Journal
CLINICA CHIMICA ACTA
Volume 437, Issue -, Pages 93-100Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.cca.2014.07.010
Keywords
Oxysterols; Dimethylaminobutyrate ester derivative; Liquid chromatography-tandem-mass spectrometry; Electrospray ionization analysis; Niemann-Pick C disease
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Two oxysterols, cholestan-3 beta,5 alpha,6 beta-triol (C-triol) and 7-ketocholesterol (7-KC), have been recently proposed as diagnostic markers of Niemann-Pick type C (NP-C) disease, representing a potential alternative diagnostic tool to the more invasive and time consuming filipin test in cultured fibroblasts. Usually, the oxysterols are detected and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using atmospheric pressure chemical ionization (APCI) or electro-spray-ionization (ESI) sources, after a variety of derivatization procedures to enhance sensitivity. We developed a sensitive LC-MS/MS method to quantify the oxysterols in plasma as dimethylaminobutyrate ester, suitable for ESI analysis. This method, with an easy liquid-phase extraction and a short derivatization procedure, has been validated to demonstrate specificity, linearity, recovery, lowest limit of quantification, accuracy and precision. The assay was linear over a concentration range of 0.5-200 ng/mL for C-triol and 1.0-200 ng/mL for 7-KC. Intra-day and inter-day coefficients of variation (CV%) were <15% for both metabolites. Receiver operating characteristic analysis estimates that the area under curve was 0.998 for C-triol, and 0.972 for 7-KC, implying a significant discriminatory power for the method in this patient population of both oxysterols. In summary, our method provides a simple, rapid and non-invasive diagnostic tool for the biochemical diagnosis of NP-C disease. (C) 2014 Elsevier B.V. All rights reserved.
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