Development of Two-Dimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Extracted Proteins of Rice Leaves: Hydrophilic Interaction-Reversed-phase Approach
文献类型: 外文期刊
第一作者: Gao Huan-Huan
作者: Gao Huan-Huan;Chen Ming-Xue;Ma You-Ning;Lin Xiao-Yan;Chai Shuang-Shuang;Qin Mei-Ling;Zhang Han-Tong;He Qiao;Chen Ming-Xue
作者机构:
关键词: Hydrophilic interaction-reversed-phase two-dimensional liquid chromatography; Protein; Rice leaves; Nanoflow reversed-phase liquid chromatography-tandem mass spectrometry; Orthogonality
期刊名称:CHINESE JOURNAL OF ANALYTICAL CHEMISTRY ( 影响因子:1.134; 五年影响因子:0.909 )
ISSN: 0253-3820
年卷期: 2018 年 46 卷 5 期
页码:
收录情况: SCI
摘要: A two-dimensional liquid chromatography method was developed for the analysis of rice leaves proteomics based on the coupling of hydrophilic interaction liquid chromatography-reversed-phase liquid chromatography with online tandem mass spectrometry. The influence of pH value of chromatographic mobile phase on the orthogonality of the hydrophilic interaction-reversed-phase two-dimensional liquid chromatography was evaluated by the changes of standard peptide retention. The results indicated that the better orthogonality (R-2 = 0.34113) was achieved from the system with hydrophilic interaction columns (pH 9.3) in the first and C-18 columns (pH 3.3) in the second LC dimension. Coupled with multiple fraction concatenation strategy, the orthogonality of two-dimensional liquid chromatography was further evaluated in the analysis of complex rice leaf proteins. The results showed that more than 50% of the total peptides were identified less than two times, and the peptides obtained from first-dimension were well distributed across the elution window, indicating that the method showed significant orthogonality in the identification of complex rice leaf proteins. Based on the proteome discoverer software, 207345 peptides belonged to 2930 protein clusters were identified.
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