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G005819

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

application for HPLC, application for SPE

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About This Item

技术

HPLC: suitable
solid phase extraction (SPE): suitable

试验参数

sample preparation: SPE (Solid Phaes Extraction)
sample: rat plasma was spiled directly from stock standard to a level of 400 ng/mL
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)
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)
column temp.: 35 °C
detector: ESI(+), full scan, m/z 100-1000
injection: 0.5 μL
sample: plasma extract, analyte concentration of final sample work-up is equivalent to 100 ng/mL

适用性

application for HPLC
application for SPE

应用

clinical

分析说明

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.

法律信息

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany

分析物

说明
    Amiodarone hydrochloride solution, 1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®
    N-Desethylamiodarone hydrochloride solution, 1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®
    Lidocaine solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
    (±)-Flecainide solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

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