反相液相色谱对多肽的分离、纯化与制备

用反相高效液相色谱（RPLC）对两种化学合成多肽－－32肽和21肽进行了分离、纯化和制备。在用分析型RPLC色谱柱对多肽样品的制备过程中，对其进样量和洗脱梯度进行了选择。每次进样量为5mg，在最优化色谱条件下，用RPLC一步就可对21肽进行分离纯化，其纯度达到98.6%。而32肽由于样品组分更加复杂，RPLC一步纯化后其纯度仅有80％。通过对色谱分离条件的再次优化，对32肽进行二次分离纯化，纯度达杂，RPLC一步纯化后其纯度仅有80％。通过对色谱分离条件的再次优化，对32肽进行二次分离纯化，纯度达到96.4%。在其最优化条件下，通过多次样品收集和冷冻干燥，分别制备了高纯度的21肽和32肽各100mg。

The Purification of Peptide in Preparative Scale with Reversed Phase High Performance Liquid Chromatography

Two kinds of synthesized peptides with 32 and 21 amino acid residues were purified in preparative scale with reversed phase high performance liquid chromatography (RPLC). With an analytical-type RPLC column, the sample size and chromatographic conditions were selected and optimized. The size was 5 mg each time. Under the optimum separation conditions, 21-peptide could be separated and purified with RPLC by one step to obtain the purity being 98.6%. However, the purity of 31-peptide was only about 80% by one step because of the presences of the more complex components in sample. With selecting the optimum separation condition, its purity was raised to 96.4% by re-purification. Under optimum separation conditions, 100 mg of the purified 21-peptied and 32 peptide were obtained through separation and collection of samples for many times and freeze-drying, respectively.