Development Validation and Application of Bioanalytical Method for Therapeutic Agent Employing Dried Matrix Microsampling Techniques

Abstract

Microsampling is a method used to collect small sample volumes (lt 50 and#956;L) from animals and humans to assess drug and chemical exposure in biological matrix. This technique can be used for routine tests or measuring concentrations of analytes, metabolites, biomarkers, and other endogenous molecules. Microsamples can be collected as dried or liquid matrix microsamples,with dried matrix samples using spotting-based techniques, dried matrix spots, or volumetric tip microsampling techniques, and liquid samples via capillary sampling or touch-activated newlinephlebotomy devices. Dried matrix spot samples are applied directly onto cellulose-based paper and dried, while volumetric absorption techniques provide accurate fixed volume sampling. newlineThe study aimed to develop a new application for dried microsampling techniques (Dried newlineBlood Spot (DBS) and Volumetric Absorptive Microsampling (VAMS)) to support the use of newlinemicrosampling in regulated bioanalysis. The project aimed to provide scientific data to newlinecompare and support the acceptance of DBS and VAMS microsampling as an alternative to newlineconventional sampling for drug discovery, animal and clinical pharmacokinetics, preclinical toxicokinetics, and therapeutic drug monitoring. newlineAn LC-MS/MS method for quantifying Lurasidone in plasma was developed and validated. A newlinestep-by-step optimization was performed to develop the best suitable, simple, and economic method. The analyte and internal standard were extracted from rat plasma using a liquid-liquid extraction method. The accuracy and precision results met the acceptance criteria. The optimum and consistent recovery was established using tertiary butyl methyl ether (TBME).The mobile phase was composed of acetonitrile, methanol, and water (10:70:20) with 0.1% newlineheptafluorobutyric acid (HFBA), an ion pairing agent. The detection was performed on tandem mass spectrometry (API 2000) in positive ion multiple reaction monitoring (MRM) mode. All parameters were validated, including selectivity, specificity, carryover effect, linearity, precision,