High‐Throughput and Eco‐friendly Supercritical Fluid Chromatography‐ Tandem Mass Spectrometry Assay With Single‐Step Acetone Precipitation and Post‐Column Makeup for Nitrendipine Quantification in Beagle Plasma

ABSTRACT

Nitrendipine (NTD), a BCS class II calcium channel blocker, poses challenges for conventional dissolution prediction. This study focused on developing a high‐throughput supercritical fluid chromatography‐tandem mass spectrometry (SFC‐MS/MS) assay to quantify NTD in beagle plasma and evaluating the biorelevance of a 0.2% sodium dodecyl sulfate (SDS) dissolution medium via in vitro‐in vivo correlation. Chromatographic separation utilized an Acquity UPC ² HSS C18 SB column with a mobile phase of CO ₂ and methanol containing 0.1% formic acid (FA), and post‐column methanol with 1% FA to enhance sensitivity. Positive multiple reaction monitoring mode was employed for detection. Plasma samples underwent a single‐step acetone precipitation process, followed by direct injection of the supernatant. The method exhibited linearity over a concentration range of 1–500 ng/mL, with acceptable precision, accuracy, extraction recovery, and matrix effect. The method achieved a total run time of 2.0 min, significantly minimizing organic solvent use compared to traditional liquid chromatography. Upon oral administration of three commercial tablet formulations to beagles, systemic exposure was observed in the order of R > A > B. This ranking aligns with the discriminatory dissolution profiles observed in 0.2% SDS, despite all formulations meeting current regulatory standards. The developed SFC‐MS/MS method is suitable for efficient bioanalysis of NTD. Furthermore, the 0.2% SDS dissolution medium appears to offer a more biorelevant surrogate for the quality control of NTD tablets.