G005849

LC/MS/MS Analysis of Warfarin Enantiomers in Plasma on Astec^® CHIROBIOTIC^® V after SPE using HybridSPE^®-Phospholipid

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: 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
• suitability: application for HPLC
• application(s): clinical; pharmaceutical (small molecule)

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

Analyte

(R)-(+)-Warfarin, ≥97% (HPLC)
(S)-(−)-Warfarin, ≥97% (HPLC)