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EC number: 944-561-8
CAS number: 1465004-85-6
Table 3 Verification of Test Concentrations
Nominal Test Concentration
(% v/v Saturated Solution)
A study was performed to assess the acute toxicity of the test item to
Daphnia magna. The method followed was designed to be compatible with
the OECD Guidelines for Testing of Chemicals (April 2004) No 202,
"Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of
Commission Regulation (EC) No. 440/2008.
Preliminary solubility work conducted indicated that it was not possible
to obtain a testable solution of the test item using traditional methods
of preparation e.g. ultrasonication and high shear mixing.
A preliminary media preparation trial indicated that a dissolved test
item concentration of approximately 29 mg/L was obtained from a
saturated solution method of preparation indicating this to be the limit
of water solubility of this item under test conditions.
Following a preliminary range-finding test, twenty daphnids (4
replicates of 5 animals) were exposed to an aqueous solution of the test
item at nominal concentrations of 1.0, 3.2, 10, 32 and 100% v/v
saturated solution for 48 hours at a temperature of 20 C to 22 C under
static test conditions. The test item solutions were prepared by
stirring an excess (50 mg/L) of test item in test water using a
propeller stirrer at approximately 1500 rpm for 24 hours. After the
stirring period any undissolved test item was removed by filtration (0.2
µm Sartorius Sartopore filter, first approximate 2 liters discarded in
order to pre-condition the filter) to produce a 100% v/v saturated
solution of the test item. This saturated solution was then further
diluted as necessary, to provide the remaining test concentrations.
Immobilization and any adverse reactions to exposure were recorded
after 24 and 48 hours.
Chemical analysis of the test preparations at 0 and 48 hours showed
measured test concentrations to range from 0.39 to 41 mg/L and 0.41 to
36 mg/L respectively. Given the slight variability in results it was
considered appropriate to calculate the results based on the geometric
mean measured test concentrations which were determined to be 0.40, 1.3,
4.1, 13 and 38 mg/L.
Exposure of Daphnia magna to the test item gave the following results
based on the geometric mean measured test concentrations:
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