Abstract
We have investigated various sample chromatographic extraction and sample preparation methods for liquid chromatography mass spectrometry analysis in order to increase the throughput of various in vivo and in vitro assays in support of drug discovery. The results indicated that direct plasma injection, although certainly faster than conventional protein precipitation for sample preparation, had problems associated with column longevity and overall robustness. Frequently a single study could not be completed without column replacement. On-line solid phase extraction, on the other hand, compared well with off-line solid phase extraction, using our LC extraction column design, as contamination of the extraction column was minimized by back flushing using the Gilson syringe pump. Finally, on-line solid phase extraction for support of Caco-2 permeability studies worked very well for both single components and mixtures as the matrix was much simpler, presenting fewer contamination problems.
Current Topics in Medicinal Chemistry
Title: The Use of On-line and Off-line Chromatographic Extraction Techniques coupled with Mass Spectrometry for Support of In Vivo and In Vitro Assays in Drug Discovery
Volume: 1 Issue: 5
Author(s): John A. Masucci, Gary W. Caldwell, William J. Jones, Stephen J. Juzwin, Patrick J. Sasso and Mary Evangelisto
Affiliation:
Abstract: We have investigated various sample chromatographic extraction and sample preparation methods for liquid chromatography mass spectrometry analysis in order to increase the throughput of various in vivo and in vitro assays in support of drug discovery. The results indicated that direct plasma injection, although certainly faster than conventional protein precipitation for sample preparation, had problems associated with column longevity and overall robustness. Frequently a single study could not be completed without column replacement. On-line solid phase extraction, on the other hand, compared well with off-line solid phase extraction, using our LC extraction column design, as contamination of the extraction column was minimized by back flushing using the Gilson syringe pump. Finally, on-line solid phase extraction for support of Caco-2 permeability studies worked very well for both single components and mixtures as the matrix was much simpler, presenting fewer contamination problems.
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Cite this article as:
John A. Masucci , Gary W. Caldwell , William J. Jones , Stephen J. Juzwin , Patrick J. Sasso and Mary Evangelisto , The Use of On-line and Off-line Chromatographic Extraction Techniques coupled with Mass Spectrometry for Support of In Vivo and In Vitro Assays in Drug Discovery, Current Topics in Medicinal Chemistry 2001; 1 (5) . https://dx.doi.org/10.2174/1568026013394985
DOI https://dx.doi.org/10.2174/1568026013394985 |
Print ISSN 1568-0266 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4294 |
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