Quantitative analysis of a Glyburide Analogue, a potential NLRP3 Inhibitor, using micro- sampling, hybrid solid phase extraction and Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)

Introduction: Recently, dried blood spot (DBS) sampling method has turned out to be a promising microsampling technique for use in preclinical studies of animals and humans in order to obtain matrix samples. With this technique, small-molecule PK characterization with LC-MS/MS has become simpler and faster. Blood sample volumes as low as 15-20 ul containing the drug are collected and applied to specialty paper, dried and then punched for analyte extraction for LCMS/MS quantification. In comparison with the standard blood collection methods used for bioanalysis, DBS sampling method has low sample volume requirements in addition to easier sample collection, handling, and storage (1). A LC-MS/MS method using DBS as sampling technique is under way in our lab for quantification of (5-chloro-2-methoxy-N-[2-(4sulfamoylphenyl) ethyl] benzamide, a novel small-molecule therapeutic drug candidate for inhibition of the NLRP3 inflammasome.