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Quantitative Analysis of Monohydroxy Sterols and Vitamin E in Human Plasma Combining Derivatization and RP-UHPLC/MS/MS in Negative Ion Mode
Year: 2025
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Quantitative Analysis of Monohydroxy Sterols and Vitamin E in Human Plasma Combining Derivatization and RP-UHPLC/MS/MS in Negative Ion Mode Cholesterol plays a crucial role in mammals and it is a major precursor to other biomolecules like bile acids, oxysterols, and steroid hormones. Moreover, dysregulation of its biosynthesis contributes to cardiovascular diseases, Alzheimer’s disease, and cancer. The determination of sterol concentrations can help to understand metabolic alterations and pathogenesis of serious diseases. Mass spectrometry (MS) coupled with chromatography is the key approach for sterol identification and quantification. Although gas chromatography was once dominant, liquid chromatography (LC) has become the most preferred method. Due to low ionization efficiency and high in-source fragmentation, sterols often produce intense fragment ions while the molecular ion is weak or absent, resulting in low sensitivity for its detection. However, this problem can be solved by the derivatization of free hydroxy group. We have previously optimized derivatization method using 3-(chlorosulfonyl)benzoic acid (Cl-SBA), enabling detection of acylglycerols, free sterols, and prenols by reversed-phase (RP) LC and multiple reaction monitoring (MRM) transitions in negative - ion mode on a QqQ instrument. This approach combines RP-UHPLC/MS/MS and the use of specific fragmentation patterns, significantly reduces limits of detection, reaching 15 – 25 pmol/mL for free sterols in plasma. In this study, we focused on the quantitation of monohydroxy sterols and vitamin E, enabling significant reduction of analysis time. Quantitative Analysis; Sterols; Vitamin E; Human Plasma; Derivatization; RP-UHPLC/MS/MS

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