LC/MS/MS Analysis of Drugs in Plasma on Ascentis® Express C18 after Extraction using SPME LC Tips

LC/MS/MS Analysis of Drugs in Plasma on Ascentis® Express C18 after Extraction using SPME LC Tips

Conditions

sample preparation Solid Phase Microextraction
sample/matrix rat plasma stabilized with K2EDTA and spiked at 100 ng/mL with each analyte
desorption process acetonitrile, 10 minutes
extraction immersion with agitation, 30 minutes
SPME fiber SPME LC Tips, C18 chemistry (57234-U)
column Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 µm particles (53822-U)
column temp. 40 °C
mobile phase [A] 5 mM ammonium formate in water; [B] 5 mM ammonium formate in acetonitrile:water (90:10 )
gradient 5% to 70% B in 3 min, to 90%B in 0.1 min, held at 90%B for 0.9 min
flow rate 0.5 mL/min
pressure 4641 psi (320 bar)
injection 2 µL
detector MS/MS ESI+, MRM m/z

Description

Analysis Note Determination of free circulating drug is important in establishing the drug′s pharmacokinetic activity.  In most cases, drug-protein complexes are formed that affect the active level of circulating drugs. In this study, biocompatible SPME was used as a rapid means of determining drug-protein binding affinities from plasma. HPLC separation was on an Ascentis Express C18 column packed with Fused-Core particles.
Categories Analytical Chromatography, Drugs, miscellaneous
Featured Industry Clinical
Pharmaceutical (small molecule)
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS, application for SPME

Materials

     
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