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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:
20 Aug. - 08 Oct. 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
Samples for TOC-analysis were taken at 0 h and 72 h: TOC-Analyser (DIMA-TOC 100).
Measurements of the cell concentrations per mL: at time 24 h, 48 h and 72 h by optical densitometry at 525 nm
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) prepared by stirring various amounts of the test item in Alga-medium
for 48 h with magnetic stirrers (open conditions). The WAFs were prepared in 2-L glass beakers.
The beakers were equipped with a glass tube that allowed the separation of the water phase by siphoning.
The test loading rate of Novares LA 100 was weighed in a weighing scoop, that afterwards was placed into the glass beaker.
The beakers were then filled with Alga-medium.
Mixing was carried out at a speed that was slow enough not to cause dispersion or emulsification of the undissolved fraction
of the test item. To ensure this, the vortex developed at the surface by stirring was set at ~ 10 % of the water depth.
After stirring for 48 h the WAFs were allowed to stand for 1 h before use to facilitate phase separation.
The extracts gained with this method were clear.

- Pre-conditioning: The test vessels of the Definite Alga-Test were rinsed with control media and WAFs for ca. 0.5 h before the definite
start of the tests to facilitate adsorption of the test item to the glass walls of the beakers.
Control media and WAFs were then discarded, and the test vessels were refilled again with control media and WAFs, respectively.
- Controls: without test material (blank; Alga medium)
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM
- Common name: unicellular green alga
- Strain: SAG 86.81
- Source: Albrecht von Haller Institut für Pflanzenwissenschaften der Universität Göttingen

- Age of inoculum (at test initiation): ca. 72 h
[Pre-culture: Incubation in 250 mL Erlenmeyer flasks at ± 25 °C on a shaker
(80 - 120 rpm), ca. 72 h, ca. 10000 lux (Gro-Lux fluorescent tubes), permanent illumination]

- Method of cultivation: Sterile on agar tubes (OECD-Medium, 0.8 % Agar), transferred to a new agar tube every 2 months


Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h
Post exposure observation period:
none
Hardness:
no data specified: Compositions of stock solutions given.
Test temperature:
23.1 - 24.4 °C (25 +-2°C)
pH:
initial (with algae): 8.0 - 8.1
final (with algae): 10.2 (control); 9.4 (5 mg/L); 9.0 (10 mg/L); 8.7 (25 mg/L); 8.5 (50 mg/L); 8.4 (100 mg/L)
Salinity:
no data specified: Compositions of stock solutions given.
Nominal and measured concentrations:
Pre-test:
Time [h] Control 100 mg/L loading
==========================================
24 (stirring) 0.1 3.1 mg C/mL (TOC of WAFs)
48 (stirring) 1.0 4.5 mg C/mL (TOC of WAFs)
==========================================

Main test series:
Time [h] Control 5 mg/L 10 mg/L 25 mg/L 50 mg/L 100 mg/L (loading)
==================================================================
0 7.6 1.8 n.d. 3.4 n.d. 4.7 mg C/L (TOC of WAF)
72 1.2 1.6 n.d 2.7 n.d. 4.4 mg C/L (TOC of WAF)
===================================================================
Note: Control value of time 0 is considered to be an artefact. TOC values include background values.
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL-Erlenmeyer
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 100 mL
- Aeration: no
- Initial cells density: approx. 5000 cells/mL
- Control end cells density: approx. 278800 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


GROWTH MEDIUM
- Standard medium used: yes


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted from deionised water and mineral stock solutions


OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: permanent
- Light intensity and quality: approx. 10000 Lux (Gro-Lux fluorescent tubes)


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: densitometrically at 525 nm (turbidity)
- Chlorophyll measurement: no


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0 - 2.5
- Range finding study
- Results used to determine the conditions for the definitive study: based on results from daphnia test
Key result
Duration:
72 h
Dose descriptor:
EL10
Remarks:
effective loading
Effect conc.:
ca. 6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
growth rate
Key result
Duration:
72 h
Dose descriptor:
EL50
Remarks:
effective loading
Effect conc.:
15 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
growth rate
Duration:
72 h
Dose descriptor:
EL10
Remarks:
effective loading
Effect conc.:
>= 3 - <= 4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
EL50
Remarks:
effective loading
Effect conc.:
8 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
< 5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
other: biomass and growth rate
Duration:
72 h
Dose descriptor:
LOELR
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 100
Basis for effect:
other: biomass and growth rate
Details on results:
- Exponential growth in the control (for algal test): yes
- Any stimulation of growth found in any treatment: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no

