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Staggered multistep elution solid‐phase extraction capillary electrophoresis/tandem mass spectrometry: a high‐throughput approach in protein analysis
Author(s) -
Lee WeiHan,
Wang CheWei,
Her GuorRong
Publication year - 2011
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.5091
Subject(s) - chemistry , chromatography , elution , solid phase extraction , capillary electrophoresis , tandem mass spectrometry , mass spectrometry , capillary electrophoresis–mass spectrometry , electropherogram , analytical chemistry (journal) , electrospray ionization
An approach based on staggered multistep elution solid‐phase extraction (SPE) capillary electrophoresis/tandem mass spectrometry (CE/MS/MS) was developed in the analysis of digested protein mixtures. On‐line coupling of SPE with CE/MS was achieved using a two‐leveled two‐cross polydimethylsiloxane (PDMS)‐based interface. Multistep elution SPE was used prior to CE to provide an additional dimension of separation, thus extending the separation capacity for the peptide mixture analysis. By decreasing in the number of co‐eluting peptides, problems stemming from ionization suppression and finite MS/MS duty cycle were reduced. As a result, sequence coverage increased significantly using multistep elution SPE‐CE/MS/MS compared to one‐step elution SPE‐CE/MS/MS in the analysis of a single protein tryptic digest (49% vs. 18%) and a six protein tryptic digest (22–71% vs. 10–44%). A staggered CE method was incorporated to increase the throughput. The electropherograms of consecutive CE runs were partially overlapped by injecting the sample plug at a fixed time interval. With the use of a 5 min injection interval, slightly poor results were obtained in comparison with the sequential CE method while the total analysis time was reduced to 28%. Copyright © 2011 John Wiley & Sons, Ltd.