Non-aqueous capillary electrophoretic separation of cholesterol and 25-hydroxycholesterol after derivatization with Girard P reagent

0480519 - ÚIACH 2018 RIV IE eng J - Článek v odborném periodiku
Greguš, M. - Roberg-Larsen, H. - Lundanes, E. - Foret, František - Kubáň, Petr - Wilson, S.R.
Non-aqueous capillary electrophoretic separation of cholesterol and 25-hydroxycholesterol after derivatization with Girard P reagent.
Chemistry and Physics of Lipids. Roč. 207, OCT (2017), s. 87-91. ISSN 0009-3084. E-ISSN 1873-2941
Institucionální podpora: RVO:68081715
Klíčová slova: cholesterol * 25-hydroxycholesterol * UV detection
Obor OECD: Analytical chemistry
Impakt faktor: 2.766, rok: 2017

Capillary electrophoresis (CE) can provide high separation efficiency with very simple instrumentation, but has yet to be explored regarding oxysterols/cholesterol. Cholesterol and 25-hydroxycholesterol (both are 4-ene-3-ketosteroids) were quantitatively transformed into hydrazones using Girard P reagent after enzymatic oxidation by cholesterol oxidase. Separation was achieved using non-aqueous capillary electrophoresis with UV detection at 280 nm, the charge-tagging Girard P reagent ensured both charge and chromophore (which are requirements for CE-UV). Excess reagent was also separated from the two analytes, eliminating the need for removal prior to the analysis. The compounds were separated in less than 5 min with excellent separation efficiency, using separation electrolytes fully compatible with mass spectrometry. The CE-UV method was used to optimize steps for charge-tagging, revealing that the procedure is affected by the analyte/reagent ratio and reaction time, but also the analyte structure.
Trvalý link: http://hdl.handle.net/11104/0276290