- Observation of abnormalities (for algal test):
Observations by microscopy at the end of the testing period of 72 h:

Control: Appearance of alga cells was normal, cell proliferations could be observed regularly.
5 mg/L: Comparable to control.
10 mg/L: More medium-sized and large alga cells compared to the control, cell proliferations could be observed regularly
but lower number of cell proliferations.
25 mg/L: Appearance of alga cells comparable to 10 mg/L but cell proliferations could not be observed.
50 mg/L: Very low number of alga cells, cells partly deformed, cell proliferations could not be observed.
100 mg/L: Comparable to 50 mg/L.
Reported statistics and error estimates:
standard deviations, coefficient of variance

Growth rates are listed below, from 0 -24, 24 - 48, and 48 - 72 h (Table 1) and average from 0 - 72 h (Table 2):

Table 1: Loading rates of the Test item, cells/mL in the test flasks at 24, 48 and 72 h with mean value,

section-by-section average specific growth rate [μ] during observation times 0-24 h, 24-48 h

and 48-72 h, mean value of the average specific growth rate [μ], standard deviation and

coefficient of variation Vc [%], inhibition [%] and mean value of inhibition of the average

specific growth rate [%] during observation times 0-24 h, 24-48 h and 48-72 h (Rep. Tab. 4)

[Note: Negative inhibition means increased growth, apparently stimulation]

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

24 h

Control 1

18746

19836

1.32

1.38

 

0.09

 

6.75

 

Control 2

22015

1.48

Control 3

18746

1.32

5

6083

7118

0.20

 

0.33

 

 

0.24

 

 

71.35

 

85.74

 

75.74

 

5

6083

0.20

85.74

5

9187

0.61

55.76

10

6083

7118

0.20

 

0.33

 

 

0.24

 

 

71.35

 

85.74

 

75.74

 

10

9187

0.61

55.76

10

6083

0.20

85.74

25

6083

4042

0.20

 

-0.27

 

 

0.40

 

 

149.40

 

85.74

 

119.67

 

25

3021

-0.50

136.64

25

3021

-0.50

136.64

50

3021

4042

-0.50

 

-0.27

 

 

0.40

 

 

149.40

 

136.64

 

119.67

 

50

6083

0.20

85.74

50

3021

-0.50

136.64

100

3021

3021

-0.50

-0.50 

136.64

136.64 

100

3021

-0.50

136.64

100

3021

-0.50

136.64

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

48 h

Control 1

46053

47253

0.90

0.87

 

0.12

 

 

13.85

 

 

 

 

Control 2

46053

0.74

Control 3

49653

0.97

5

46053

43681

2.02

 

1.83

 

 

0.26

 

 

14.42

 

-132.58

 

-110.64

 

5

42495

1.94

-123.34

5

42495

1.53

-75.98

10

42495

44867

1.94

 

1.86

 

 

0.22

 

 

11.75

 

-123.34

 

-113.71

 

10

46053

1.61

-85.22

10

46053

2.02

-132.58

25

32069

34359

1.66

 

2.20

 

 

0.46

 

 

21.07

 

-91.00

 

-152.41

 

25

35503

2.46

-183.11

25

35503

2.46

-183.11

50

15518

16594

1.64

 

1.47

 

 

0.47

 

 

31.94

 

-88.02

 

-68.45

 

50

15518

0.94

-7.60

50

18746

1.83

-109.73

100

18746

13422

1.83

1.45 

0.36 

24.75 

-109.73

-66.38 

100

9187

1.11

-27.79

100

12332

1.41

-61.62

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

72 h

Control 1

305342

278840

1.89

 

1.77

 

 

0.14

 

 

8.14

 

 

 

 

 

 

 

Control 2

282272

1.81

Control 3

248906

1.61

5

206731

229445

1.50

 

