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Determination of the lipid profile in biological material by the method HPLC-ELSD
Vaindlová, Petra ; Pacáková, Věra (advisor) ; Feltl, Ladislav (referee)
A method of high-performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD) has been optimalized for the determination of neutral and polar lipids. Column filled by silica with chemically bonded diol has been used as stationary phase. As mobile phase, a ternary gradient composed from A: hexan-tetrahydrofuran 99:1 (v/v), B: isopropanol-chloroform-acetic acid 82:20:0,01 (v/v/v), C: isopropanol-water-triethylamine 47:47:6 (v/v/v) was used. Calibration curves have been measured in the range 2-200 μg of the injected amount; for individual lipid classes, optimal interlay of experimental data corresponded to the following functions: triacylglycerols - third order polynom (R=0,998), cholesterol esters - exponential dependence (R=0,998), free cholesterol - third order polynom (R=0.9998), ceramid - exponential dependence (R=0,992), cardiolipin - square dependence (R=0,998), phosphatidylethanolamine - exponential dependence (R=0,999), phosphatidylcholine - square dependence (R=0,997), phosphatidylserine - third order polynom (R=0,9985), sphingomyelin - third order polynom (R=0,9997), lysophosphatidylcholine - exponential dependence (R=0,9986). Analysis of the synthetic control sample showed recovery in the range of 82-95%. On the basis od these measurements, concentration of...
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