LC/MS/MS Analysis of Warfarin Enantiomers in Plasma on Astec® CHIROBIOTIC® V 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-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 analyze directly
column
Astec CHIROBIOTIC® V, 10 cm x 4.6 mm I.D., 5 μm particles (11022AST)
mobile phase
[A] 0.1% formic acid (pH unadjusted); [B] acetonitrile, 75:25 (A:B)
flow rate
1 mL/min
column temp.
35 °C
detector
ESI+, 100-1000 m/z
injection
2 μL
Description
Analysis Note
The R- and S- forms of warfarin, a commonly prescribed anticoagulant, are cleared by the body via different pathways and have different pharmacokinetic and pharmacodynamic effects. An Astec CHIROBIOTIC V HPLC column provided rapid resolution of the enantiomers under conditions that are compatible with mass spectrometric (MS) detection. By using HybridSPE-Phospholipid for the sample prep, ion-suppressing phospholipids were removed for improved sensitivity and longer column lifetime. 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.
Legal Information
Astec is a registered trademark of Merck KGaA, Darmstadt, Germany
CHIROBIOTIC is a registered trademark of Sigma-Aldrich Co. LLC
HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany
null