Fatty acids C16-18 (even numbered), mono and di esters with Sucrose

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

22 - 25 Aug 2016

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no chemical analysis

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 100 mg of test item was dissolved in 1000 mL test medium to prepare a stock solution of 100 mg/L. The solution was stirred fro 48 h. Afterwards it was devided into each test vessel.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: ATCC 22662
- Source (laboratory, culture collection): American Type Culture Collection (supplied 30 Jun 1995)

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test temperature:

22.5 - 22.8 °C

pH:

7.9 - 8.0 (control)
7.7 - 8.3 (treatment)

Nominal and measured concentrations:

nominal: control, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Sterilized 300 mL Erlenmeyer flask (with gas-permeable stopper); fill volume: 100 mL
- Aeration: constantly shaken (approx. 100 rpm)
- Initial cells density: 0.75 E+04 cells/mL (equivalent to 2.8 Chlorophyll fluorescence values)
- Control end cells density: Cell density not given; 260 Chlorophyll fluorescence value
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified water according to guideline
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continuous illumination
- Light intensity and quality: 88-91 µmol*m-2*s-1 (wavelength: 400-700 nm)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
- Chlorophyll measurement: Chlorophyll was measured using a spectrophotometer every 24 h.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: control, 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: No concentration dependant inhibition of growth was recorded. At 100 mg/L a maximum inhibition of growth of 2.8% was measured.

Reference substance (positive control):

yes

Remarks:

potassium dichromate (tested periodically)

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

- Exponential growth in the control (for algal test): yes
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No precipitation of test item was recorded.
- Any stimulation of growth found in any treatment: Stimulation of growth was recorded at 100 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 0.78 mg/L
- Other: EC50 is within the normal range of the reference substance in the laboratory [mean ± S.D.: 1.0 ± 0.19 mg/L (n=29)].

Reported statistics and error estimates:

F test was done to determine homogeneity of variances. Student’s t-test was used to estimate the significant difference in comparison with the control.

Table 1: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72-hour test period.	> 71	yes
The mean coefficient of variation for section-by-section specific growth rates (days 0-1, 1-2 and 2-3, for 72-hour tests) in the control cultures must not exceed 35%	7.9%	yes
The coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% in tests with Pseudokirchneriella subcapitata and Desmodesmus subspicatus. For other less frequently tested species, the value should not exceed 10%.	2.3%	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

22 - 25 Aug 2016

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no chemical analysis

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 100 mg of test item was dissolved in 1000 mL test medium to prepare a stock solution of 100 mg/L. The solution was stirred fro 48 h. Afterwards it was devided into each test vessel.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: ATCC 22662
- Source (laboratory, culture collection): American Type Culture Collection (supplied 30 Jun 1995)

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test temperature:

22.5 - 22.8 °C

pH:

7.9 - 8.0 (control)
7.7 - 8.1 (treatment)

Nominal and measured concentrations:

nominal: control, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Sterilized 300 mL Erlenmeyer flask (with gas-permeable stopper); fill volume: 100 mL
- Aeration: constantly shaken (approx. 100 rpm)
- Initial cells density: 0.75 E+04 cells/mL (equivalent to 2.6 Chlorophyll fluorescence values)
- Control end cells density: Cell density not given; 180 Chlorophyll fluorescence value
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified water according to guideline
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continuous illumination
- Light intensity and quality: 89 µmol*m-2*s-1 (wavelength: 400-700 nm)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
- Chlorophyll measurement: Chlorophyll was measured using a spectrophotometer every 24 h.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: control, 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: No concentration dependant inhibition of growth was recorded. At 100 mg/L a maximum inhibition of growth of 2.8% was measured.

Reference substance (positive control):

yes

Remarks:

potassium dichromate (tested periodically)

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

- Exponential growth in the control (for algal test): yes
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No precipitation of test item was recorded.
- Any stimulation of growth found in any treatment: Stimulation of growth was recorded at 100 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 0.78 mg/L
- Other: EC50 is within the normal range of the reference substance in the laboratory [mean ± S.D.: 1.0 ± 0.19 mg/L (n=29)].

