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

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)
column temp. 35 C
mobile phase [A] 0.1% formic acid water; [B] acetonitrile, (75:25, A:B)
flow rate 1 ml/min
injection 2 µl
detector ESI+, 100-1000 m/z

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.
Categories Analytical Chromatography, SPE Applications, Clinical, Chiral Applications, anticoagulants, Anticoagulants, Clinical, Forensic & Toxicology, Therapeutic Drug Monitoring
Featured Industry Clinical
Pharmaceutical (small molecule)
Legal Information HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC, Warfarin is a trademark of Wisconsin Alumni Research Foundation, CHIROBIOTIC is a registered trademark of Sigma-Aldrich Co. LLC, Astec is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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