Abstract
A segment flow protocol has been proposed for coupling nano-liquid chromatography (nano-LC) with mass spectrometry (MS) in peptide-based proteomics research. LC effluent was segmented by perfluorodecalin oil and stored as segments in sequence to carry out the following off-line MSn analysis. In this way, incomplete identification of co-eluted peptides resulting from the MS scan rate, relatively slower than the LC elution rate, was solved to some extent for on-line coupling LC-MSn analysis. The complex proteins from a sample of T. tengcongensis were successfully identified with one run using segment as the interface between nano-LC and MSn analysis, compared to the total identified proteins with six runs using routine on-line nano-LC-MSn analysis. This segment flow approach could eliminate detection loss and sample cross-contamination, providing a new method for peptide-based proteomic research, especially when the amount of sample is limited.
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