The application of multiplexed,multi‐dimensional ultra‐high‐performance liquid chromatography/tandem mass spectrometry to the high‐throughput screening of lysosomal storage disorders in newborn dried bloodspots |
| |
| Authors: | David C Kasper Joseph Herman Victor R De Jesus Thomas P Mechtler Thomas F Metz Bori Shushan |
| |
| Institution: | 1. Medical University of Vienna, Department of Pediatrics and Adolescent Medicine, W?hringer Gürtel 18–20, Vienna, Austria;2. Thermo Fisher Scientific, 101 Constitution Boulevard, Franklin, MA 02038, USA;3. Newborn Screening and Molecular Biology Branch, Centers for Disease Control and Prevention, 4770 Buford Highway, NE, Atlanta, GA 30341, USA;4. Clinical Mass Spec Consultants, 164 Glen Road, Toronto, ON, Canada M4W 2W6 |
| |
| Abstract: | Lysozomal storage disorders are just beginning to be routinely screened using enzyme activity assays involving dried blood spots and tandem mass spectrometry (MS/MS). This paper discusses some of the analytical challenges associated with published assays including complex sample preparation and potential interference from excess residual substrate. Solutions to these challenges are presented in the form of on‐line two‐dimensional chromatography to eliminate off‐line liquid‐liquid extraction (LLE) and solid‐phase extraction (SPE), the use of ultra‐high‐performance liquid chromatography (UHPLC) to separate excess substrate from all other analytes and multiplexed sample introduction for higher throughput required of a population screening assay. High sensitivity, specificity and throughput were demonstrated using this novel method. Copyright © 2010 John Wiley & Sons, Ltd. |
| |
| Keywords: | |
|
|
