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LC/MS/MS Analysis of Warfarin™ in Plasma on Ascentis® Express C18 after SPE using HybridSPE®-Phospholipid

LC/MS/MS Analysis of Warfarin™ in Plasma on Ascentis® Express C18 after SPE using HybridSPE®-Phospholipid

Conditions

sample/matrix rabbit plasma, unfiltered K2-EDTA, spiked with warfarin at 100 ng/mL (3:1, plasma:1% formic acid in acetonitrile)
SPE tube/cartridge HybridSPE(R)-Phospholipid, 96-well plate (575656-U)
sample addition to each well add 100 µl plasma, followed by a 300 µl of 1% formic acid in acetonitrile, agitate on orbital shaker for 4 minutes
elution collect filtrate and anlyze directly
column Ascentis(R) Express C18, 10 cm x 2.1 mm I.D., 2.7 µm particles (53823-U)
column temp. 35 C
mobile phase [A] 5mM ammonium formate, pH 4.2 with formic acid; [B] 5mM ammonium formate in 95:5 acetonitrile:water, 50:50 (A:B)
flow rate 0.3 ml/min
injection 2 µl
detector ESI+, 100-1000 m/z

Description

Analysis Note Warfarin is a widely prescribed oral anticoagulant for treatment and prevention of thrombosis and thromboembolism. Here we show the LC/MS/MS analysis of warfarin in plasma samples by reversed-phase chromatography with effective sample prep to remove endogenous phospholipids using HybridSPE-Phospholipid. UHPLC analysis utilized an Ascentis Express C18 column. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.
Categories Analytical Chromatography, Clinical, SPE Applications, Anticoagulants, anticoagulants
Featured Industry Clinical
Pharmaceutical (small molecule)
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC, Warfarin is a trademark of Wisconsin Alumni Research Foundation
suitability application for HPLC

Materials

     
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