An Automated High Performance Capillary Liquid Chromatography Fourier Transform Ion Cyclotron Resonance Mass Spectrometer for High-Throughput Proteomics

Journal Article · Sun Feb 01 04:00:00 EST 2004 · Journal of the American Society for Mass Spectrometry, 15(2):212-232

DOI:https://doi.org/10.1016/j.jasms.2003.09.008· OSTI ID:15010460

Belov, Mikhail E; Anderson, Gordon A; Wingerd, Mark A; Udseth, Harold R; Tang, Keqi; Prior, David C; Swanson, Kenneth R; Buschbach, Michael A; Strittmatter, Eric F; Moore, Ronald J; Smith, Richard D

We report on a fully automated 9.4 tesla Fourier transform ion resonance cyclotron (FTICR) mass spectrometer coupled to reverse-phase chromatography for high-throughput proteomic studies. Modifications made to the front-end of a commercial FTICR instrument – a dual-ESI-emitter ion source; dual-channel electrodynamic ion funnel; and collisional-cooling, selection and accumulation quadrupoles – significantly improved the sensitivity, dynamic range and mass measurement accuracy of the mass spectrometer. A high-pressure capillary liquid chromatography (LC) system was incorporated with an autosampler that enabled 24 h/day operation. A novel method for accumulating ions in the ICR cell was also developed. Unattended operation of the instrument revealed the exceptional reproducibility (1-5% deviation in elution times for peptides from a bacterial proteome), repeatability (10-20% deviation in detected abundances for peptides from the same aliquot analyzed a few weeks apart) and robustness (high-throughput operation for 5 months without downtime) of the LC/FTICR system. When combined with modulated-ion-energy gated trapping, the internal calibration of FTICR mass spectra decreased dispersion of mass measurement errors for peptide identifications in conjunction with high resolution capillary LC separations to < 5 ppm over a dynamic range for each spectrum of 10 3.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 15010460
• Report Number(s): PNNL-SA-38749; KP1601010
• Journal Information: Journal of the American Society for Mass Spectrometry, 15(2):212-232, Journal Name: Journal of the American Society for Mass Spectrometry, 15(2):212-232
• Country of Publication: United States
• Language: English

Related Subjects

43 PARTICLE ACCELERATORS