title: Liposomes as potential masking agents in sport doping. Part 1: analysis of phospholipids and sphingomyelins in drugs and biological fluids by aqueous normal-phase liquid chromatography-tandem mass spectrometry.
abstract: In the present work, aqueous normal-phase liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), in different acquisition modes, was employed for the direct analysis and profiling of nine phospholipid classes (phosphatidic acids, phosphatidylserines, phosphatidylethanolamines, lysophosphatidylethanolamines, phosphatidylglycerols, phosphatidylinositols, phosphatidylcholines, lysophosphatidylcholines, and sphingomyelins) in biological and pharmaceutical matrices. After chromatographic separation by a diol column, detection and elucidation of phospholipid and sphingomyelin classes and molecular species were performed by different scan acquisition modes. For screening analysis, molecular ions [M + H]+were detected in positive precursor ion scan of m/z 184 for the classes of phosphatidylcholines, lyso-phosphatidylcholines and sphingomyelins; while phosphatidylethanolamines and lyso-phosphatidylethanolamines were detected monitoring neutral loss scan of 141 Da; and phosphatidylserines detected using neutral loss scan of 184 Da. Molecular ions [M-H]-were instead acquired in negative precursor ion scan of m/z 153 for the classes of phosphatidic acids and phosphatidylglycerols; and of m/z 241 for the phosphatidylinositols. For the identification of the single molecular species, product ion scan mass spectra of the [M + HCOO]-ions for phosphatidylcholines and [M + H]+ions for the other phospholipids considered were determined for each class and compared with the fragmentation pattern of model phospholipid reference standard. By this approach, nearly 100 phospholipids and sphingomyelins were detected and identified. The optimized method was then used to characterize the phospholipid and sphingomyelin profiles in human plasma and urine samples and in two phospholipid-based pharmaceutical formulations, proving that it also allows to discriminate compounds of endogenous origin from those resulting from the intake of pharmaceutical products containing phospholipidic liposomes. Copyright © 2016 John Wiley & Sons, Ltd.
authors: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.
authors: Simone
authors: S
authors: Esposito
authors: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.|Dipartimento di Chimica e Tecnologie del Farmaco, 'Sapienza'- Università di Roma, Piazzale Aldo Moro, 5, 00185, Rome, Italy.
authors: Sonia
authors: S
authors: Colicchia
authors: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.
authors: Xavier
authors: X
authors: de la Torre
authors: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.
authors: Monica
authors: M
authors: Mazzarino
authors: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.|Dipartimento di Medicina Sperimentale, 'Sapienza' Università di Roma, Viale Regina Elena 324, 00161, Rome, Italy.
authors: Francesco
authors: F
authors: Botrè
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D004300:Doping in Sports; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008081:Liposomes / Q000032:analysis / Q000097:blood* / Q000652:urine*; D008297:Male; D004364:Pharmaceutical Preparations / Q000097:blood / Q000737:chemistry / Q000652:urine; D010743:Phospholipids / Q000032:analysis / Q000097:blood* / Q000652:urine*; D013109:Sphingomyelins / Q000032:analysis / Q000097:blood* / Q000652:urine*; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008081:Liposomes; D004364:Pharmaceutical Preparations; D010743:Phospholipids; D013109:Sphingomyelins
keywords: aqueous normal-phase liquid chromatography; doping masking agents; liposomes; phospholipids; sphingomyelins; tandem mass spectrometry