Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

UHPLC/MS Determination of Oxylipins in Clinical Samples

Autoři: Chocholoušková Michaela | Jirásko Robert | Holčapek Michal

Rok: 2018

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	UHPLC/MS Determination of Oxylipins in Clinical Samples	Oxylipins, including eicosanoids, octadecanoids and docosanoids, comprise a class of bioactive lipids. They are generated from polyunsaturated fatty acids (PUFA), such as arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, by enzymatic or non-enzymatic oxidation. The enzymatic way is implemented due to the cyclooxygenase (COX), the lipooxygenase (LOX) and the cytochrome P450 enzymes. This enzymatic process forms a lot of oxylipins with similar structures, but different biological activity. Their concentrations are the lowest among all endogenous lipids in the human body, so their analysis, especially quantitation, is very demanding, but they are very important due to their roles in the inflammation and other diseases. Many oxylipins are related to ongoing inflammation and chronic diseases including cancer. An ultrahigh-performance liquid chromatography (UHPLC) coupled to electrospray ionization mass spectrometry using triple quadrupole analyzer is the best and the most useful for the analysis of oxylipins. Analytes are separated in reversed-phase mode, especially on octadecylsilica (C18) columns and metabolites are detected by selected reaction monitoring (SRM), because the SRM scan provides high sensitivity and specificity. The mass spectrometer is operated in the negative-ion mode due to the presence of carboxylic group. The final UHPLC/MS method was developed for analysis oxylipins in biological samples, especially in human plasma. The development of this method involves the preparation of samples by solid-phase extraction using C18 column, where deproteination is the most important step, also the optimization analytical conditions, such as collision energy of each precursor/product ion pairs used for SRM detection, and the full validation including calibration curve, recovery rate, accuracy, precision, matrix effect, selectivity and quality control with the selection of appropriate internal standards. A deuterated analog of the analyte is used as inte	UHPLC/MS; Oxylipins; Plasma