1.66

 

 

0.13

 

 

8.14

 

15.27

 

6.51

 

5

243490

1.75

1.50

5

238115

1.72

2.76

10

134777

98229

1.15

 

0.75

 

 

0.35

 

 

46.84

 

34.87

 

57.58

 

10

76004

0.50

71.73

10

83905

0.60

66.15

25

9187

8153

-1.25

 

-1.46

 

 

0.27

 

 

18.70

 

170.54

 

182.12

 

25

9187

-1.35

176.28

25

6083

-1.76

199.54

50

6083

4042

-0.94

 

-1.47

 

 

0.47

 

 

31.94

 

152.84

 

182.72

 

50

3021

-1.64

192.34

50

3021

-1.83

203.00

100

3021

3021

-1.83

-1.45 

0.36 

24.75 

203.00

181.71 

100

3021

-1.11

162.76

100

3021

-1.41

179.37

Table 2: Loading rate of the Test item, cells/mL in the test flasks at 72 h, average specific growth

rate [μ] over the whole testing period 0-72 h, mean value of the average specific growth

rate [μ], standard deviation and coefficient of variation Vc [%], inhibition [%] and mean value

of inhibition [%] of the average specific growth rate [μ] (Rep. Tab. 5)

Loading rate

[mg/L]

Cells / mL

Average

Specific

growth rate

[µ]

Average

Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation

Vc [%]

Inhibition of Average

Specific

growth rate

[%]

Inhibition of Average

Specific

growth rate

Mean value

[%]

72 h

Control 1

305342

1.37

1.34

0.03

2.57

Control 2

282272

1.34

Control 3

248906

1.30

5

206731

1.24

 

1.27

 

 

0.03

 

 

2.32

 

7.36

 

4.83

 

5

243490

1.30

3.29

5

238115

1.29

3.84

10

134777

1.10

 

0.98

 

 

0.10

 

 

10.40

 

18.01

 

26.69

 

10

76004

0.91

32.27

10

83905

0.94

29.80

25

9187

0.20

 

0.16

 

 

0.08

 

 

50.54

 

84.86

 

88.28

 

25

9187

0.20

84.86

25

6083

0.07

95.12

50

6083

0.07

 

-0.09

 

 

0.13

 

 

149.40

 

95.12

 

106.73

 

50

3021

-0.17

112.54

50

3021

-0.17

112.54

100

3021

-0.17

-0.17 

112.54

112.4 

100

3021

-0.17

112.54

100

3021

-0.17

112.54

Validity criteria fulfilled:
yes
Remarks:
1) Biomass in controls increased 56x in 72 h (nom. >= 16x). 2) Mean var-coeff. for section-spec. growth rates: 33.9 % (nom. ≤ 35%). 3) Var-coeff. of av. spec. growth rates for the whole period in replicate controls: 2.6% (nom. value ≤ 7%).
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 Sep. - 09 Oct. 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Name of test material (as cited in study report): Novares LA 700, (phenol, methylstyrenated)
- Lot/batch No.: 23861
- Composition of test material: composition is specified in IUCLID Sect. 13 - Assessment reports under Certificate of Analysis_Novares LA 700_phenol, methylstyrenated
- Stability under test conditions: no measured data; based on chemical structure assumed to be stable
- Storage condition of test material: room temperature, exclusion of light
Analytical monitoring:
yes
Details on sampling:
Samples for TOC-analysis were taken at 0 h and 72 h: TOC-Analyser (DIMA-TOC 100).
Measurements of the cell concentrations per mL: at time 24 h, 48 h and 72 h by optical densitometry at 525 nm
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fractions (WAFs) prepared by stirring various amounts of the test item in Alga-medium
for 24 h with magnetic stirrers (open conditions). The WAFs were prepared in 2-L glass beakers.
The beakers were equipped with a glass tube that allowed the separation of the water phase by siphoning.
The test loading rate of Novares LA 700 was weighed in a weighing scoop, that afterwards was placed into the glass beaker.
The beakers were then filled with Alga-medium.
Mixing was carried out at a speed that was slow enough not to cause dispersion or emulsification of the undissolved fraction
of the test item. To ensure this, the vortex developed at the surface by stirring was set at ~ 10 % of the water depth.
After stirring for 24 h the WAFs were allowed to stand for 1 h before use to facilitate phase separation.
The extracts gained with this method were clear.

