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EC number: 617-081-9
CAS number: 80473-92-3
All test and control cultures were inspected microscopically at 72
hours. After 72 hours there were no abnormalities detected in the
control or test cultures at 0.47, 1.7, 5.8 and 18 mg/L, however no
intact cells were observed to be present in the test cultures at 58 mg/L.
A study was performed to assess the effect of the test item on the
growth of the green alga Pseudokirchneriella subcapitata. The method
followed that described in the OECD Guidelines for Testing of Chemicals
(2006) No 201, "Freshwater Alga and Cyanobacteria, Growth Inhibition
Test" referenced as Method C.3 of Commission Regulation (EC) No 761/2009.
Preliminary solubility work conducted indicated that the test item was
not readily soluble in water 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 118 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, Pseudokirchneriella
subcapitata was exposed to solutions of the test item at nominal
concentrations of 1.0, 3.2, 10, 32 and 100% v/v saturated solution
(three replicate flasks per concentration) for 72 hours, under constant
illumination and shaking at a temperature of 24 ± 1 °C. The test item
solutions were prepared by stirring an excess (100 mg/L) of test item in
culture medium using a propeller stirrer at approximately 1500 rpm for
48 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 precondition 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
Samples of the algal populations were removed daily and cell
concentrations determined for each control and treatment group, using a
Coulter® Multisizer Particle Counter.
Analysis of the test preparations at 0 hours showed measured test
concentrations to range from 0.56 to 67 mg/L. A decline in measured test
concentration was observed at 72 hours to between 0.39 and 49 mg/L (67%
to 78% of the 0-Hour measured test concentrations) and hence it was
considered appropriate to calculate the results based on the geometric
mean measured test concentration only in order to give a “worst case”
analysis of the data.
Exposure of Pseudokirchneriella subcapitata to the test item gave the
following results based on the geometric mean measured test
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