Buspirone is an anxiolytic psychoactive drug of the azapirone chemical class, and is primarily used to treat generalized anxiety disorder (GAD). Buspirone is a common model compound for xenobiotic metabolism. Here we describe the application of a Spectral Similarity Correlation methodology (‘S Score’) to the analysis of data generated from a human microsomal incubation of Buspirone. Non-targeted data-dependent MSMS data was collected from a human microsomal incubate using a Shimadzu LCMS IT-TOF system. The 'S scores’were derived from the correlation of fragment ion and neutral loss species of unknown chromatographic peaks to those of Buspirone. This approach was unbiased and success was not influenced any user interaction.