- Pre-conditioning: The test vessels of the Definite Alga-Test were rinsed with control media and WAFs for ca. 0.5 h before the definite
start of the tests to facilitate adsorption of the test item to the glass walls of the beakers.
Control media and WAFs were then discarded, and the test vessels were refilled again with control media and WAFs, respectively.
- Controls: without test material (blank; Alga medium)
Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM
- Common name: unicellular green alga
- Strain: SAG 86.81
- Source: Albrecht von Haller Institut für Pflanzenwissenschaften der Universität Göttingen

- Age of inoculum (at test initiation): ca. 72 h
[Pre-culture: Incubation in 250 mL Erlenmeyer flasks at ± 25 °C on a shaker
(80 - 120 rpm), ca. 72 h, ca. 10000 lux (Gro-Lux fluorescent tubes), permanent illumination]

- Method of cultivation: Sterile on agar tubes (OECD-Medium, 0.8 % Agar), transferred to a new agar tube every 2 months


Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h
Post exposure observation period:
none
Hardness:
no data specified: Compositions of stock solutions given.
Test temperature:
23.8 - 24.3 °C (25 +-2°C)
pH:
initial (with algae): 7.8 - 8.2
final (with algae): 10.1 (control); 10.3 (5 mg/L); 10.3 (10 mg/L); 10.4 (25 mg/L); 10.1 (50 mg/L); 10.3 (100 mg/L); 9.7 (250 mg/L)
Salinity:
no data specified: Compositions of stock solutions given.
Nominal and measured concentrations:
Time [h] Control 5 mg/L 10 mg/L 25 mg/L 50 mg/L 100 mg/L 250 mg/L (loading)
============================================================================
0 1.7 2.2 n.d. n.d. 2.8 n.d. 3.6 mg C/L (TOC of WAF)
72 1.8 2.2 n.d n.d. 2.7 n.d. 3.5 mg C/L (TOC of WAF)
===================================================================
Note: TOC values include background values. / n.d. = not determined
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL-Erlenmeyer
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 100 mL
- Aeration: no
- Initial cells density: approx. 5000 cells/mL
- Control end cells density: approx. 740,000 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


GROWTH MEDIUM
- Standard medium used: yes


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted from deionised water and mineral stock solutions


OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: permanent
- Light intensity and quality: approx. 10000 Lux (Gro-Lux fluorescent tubes)


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: densitometrically at 525 nm (turbidity)
- Chlorophyll measurement: no


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0 - 2.5
- Range finding study
- Results used to determine the conditions for the definitive study: based on results from daphnia test
Key result
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
> 250 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 700
Basis for effect:
growth rate
Key result
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
25 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 700
Basis for effect:
other: growth rate and biomass
Duration:
72 h
Dose descriptor:
EL50
Effect conc.:
178 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 700
Basis for effect:
biomass
Duration:
72 h
Dose descriptor:
LOELR
Effect conc.:
50 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
Novares LA 700
Basis for effect:
other: biomass
Details on results:
- Exponential growth in the control (for algal test): yes
- Any stimulation of growth found in any treatment: yes, at 5 mg/L
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no

- Observation of abnormalities (for algal test):
Observations by microscopy at the end of the testing period of 72 h:

Control: Appearance of alga cells was normal, cell proliferations could be observed regularly.
5 mg/L: Comparable to control.
10 mg/L: Comparable to control.
25 mg/L: Comparable to control.
50 mg/L: More medium-sized and large alga cells compared to the control.
100 mg/L: Comparable to 50 mg/L.
250 mg/L: Appearance of alga cells comparable to 50 mg/L, but lower number of cells.

