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

摘要

Lysozomal storage disorders are just beginning to be routinely screened using enzyme activity assaysinvolving dried blood spots and tandem mass spectrometry (MS/MS). This paper discusses some ofthe analytical challenges associated with published assays including complex sample preparationand potential interference from excess residual substrate. Solutions to these challenges are presentedin 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 introductionfor higher throughput required of a population screening assay. High sensitivity, specificity andthroughput were demonstrated using this novel method.