title: High-throughput and sensitive quantitation of plasma catecholamines by ultraperformance liquid chromatography-tandem mass spectrometry using a solid phase microwell extraction plate.
abstract: Plasma catecholamines provide a reliable biomarker of sympathetic activity. The low circulating concentrations of catecholamines and analytical interferences require tedious sample preparation and long chromatographic runs to ensure their accurate quantification by HPLC with electrochemical detection. Published or commercially available methods relying on solid phase extraction technology lack sensitivity or require derivatization of catecholamine by hazardous reagents prior to tandem mass spectrometry (MS) analysis. Here, we manufactured a novel 96-well microplate device specifically designed to extract plasma catecholamines prior to their quantification by a new and highly sensitive ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Processing time, which included sample purification on activated aluminum oxide and elution, is less than 1 h per 96-well microplate. The UPLC-MS/MS analysis run time is 2.0 min per sample. This UPLC-MS/MS method does not require a derivatization step, reduces the turnaround time by 10-fold compared to conventional methods used for routine application, and allows catecholamine quantification in reduced plasma sample volumes (50-250 μL, e.g., from children and mice).
authors: Service de Biomédecine, Laboratoire des Catécholamines et Peptides, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
authors: Marielle
authors: M
authors: Dunand
authors: Danilo
authors: D
authors: Gubian
authors: Maxime
authors: M
authors: Stauffer
authors: Karim
authors: K
authors: Abid
authors: Eric
authors: E
authors: Grouzmann
mesh_terms: D000328:Adult; D002395:Catecholamines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004867:Equipment Design; D005260:Female; D057166:High-Throughput Screening Assays / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D018401:Sample Size; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000295:instrumentation* / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D055815:Young Adult
chemical_list: D002395:Catecholamines