Reported statistics and error estimates:

F test was done to determine homogeneity of variances. Student’s t-test was used to estimate the significant difference in comparison with the control.

Table 1: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72-hour test period.	> 65	yes
The mean coefficient of variation for section-by-section specific growth rates (days 0-1, 1-2 and 2-3, for 72-hour tests) in the control cultures must not exceed 35%	10%	yes
The coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% in tests with Pseudokirchneriella subcapitata and Desmodesmus subspicatus. For other less frequently tested species, the value should not exceed 10%.	2.3%	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

27 Feb - 02 Mar 2017

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no chemical analysis22

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The necessary amount of test item was weighed and the medium was added to obtain the different test concentrations. The solutions were stirred for 48 h. Afterwards they were distributed into each test vessel.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: ATCC 22662
- Source (laboratory, culture collection): American Type Culture Collection (supplied 30 Jun 1995)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

23.5 - 23.6 °C

pH:

7.8 - 7.9 (control)
7.8 - 8.2 (treatment)

Nominal and measured concentrations:

nominal: control, 6.3, 13, 25, 50 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Sterilized 300 mL Erlenmeyer flask (with gas-permeable stopper); fill volume: 100 mL
- Aeration: constantly shaken (approx. 100 rpm)
- Initial cells density: 0.75 E+04 cells/mL (equivalent to 81 Chlorophyll fluorescence values)
- Control end cells density: Cell density not given; 4500 Chlorophyll fluorescence value (mean)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified water according to guideline
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continuous illumination
- Light intensity and quality: 88-91 µmol*m-2*s-1 (wavelength: 400-700 nm)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
- Chlorophyll measurement: Chlorophyll was measured using a spectrophotometer every 24 h.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: control, 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: No concentration dependant inhibition of growth was recorded. At 100 mg/L a maximum inhibition of growth of 2.8% was measured.

Reference substance (positive control):

yes

Remarks:

potassium dichromate (tested periodically)

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

- Exponential growth in the control (for algal test): yes
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No precipitation of test item was recorded.
- Any stimulation of growth found in any treatment: Stimulation of growth was recorded at 6.3, 13 and 25 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 1.3 mg/L
- Other: EC50 is within the normal range of the reference substance in the laboratory [mean ± S.D.: 0.99 ± 0.19 mg/L (n=29)].

Reported statistics and error estimates:

Homogeneity of variances was tested using Bartlett’s test. One way ANOVA and Dunnett’s multiple comparison were used to estimate the significant difference between each exposure level and control.

Table 1: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
The biomass in the control cultures should have increased exponentially by a factor of at least 16 within the 72-hour test period.	> 43	yes
The mean coefficient of variation for section-by-section specific growth rates (days 0-1, 1-2 and 2-3, for 72-hour tests) in the control cultures must not exceed 35%	18%	yes
The coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% in tests with Pseudokirchneriella subcapitata and Desmodesmus subspicatus. For other less frequently tested species, the value should not exceed 10%.	3.6%	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Description of key information

EC50 (72 h) > 100 mg/L (nominal, OECD 201)
NOEC (72 h) ≥ 100 mg/L (nominal, OECD 201)

Additional information

Three experimental studies are available investigating the effects of Fatty acids C16-18 (even numbered), mono and diesters with sucrose (no CAS) to the freshwater algae Pseudokirchneriella subcapitata. All studies were performed according to OECD 201 (GLP). Two studies were performed as a limit test with a concentration of 100 mg/L whereas one study was performed as a dose-response study with five concentrations up to 100 mg/L. The test item was added to the test vessels at appropriate amounts and the solutions were stirred for 48 h to ensure complete dissolution. No precipitation of undissolved test material was recorded in any of the test vessels. After 72 h of exposure the growth of algae cells was not inhibited by the test item resulting in a EC50 (72 h) > 100 mg/L and a NOEC (72 h) ≥ 100 mg/L. All effect concentrations were based on the nominal concentrations since no chemical analysis was performed. However, the chemical analysis in the acute fish study indicated that the substance is stable in aqueous solutions.