Study on Fluorescence labeling and Determination of Polypeptide (PC2～PC6) by High Performance Liquid Chromatography
LI Jing-xi1, 3, GAO Li-jie1, 2, CAO Wei1, ZHENG Li1, CHEN Jun-hui1, XU Xiu-li2, WANG Xiao-ru1
1. Marine Ecology Research Center, First Institute of Oceanography of State Oceanic Administration, Qingdao 266061, China 2. School of Ocean Sciences, China University of Geosciences(Beijing)，Beijing 100083, China 3. College of Chemistry and Chemical Engineering of Ocean University of China, Qingdao 266100, China
Abstract：This study was based on the thiol groups(-SH) of PC2～PC6, which could be reacted with the Monobromobimane(mBBr), in order to get polypeptide derivatives with fluorescent signal. A new method was developed for measuring the Polypeptides by high performance liquid chromatography with fluorescence detector, then the chromatographic conditions of HPLC was optimized; meawhile the reaction proportion of PCs and mBBr was identified by Trap-MS. The results showed that, the reaction proportion of PCs and mBBr was 1∶1, the polypeptide derivatives had good stability; the five compounds separation was better, and the peak time focused on the 16.6～22.0 min; the linear correlation coefficient of PC2, PC3, PC4, PC5 and PC6 was >0.999 1, and the limits of quantification were 0.3, 0.05, 0.3, 0.5 and 0.8 mg·L-1 respectively, the recovery rate was 83.0%～102.0%; the method was reproducible, RSD<2%, this method for measuring the peptide compounds was rapid and accurate.
李景喜1, 3，高丽洁1, 2，曹 为1，郑 立1，陈军辉1，徐秀丽2，王小如1 . 荧光标记-高效液相色谱测定多肽(PC2～PC6)的研究 [J]. 光谱学与光谱分析, 2014, 34(12): 3301-3306.
LI Jing-xi1, 3, GAO Li-jie1, 2, CAO Wei1, ZHENG Li1, CHEN Jun-hui1, XU Xiu-li2, WANG Xiao-ru1 . Study on Fluorescence labeling and Determination of Polypeptide (PC2～PC6) by High Performance Liquid Chromatography. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2014, 34(12): 3301-3306.
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