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Updated: May 6, 2026
15:35
Analyzing Large Protein Complexes by Structural Mass Spectrometry
Published on: June 19, 2010
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Complete large-molecule high-resolution mass spectra from 50-femtomole microvolume injection
N L Kelleher1, M W Senko, D P Little
1Department of Chemistry, Cornell University, Ithaca, New York, USA.
Journal of the American Society for Mass Spectrometry
|November 12, 2013
Summary
Direct injection into Fourier-transform mass spectrometry (FTMS) provides high signal-to-noise ratios. Gated trapping significantly reduces sample consumption for FTMS analysis.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Spectroscopy
Background:
- Electrospray ionization (ESI) is a crucial technique for mass spectrometry.
- Fourier-transform mass spectrometry (FTMS) offers high resolution and mass accuracy.
- Efficient sample introduction is vital for sensitive analysis in FTMS.
Purpose of the Study:
- To evaluate direct injection for ESI-FTMS.
- To assess the performance of gated trapping for ESI-FTMS.
- To determine the minimum sample quantity required for FTMS analysis.
Main Methods:
- Direct injection of ubiquitin into the ESI solvent stream.
- Utilizing a microvolume sample valve for precise delivery.
- Implementing gated ion trapping for enhanced sensitivity.
Main Results:
- Direct injection of 5×10(-14) mol ubiquitin yielded a spectrum with 85:1 S/N, 2-ppm mass accuracy, and isotopic resolution.
- Gated trapping consumed only 5×10(-18) mol of ubiquitin.
- Gated trapping produced a spectrum with 23:1 S/N and a resolving power of ~3×10(5).
Conclusions:
- Direct injection is effective for sensitive ESI-FTMS analysis.
- Gated trapping dramatically reduces sample requirements for FTMS.
- This method enables high-performance mass spectrometry with minimal sample.
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