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A simple micro‐extraction plate assay for automated LC‐MS/MS analysis of human serum 25‐hydroxyvitamin D levels
Author(s) -
Geib Timon,
Meier Florian,
Schorr Pascal,
Lammert Frank,
Stokes Caroline S.,
Volmer Dietrich A.
Publication year - 2015
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.3522
Subject(s) - chromatography , chemistry , triple quadrupole mass spectrometer , extraction (chemistry) , mass spectrometry , electrospray , calibration , calibration curve , sample preparation , immunoassay , selected reaction monitoring , detection limit , tandem mass spectrometry , statistics , mathematics , biology , immunology , antibody
This short application note describes a simple and automated assay for determination of 25‐hydroxyvitamin D (25(OH)D) levels in very small volumes of human serum. It utilizes commercial 96‐well micro‐extraction plates with commercial 25(OH)D isotope calibration and quality control kits. Separation was achieved using a pentafluorophenyl liquid chromatography column followed by multiple reaction monitoring‐based quantification on an electrospray triple quadrupole mass spectrometer. Emphasis was placed on providing a simple assay that can be rapidly established in non‐specialized laboratories within days, without the need for laborious and time consuming sample preparation steps, advanced calibration or data acquisition routines. The analytical figures of merit obtained from this assay compared well to established assays. To demonstrate the applicability, the assay was applied to analysis of serum samples from patients with chronic liver diseases and compared to results from a routine clinical immunoassay. Copyright © 2015 John Wiley & Sons, Ltd.