著者

Ikuma KURODA Yukihiro SHINTANI Masanori MOTOKAWA Shuzo ABE Masahiro FURUNO

出版者

The Japan Society for Analytical Chemistry

雑誌

Analytical Sciences (ISSN:09106340)

巻号頁・発行日

vol.20, no.9, pp.1313-1319, 2004 (Released:2005-01-21)

参考文献数

14

被引用文献数

80 92

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A methodology of phosphopeptide-selective analysis coupled with column-switching HPLC utilizing titania as precolumn media is presented. Phosphopeptides were selectively enriched on titania packing within a protein/peptide mixture without any additional procedure, and analyzed by column-switching high-performance liquid chromatography. First, phospho-compounds were separated from complex mixtures by trapping them under acidic conditions on a titania packing, where non-phosphorylated compounds were effused out of the precolumn. Subsequently, phospho-compounds were desorbed from the titania column under a specific condition and analyzed. The behavior of phospho-compounds on a titania surface, especially adsorption/desorption, was precisely examined and optimized. A phosphoric buffer was successively employed for the elution of phosphopeptides on a titania surface by competition with the free phosphate group. From the successes of a selective concentration/analysis of phosphopeptides with column-switching HPLC with a titania precolumn, a novel phosphopeptide-selective RP-HPLC analysis has been shown to have an application possibility as a tool for phosphoproteomics.