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Improving liquid chromatography-mass spectrometry sensitivity using a subambient pressure ionization with
Keqi Tang1, Jason S Page, Ioan Marginean
1Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Journal of the American Society for Mass Spectrometry
|September 29, 2011
Summary
Subambient pressure ionization with nanoelectrospray (SPIN) enhances liquid chromatography-mass spectrometry sensitivity for complex samples. This method offers a 5- to 12-fold improvement for peptide detection compared to conventional methods.
Area of Science:
- Analytical Chemistry
- Mass Spectrometry
- Chromatography
Background:
- Conventional nanoelectrospray ionization (ESI) can be limited in sensitivity for complex sample analysis.
- Optimizing ion source parameters is crucial for improving detection limits in liquid chromatography-mass spectrometry (LC-MS).
Purpose of the Study:
- To evaluate the compatibility and performance of the subambient pressure ionization with nanoelectrospray (SPIN) source for gradient reversed-phase LC-MS.
- To assess the sensitivity improvements offered by the SPIN source for complex biological samples.
Main Methods:
- Utilized a SPIN ion source operating at ~15-30 Torr with gradient reversed-phase liquid chromatography.
- Analyzed complex samples including a protein tryptic digest and a whole cell lysate.
- Employed low liquid chromatographic flow rates (100-400 nL/min) to ensure stable electrospray.
Main Results:
- Demonstrated successful application of SPIN for gradient LC-MS analysis of complex samples.
- Achieved stable electrospray at low flow rates, preventing electrical breakdown.
- Increased operating pressure in the SPIN source prevented solvent freezing and improved desolvation.
- Obtained a 5- to 12-fold increase in sensitivity for detected peptides compared to conventional atmospheric pressure nanoelectrospray ionization (ESI).
Conclusions:
- The SPIN ion source is a viable and highly sensitive interface for gradient reversed-phase LC-MS.
- SPIN technology significantly enhances peptide detection sensitivity, particularly for challenging complex samples.
- Optimized operating pressure in SPIN sources improves performance and overcomes limitations of conventional ESI.
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