数字农科院2.0

Development Of Two-Dimensional Liquid Chromatography C.oupled With Tandem Mass S pectrometry For Identification Of Extracted Proteins Of Rice Leaves: Hydrophilic Interaction-Reversed-Phase Approach

文献类型： 外文期刊

作者： Gao Huan-Huan;Chen Ming-Xue;Chai Shuang-Shuang;He Qiao;Lin Xiao-Yan;Ma You-Ning;Qin Mei-Ling;Zhang Han-Tong;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

ISSN： 0253-3820

年卷期： 2018 年 46 卷 5 期

页码：

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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