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Strategies to Identify and Confirm Phase I Metabolites of Glyburide Using the 4000 Q TRAP System

Purpose

In this application note, several different metabolite identification approaches are evaluated for their merits. A comprehensive examination of four Information Dependent Acquisition (IDA) survey modes was performed to illustrate data quality and types of metabolites found. A significant number of metabolites were found with each mode of operation to create a thorough investigation of the metabolites derived from glyburide.

Overview

A common strategy for metabolite analysis consists of using two mass spectrometry (MS) systemsa triple quadrupole MS system to identify metabolites, and a separate ion trap MS system to characterize the metabolites. The sample is split and run on the separate instruments in a sequential manner with the metabolite identification information from the triple quadrupole used to drive the characterization analysis on the 3D ion trap. Alternatively, methods have been proposed that split the HPLC flow into the two separate MS systems in an effort to get the information at the same time. Either way, the two separate systems approaches are not ideal. Triple quadrupole scan modes such as precursor ion and neutral loss provide a very selective method for the identification of structurally similar metabolites even in the presence of major background contaminants from complex biological matrices. With these selective scan modes, conventional triple quadrupoles can effectively identify metabolite candidates for further characterization analysis. Unfortunately, due to their inherent poor MS/MS full scan sensitivity, triple quadrupoles are not ideal tools to acquire MS/MS characterization data on critical low-level metabolites.

Also 3D ion trap systems alone cannot provide complete metabolite investigation. In this single-system approach, a single MS survey scan is commonly followed by a dependent MS/MS scan for structural characterization
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