Reported statistics and error estimates:
standard deviations, coefficient of variance

Growth rates are listed below, from 0 - 24, 24 - 48, and 48 - 72 h (Table 1) and average from 0 - 72 h (Table 2):

Table 1: Loading rates of the Test item, cells/mL in the test flasks at 24, 48 and 72 h with mean value,

section-by-section average specific growth rate [μ] during observation times 0-24 h, 24-48 h

and 48-72 h, mean value of the average specific growth rate [μ], standard deviation and

coefficient of variation Vc [%], inhibition [%] and mean value of inhibition of the average

specific growth rate [%] during observation times 0-24 h, 24-48 h and 48-72 h (Rep. Tab. 4)

[Note: Negative inhibition means increased growth, apparently stimulation]

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

24 h

Control 1

25325

32124

1.62

1.84

 

0.22

 

11.71

 

Control 2

38979

2.05

Control 3

32069

1.86

5

38979

37820

2.05

 

2.02

 

0.05

 

 

2.67

 

-11.32

 

-9.63

 

5

35503

1.96

-6.25

5

38979

2.05

-11.32

10

35503

32083

1.96

 

1.86

 

0.11

 

 

5.76

 

-6.25

 

-0.56

 

10

28677

1.75

5.32

10

32069

1.86

-0.74

25

32069

34359

1.86

 

1.93

 

0.06

 

 

3.05

 

-0.74

 

-4.42

 

25

35503

1.96

-6.25

25

35503

1.96

-6.25

50

32069

32083

1.86

 

1.86

 

0.11

 

 

5.76

 

-0.74

 

-0.56

 

50

35503

1.96

-6.25

50

28677

1.75

5.32

100

28677

28690

1.75

 

1.74

 

0.12

 

 

6.78

 

5.32

 

5.55

 

100

32069

1.86

-0.74

100

25325

1.62

12.06

250

25325

24222

1.62

1.58 

0.08

5.13 

12.06

14.59 

250

25325

1.62

12.06

250

22015

1.48

19.65

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

48 h

Control 1

186635

219590

2.00

1.93

 

0.07

 

 

3.49

 

 

 

 

Control 2

265403

1.92

Control 3

206731

1.86

5

222237

227915

1.74

1.79

 

 

0.09

 

 

5.10

 

9.64

 

7.00

 

5

201645

1.74

9.84

5

259863

1.90

1.52

10

130311

147198

1.30

1.52

 

 

0.20

 

 

12.94

 

32.50

 

21.09

 

10

139285

1.58

17.96

10

171997

1.68

12.81

25

153057

148852

1.56

1.46

 

 

0.20

 

 

13.49

 

18.87

 

24.36

 

25

121502

1.23

36.13

25

171997

1.58

18.09

50

134777

141036

1.44

1.48

 

 

0.04

 

 

2.87

 

25.47

 

23.25

 

50

162444

1.52

21.06

50

125886

1.48

23.21

100

91970

108777

1.17

1.33

 

 

0.16

 

 

12.36

 

39.50

 

30.93

 

100

121502

1.33

30.85

100

112858

1.49

22.43

250

79934

77328

1.15

1.16

0.12 

10.50 

40.33

39.69 

250

72116

1.05

45.68

250

79934

1.29

33.06

Loading rate

[mg/L]

Cells / mL

Cells / mL

Mean value

Average Specific

growth rate

[µ]

Average Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation Vc [%]

Inhibition of Average Specific
growth rate

[%]

Inhibition of Average Specific

growth rate

Mean value

[%]

72 h

Control 1

689559

745202

1.31

 

1.23

 

 

0.17

 

 

13.90

 

 

 

 

 

 

 

Control 2

747421

1.04

Control 3

798628

1.35

5

697701

906541

1.14

 

1.36

 

 

0.36

 

 

26.27

 

7.08

 

-10.55

 

5

1188331

1.77

-44.06

5

833591

1.17

5.33

10

513688

658879

1.37

 

1.45

 

 

0.24

 

 

16.82

 

-11.41

 

-18.17

 

10

492359

1.26

-2.55

10

970591

1.73

-40.54

25

513688

740153

1.21

 

1.58

 

 

0.43

 

 

26.93

 

1.66

 

-28.62

 

25

942447

2.05

-66.38

25

764324

1.49

-21.14

50

789990

587766

1.77

 

1.40

 

 

0.32

 

 

22.69

 

-43.63

 

-13.83

 

