G005818

LC/MS (TOF) Analysis of Antiarrhythmic Drugs and Metabolites in Plasma on Ascentis^® Express HILIC after Sample Prep using HybridSPE^®-Phospholipid

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: sample preparation: SPE (Solid Phase Extraction); sample/matrix: rat plasma stabilized with K₂EDTA was spiked 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
• suitability: application for HPLC
• application(s): clinical

Description

Analysis Note

The basic characteristics of these compounds 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 facilitates increased analyte response in ESI(+) MS detections. This application used HybridSPE-Phospholipid sample prep to remove phospholipids and precipitated proteins prior to LC/MS analysis on an Ascentis Express HILIC Fused-Core HPLC column. The highest grade solvents provided clean, robust operation. Cerilliant CRMs provided reliable quantification. The resulting method was robust and sensitive.

Legal Information

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

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

Analyte

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^®