Chinese Journal of Chromatography

Chinese Journal of Chromatography ›› 2013, Vol. 31 ›› Issue (8): 763-768.DOI: 10.3724/SP.J.1123.2013.03003

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Influence of pH value of mobile phase on phosphopeptide enrichment selectivity under hydrophilic interaction liquid chromatography mode by using Click OEG-CD matrix

ZHAO Yanyan1, GUO Zhimou2, LI Xiuling2, LIANG Xinmiao2   

  1. 1. College of Pharmacy, Dalian Medical University, Dalian 116044, China;
    2. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
  • Received:2013-03-01 Revised:2013-03-28 Online:2013-08-28 Published:2013-09-29
  • Contact: O658

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Abstract:

The digest of the standard proteinα-casein was used as the objective to investigate the influence of pH value of mobile phase on phosphopeptide enrichment selectivity by using Click OEG-CD matrix. Disodium phenyl phosphate was primarily selected as the model sample. The results indicated that phosphate group was difficultly to be ionized when the pH value of mobile phase was below its pKa value. The retention of disodium phenyl phosphate on the Click OEG-CD matrix was not based on ion exchange interaction, leading to the weak retention. The influence of pH value (2, 4 and 6) of mobile phase under hydrophilic interaction liquid chromatography (HILIC) mode on phosphopeptide enrichment selectivity was investigated taking the digest of α-casein as a sample. Phosphopeptides could not be enriched on the matrix with the pH value of 2; phosphopeptides could be enriched on the matrix and the elution window was narrow with the pH value of 4; phosphopeptides could be enriched on the matrix while the elution window was broad with the pH value of 6. According to the investigation, Click OEG-CD matrix could be better applied in phosphopeptide enrichment.

Key words: Click OEG-CD matrix, pH value of mobile phase, phosphopeptide

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