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Toxicity to aquatic algae and cyanobacteria

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Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
other: published data
Adequacy of study:
key study
Study period:
2005-01-10 to 2005-01-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
GLP Code: SDA-003/4-30
Details on sampling:
- Concentrations: all nominal concentrations were tested at time 0, 24 and 72 h.

- Sampling method: samples were taken from the vessels with algae and vessels prepared without algae.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: A stock solution was prepared by adding 180 mg of the test substance to sterilised test medium and adjusting the volume to 1 L. The solution was stirred for 24 hours. The stock solution was diluted with sterilised growth medium to obtain the nominal test concentrations under sterile conditions: 7.7, 17, 37, 82 and 180 mg hexanol/L.

- Controls: standard control using dilution water.
Test organisms (species):
Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM

- Common name: freshwater green alga

- Strain: Pseudokirchneriella subcapitata

- Source (laboratory, culture collection): SAG, Culture Collection of Algae at Pflanzenphysiologisches Institut of the University at Göttingen, Albrecht von Haller Institut, Untere Klarspüle 2, D-37073 Göttingen, Catalog No 61.81.
ACCLIMATION

- Culturing media and conditions: The stock cultures were maintained fulfilling the criteria of the OECD guidelines. Prior to testing a pre-culture was established in test medium to obtain exponentially growing algae for the test.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Post exposure observation period:
None
Hardness:
N.R.
Test temperature:
22.0 - 22.2 °C
pH:
8.52 - 8.58 (Controls), 8.08 - 8.40 (test concentrations)
Dissolved oxygen:
N.R.
Salinity:
N.A.
Nominal and measured concentrations:
Nominal: 7.7, 17, 37, 82 and 180 mg hexanol/L
Mean measured (with algae): 4.72, 11.3, 31.2, 50.1 and 111.2 mg hexanol/L
Mean measured (without algae present): 5.48, 12.3, 29.3, 54.9 and 105.5 mg hexanol/L
Details on test conditions:
TEST SYSTEM

- Test vessel: glass flask, with cultures being resuspended continuously by shaking on a laboratory shaker (Incubation Shaker Multitron ®, INFORS, Switzerland).

- Type: closed

- Material, size, headspace, fill volume: Test vessels are 250 mL conical glass flasks covered with silicone-sponge caps. The vessels and caps were sterilized prior to use (autoclaving or heating).

- Aeration: none

- Initial cells density: 10,000 cells/mL

- Control end cells density: 143.3x10^4 cells/mL

- No. of vessels per concentration: 3
- No. of vessels per control: 6


GROWTH MEDIUM

- Standard medium used: yes


TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: A sterilized synthetic growth medium according to OECD 201

- Culture medium different from test medium: no

- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS

- Sterile test conditions: yes

-Adjustment of pH: none

- Photoperiod: continuous

- Light intensity and quality: 8000 Lux
EFFECT PARAMETERS MEASURED:

- Determination of cell concentrations: Cell concentrations were determined using an electronic particle counter (CASY 1 Model TT Schärfe System
Reutlingen, Germany) from time 0h to 72h every 24h. The correctness of the electronic counts was checked by microscopically counting following internal operation procedures.


TEST CONCENTRATIONS

- Spacing factor for test concentrations: five concentrations arranged in a geometric series, spaced by a factor of 2.2

- Range finding study: yes, details not reported.
Reference substance (positive control):
no
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
11.3 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
31.2 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
19.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
79.7 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
NOEC
Effect conc.:
< 4.72 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
LOEC
Effect conc.:
< 4.72 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
4.97 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
20.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
biomass
Details on results:
- Exponential growth in the control: yes
Reported statistics and error estimates:
Calculation of the percent inhibition of growth rate, yield (cell counts) and biomass integral was performed according to the OECD guideline.
A concentration-effect relationship was observed and statistically analyzed to obtain effect concentrations.
The test results were statistically analyzed to determine EC10 and EC50 values together with 95 % confidence intervals using Probit-analysis assuming lognormal distribution of the values by using the computer program ToxRat.
The NOEC and LOEC values for yield and biomass were determined by the Welch t-test.

