Pataj, Zoltan and Liebisch, Gerhard and Schmitz, Gerd and Matysik, Silke (2016) Quantification of oxysterols in human plasma and red blood cells by liquid chromatography high-resolution tandem mass spectrometry. JOURNAL OF CHROMATOGRAPHY A, 1439. pp. 82-88. ISSN 0021-9673, 1873-3778
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Oxysterols are important intermediates in numerous metabolic and catabolic pathways and their biological significance is also proven. The present paper describes a reliable and short liquid chromatography-high-resolution mass spectrometry method (LC-MS/HR-MS) for the quantification of 8 different oxysterols (24(S)-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, 4 beta-hydroxycholesterol, 7 alpha-hydroxycholesterol, 7 beta-hydroxycholesterol, 7-ketocholesterol and cholestan-3 beta,5 alpha,6 beta-triol) in human plasma and red blood cells. Oxysterols were extracted with iso-octane after saponification of esterified sterols. Due to the poor ionization efficiency of the target compounds in electrospray ionization (ESI) derivatization of the molecules has been performed with N,N-dimethylglycine (DMG). Within less than 8 min we were able to achieve baseline separation of the isobaric 24(S)-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, 4 beta-hydroxycholesterol, 7 alpha-hydroxycholesterol and 7 beta-hydroxycholesterol. Moreover, high mass resolution was advantageously applied to resolve quasi-isobaric interferences. The method was validated based on the recommendations of US Food and Drug Administration and the European Medicines Agency guidelines. Oxysterol concentrations were determined in human plasma and red blood cells from healthy volunteers. Furthermore, the applicability for clinical use has been proven by the analysis of oxysterols as biomarkers in Niemann-Pick type C or cerebrotendinous xanthomatosis patients. (C) 2015 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HPLC-ESI-MS/MS; CHOLESTEROL PRECURSORS; STEROL 27-HYDROXYLASE; OXIDATION-PRODUCTS; BIOLOGICAL SAMPLES; PLANT STEROLS; SERUM SAMPLES; C DISEASE; BIOSYNTHESIS; VALIDATION; HR-MS (high-resolution mass spectrometry); Orbitrap; Oxysterols; Human plasma; Red blood cells; Niemann-Pick type C |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 03 Apr 2019 10:23 |
| Last Modified: | 03 Apr 2019 10:23 |
| URI: | https://pred.uni-regensburg.de/id/eprint/3223 |
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