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G005827

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

application for HPLC

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technique(s)

HPLC: suitable

test parameters

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

suitability

application for HPLC

application(s)

clinical
pharmaceutical (small molecule)

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

Analyte

Description
    Warfarin solution, 1.0 mg/mL in acetonitrile, ampule of 1 mL, certified reference material, Cerilliant®

SPE tube or plate

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analytical column

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