Quantification of Steroids and 25-OH Vitamin D3 in Dried Blood Spots and Serum Using Liquid Chromatography Tandem Mass Spectrometry

摘要

Steroids are important biomarkers for a number of clinical conditions. Most clinical assays for steroids are based on immunoassays which have problems quantifying steroids due to cross-reactivity problems. A comprehensive steroid panel is challenging due to the need to separate structurally related compounds in a rapid manner that is capable of quantifying low concentrations present in complex biological specimens. Most clinical assays are based on serum or plasma, however, dried blood spot (DBS) samples are convenlently obtained from a prick of the finger, or heel and spotted onto filter paper. The goal of this project is to develop a simple extraction method (without derivatization) that is capable of measuring a variety of clinically important steroids from DBS using LC-MS/MS. The following steroids: cortisol, 17-alpha hydroxy hydroxyl progesterone, progesterone, testosterone and mono-hydroxy metabolites of vitamin D3 are extracted from DBS using the liquid- liquid extraction method, prior to analysis with LC-MS/MS. A sensitive and well separated LC-MS/MS method was developed on a triple quadrupole mass spectrometer.