Linseed oil, polymerized

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:

08 March 2010-11 March 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Linseed oil, polymerized

EC Number:

614-114-9

Cas Number:

67746-08-1

Molecular formula:

Not applicable for UVCB

IUPAC Name:

Linseed oil, polymerized

Details on test material:

- Name of test material (as cited in study report): Standolized linseed oil
- Substance type: Hazy yellow/brown viscous liquid
- Physical state: liquid
- Analytical purity: treated as 100% pure
- Lot/batch No.: 09.317.013
- Expiration date of the lot/batch: 21 December 2010
- Stability under test conditions: stable
- Storage condition of test material: At room temperature in the dark
- Other: -

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:TOC-analysis from the WAF prepared at a loading rate of 100 mg/l and the untreated control from vessels incubated without algae
- Sampling method:
Frequency at t=0 h, t=24 h and t=72 h
Volume 40 ml
Storage Samples were stored in a refrigerator until analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Preparation of the test solutions started with loading rates prepared individually at 1.0, 10 and 100 mg/l applying 48 hours of magnetic stirring to achieve maximum dissolution in test medium. The magnetic stirring was followed by a settlement period of 2 hours. This resulted in clear and colourless solutions with a floating layer and a precipitate. The Water Accommodated Fractions (WAFs) were then collected by siphoning the water phase. The final test solutions were all clear and colourless.

Test organisms

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
- Age of inoculum (at test initiation): 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 10E4 cells/ml.
- 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.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test conditions

Hardness:

0.24mmol/l (24 mg CaCO3/l)

Test temperature:

between 22.8 and 23.6°C

pH:

At t=0 h: 7.7-8.2
At t=72 h:8.2

Salinity:

not applicable

Nominal and measured concentrations:

Nominale concentration: WAFs prepared at loading rates of 1.0, 10 and 100 mg/l
Measured concentration:During the test samples for TOC-analysis were taken from the WAF prepared at a loading rate of 100 mg/l and the untreated control. The TOC concentrations measured in the treated samples were only slightly exceeding those measured in the control samples, indicating that the actual test substance concentration was very low. The actual measured concentration was between approximately 0.3 and 0.5 mg C/l when corrected for the carbon measured in the control samples. The TOC content of Standolized linseed oil was determined to be 77.88% . Thus, the actual Standolized linseed oil concentration in solution was expected to be between 0.4 and 0.6 mg/l.

Details on test conditions:

TEST SYSTEM
- Test vessel:100 ml, all-glass, containing 50 ml of test solution
- Type (delete if not applicable): open
- Aeration: no
- Initial cell density:10000 cells/ml
-Control end cell density: 1597000 cells/ml
- No. of vessels per concentration (replicates):6 replicates of the control and the WAF 100 mg/l;3 replicates of the 1.0 and 10 mg/l WAFs

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: continuous
- Light intensity and quality: TLD-lamps of the type ‘Cool-white’ of 30 Watt, with a light intensity within the range of 63 to 71 µE/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 Varian Cary 50 single beam spectrophotometer with cuvettes (pathlength =10 mm). Algal medium was used as blank and the extra replicates without algae as background for the treated solutions.
- Other: 72 h NOErC, 72 h NOEbC, 72 h ErC50, 72 h EbC50


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: WAF prepared at 1.0, 10 and 100mg/l

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: waf prepared at 100 mg/l

Effect concentrations 2

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: > waf prepared at 100 mg/l

Effect concentrations 3

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: yield

Remarks on result:

other: waf prepared at 100 mg/l

Effect concentrations 4

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: yield

Remarks on result:

other: > waf prepared at 100 mg/l

Effect concentrations 5

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.4 - 0.6 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: TOC

Basis for effect:

growth rate

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 0.4 - 0.6 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: TOC

Basis for effect:

growth rate

Effect concentrations 7

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.4 - 0.6 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: TOC

Basis for effect:

other: Yield

Effect concentrations 8

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 0.4 - 0.6 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

other: TOC

Basis for effect:

other: Yield

Details on results:

-No significant differences were recorded between the values for growth rate or yield at any of the test concentrations when compared to the control group.
Microscopic observations at the end of the test in the WAF prepared at 100 mg/l revealed a normal and healthy appearance of the exposed cells when compared to the control

Results with reference substance (positive control):

- Results with reference substance valid: yes
-The EC50 for growth rate reduction (ERC50: 0-72h) was 1.2 mg/l with a 95% confidence interval ranging from 0.77 to 2.0 mg/l.
-The EC50 for yield inhibition (EYC50: 0-72h) was 0.44 mg/l with a 95% confidence interval ranging from 0.37 to 0.54 mg/l

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present study with Pseudokirchneriella subcapitata, no reduction of growth rate or inhibition of yield was recorded at a WAF prepared at a Standolized linseed loading rate of 100 mg/l (NOEC).
Hence, both the EC50 for growth rate reduction (ERC50: 0-72h) and the EC50 for yield inhibition (EYC50: 0-72h) exceeded a WAF prepared at a loading rate of 100 mg/l, which can be considered to represent the maximum soluble concentration in test medium. TOC-analyses showed that the actual concentration in solution was expected to be between 0.4 and 0.6 mg/l