Ezchrom elite software

MDA was determined according to a previously described HPLC method after derivatization with 2,4-dinitrophenylhydrazine (DNPH) [30 (link)]. Protein-bound MDA was hydrolyzed and deproteinized as described previously [25 (link)]. The supernatant was mixed with 12.5 µL DNPH solution and injected into the HPLC system (injection volume: 40 µL). The MDA standard was prepared as previously described [31 (link),32 (link)]. The DNPH derivatives (hydrazones) were isocratically separated, and the utilized HPLC consisted of an L-2200 autosampler, L-2130 HTA pump, and L-2450 diode array detector (all: VWR Hitachi; Vienna; Austria). Detector signals (absorbance at 310 nm) were recorded, and the EZchrom Elite software (VWR) was used for data acquisition and analysis.

In order to determine the soluble fraction of the investigated compound in biological media, HPLC–DAD analysis was performed. VWR Hitachi HPLC–DAD systems (containing the L-2130 HTA-pump, L-2130 degasser, L-2200 autosampler, L-2300 column oven, L-2450 diode array-detector and the EZChrom Elite software, VWR, Darmstadt, Germany) was used. The separation was performed on Gemini C₁₈ column (150 × 4.6 mm I.D., 5 μm pore size, Phenomenex Inc., Torrance, CA). A mixture of A–ACN and B–H₂O was used as a mobile phase (gradient conditions, 30 % A in 0 min, 60 % A in 5 min and 30 % A in 10 min). The injection volume of the sample was 50 μL. A detection wavelength of 300 nm was used for the quantification. The analysis time was 15 min.
The concentrations of stock solution of FLU and FEN in media solutions used in four ecotoxicity tests were as follows: 1 mg L⁻¹ for L. minor and S. vacuolatus (the solution consisted 0.2 % DMSO), 0.05 mg L⁻¹ (0.01 % DMSO) for FEN and 0.075 mg L⁻¹ (0.015 % DMSO) for FLU for D. magna and 0.3 mg L⁻¹ for V. fischeri (0.06 % DMSO). In order to calculate the bioavailability, the peak area of the signals obtained from the analysis of the above-mentioned mixtures were compared to the peak area of the signals obtained from the analysis of the same concentrations of FLU and FEN prepared in ACN.