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Use of On-line Size Exclusion Chromatography (SEC) for Sample Clean-up in LC/MS Analyses

Chromatography and Mass Spectrometry Application Report

Tania A. Sasaki and Paul Shieh, Thermo Electron Corporation, San Jose, CA

Introduction
Use of LC/MSn has experienced a dramatic increase in applications throughout the drug discovery and development process, from high-throughput screening to ADME/Tox studies. The Finnigan LCQ series ion trap mass spectrometers meet the demands for rapid screening of large numbers of drug candidates and their metabolites because of their sensitivity, high selectivity of MSn, ease of use, and ability to tolerate complex biological matrices. Often, compounds analyzed for drug discovery are present in biological matrices, such as plasma and serum. The task of monitoring target drug compounds and/or metabolites in these matrices can present considerable challenges when trying to achieve optimum separation, identification, and detection. It is often necessary to incorporate a time-consuming sample clean-up step into the analytical method. However, because of the growing emphasis on rapid method development and analysis in drug discovery and development, it is desirable to minimize the number of steps and overall time required in the analytical method without compromising data quality.

Goal
This report demonstrates the use of SEC as a quick online clean-up step that improves both the ruggedness of LC/MS methods and the quality of data obtained. On-line SEC also decreases HPLC column degradation, adding to the robustness of the overall analytical method. Furthermore, on-line clean-up reduces sample preparation, which saves time and minimizes potential analyte loss.

Experimental
All analyses were performed using a Finnigan Surveyor MS Pump and Autosampler and a Finnigan LCQ Deca XP Plus mass spectrometer operated in both full-scan and
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