Table 1. Cell densities, biomass and growth rate, and percentage inhibition of the test organism treated at a range of concentrations of the test substance. Biomass and growth rate are the mean at 72 h, the percentage inhibition are expressed relative to the period 0 -72 h.

      Treatment concentration (measured concentration average mg/L)

 Cell density            

 Biomass

 Biomass % inhibition

 Growth rate

Growth rate % inhibition 

 Control 0

 Replicate #

 24 h

48 h 

 72 h

 

-

 -

 -

 

 1

 5.0

 27.0

 136.3

  -

 -

 -

 -

 

 2

 4.5

 27.0

 138.6

  -

-

 -

 -

 

 3

 4.6

 29.3

 148.6

 -

 -

 -

 -

 

 4

 4.9

 30.2

 148.1

 -

 -

 -

 -

 

 5

 4.6

 29.7

 140.4

 -

 -

 -

 -

 

 6

 5.1

 27.8

 147.7

 -

 -

 -

 -

 Mean (± SD)

 -

 4.8 ± 0.2

 28.5 ± 1.4

143.3 ± 5.5 

 102.4 ±3.8

 0

 1.655 ± 0.013

 0

 4.72 mg/L

 1

 3.7

 26.2

 154.5

 -

 -

 -

 -

 

 2

 3.1

 21.6

 137.8

 -

 -

 -

 -

 

 3

 1.4

 12.1

 72.4

 -

 -

 -

 -

 Mean (± SD)

 

 2.7 ± 1.2

 20.0 ± 7.2

 121.6 ± 43.4

 81.0 ± 30.0

 20.9

 1.583 ± 0.136

4.3 

 11.3 mg/L

 1

 3.0

 23.8

 145.7

 -

 -

 -

 -

 

 2

 3.0

 17.5

 118.5

 -

 -

 -

 -

 

 3

 2.6

 16.6

 117.0

 -

 -

 -

 -

 Mean (± SD)

 

 2.9 ± 0.2

 19.3 ± 3.9

 127.1 ± 16.2

 83.2 ± 12.1

18.7

 1.613 ± 0.041

 2.5

 31.2 mg/L

 1

 1.6

 2.2

 78.5

 -

 -

 -

 -

 

 2

 1.7

 6.5

 43.5

 -

 -

 -

 -

 

 3

 2.0

 8.8

 27.5

 -

 -

 -

 -

 Mean ± SD

 

 1.7 ± 0.2

 5.8 ± 3.4

 49.8 ± 26.1

 30.0 ± 9.5

 70.7

 1.272 ± 0.175

 23.1

 50.1 mg/L

 1

 1.8

 7.4

32.6 

 -

 -

 -

 -

 

 2

 2.0

 7.7

 36.0

 -

 -

 -

 -

 

 3

 1.9

 7.1

 32.6

 -

 -

 -

 -

 Mean (± SD)

 -

 1.9 ± 0.1

 7.4 ± 0.3

 33.8 ± 2.0

23.6 ± 1.4

 76.9

 1.173 ± 0.019

 29.1

 111.2 mg/L

 1

 1.6

 4.0

 7.8

 -

 -

 -

 -

 

 2

 1.7

 2.4

 4.8

 -

 -

 -

 -

 

 3

 1.5

 3.5

 6.0

 -

 -

 -

 -

Mean (± SD)

 -

 1.6 ± 0.1

 3.3 ± 0.8

 6.2 ± 1.5

 5.5 ± 1.5

94.6

 0.063 ± 0.079

 63.6

 

Growth rate was significantly reduced between 50.1 and 111.2 mg/L. The calcuated ErL50 was 79.7 mg/L.

Table 2. Results of analytical monitoring (mg/L).

