean: 52.582 66.843 59.630 65.065 37.408 14.719
St. Dev.: 4.175 8.847 9.740 4.286 3.687 1.746
CV: 7.900 13.200 16.300 6.600 9.900 11.900
Validity criteria fulfilled:
yes
Remarks:
> 16-fold increase in controls, coefficients of variation for section-by-section specific growth rates and average specific growth rates were 9.8 and 2.0% respectively
Conclusions:
The acute toxicity (72h-ELr50) of Blown linseed oil towards Pseudokirchneriella subcapitata is > 100 mg/L.
Executive summary:

The acute toxicity of Blown linseed oil towards Pseudokirchnerialla subcapitata was investigated according to OECD guideline 201 under GLP. Algal cells were exposed to nominal WAFs of 10, 18, 32, 56 and 100 mg/L and were observed for 72 hours. Based on nominal loading rates the 72h-ErL10, 72h-EbL10, 72h-ErL50 and 72h-EbL50 were found to be 61, 41, >100 and 75 mg/L respectively.

Description of key information

In a WAF study with Linseed oil, oxidized according to the guideline 201, the EL50 based on the growth rate was observed to be >100 mg/L. The study is used for read across to this dossier with Rape oil, oxidized

Key value for chemical safety assessment

EC50 for freshwater algae:
100 mg/L
EC10 or NOEC for freshwater algae:
61 mg/L

Additional information

One study is available for the toxicity towards algae with Blown Rape oil. The study of Kroon (1992) is disregarded because effects are observed at levels exceeding the solubility limit of the substance and exposure solutions were opaque and inhomogenous. Undissolved material has disturbed the measurements of algal cell density.

The WAF study of Tobor-Kaplon (2013) with Blown Linseed oil is conducted according to the guideline 201 and fulfills all validity criteria. The EL50 based on the growth rate was >100 mg/L. This study is used for read across to this dossier with Rape oil, oxidized. A justification is provided in the read-across justification document.