Development of Two-Dimensional Liquid Chromatography Coupled with Tandem Mass Spectrometry for Identification of Extracted Proteins of Rice Leaves: Hydrophilic Interaction-Reversed-phase Approach
10.11895/j.issn.0253-3820.171398
- VernacularTitle:亲水作用-反相二维液相色谱串联质谱法鉴定水稻蛋白质
- Author:
Huan-Huan GAO
1
;
You-Ning MA
;
Xiao-Yan LIN
;
Shuang-Shuang CHAI
;
Mei-Ling QIN
;
Han-Tong ZHANG
;
Qiao HE
;
Ming-Xue CHEN
Author Information
1. 浙江工业大学化学工程学院
- Keywords:
Hydrophilic interaction-reversed-phase two-dimensional liquid chromatography;
Protein;
Rice leaves;
Nanoflow reversed-phase liquid chromatography-tandem mass spectrometry;
Orthogonality
- From:
Chinese Journal of Analytical Chemistry
2018;46(5):650-657
- CountryChina
- Language:Chinese
-
Abstract:
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 C18columns(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.