 Nominal conc. (mg/L)

 Sample

Replicate #

 0h (initial)

24 h

 24 h % of initial

 72 h

 72 h % of initial

 Avergae of all measurements

 7.7

 with algae

 1

 4.38

 5.4

 123

 3.7

 85

 

 

 

 2

 

 5.5

 126

 3.7

 85

 

 

 

 3

 

 6.0

 136

 5.0

 115

 

 

 

 Mean

 

 5.6

 128

 4.1

 95

 4.72

 

 without algae

 4

 5.34

 5.5

 102

 5.2

 97

 

 

 

 5

 

 6.00

 111

 5.7

 106

 

 

 

 mean

 

 5.7

 107

 5.4

 102

 5.48

 17

 with algae*

 1

 13.6

 11.9

 88

 7.9

 58

 

 

 

 2

 

 12.9

 95

 8.7

 64

 

 

 

3

 

 13.6

 100

 7.53

 56

 

 

 

 mean

 

 12.8

 94

 7.53

 56

 11.3

 

 without algae

 4

 13.6

13.3 

 98

 8.0

 59

 

 

 

 5

 

 16.8

 124

 8.6

 64

 

 

 

 mean

 

 15.0

 120

 24.1

 61

 12.3

 37

 with algae**

1

 32.5

 39.0

 120

 24.1

 74

 

 

 

 2

 

 24.4

 75

 30.1

 94

 

 

 

 3

 

 40.4

 124

 26.5

 82

 

 

 

 mean

 

34.6

 107

 26.5

 81.6

 31.2

 

 without algae

 4

 32.5

 26.3

 81

 25.7

 79.2

 

 

 

 5

 

 39.4

 121

 19.4

 60

 

 

 

 mean

 

 32.8

 101

 22.5

 69

 29.3

 82

with algae

 1

 54.7

 51.0

 92

 28.7

 26

 

 

 

 2

 

62.2

 113

 31.5

 58

 

 

 

 3

 

 71.1

 130

 34.2

 63

 

 

 

 mean

 

 61.3

 112

 34.2

 63

 50.1

 

 without algae

 4

 52.5

 78.8

 150

 42.9

 82

 

 

 

 5

 

 64.6

 123

 38.4

 73

 

 

 

 mean

 

 71.7

 137

 40.7

 77

 54.9

 180

 with algae

 1

 112.9

 120.9

 107

 104.3

 92

 

 

 

 2

 

 86.5

 77

 113.6

 101

 

 

 

 3

 

 84.9

 75

 152.2

 135

 

 

 

 mean

 

 97.4

 86

 123.4

 109

 111.2

 

 without algae

 4

 120.0

 70.6

 59

 121.9

 102

 

 

 

 5

 

 75.6

 63

 124.7

 104

 

 

 

 mean

 

 73.1

 61

 123.3

 103

 105.5

* 0 h measured: 32.0 mg/L; regarded as outlier, omitted from evaluation.

** 0 h measured : 13.5 mg/L; regarded as outlier, omitted from evaluation.

Above are results if the analytical monitoring of the test substance (1 -hexanol) over the experimental duration in all test concentrations with and without algae. The mean measured test concentrations varied in some cases by more than +/20% of the nominal values. 

The results are therefore interpreted with respect to the mean measured values. The values in bold are the values effectively used. All values at time 0 h were taken from the test solution prior to the addition of algae. Samples at time 48h and 72h were taken from the exposure vessels.

Validity criteria fulfilled:
yes
Conclusions:
A 72h EC50 value of 79.7 mg/L and NOEC value of 11.3 mg/L have been determined for the effect of the test substance on growth rate of Pseudokirchnerella subcapitata.
Executive summary:

The reliable data were obtained in a test with hexanol . A 72-h ErC50value of 80 mg/L and a NOEC of 11 mg/L were reported for hexanol based on measured exposure concentrations. These values give an acute to chronic ratio of 7.3.

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
(Q)SAR
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification
Justification for type of information:
QSAR prediction: US EPA accepted QSAR method for chemicals properties assessment.
Qualifier:
according to guideline
Guideline:
other: QSAR model
Principles of method if other than guideline:
ECOSAR Program(v1.00)
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
other: Green Algae
Details on test organisms:
QSAR model-Green Algae
Test type:
other: QSAR model
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
not applicable. QSAR model
pH:
not applicable. QSAR model
Dissolved oxygen:
not applicable. QSAR model
Salinity:
not applicable. QSAR model
Nominal and measured concentrations:
not applicable. QSAR model
Details on test conditions:
CAS Number:80206-82-2
SMILES : CCCCCCCCCCCCO
CHEM : Alcohols, C12-14
MOL FOR: C12 H26 O1
MOL WT : 186.34
Log Kow: 4.77 (KowWin estimate)
Wat Sol: 4 mg/L (experimental database)

Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
0.704 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: (Q)SAR prediction
Duration:
30 d
Dose descriptor:
NOEC
Effect conc.:
0.427 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: (Q)SAR prediction
Details on results:
A 96 h LC50 value of 0.704 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.
The data show the multi-component substances containing alcohols with carbon numbers in the ranges C12-16 to be acutely toxic at concentrations of between 0.03-0.1 mg/L. At these concentrations all the components are likely to have been fully dissolved.
This result signifies that the test substance is expected to be toxic.

