1 research outputs found
High-Throughput and Sensitive Quantitation of Plasma Catecholamines by Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Using a Solid Phase Microwell Extraction Plate
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)