This application note describes a method for determining the metabolic stability profile of buspirone using an Agilent 1200 Series Rapid Resolution LC/TOF system on a short Rapid Resolution High Throughput (RRHT) column. Simultaneously, the main metabolites are identified in the same experiment by the automated use of a long RRHT column, which provides the necessary high resolution.
The document details a method for analyzing drug metabolism where metabolic stability is assessed on a short Rapid Resolution column, while main metabolites are identified simultaneously using a long high-resolution column. This approach utilizes the Agilent 1200 Series binary pump SL, autosampler SL, and thermostatting capabilities to handle fast LC/MS analysis.
The study focuses on buspirone, demonstrating that approximately 80% of the drug is metabolized within 20 minutes under specific incubation conditions. The method employs an Agilent 6210 Series TOF mass spectrometer with ESI positive mode and dual sprayer interface to acquire accurate molecular masses for identifying hydroxy buspirone metabolites.
The reaction was carried out at 37 °C for 380 minutes.
The flow rate for the fast LC method is set to 1 mL/min.