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介绍了一种基于小多肽库的高效液相色谱手性固定相(CSP)的制备.胰蛋白酶酶解牛血清白蛋白(BSA)的优化条件为:在pH8.2,37℃下,酶解6 h,所得到的小多肽库以最大截留分子量(MWCO)为6 000的超滤膜过滤,滤液为含分子量小于6000的多肽分子库,以羰基咪唑法将该小多肽库键合至氨基化硅胶上。与完整蛋白-CSP相比,该多肽-CSP拥有更高的多肽键合密度以及更好的稳定性。对流动相组成以及pH对保留和立体选择性的影响也进行了考察,在优化的条件下,D,L-色氨酸、D,L-酪氨酸以及手性药物R,S-盐酸苯海索在该新型多肽-CSP上均得到了很好的分离。
A high performance liquid chromatographic chiral stationary phase (CSP) based on small peptide library was introduced in this paper.The optimal conditions of tryptic digestion of bovine serum albumin (BSA) were as follows: enzymatic hydrolysis After 6 h, the obtained peptide library was filtered with an ultrafiltration membrane with a MWCO of 6 000. The filtrate was a molecular library containing peptides with a molecular weight of less than 6,000. The peptide library was bonded to amination by carbonyl imidazole On silica gel. Compared to the intact protein-CSP, the polypeptide-CSP possesses a higher polypeptide binding density and better stability. The effects of mobile phase composition and pH on retention and stereoselectivity were investigated. Under optimal conditions, D, L-tryptophan, D, L-tyrosine and chiral drugs R, S- Hassow was well separated on the novel polypeptide-CSP.