Materials and Methods- Toxicity – SESAMULS® OW
Test Item
Designation in Test Facility: | 14121110N |
Name |
SESAMULS® OW (Polyglyceryl-3 Cetyl Ether, Sesame Oil, Malic Acid) |
Batch No. | C0092 |
Storage
The test item was stored in a closed vessel dark and dry at room temperature.
Preparation
Due to the poor solubility of the test item, the water-accommodated fraction (WAF) of the concentrations to be tested was prepared. This was done by weighing the nominal loads (1 mg/L, 10 mg/L and 100 mg/L), adding the appropriate amount of nutrient medium (demineralised water enriched with minerals but without algae) and shaking for 24 hours. The resulting solutions were filtrated through 0.45 μm filters.
Positive Control
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a separate reference test (GLP study).
Test System
Specification
Unicellular freshwater green alga.
Genus | Desmodesmus |
Species | subspicatus |
SAG Strain Number | 86.81 |
Taxonomic position |
Chlorophyta – Chlorophyceae |
Origin and Culture
The culture of Desmodesmus subspicatus was obtained in February 2019 by MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen). The algae are kept as stock culture on solid agar at 7 °C. From the stock culture, a permanent culture was prepared. From an aliquot of the permanent culture, the pre-culture was prepared.
Chemicals
All solutions were sterilised before use.
Composition of the solutions:
Stock solution I | |
NH4Cl |
1500 mg |
MgCl2*6H2O |
1200 mg |
CaCl2*2H2O |
1800 mg |
MgSO4*7H2O |
1500 mg |
KH2PO4 |
180 mg |
H2O deionized |
ad 1000 mL |
Stock solution II | |
FeCl3*6H2O |
64 mg |
Na2EDTA*2H2O |
100 mg |
H2O deionized |
ad 1000 mL |
Stock solution III | |
H3BO3 |
185 mg |
MnCl2*4H2O |
415 mg |
ZnCl2 |
3 mg |
CoCl2*6H2O |
1.5 mg |
CuCl2*2H2O |
0.01 mg |
Na2MoO4*2H2O |
7 mg |
H2O deionized |
ad 1000 mL |
Stock solution IV | |
NaHCO3 |
50 mg |
H2O deionized |
ad 1000 mL |
Pre-culture-medium | |
Stock solution I | 10.0 mL |
Stock solution II | 1.0 mL |
Stock solution III | 1.0 mL |
Stock solution IV | 1.0 mL |
H2O deionized |
ad 1000 mL |
Nutrient-medium (10-fold concentrated) | |
Stock solution I | 100 mL |
Stock solution II | 10 mL |
Stock solution III | 10 mL |
Stock solution IV | 10 mL |
H2O deionized |
ad 1000 mL |