A NOEC value of 0.427 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.
This result signifies that the test substance is expected to be toxic.

Overview of QSARs estimations onaquatic toxicity of Alcohols, C12-14 to Green algae

 

Method

 

Results

Remarks

Reference

QSAR

estimate

Green Algae, 96-hr EC50 =0.704 mg/L (ppm)

Green Algae, 30-day, NOEC cited as a ChV value = 0.427 mg/L (ppm)

 

Reliable with restrictions,

QSAR estimated data

ECOSAR Program (v1.00)

 

Green Algae, 96-hr EC50 =0.704 mg/L (ppm)

Green Algae, 30-day, NOEC cited as a ChV value = 0.427 mg/L (ppm)

Validity criteria fulfilled:
yes
Remarks:
method for chemicals properties assessment.
Conclusions:
A 96 h LC50 value of 0.704 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.
The data show the multi-component substances containing alcohols with carbon numbers in the ranges C12-16 to be acutely toxic at concentrations of between 0.03-0.1 mg/L. At these concentrations all the components are likely to have been fully dissolved.
This result signifies that the test substance is expected to be toxic.

A NOEC value of 0.427 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.
This result signifies that the test substance is expected to be toxic.




Executive summary:

A 96 h LC50 value of 0.704 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.

The data show the multi-component substances containing alcohols with carbon numbers in the ranges C12-16 to be acutely toxic at concentrations of between 0.03-0.1 mg/L. At these concentrations all the components are likely to have been fully dissolved.

This result signifies that the test substance is expected to be toxic.

A NOEC value of 0.427 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.

This result signifies that the test substance is expected to be toxic.

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Principles of method if other than guideline:
The acute toxicity of LOROL SPEZIAL to R. subcapitata was determined in sealed 72 h growth inhibition toxicity tests.
Algal inocula were exposed to test solutions which were not renewed during the test. The tests were repeated three times.
The test material was directly added to the WAF preparation vessels using ethanol as a carrier (0.1 mL/L).
The vessels were then stirred at approximately 100 rpm for a period of approximately 24 hours, before running off the WAFs for use as the test media.
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
TEST SYSTEM
Loading rates: 0.003, 0.01, 0.03, 0.1, 0.3, and 1 mg/l
Renewal of test solution: None
Exposure vessel type: 287 ml Erlenmeyer flasks
Number of replicates: 3
Initial cell concentration: 5000 cells/ml

