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EC number: 218-664-7
CAS number: 2212-81-9
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)
Following a preliminary range-finding test and initial experiment, Pseudokirchneriella
subcapitata was exposed to a solution of the test item at a nominal
concentration of 100% v/v saturated solution (six replicate flasks) for
72 hours, under constant illumination and shaking at a temperature of
24 ±1 °C. The test item solution was prepared by stirring an excess
(10 mg/L) of test item in culture medium using a magnetic stirrer for
23 hours. After 23 hours the stirring was stopped and
the mixture was allowed to stand for 1-Hour, prior to removal of the
aqueous phase by mid-depth siphoning, to give a 100% v/v saturated
solution of the test item.
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 100% v/v saturated solution test preparation at
0 hours showed a measured test concentration of 0.0078 mg/L was
obtained. A decline in measured test concentration was observed after
each 24-Hour period in the range of 0.0045 mg/L at 24 hours, 0.0021 mg/L
at 48 hours and less than the limit of quantification (LOQ) of the
analytical method employed, determined to be 0.00031 mg/L at 72 hours.
Analysis of an additional 100% v/v saturated solution test
preparation containing no algal cells which was incubated alongside the
test showed a measured test concentration of 0.0059 mg/L was obtained at
72 hours (75% of the 0-Hour measured test concentration). This result
confirms that the majority of losses observed over the test period were
due to adsorption of the test item to the algal cells present rather
than instability of the test item. It was therefore considered
appropriate to calculate the results based on the geometric mean of the
0-Hour measured test concentration and the 72-Hour measured
concentration obtained from the preparation containing no algal cells as
this gave a more representative measurement of the concentration of test
item to which the algae were exposed.
Exposure of Pseudokirchneriella subcapitata to the test item gave
EC50values based on the geometric mean measured test
concentration of greater than 0.0068 mg/L. The
No Observed Effect Concentration (NOEC) was determined to be 0.0068 mg/L.
This study showed that there were no toxic effects at the limit of
water solubility (100% v/v saturated solution).
The analgous test substance CAS 25155-25-3
[1,3-phenylenebis(1-methylethylidene)]bis[tert-butyl] peroxide was not
found to be toxic to Pseudokirchneriella subcapitata at a
nominal loading rate of 1.0 mg/L, equivalent to an initial mean measured
concentration of 0.0471 mg/L.
A study with the target substance (CAS 2212 -81 -9) is was performed to
strengthen the Read Across.
The effect of the test item on the growth of Pseudokirchneriella
subcapitata has been investigated and gave EC50 values of greater
than 0.0068 mg/L based on the geometric mean measured test
concentration. The No Observed Effect Concentration was 0.0068 mg/L.
This study showed that there were no toxic effects at the limit of water
solubility (100% v/v saturated solution).
Both studies show that the test substances did not give an effect at the
limit of water solubility.
The toxicity of analogous CAS 25155 -25 -3 [1,3(or
1,4)-phenylenebis(1-methylethylidene)]bis[tert-butyl] peroxide was
evaluated with Pseudokirschneriella subcapitata in a study
performed in accordance with OECD testing guideline 201 and GLP
The algae were exposed to nominal test concentrations of 1 mg/L
under static conditions during 72 hours. The mean measured
concentrations were not maintained between 80 and 120 % of nominal.
pH range and dissolved oxygen concentration were reported and in
agreement with the guideline recommendations. Quality criteria were
The test substance was not found to be toxic to Pseudokirchneriella
subcapitata at a nominal loading rate of 1.0 mg/L, equivalent to an
initial mean measured concentration of 0.0471 mg/L.
Exposure to the test item gave EC50 values of greater than 0.0068 mg/L
based on the geometric mean measured test concentration. The No
Observed Effect Concentration was 0.0068 mg/L.
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