title: Simultaneous quantitation of sphingoid bases by UPLC-ESI-MS/MS with identical13C-encoded internal standards.
abstract: Free sphingoid bases (lysosphingolipids) of primary storage sphingolipids are increased in tissues and plasma of several sphingolipidoses. As shown earlier by us, sphingoid bases can be accurately quantified using UPLC-ESI-MS/MS, particularly in combination with identical13C-encoded internal standards. The feasibility of simultaneous quantitation of sphingoid bases in plasma specimens spiked with a mixture of such standards is here described. The sensitivity and linearity of detection is excellent for all examined sphingoid bases (sphingosine, sphinganine, hexosyl-sphingosine (glucosylsphingosine), hexosyl2-sphingosine (lactosylsphingosine), hexosyl3-sphingosine (globotriaosylsphingosine), phosphorylcholine-sphingosine) in the relevant concentration range and the measurements show very acceptable intra- and inter-assay variation (<10% average). Plasma samples of a series of male and female Gaucher Disease and Fabry Disease patients were analyzed with the multiplex assay. The obtained data compare well to those earlier determined for plasma globotriaosylsphingosine and glucosylsphingosine in GD and FD patients. The same approach can be also applied to measure sphingolipids in the same sample. Following extraction of sphingolipids from the same sample these can be converted to sphingoid bases by microwave exposure and subsequently quantified using13C-encoded internal standards.
authors.affiliation: Dept. Medical Biochemistry, Leiden Institute of Chemistry (LIC), Leiden University, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Mirzaian
authors.affiliation: Dept. Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Wisse
authors.affiliation: Dept. Medical Biochemistry, Leiden Institute of Chemistry (LIC), Leiden University, The Netherlands.
authors.forename: M J
authors.initials: MJ
authors.lastname: Ferraz
authors.affiliation: Dept. Biochemistry, Christian-Albrechts-Universität, Kiel, Germany.
authors.forename: A R A
authors.initials: ARA
authors.lastname: Marques
authors.affiliation: Newborn Screening Metabolism & Genetics Unit, Human Genetics Department, National Institute of Health Dr Ricardo Jorge, Porto, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Gaspar
authors.affiliation: Dept. Medical Biochemistry, Leiden Institute of Chemistry (LIC), Leiden University, The Netherlands.
authors.forename: S V
authors.initials: SV
authors.lastname: Oussoren
authors.affiliation: Dept. Medical Biochemistry, Leiden Institute of Chemistry (LIC), Leiden University, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Kytidou
authors.affiliation: Dept. Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, The Netherlands.
authors.forename: J D C
authors.initials: JDC
authors.lastname: Codée
authors.affiliation: Dept. Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, The Netherlands.
authors.forename: G
authors.initials: G
authors.lastname: van der Marel
authors.affiliation: Dept. Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, The Netherlands.
authors.forename: H S
authors.initials: HS
authors.lastname: Overkleeft
authors.affiliation: Dept. Medical Biochemistry, Leiden Institute of Chemistry (LIC), Leiden University, The Netherlands. Electronic address: j.m.f.g.aerts@lic.leidenuniv.nl.
authors.forename: J M
authors.initials: JM
authors.lastname: Aerts
mesh_terms: D002247:Carbon Isotopes / Q000592:standards; D002851:Chromatography, High Pressure Liquid; D000795:Fabry Disease / Q000097:blood; D005260:Female; D005776:Gaucher Disease / Q000097:blood; D006801:Humans; D008297:Male; D012015:Reference Standards; D013106:Sphingolipidoses / Q000097:blood*; D013107:Sphingolipids / Q000032:analysis* / Q000097:blood; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D013107:Sphingolipids
keywords: (13)C-encoded internal standards; Fabry disease; Gaucher disease; Mass spectrometry; Quantification; Sphingoid bases