The use of liquid phase deposition prepared phosphonate grafted silica nanoparticle-deposited capillaries in the enrichment of phosphopeptides
✍ Scribed by Jian-Hong Wu; Yong Zhao; Ting Li; Cong Xu; Kuang Xiao; Yu-Qi Feng; Lin Guo
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 424 KB
- Volume
- 33
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
Abstract
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 Fe^3+^, Zr^4+^ or Ti^4+^. The enrichment capacities of three different OT‐IMAC capillary columns were compared by using tryptically digested α‐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‐Zr^4+^ capillary column can be used to selectively isolate phosphopeptides from real samples, and can enrich for β‐casein phosphopeptides from concentrations as low as 1.7×10^−9^ M.