LC/MS (TOF) Analysis of Warfarin Enantiomers in Plasma on Astec® CHIROBIOTIC® V after SPE using HybridSPE®-Phospholipid
CONDITIONS
sample/matrix
Rat plasma stabilized with K2EDTA was acquired from Lampire Biological Laboratories, (Pipersville PA). (Plasma was spiked directly from stock standard to a level of 100 ng/mL in 3:1 (plasma:1% formic acid acetonitrile).)
SPE tube/cartridge
Pk1 50 mg/96-Well HybridSPE-Phospholipid Plate (575656-U)
condition
Apply 100 μL of plasma to plate, followed by 300 μL of 1% formic acid acetonitrile. Agitate via vortex for 4 min.
elution
Place on vacuum manifold and apply 10” Hg vacuum for 4 minutes. Collect filtrate and analyze directly.
column
CHIROBIOTIC V,10 cm x 4.6 mm I.D., 5 μm particles (11022AST)
mobile phase
[A] 0.1% formic acid water; [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 reference material grade standards provided reliable 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
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