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EC number: 948-091-4 | CAS number: -
Table 1: Measured Concentrations in Samples taken during the Final Test and Calculated Time Weighted Average Concentrations
Loading rate (mg/L)
Measured concentration (µg/L)
TWA conc. (µg/L)
(a) Incubated without algae; (b) estimated by extrapolation of the calibration curve; (c) actual concentration was taken as half of the LOD, which was 0.31 µg/L on the day of analysis.
Table 2: Effect Parameters based on Loading Rates (mg/L)
cl – confidence limit; * based on statistical significance; #based on biological relevance.
Table 3: Effect Parameters based on Time Weighted Average Concentrations (µg/L)
Parameter (µg/L) (a)
(a) Effect parameters are indicative and should be interpreted with caution as it is unknown if the analytically monitored constituent is responsible for the recorded effects; cl – confidence limit; * based on statistical significance; #based on biological relevance.
Table 4: Growth Rate and Percentage Inhibition for the Total Test Period
Loading rate (mg/L)
0.010 [1.8 ]
[ ] – TWA concentration (µg/L), * Effect was statistically significant,#effect was statistically significant but biologically not relevant (<10%).
Table 5: Growth Rate and Percentage Inhibition at Different Time Intervals
0 – 24 h
24 – 48 h
48 – 72h
[ ] – TWA concentration (µg/L)
The objective of the study was to evaluate BEROL 1872 for its ability to generate toxic effects in Raphidocelis subcapitata (formerly known as Pseudokirchneriella subcapitata) during an exposure period of 72 hours and, if possible, to determine the NOELR, EL10, EL20 and EL50 for both inhibition of growth rate and inhibition of yield.
The study procedures described in this report were based on the OECD guideline No. 201, 2006; Annex 5 corrected 28 July 2011. In addition, procedures were based on the test methods described in the OECD series on testing and assessment number 23, 2018.
The batch of BEROL 1872 tested was a light yellow liquid UVCB and was not completely soluble in test medium at the loading rates initially prepared. Water Accommodated Fractions (WAFs) were individually prepared at loading rates in the range of 0.010 to 1.0 mg/L and used as test concentrations. Test solutions were prepared by using stock solutions in acetonitrile (ACN). Since the test item is an UVCB, the solvent was completely evaporated prior to addition of the test medium to avoid modification of WAF-composition due to presence of a water-miscible solvent.
A final test was performed based on the results of a preceding combined limit/range-finding test and range-finding test. Six replicates of exponentially growing algal cultures per group were exposed to a blank and a solvent control. Three replicates per test concentration were exposed to WAFs individually prepared at loading rates of 0.010, 0.032, 0.10, 0.32 and 1.0 mg/L. The initial algal cell density was 10^4 cells/mL. The total exposure period was 72 hours and samples for analytical confirmation of exposure concentrations were taken at the start, after 24 and 72 hours of exposure.
Growth rate and yield inhibition increased with increasing loading rate of BEROL 1872 from 0.010 mg/L upwards, resulting in 96% growth rate and 100% yield inhibition at the highest test concentration. Statistically significant growth rate and yield inhibition was found at all test concentrations. However, inhibition of growth rate at loading rates of 0.010 and 0.032 mg/L was below 10% and therefore considered to be biologically not relevant. Samples taken from all test concentrations and the control were analysed by following a fragment ion (m/z 88.8) obtained from the most abundant ion trace (m/z 420.3). The measured concentrations increased with increasing loading rate throughout the test, which indicates proper preparation of the test solutions. In addition, the analysed fraction of the test item correlated to the recorded effects on algal growth. Since the test item is a UVCB, effect parameters were expressed in terms of loading rates and Time Weighted Average (TWA) concentrations, which are indicative for the analytically monitored fraction of the test item. The study met the acceptability criteria prescribed by the study plan and was considered valid.
The effect parameters (based on loading rates) obtained in this study are summarized in the table below.
cl – confidence limit, * based on statistical significance,#based on biological relevance.
Table: Effect Parameters based on Time Weighted Average Concentrations (µg/L)
(a)Effect parameters are indicative and should be interpreted with caution as it is unknown if the analytically monitored constituent is responsible for the recorded effects; cl – confidence limit; * based on statistical significance; #based on biological relevance.
It should be noted that the CMC (used here as water solubility) reported is 310 mg/L and that it was decided to follow the WAF approach because the test item is a UVCB and was observed to be not completely soluble in test medium.
These observations in relation to the solubility appear to be contradictory and therefore as worst-case the effect data based on geometric mean measured values will be used in stead. The highest nominal WAF concentration of 1 mg/L is about a factor 310 below the measured water solubility and it is therefore assumed that the test substance was actually completely dissolved and that the measured concentration for the most abundant ion trace (m/z 420.3) is representative for the availability of the whole test item.
The 72h-ErC50 and ErC10 based time weighted average measured concentrations are 25 (19 - 35) and 3.3 (1.6 - 5.4) µg/L. These values will be used as key value for chemical safety assessment.
The 72h-EL50 of the substance for growth rate inhibition (ErL50) was 0.12 mg/L (95% confidence interval: 0.094 - 0.14 mg/L). The 72h-ErL10 was 0.033 mg/L (95% confidence interval: 0.019 - 0.045 mg/L). The 72h-NOEL was 0.032 mg/L.
The objective of the study was to evaluate BEROL 1872 for its ability to generate toxic effects in Raphidocelis subcapitata (formerly known asPseudokirchneriella subcapitata) during an exposure period of 72 hours and, if possible, to determine the NOELR, EL10, EL20 and EL50 for both inhibition of growth rate and inhibition of yield.
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