LC/MS (TOF) Analysis of Antiarrhythmic Drugs and Metabolites on Ascentis® Express HILIC: Improvement in Analyte Response after Solid Phase Extraction (SPE) using HybridSPE®-Phospholipid

LC/MS (TOF) Analysis of Antiarrhythmic Drugs and Metabolites on Ascentis® Express HILIC: Improvement in Analyte Response after Solid Phase Extraction (SPE) using HybridSPE®-Phospholipid

Conditions

sample preparation SPE (Solid Phaes Extraction)
SPE tube/cartridge HybridSPE-Phospholipid 96-Well Plate, 50 mg/2 mL (575656-U)
sample addition 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 Ascentis Express HILIC, 10 cm x 2.1 mm I.D., 2.7 µm particles (53939-U)
column temp. 35 C
mobile phase [A] 5 mM ammonium formate; [B] 5 mM ammonium formate in acetonitrile; (5:95, A:B, pH 7.0 with formic acid)
flow rate 0.4 ml/min
pressure 1305 psi (90 bar)
sample rat plasma was spiled directly from stock standard to a level of 400 ng/mL
sample plasma extract, analyte concentration of final sample work-up is equivalent to 100 ng/mL
injection 0.5 µl
detector ESI(+), full scan, m/z 100-1000

Description

Analysis Note The basic characteristics of antiarrhythmic cardiac drugs and associated metabolites make them targets for HILIC chromatographic separation. HILIC mobile phases consist of a high composition of acetonitrile which facilitates the direct analysis of precipitated plasma samples without the need for additional sample solvent exchange. In most cases, the high organic mobile phase also increases analyte response with ESI(+) MS detection. This application used HybridSPE-Phospholipid sample prep to remove phospholipids and precipitated proteins prior to LC/MS analysis on Ascentis Express HILIC. These chromatograms compare the response with HybridSPE-Phospholipid vs. standard protein precipitation sample prep methods. The highest grade solvents provided clean, robust operation. Cerilliant CRMs provided reliable quantification.
Categories Analytical Chromatography, SPE Applications, Pharmaceutical Analysis, Clinical, Antiarrythmic, cardiac drugs, Antiarrhythmic drugs, Cardiac Drugs, Cardiac drugs, antiarrhythmic drugs
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for SPE, application for HPLC

Materials

     
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