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Citation: GAO Huan-Huan, MA You-Ning, LIN Xiao-Yan, CHAI Shuang-Shuang, QIN Mei-Ling, ZHANG Han-Tong, HE Qiao, CHEN Ming-Xue. Development of Two-Dimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Extracted Proteins of Rice Leaves: Hydrophilic Interaction-Reversed-phase Approach[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(5): 650-657,734. doi: 10.11895/j.issn.0253-3820.171398 shu

Development of Two-Dimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Extracted Proteins of Rice Leaves: Hydrophilic Interaction-Reversed-phase Approach

¹ College of Chemical Engineering, Zhejiang University of Technology, Hangzhou 310014, China;
² Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China

Received Date: 6 November 2017
Revised Date: 16 March 2018

Fund Project: This work was supported by the National Natural Science Foundation of China (No.31100405)

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 (R²=0.34113) was achieved from the system with hydrophilic interaction columns (pH 9.3) in the first and C₁₈ 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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Development of Two-Dimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Extracted Proteins of Rice Leaves: Hydrophilic Interaction-Reversed-phase Approach

南开大学师唯/华北电力大学（保定）刘景维：二维配位聚合物中有序的亲锂冠醚位点用于无枝晶锂沉积

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通讯作者: 陈斌, bchen63@163.com