G005855

LC/MS Analysis of Antiarrhythmic Drugs and Metabolites on Ascentis^® Express HILIC

application for HPLC

Properties

• technique(s): HPLC: suitable
• test parameters: 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), adjusted to pH 7.0 with formic acid; flow rate: 0.4 mL/min; pressure: 1305 psi (90 bar); column temp.: 35 °C; detector: ESI+, 100-1000 m/z; injection: 0.5 μL; sample: each compound, 300 ng/mL in 75:25 (1% formic acid acetonitrile:water)
• suitability: application for HPLC
• application(s): clinical

Description

Analysis Note

The basic nature of these antiarrhythmic agents makes them ideal candidates for HILIC (hydrophilic interaction) chromatographic separation. The benefit of HILIC over traditional reversed-phase chromatography is two-fold for both sample introduction and analyte detection. First, the high acetonitrile concentration of HILIC mobile phases allows for direct analysis of precipitated plasma samples without the need for additional sample solvent exchange. Second, the high acetonitrile content provides increased analyte response in positive ESI MS detection. Of the various HILIC-mode columns tested, method development for this assay determined the Ascentis Express HILIC, 2.7 μm particles, provided the best chromatographic resolution of the antiarrhythmic drugs while maintaining high peak efficiency for enhanced detection levels.

Legal Information

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

Analyte

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