Vehicle:
yes
Details on test solutions:
STOCK AND TEST SOLUTION AND THEIR PREPARATION
Test medium: Water accommodated fractions
Vehicle, solvent: Ethanol
Concentration of vehicle, solvent: 0.1 ml/l
STABILITY OF TEST CHEMICAL SOLUTIONS:
Considered stable
DILUTION WATER
Source: Not reported
Aeration: Not reported
Alkalinity: Not reported
Hardness: Not reported
Conductance: Not reported
Test organisms (species):
Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISMS
Strain: Raphidocelis subcapitata
Supplier: Institute of Freshwater Ecology, Windermere
Pretreatment: Not reported
Controls: 2 sets of controls (6 replicates and 1 blank flask), one set containing ethanol at 0.1 ml/L
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Test temperature:
Test temperature: 21.8-24.2
pH:
pH mean: 7.3-9.4
Dissolved oxygen:
Not reported
Nominal and measured concentrations:
Loading rates: 0.003, 0.01, 0.03, 0.1, 0.3, and 1 mg/l
Reference substance (positive control):
not specified
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
0.1 - 0.3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
0.03 - 0.3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
other: NOEL
Effect conc.:
0.003 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
Result:
RESULTS: EXPOSED
EbL50 = 0.03 - 0.3 mg/l
ErL50 = 0.1 - 0.3 mg/l
Based on loading rates
For LOROL SPEZIAL, the ranges for the EbL50 and ErL50 were 0.1 to 0.3 mg/L respectively with one exception; the EbL50 in Test 3 was in the range 0.03 to 0.1 mg/L.
Validity criteria fulfilled:
yes
Conclusions:
A 72h EL50 value of 0.1-0.3 mg/L and NOEL value of 0.003 mg/L have been determined for the effect of the test substance on growth rate of Raphidocelis subcapitata
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Qualifier:
according to guideline
Guideline:
other: EU Commission Directive 92/69/EEC of July 31, 1992
Principles of method if other than guideline:
EU Commission Directive 92/69/EEC of July 31, 1992
GLP compliance:
yes
Analytical monitoring:
not specified
Vehicle:
not specified
Details on test solutions:
STOCK AND TEST SOLUTION AND THEIR PREPARATION
Test medium: Water accommodated fractions
STABILITY OF TEST CHEMICAL SOLUTIONS:
Considered stable
DILUTION WATER
Source: Not reported
Aeration: Not reported
Alkalinity: Not reported
Hardness: Not reported
Conductance: Not reported
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISMS
Species: Scenedesmus subspicatus (Algae)
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Test temperature:
Test temperature: 21.8-25
pH:
pH mean: 7.3-9.4
Dissolved oxygen:
Not reported
Nominal and measured concentrations:
Loading rates: 0.3, 1.2, and 1.9 mg/l
Reference substance (positive control):
not specified
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
1.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to biomass growth
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
1.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to growth rate
Duration:
72 h
Dose descriptor:
EL0
Effect conc.:
0.3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to biomass growth
Details on results:
Result:
EbL0 : = 0.3
EbL50 : = 1.2
ErL50 : = 1.9
The reported values represent the ErC50 (related to growth rate) and the EbC50 (related to biomass growth)
Validity criteria fulfilled:
yes
Conclusions:
A 72h EL50 value of 1.2-1.9 mg/L have been determined for the effect of the test substance on growth rate and biomass growth of Scenedesmus subspicatus (Algae)
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
WAFs were prepared by loading test medium with the respective amount of the test item. After stirring the contents were left to settle for 24h. The WAFs were filtered prior to use through a non polar filter (0.2 um Millex FG, 50 mm, Millipore).
Vehicle:
not specified
Details on test solutions:
STOCK AND TEST SOLUTION AND THEIR PREPARATION
Test medium: Water accommodated fractions
STABILITY OF TEST CHEMICAL SOLUTIONS:
Considered stable
DILUTION WATER
Source: Not reported
Aeration: Not reported
Alkalinity: Not reported
Hardness: Not reported
Conductance: Not reported
Test organisms (species):
Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISMS
Species: Pseudokirchneriella subcapitata
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
72 h
Test temperature:
Test temperature: 21.8-25
pH:
pH mean: 7.3-9.4
Dissolved oxygen:
Not reported
Nominal and measured concentrations:
Loading rates: 0.6, 0.8,1.2,1.4,2.4 and 2.9 mg/l
Reference substance (positive control):
not specified
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
1.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to Growth rate
Duration:
72 h
Dose descriptor:
LOELR
Effect conc.:
2.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to growth rate
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
0.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to Growth rate
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
2.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: related to Growth rate
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
0.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: related to biomass
Duration:
72 h
Dose descriptor:
LOELR
Effect conc.:
1.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: related to biomass
Duration:
72 h
Dose descriptor:
EL10
Effect conc.:
0.8 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: related to biomass
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
1.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: related to biomass
Details on results:
Result:
RESULTS: EXPOSED
Growth rate
NOEL = 1.2 mg/l
LOEL = 2.4 mg/l
ErL10 = 0.6 mg/l
ErL50 = 2.9 mg/l
Biomass
NOEL = 0.6 mg/l
LOEL = 1.2 mg/l
EbL10 = 0.8 mg/l
EbL50 = 1.4 mg/l
Based on loading rates

