Bioanalytical method development and validation for estimation of finerenone in spiked human plasma using RP-HPLC technique and application of statistical tool.

ABSTRACT: A simple approach was developed for the rapid, selective and sensitive estimation of finerenone in spiked human plasma utilizing the high-performance liquid chromatography (HPLC) method. The protein precipitation technique was adopted for sample preparation, subsequently followed by separation and detection using the HPLC system. The quantification of finerenone in spiked human plasma was validated in compliance with regulatory guidelines (USFDA, EMEA and ICH). A mixture consisting of acetonitrile and water was utilized as the mobile phase. The separation was accomplished using a Phenomenex C18 column. The concentration range achieved was from 150 ng/ml to 6000 ng/ml. The accuracy obtained was 95 to 103% for calibration standards and 92 to 102 % for quality control (QC) samples. The precision was < 4% and recovery study showed 88% recovery. The stability study was conducted and the proposed method was determined to be stable. The application of one-way ANOVA revealed no significant variability in intra- and inter-day precision (p > 0.05), thereby confirming the consistency of the method. The method was effectively developed and validated, demonstrating simplicity, accuracy, rapidity, sensitivity and cost effectiveness.