酪蛋白多肽的制备和色谱分离方法-聚合物分子工程国家重点试验室.PDF

2010 7 Vol. 28 No. 7 July 2010 Ch inese J ourna l of Chrom atog rap hy 637~ 643 研究论文 DOI: 10. 3724/SP. J. 1123. 2010. 00637 蔡焕新, 殷宝茹, 姚 萍* (, , 200433) : , (RPHPLC/ESIM S), , ; RPHPLC , ESIMS , , , ,, : ; ;; ;; : O658 : :2010 Preparation and high performance liquid chromatographic separation of casein peptides * C IHuanxin, YIN Baoru, Y O Ping (The Key Laboratory of Molecu lar E gi eer i g of Polym er of M i istry of Edu catio , Departm e t of Macrom olecu lar Scie ce, Fuda U iversity, Sha ghai 20043 3, Ch i a ) Abstract: preparative reversedphase high perform ance liquid chromatographyelectrospray ionizationm ass spectrometry (RPHPLC/ESIMS) method to obtain low cost casein peptides was developed. Casein was hydrolyzed by trypsin and the hydrolysate was analyzed by RP HPLC/ESIMS firstly. The gradient elution program in an analytical column was optmi ized to achieve a better separation. The effect of pH of the m obile phase on the separation was also investigated. Then, the optmi al analytical conditions were applied in a preparative column directly. In each loading of the hydrolysate, severalpure peptide fractionswere collected auto matically by the inductions of ultraviolet absorbance and mass spectrometry signals together. The process is smi ple and effective. The influence of the loading amount in a preparative column was investigated intensively. For the collection consisting of hydrophilic peptides that were poorly separated in the prmi ary process, secondary separation was performed. By