Fatty acids, vegetable-oil, polymd., unreacted fraction

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 June to 10 July 2017 (experimental dates)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

2006

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Version / remarks:

2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, vegetable-oil, polymd., unreacted fraction

EC Number:

273-100-7

EC Name:

Fatty acids, vegetable-oil, polymd., unreacted fraction

Cas Number:

68937-92-8

Molecular formula:

Substance is a UVCB - no structural or molecular formula is available.

IUPAC Name:

5-[(5E)-icos-5-enoyloxy]icosanoic acid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot No.of test material: 1367Q13601
- Expiration date of the lot/batch: 17/09/2017
- Purity test date: 16/05/2014

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: Not specified
- Solubility and stability of the test substance in the solvent/vehicle: Not specified

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control and 100 mg/L nominal loading rate WAF
- Sampling method: 0h from bulk test solution, 72h from incubated pooled replicates
- Sample storage conditions before analysis: Frozen until required for analysis

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test item (200 mg) was added to surface of culture medium (2 L) to give nominal loading rate 100 mg/L. Culture medium was then stirred by magnetic stirrer for 23 hours, after which mixture allowed to settle for 1 hour. Aqueous phase (WAF) was removed by mid-depth siphoning.
- Eluate: Culture medium
- Differential loading: No
- Controls: Culture medium only
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Micorscopic inspection of WAF showed no micro-dispersion or undissolved test material.

Test organisms

Test organisms (species):

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

Details on test organisms:

TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: CCAP 278/4
- Source (laboratory, culture collection): Culture Collection of Alage and Protozoa (CCAP), SAMS Reasearch Services Ltd, Scottish Marine Institute, Oban, Scotland
- Age of inoculum (at test initiation): Not specified

ACCLIMATION
- Acclimation period: No
- Culturing media and conditions (same as test or not): Same as test
- Any deformed or abnormal cells observed: All test and control cultures inspected microscopically at 72 hours (end of test). No abnormalities observed in control or test cultures

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test conditions

Hardness:

No data

Test temperature:

24 +/- 1oC

pH:

7.8 to 8.0

Dissolved oxygen:

No data

Salinity:

No data

Conductivity:

No data

Nominal and measured concentrations:

Nominal concentrations: Control and 100 mg/L loading rate WAF
Measured concentrations: >LoQ for both control and 100 mg/L loading rate WAF. LoQ calculated to be 0.042 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 mL glass conical flask
- Type (delete if not applicable): open / closed: Test vessels plugged with polyeurethane foam bung to reduce evaporation
- Material, size, headspace, fill volume: Glass, 250 mL, fiilled with 100 mL test solution
- Aeration: No
- Initial cells density: 5.0 x 10.3 cells/mL
- Control end cells density: 1.4 x 10.6 cells/mL
- 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:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity:
- Culture medium different from test medium:
- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality:
- Salinity (for marine algae):

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]
- Chlorophyll measurement:
- Other:

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Results and discussion

Effect concentrations

Effect concentrations 1

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

Effect concentrations 2

Key result

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

72 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

Yield

Effect concentrations 4

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

Yield

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): None observed
- Effect concentrations exceeding solubility of substance in test medium: Yes

Any other information on results incl. tables

Nominal loading rate WAF (mg/L	Growth rate		Yield
	0-72h mean growth rate (cells/mL/h)	% inhibition	0-72h mean yield (cells/mL)	% inhibition
Control	0.079	-	1.45 x 10^6	-
100	0.079	0	1.47 x 10^6	0

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 72 hour toxicity results are based on the test item as a whole and the results have been reported based on nominal loading rates only. Under the conditions of the test the 72 hour EL50* for growth rate inhibition was >100 mg/L loading rate WAF and the NOELR* was 100 mg/L loading rate WAF. The toxicity of the test item can be considered as being beyond the range tested and exceeded the maximum soluble concentration of the test item in the test medium used for the test i.e. the EL50 exceeded the limit of quantification (LoQ). The LoQ was calculated to be 0.042 mg/L.
* NOELR - No Observed Effect Loading Rate
* EL - Effective Loading Rate

Executive summary:

A 72- hour toxicity study with the green alga, Pseudokirchneriella subcapitata, was conducted in accordance with the OECD test guideline 201 and in compliance with GLP. The test was performed as a limit test at a nominal loading rate of 100mg/L WAF (Water Accommodated Fraction), with the single test solution prepared from a 100 mg/L saturated solution. The 72 hour measured concentration was <LoQ (Limit of Quantification) of the analytical method. Under the conditions of the test the EL50 for growth rate inhibition was >100 mg/L loading rate WAF and the NOELR was 100 mg/L loading rate WAF. This ecotoxicity study is considered acceptable and satisfies the guideline requirement for toxicity to aquatic algae.

* NOELR - No Observed Effect Loading Rate

* EL - Effective Loading Rate