Journal
JOURNAL OF SEPARATION SCIENCE
Volume 33, Issue 12, Pages 1806-1815Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.201000029
Keywords
Enrichment; Liquid phase deposition; Matrix-assisted laser desorption ionization time of flight mass spectrometry; Phosphopeptide; Silica nanoparticles
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Funding
- National Science Fund for Distinguished Young Scholars [20625516]
- Science Fund for Creative Research Groups [20921062]
- National 973 project of China [2007CB914200, 2006CB504300, 2006CB910301]
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In our current work, we describe how open tubular-immobilized metal-ion affinity chromatography (OT-IMAC) capillary columns connected to a solid phase microextraction (in-tube SPME) device can be used for the enrichment of phosphopeptides. A phosphonate modified silica nanoparticle (NP)-deposited capillary was prepared by liquid phase deposition (LPD), and used for the immobilization of Fe3+, Zr4+ or Ti4+. The enrichment capacities of three different OT-IMAC capillary columns were compared by using tryptically digested a-casein as sample. The improved extraction efficiency in our technique was demonstrated by comparing to a directly modified capillary, and a comparison of phosphopeptide extraction from simple and complex samples was tested for both modes. Our results show that the NP-IMAC-Zr4+ capillary column can be used to selectively isolate phosphopeptides from real samples, and can enrich for beta-casein phosphopeptides from concentrations as low as 1.7 x 10(-9) M.
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