Validity criteria fulfilled:
yes
Conclusions:
A 72h NOEL value of 0.6-1.2 mg/L have been determined for the effect of the test substance on growth rate and biomass growth of Pseudokirchneriella subcapitata
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
other: published data
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: German Industry Standard: DIN 38412, Part 9
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Deviations:
yes
Remarks:
no analytical monitoring, no range finding test, no reporting of water quality parameters.
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: unfiltered test solutions achieved nominal concentrations by direct addition of the test material into the test vessels. The filtered solutions were made from serial dilutions of a 1 g/L filtered stock solution.
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM

- Strain: SAG 8681

- Source (laboratory, culture collection): Institute for plant physiology, University Gottingen

- Age of inoculum (at test initiation): 3 days

- Method of cultivation: not reported


ACCLIMATION - not reported
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
range: 22-23.5 degC
Nominal and measured concentrations:
Nominal Loading Rate: 10, 30, 100 and 300 mg/L (Unfiltered)
Nominal Loading Rate: 0.1, 0.3, 1.0, 3.0, 10.0 mg/L (Filtrate)
Details on test conditions:
TEST SYSTEM

- Test vessel: Erlenmeyer flasks

- Material, size, headspace, fill volume: 300 ml with 100 ml fill volume

- Initial cells density: 1*10^4 cells/ml

- Control end cells density: 2.7*10^6

- No. of vessels per concentration (replicates): 3

- No. of vessels per control (replicates): 3



GROWTH MEDIUM

- Standard medium used: the medium was defined as Algae 10.10 / TFB Central laboratory soil nutrients.


TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: not reported

- Culture medium different from test medium: not reported

- Intervals of water quality measurement: not reported


OTHER TEST CONDITIONS

- Adjustment of pH: not reported

- Photoperiod: continuous

- Light intensity and quality: 2000 lux


EFFECT PARAMETERS MEASURED

- Determination of cell concentrations: cell numbers were determined at 24, 48, 72 and 96 h via the Thoms-Kammer method


TEST CONCENTRATIONS

- Spacing factor for test concentrations: 3
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: non-toxic at the limit of solubility
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
> 21 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
96 h
Dose descriptor:
EC0
Effect conc.:
< 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: due to variability in cell density, the EL0 could not be reliably determined.
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
250 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: non-toxic at the limit of solubility
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
26 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Duration:
96 h
Dose descriptor:
EC0
Effect conc.:
< 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: due to variability in cell density, the EL0 could not be reliably determined.
Duration:
96 h
Dose descriptor:
EC0
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution with test material
Basis for effect:
growth rate
Remarks on result:
other: non-toxic at the limit of solubility
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution with test material
Basis for effect:
growth rate
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
0.64 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution
Basis for effect:
growth rate
Duration:
96 h
Dose descriptor:
EC0
Effect conc.:
<= 0.1 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution with test material
Basis for effect:
biomass
Remarks on result:
other: non-toxic at the limit of solubility
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
0.1 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution with test material
Basis for effect:
biomass
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
> 10 mg/L
Nominal / measured:
nominal
Conc. based on:
other: filtered test solution with test material
Basis for effect:
biomass
Details on results:
- Exponential growth in the control (for algal test): yes

- Effect concentrations exceeding solubility of substance in test medium: the test was conducted at loading rates that were all much higher than the limit of solubility of the test substance (ca. 0.0011 mg/L).
Reported statistics and error estimates:
The analysis of the data was carried out in accordance with DIN regulation with a computer program by Mr. Holtmann, TFA / mathematical methods, Henkel KGaA.

Table 1. Summary of findings (average of 3 replicates).

  Nominal Loading Rates (mg/L) 

   Cell concentrations (cells/ml)           

