Determination of lercanidipine in human plasma by an improved UPLC-MS/MS method for a bioequivalence study

Abstract

An improved and reliable ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for the determination of lercanidipine in human plasma. Plasma samples with lercanidipine-d3 as an internal standard (IS) were prepared by solid phase extraction on Phenomenex Strata-X cartridges using 100 µL of human plasma. Chromatographic analysis was performed on UPLC BEH C₁₈ (50 mm×2.1 mm, 1.7 µm) column under isocratic conditions. Linear calibration curves were obtained over a wide dynamic concentration range of 0.010-20.0 ng/mL. Matrix effect was assessed by post-column infusion, post-extraction spiking and standard-line slope methods. The mean extraction recovery was >94% for the analyte and IS. Inter-batch and intra-batch precision (% CV) across five quality controls was <5.8%. Bioequivalence study was performed with 36 healthy subjects after oral administration of 10 mg of lercanidipine and the assay reproducibility was evaluated by reanalysis of 133 incurred samples.

Keywords: Bioequivalence; Human plasma; Lercanidipine; Solid phase extraction; UPLC–MS/MS.

Fig. 1

Representative MRM ion-chromatograms of (A) blank plasma with working solution of lercanidipine-d3 (m/z 615.2→283.1) (IS), (B) lercanidipine (m/z 612.2→280.1) at LLOQ and IS, and (C) lercanidipine in real subject sample at C_max and IS after administration of 10 mg dose of lercanidipine hydrochloride, LERCAN®.

Fig. 2

Injection of extracted blank human plasma during post-column infusion of (A) lercanidipine at ULOQ level and (B) lercanidipine-d3.

Fig. 3

Mean plasma concentration-time profile of lercanidipine after oral administration of 10 mg of LERCAN® and 10 mg of ZANIDIP® tablet formulations by 36 healthy Indian volunteers.