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Published on: March 14, 2013
Universal LC-MS method for minimized carryover in a discovery bioanalytical setting
John S Williams1, Stephanie H Donahue, Hong Gao
1Vertex Pharmaceuticals, Inc., 130 Waverley St, Cambridge, MA 02139, USA.
Bioanalysis
|May 23, 2012
Summary
Carryover in bioanalytical LC-MS methods can be reduced by optimizing column choice and elution gradients. A new 3-minute method effectively minimizes carryover for diverse new chemical entities in drug discovery.
Area of Science:
- Analytical Chemistry
- Mass Spectrometry
- Drug Discovery
Background:
- Carryover is a significant challenge for accurate quantitation in bioanalytical liquid chromatography-mass spectrometry (LC-MS).
- Carryover in drug discovery is often caused by multiple sources beyond the autosampler.
Purpose of the Study:
- To investigate and mitigate carryover issues in bioanalytical LC-MS.
- To develop a robust and rapid method for diverse new chemical entities.
Main Methods:
- Evaluated various columns, injector wash protocols, and elution gradient compositions.
- Systematically analyzed the impact of hardware and mobile phase conditions on carryover.
Main Results:
- Column chemistry and elution gradients significantly influenced carryover levels.
- Cycling mobile phases (high/low organic) during column washing was more effective than a continuous high-organic wash.
- Different columns exhibited distinct carryover behaviors.
Conclusions:
- Optimizing column selection and gradient elution is crucial for minimizing LC-MS carryover.
- A novel 3-minute LC-MS method was developed, integrating column, gradient, and autosampler configurations.
- This method demonstrates robustness and minimizes carryover for a wide range of new chemical entities in drug discovery settings.