filtered 

unfiltered 

 24 h

 48 h

 72 h

 96 h

 0

 -

 8.3*10^4

 5.1*10^5

 1.2*10^6

 2.7*10^6

 10

 -

 1.2*10^5

 5.2*10^5

 1.1*10^6

 2.1*10^6

 30

 -

 8.3*10^4

 5.6*10^5

 1.1*1 0^6

 2.2*10^6

 100

 -

 7.3*10^4

 5.1*10^5

 9.3*10^5

 1.9*10^6

 300

 -

 2.3*10^4

 6.7*10^4

 6.2*10^5

 1.1*10^6

 -

 0

 6.0*10^4

 3.5*10^5

 1.4*10^6

 2.6*10^6

 -

 0.1

 5.3*10^4

 3.3*10^5

 1.2*10^6

 2.4*10^6

-

 0.3

 7.7*10^4

 3.1*10^5

 1.4*10^6

 2.7*10^6

 -

 1.0

 5.0*10^4

 3.3*10^5

 1.2*10^6

 2.4*10^6

 -

 3.0

 8.0*10^4

 3.8*10^5

 1.4*10^6

 2.4*10^6

 -

 10

 7.3*10^4

 3.8*10^5

 1.5*1 0^6

 2.2*10^6

EL values are said not be accurately identified in many cases since there was large variation in the cell concentrations measured (this is not clear from the table of results reported, identical to Table 1 here). All of the concentrations tested in the unfiltered and filtered solutions were above the limit of solubility. Any effect seen at these concentrations should be taken with caution as they may represent effects deriving from physical fouling rather than true toxicity of the test substance. The inconsistent EC values and cell concentrations may well be explained by the fact that the concentrations tested are much higher than the limit of solubility.

 

Validity criteria fulfilled:
not specified
Remarks:
the validity criteria for cell growth in the control was achieved, but there is no data on pH.
Conclusions:
The 96 h EC values could not be accurately determined due to large variations in cell concentrations. The EC50 values determined are reliant on confounding factors due to the concentrations used (exceedinly higher than the limit of solubility) and therefore should be treated with caution. The water solubility of octadecanol is 0.0011 mg/L at 25oC . The results indicate that no toxic effects were present at the limit of solubility of the test substance, i.e. EC50 and NOEC >0.0011 mg/L (>LoS).
Executive summary:

EC10 = 26 mg/l              

EC50 = 250 mg/l

The water solubility of octadecanol is 0.0011 mg/L at 25oC . The results indicate that no toxic effects were present at the limit of solubility of the test substance, i.e. EC50 and NOEC >0.0011 mg/L (>LoS).

Description of key information

A 96 h LC50 value of 0.704 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.
The data show the multi-component substances containing alcohols with carbon numbers in the ranges C12-16 to be acutely toxic at concentrations of between 0.03-0.1 mg/L. At these concentrations all the components are likely to have been fully dissolved. 
This result signifies that the test substance is expected to be toxic. 
  
A NOEC value of 0.427 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group. 
This result signifies that the test substance is expected to be toxic. 
The available data for saltwater algae and cyanobacteria are limited and to obtaine the value for PNECs for saltwaters is taken account a assesment factor 10 to extrapolate to LC50 following the “Guidance on Information Requirement and Chemical Safety Assessment R.10: Characterisation of dose [concentration]- response for environment”and “Table R.10-5 Assessment factors proposed for deriving PNEC water for saltwater for different data sets”, to account for the possibility that greater taxonomic diversity in the marine environment makes it likely that some marine taxa may be more sensitive than the most sensitive freshwater taxa

Key value for chemical safety assessment

EC50 for freshwater algae:
0.704 mg/L
EC50 for marine water algae:
0.07 mg/L
EC10 or NOEC for freshwater algae:
0.427 mg/L
EC10 or NOEC for marine water algae:
0.043 mg/L

Additional information

A 96 h LC50 value of 0.704 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.

The data show the multi-component substances containing alcohols with carbon numbers in the ranges C12-16 to be acutely toxic at concentrations of between 0.03-0.1 mg/L. At these concentrations all the components are likely to have been fully dissolved.

This result signifies that the test substance is expected to be toxic.

 

A NOEC value of 0.427 mg/L has been calculated for the effects of the test substance on the mortality of the Algae group.

This result signifies that the test substance is expected to be toxic.

The available data for saltwater algae and cyanobacteria are limited and to obtaine the value for PNECs for saltwaters is taken account a assesment factor 10 to extrapolate to LC50 following the “Guidance on Information Requirement and Chemical Safety Assessment R.10: Characterisation of dose [concentration]- response for environment”and “Table R.10-5 Assessment factors proposed for deriving PNEC water for saltwater for different data sets”, to account for the possibility that greater taxonomic diversity in the marine environment makes it likely that some marine taxa may be more sensitive than the most sensitive freshwater taxa