title: 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.
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.
authors.affiliation: Medical University of Vienna, Department of Pediatrics and Adolescent Medicine, Währinger Gürtel 18-20, Vienna, Austria.
authors.forename: David C
authors.initials: DC
authors.lastname: Kasper
authors.forename: Joseph
authors.initials: J
authors.lastname: Herman
authors.forename: Victor R
authors.initials: VR
authors.lastname: De Jesus
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Mechtler
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Metz
authors.forename: Bori
authors.initials: B
authors.lastname: Shushan
mesh_terms: D001800:Blood Specimen Collection / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004796:Clinical Enzyme Tests / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D007231:Infant, Newborn; D016014:Linear Models; D016464:Lysosomal Storage Diseases / Q000097:blood*; D015997:Neonatal Screening / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*