50

542705

1.21

2.03

50

430603

1.23

0.12

100

550063

508325

1.79

 

1.53

 

 

0.30

 

 

19.93

 

-45.26

 

-24.08

 

100

602721

1.60

-30.07

100

372191

1.19

3.08

250

282272

288404

1.26

1.31 

0.07 

5.54 

-2.47

-6.67 

250

259863

1.28

-4.11

250

323078

1.40

-13.44

Table 2: Loading rate of the Test item, cells/mL in the test flasks at 72 h, average specific growth

rate [μ] over the whole testing period 0-72 h, mean value of the average specific growth

rate [μ], standard deviation and coefficient of variation Vc [%], inhibition [%] and mean value

of inhibition [%] of the average specific growth rate [μ] (Rep. Tab. 5)

Loading rate

[mg/L]

Cells / mL

Average

Specific

growth rate

[µ]

Average

Specific

growth rate

[µ]

Mean value

Standard deviation

Coefficient of variation

Vc [%]

Inhibition of Average

Specific

growth rate

[%]

Inhibition of Average

Specific

growth rate

Mean value

[%]

72 h

Control 1

689559

1.64

1.67

0.02

1.47

Control 2

747421

1.67

Control 3

798628

1.69

5

697701

1.65

 

1.73

 

 

0.09

 

 

5.24

 

1.28

 

-3.45

 

5

1188331

1.82

-9.36

5

833591

1.71

-2.28

10

513688

1.54

 

1.61

 

 

0.13

 

 

7.87

 

7.40

 

3.44

 

10

492359

1.53

8.25

10

970591

1.76

-5.32

25

513688

1.54

 

1.66

 

 

0.10

 

 

6.21

 

7.40

 

0.71

 

25

942447

1.75

-4.73

25

764324

1.68

-0.54

50

789990

1.69

 

1.58

 

 

0.10

 

 

6.47

 

-1.20

 

5.34

 

50

542705

1.56

6.30

50

430603

1.49

10.93

100

550063

1.57

 

1.53

 

 

0.09

 

 

5.56

 

6.03

 

8.03

 

100

602721

1.60

4.21

100

372191

1.44

13.84

250

282272

1.34

1.35 

0.04 

2.71 

19.37

19.02 

250

259863

1.32

21.02

250

323078

1.39

16.67

Validity criteria fulfilled:
yes
Remarks:
1) Biomass in controls increased 149x in 72 h (nom. >= 16x). 2) Mean var-coeff. for section-spec. growth rates: 23.9 % (nom. ≤ 35%). 3) Var-coeff. of av. spec. growth rates for the whole period in replicate controls: 1.5 % (nom. value ≤ 7%).

Description of key information

Due to substance properties, toxicity to aquatic algae and cyanobacteria is determined based on loadings. The lowest EL50 value of two tests using two technical products of 'Oligomerisation and alkylation reaction products of 2-phenylpropene and phenol' is selected to characterise the acute toxicity to algae and cyanobacteria.

Key value for chemical safety assessment

EC50 for freshwater algae:
15 mg/L
EC10 or NOEC for freshwater algae:
6 mg/L

Additional information

The substance 'Oligomerisation and alkylation reaction products of 2-phenylpropene and phenol' (OAPP) (previous name phenol, methylstyrenated) is a UVCB substance with a complex and within limits variable composition. Water solubility is low and not clearly defined due to the differing solubility of individual components. Depending on the quantity used, concentration of material dissolved in saturated water samples is variable. Therefore, water accommodated fractions (WAFs) have been used in aquatic toxicity tests. Thus, results are presented as loadings (EL50, EL10).

Toxicity to aquatic algae and cyanobacteria has been tested in valid GLP guideline studies (OECD TG 201) with two different technical products of OAPP (LA 100 and LA 700). The two products are composed of the same constituents and differ only in the quantitative composition of components. They are subsumed under the same EC Number (700-960-7).

The solubility of the two technical products is somewhat different. Therefore, it can be expected that aqueous effect levels differ to some degree. For reasons of comprehensive protection, the lowest EL50 / EL10 values (15 mg/L / ca. 6 mg/L, based on growth rate) (LA 100) are selected as key values for the chemical safety assessment.