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EC number: 202-297-4
CAS number: 94-04-2
IntroductionA 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.
MethodsPreliminary 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 13 mg/L was obtained from a saturated solution (shake flask) method of preparation indicating this to be the limit of water solubility of this item under test conditions.Due to the potentially volatile nature of the test item, testing was conducted in completely filled and sealed test vessels in order to minimize possible losses due to volatilization.Following a preliminary range-finding test, 20 daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at nominal concentrations of 4.5, 10, 22, 48 and 100% v/v saturated solution for 48 hours at a temperature of approximately 20 °C to 21 °C under semi-static test conditions. The test item solutions were prepared by employing a shake flask method of preparation, this method of preparation was undertaken due to the lack of test item found during a traditional saturated solution approach (due to the potential volatility of the test item). The test item solutions were prepared in four replicates by shaking in excess (100 mg/L) of the test item in test water at approximately 150 rpm, at a temperature of 24 °C for 24 hours. After the shaking period the contents of the replicate flasks were pooled and any undissolved test item was removed by filtration through a 0.2 μm Gelman Acrocap filter (first approximate 100 mL was discarded in order to pre-condition the filter) to give a 100% v/v saturated solutions of the test item. This saturated solution was then further diluted to give the 48, 22, 10 and 4.5% v/v saturated solutions of the test item.Immobilization and any adverse reactions to exposure were recorded after 24 and 48 hours.
ResultsAnalysis of the fresh test preparations at 0 and 24 hours showed measured test concentrations to range from 0.44 to 14 mg/L. Measured test concentrations of the aged test preparations at 24 and 48 hours showed measured test concentrations to range from 0.27 to 12 mg/L. As not all test concentrations were maintained within ± 20% of the measured initial test concentration it was considered appropriate to calculate the results based on the geometric mean measured test concentration in order to give a “worst case” analysis of the data. The geometric mean measured test concentrations were determined to be 0.36, 0.97, 2.3, 5.3 and 12 mg/L.Exposure of Daphnia magna to the test item gave the following results based on the geometric mean measured test concentration:
The EC50 for freshwater daphnia data is 1.2 mg/L.
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