Citation: ZHAO Yanyan, GUO Zhimou, LI Xiuling, LIANG Xinmiao. Influence of pH value of mobile phase on phosphopeptide enrichment selectivity under hydrophilic interaction liquid chromatography mode by using Click OEG-CD matrix[J]. Chinese Journal of Chromatography, ;2013, 31(8): 763-768. doi: 10.3724/SP.J.1123.2013.03003 shu

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

1.
1. College of Pharmacy, Dalian Medical University, Dalian 116044, China;

Corresponding author: LI Xiuling,
Received Date: 1 March 2013
Available Online: 28 March 2013
Fund Project: 国家自然科学基金项目(21105100,21105007). (21105100,21105007)

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 pK_a 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.
- pH value of mobile phase,
- Click OEG-CD matrix,
- phosphopeptide
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