A High-Performance Liquid Chromatography Assay Method for the Determination of Lidocaine in Human Serum.

ABSTRACT

Here we report on the development of a selective and sensitive high-performance liquid chromatographic method for the determination of lidocaine in human serum. The extraction of lidocaine and procainamide (internal standard) from serum (0.25 mL) was achieved using diethyl ether under alkaline conditions. After liquid-liquid extraction, the separation of analytes was accomplished using reverse phase extraction. The mobile phase, a combination of acetonitrile and monobasic potassium phosphate, was pumped isocratically through a C18 analytical column. The ultraviolet (UV) wavelength was at 277 nm for the internal standard, and subsequently changed to 210 for lidocaine. The assay exhibited excellent linearity (r² > 0.999) in peak response over the concentration ranges of 50-5000 ng/mL lidocaine HCl in human serum. The mean absolute recoveries for 50 and 1000 ng/mL lidocaine HCl in serum using the present extraction procedure were 93.9 and 80.42%, respectively. The intra- and inter-day coefficients of variation in the serum were <15% at the lowest, and <12% at other concentrations, and the percent error values were less than 9%. The method displayed a high caliber of sensitivity and selectivity for monitoring therapeutic concentrations of lidocaine in human serum.