Optimizing an ultrafast generic high-performance liquid chromatography/tandem mass spectrometry method for faster discovery pharmacokinetic sample throughput
Article first published online: 16 SEP 2005
Copyright © 2005 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 19, Issue 20, pages 2905–2910, 30 October 2005
How to Cite
Dunn-Meynell, K. W., Wainhaus, S. and Korfmacher, W. A. (2005), Optimizing an ultrafast generic high-performance liquid chromatography/tandem mass spectrometry method for faster discovery pharmacokinetic sample throughput. Rapid Commun. Mass Spectrom., 19: 2905–2910. doi: 10.1002/rcm.2147
- Issue published online: 16 SEP 2005
- Article first published online: 16 SEP 2005
- Manuscript Revised: 15 AUG 2005
- Manuscript Accepted: 15 AUG 2005
- Manuscript Received: 6 JUL 2005
For higher throughput screening, where the number of new chemical entities (NCEs) to test is rapidly increasing, fast sample turnaround time is essential. In order to increase efficiency a generic high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method, with a cycle time of 85 s (42 injections/h), was created. This was accomplished through the use of a 1-min ballistic gradient and the optimization of the autosampler. The gradient was optimized by varying the organic mobile phase concentration and examining its ballistic characteristics with respect to matrix ion suppression and compound retention time. The autosampler time could be reduced by optimizing several parameters and by determining the source of most of the carryover in order to reduce the number of syringe and injector washes. Finally, the reliability of the new generic method is demonstrated by comparison of sample data with a standard 2-min linear gradient method that showed that the data sets were well correlated. For plasma AUC (ng.h/mL) of 28 NCEs, the regression line had a slope of 0.92 and the R2 was 0.929. The described method was found to be useful for both rat plasma and tissue samples. Copyright © 2005 John Wiley & Sons, Ltd.