Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

The temperature in the experiment was slightly higher than stated in the guideline. Because normal growth was observed, this can be stated as uncritical. The deviation was signed and assessed by the study director on 22. Jan. 2016

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

yes

Remarks:

The temperature in the experiment was slightly higher than stated in the guideline. Because normal growth was observed, this can be stated as uncritical. The deviation was signed and assessed by the study director on 22. Jan. 2016

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

EC Number:

941-319-3

Molecular formula:

Unknown (Substance of Unknown or Variable Composition)

IUPAC Name:

Esterification products of Grape seeds, Vitis vinifera L. (Vitaceae), extract with hexadecanoyl chloride

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Appearance: brown powder
Composition: Palmitoyl grape seed extract (UVCB)
CAS No. 226994-27-0
Purity :100 % UVCB
Homogeneity : homogeneous
Volatility : unknown
Production date : 24. Nov. 2015
Expiry date :19. Oct. 2018
Storage : The test item was stored in a closed vessel dark and dry at room temperature at 15.2 – 20.8 °C.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Four days before the start of each test, an aliquot of the permanent culture was brought into nutrient medium and incubated under continuous lighting for 96 hours. The resulting culture grew exponentially.
Before usage, the pre-culture was checked for the absence of cell aggregates and the cell number of culture was determined via photometric measurement.

Test solutions

Vehicle:

yes

Remarks:

Water

Details on test solutions:

Test item: Palmitoyl grape seed extract.
Prepartion of the test solution: The water-accommodated fraction (WAF) was prepared. This was done by weighing the nominal load 100 mg/L (real load 100.5 mg/L) adding the appropriate amount of algal medium
(demineralised water enriched with minerals but without algae) and shaking for 23 hours. After membrane filtration, the solution was used to prepare the treatment.

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

Specification: unicellular freshwater green alga. SAG Strain Number 86.81
Origin and Culture: The culture of Desmodesmus subspicatus was obtained in February 2014 by MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen). The algae are kept as stock culture on solid agar at 2 - 8 °C. From the stock culture, a permanent culture was prepared. From an aliquot of the permanent culture, the pre-culture was prepared.

Study design

Test type:

static

Water media type:

freshwater

Total exposure duration:

72 h

Test conditions

Test temperature:

22.8 - 24.2 °C

Details on test conditions:

Treatments tested: 100 mg/L;
Number of replicates: 6 replicates for the control and 6 replicates for the treatment;
Vessels: glass flasks total volume 65 mL;
Lighting: 5300 Lux
Blank control: deionised water with nutrient medium and algae;
Treatments: test solution and algae.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a separate reference test.

Results and discussion

Effect concentrations

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

All validity criteria were met. No observations were made which might cause doubts concerning the validity of the study outcome. The result of the test can be considered valid.

Conclusions:

One valid experiment was performed.
The study was performed as a limit test using one concentration containing 100 mg/L nominal concentration. The treatment showed slight inhibition of algal growth.
At the beginning and at the end of the test, the content of the test item in the test solutions was estimated using DOC-determination. Because of the low solubility in test medium, no calculation from carbon content of the test item (76.54 %) and DOC measurement was possible. The measured DOC concentrations in the treatments were in a similar low range as the blank control. Therefore, the determination of the biological results was based on
the nominal concentration, which represents the nominal loading of the test item.
The 72h-EC50s of potassium dichromate were determined in a separate reference test. For the estimation of the 72h-EC50s of the positive control, the fits showed sufficient statistical correspondence of the data with the dose-response-equation. The values were within the range of the laboratory.
All validity criteria were met. No observations were made which might cause doubts concerning the validity of the study outcome. The result of the test can be considered valid.

Executive summary:

One valid experiment was performed. The study was performed as a limit test using one concentration containing 100 mg/L nominal concentration. Incubation time (test system Desmodesmus subspicatus) was 72 hours. The cell concentration of each replicate was determined by measuring the absorption of the solutions at 440 nm every 24 hours with a spectral photometer. The cell density of the cultures was calculated based on the correlation curve between the absorption and the cell density of the cultures determined with a cell counter. Growth rate μ and the yield1 were determined from the cell number at the respective observation times. The treatment showed slight inhibition of algal growth. At the beginning and at the end of the test, the content of the test item in the test solutions was estimated using DOC-determination. Because of the low solubility in test medium, no calculation from carbon content of the test item (76.54 %) and DOC measurement was possible. The measured DOC concentrations in the treatments were in a similar low range as the blank control. Therefore, the determination of the biological results was based on the nominal concentration, which represents the nominal loading of the test item. The 72h-EC50s of potassium dichromate (K2Cr2O7, CAS No. 7778-50-9) were determined in a separate reference test. The values lay within the range of the laboratory (growth rate 0.73 - 1.10 mg/L, yield 0.21 – 0.55 mg/L). The following results for the test item Palmitoyl grape seed extract were determined:

Endpoint	NOEC	LOEC	EC10	EC50
Growth Rate	< 100 mg/L	100 mg/L	>100 mg/L	>100 mg/L
Yield	< 100 mg/L	100 mg/L	< 100 mg/L	>100 mg/L