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EC number: 233-634-3
CAS number: 10287-53-3
Table 1: Cumulative
Immobilisation Data and Observations in the Definitive Test over 24 hours
0-Hour Measured Concentration(mg/L)
Cumulative Immobilised Daphnia(Initial Population: 5 Per Replicate)
1R 2T 2N
N = No sub-lethal effects observed
T = Trapping at the surface
R = Reduced mobility
= All daphnia immobilised
Table 3: Cumulative
Immobilisation Data and Observations in the Definitive Test over 48 hours
Cumulative Immobilized Daphnia(Initial Population: 5 Per Replicate)
= All daphnia immobilised
The potential toxicity of the test material to Daphnia magna was
determined in accordance with the standardised guidelines OECD 202 and
EU Method C.2 under GLP conditions, using an acute immobilisation test.
Preliminary solubility work conducted indicated that it was not possible
to obtain a testable solution of the test material using traditional
methods of preparation e.g. ultrasonication and high shear mixing. A
preliminary media preparation trial indicated that a dissolved test
material concentration of approximately 46 mg/L was obtained from a
saturated solution method of preparation indicating this to be the limit
of water solubility of this material 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
material at nominal concentrations of 1.0, 2.5, 6.4, 16 and 40 % v/v
saturated solution for 48 hours at a temperature of 21 to 22 °C under
static test conditions. The test material solutions were prepared by
stirring an excess of test material in test water using a propeller
stirrer for 24 hours; any undissolved test material was removed by
filtration to produce a 100 % v/v saturated solution of the test
material. This saturated solution was then further diluted as necessary,
to provide the required test concentrations. Immobilisation and any
adverse reactions to exposure were recorded after 24 and 48 hours.
Chemical analysis of the freshly prepared test preparations at 0 hours
showed measured test concentrations to be 0.34, 1.1, 3.1, 8.1 and 21
mg/L. There was no significant change in the measured concentrations at
48 hours and so the results are based on 0-hour measured test
Complete immobilisation was observed after 48 hours in the two highest
test concentrations; 10 % immobilisation was observed in the 3.1 mg/L
test concentration. Sub-lethal effects of exposure were observed in the
control and the 1.1, 3.1, 8.1 and 21 mg/L test concentrations. These
responses were trapping at the surface and reduced mobility.
The validity and water quality criteria of the test were both met.
Under the conditions of the study, the 48-hour EC50 value was 4.5 mg/L
with 95 % CL of 4.0 to 5.2 mg/L. The No Observed Effect Concentration
was 1.1 mg/L.
In the key study, the potential toxicity of the test material to Daphnia
magna was determined in accordance with the standardised guidelines OECD
202 and EU Method C.2 under GLP conditions, using an acute
immobilisation test. The study was awarded a reliability score of 1 in
accordance with the criteria set forth by Klimisch et al. (1997).
In the supporting study, the potential of the test material to cause
acute toxicity to Daphnia magna was determined broadly in accordance
with the standardised guideline OECD 202 under GLP conditions. The study
was awarded a reliability score of 1 in accordance with the criteria set
forth by Klimisch et al. (1997).
Daphnia magna were exposed to the test material at nominal
concentrations of 1.0, 10.0 and 100.0 mg/L alongside a vehicle (0.1 mL/L
acetone) and negative control under static conditions. The Daphnia were
in duplicate sealed conical flasks (5 per flask) for a period of 48
hours. Daphnids were observed after 24 and 48 hours and were considered
to be immobile if they did not swim during a 15 second period of
The lowest concentration causing 100 % immobilisation was 100 mg/L and
the highest concentration causing no immobilisation was 10 mg/L.
Under the conditions of this study, the 48 h EC50 was 31.8 mg/L.
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