LC/MS Analysis of Antiarrhythmic Drugs and Metabolites in Plasma on Ascentis® Express HILIC following Sample Prep using HybridSPE®-Phospholipid
Materials
analytical column
SPE tube or plate
standard
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®(±)-Flecainide solution
1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®Lidocaine solution
1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
rabbit plasma, unfiltered K2-EDTA, spiked with each compound at 100 ng/mL (3:1, plasma:1% formic acid in acetonitrile)
SPE tube/cartridge
HybridSPE-Phospholipid, 96-well plate (575656-U)
sample addition
to each well add 100 μL plasma, followed by a 300 μL of 1% formic acid in acetonitrile
elution
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), 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
Description
Analysis Note
This application shows the extraction and analysis of antiarrhythmic cardiac drugs in plasma samples using LC/MS/MS. HILIC chromatography on Ascentis Express along with a selective sample preparation using HybridSPE-Phospholipid provided sensitivity and resolution. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany