细胞色素C肽图分析  

Peptide mapping analysis of cytochrome C

作　　者：刘莉莎[1] 王鑫钰 李菲菲[2] 郝苏丽[1] 范慧红[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]北京诺华制药有限公司.上海201210

出　　处：《药物分析杂志》2014年第11期2060-2065,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:对不同种属来源的细胞色素C原料及上市细胞色素C制剂进行肽图分析,建立细胞色素C的种属鉴别方法。方法:将细胞色素C用胰蛋白酶酶解处理后,采用高效液相色谱分析,色谱柱为Grace Vydac 208TP C8(5 mm,250 mm×4.6mm),流动相A为含0.1%三氟乙酸的水溶液,流动相B为含0.1%三氟乙酸的乙腈溶液,梯度洗脱,流速0.4 m L·min-1,检测波长214 nm。结果:马心源和鸡胸肉源的细胞色素C与猪心、牛心来源的细胞色素C肽图谱差别较大,可以明显区分开来,而猪心、牛心来源的细胞色素C肽图谱基本一致;制剂肽图显示均为猪心或牛心源细胞色素C。结论:法定标准规定细胞色素C应由猪心或牛心提取,建立的肽图分析方法能够将标准规定以外的其他来源的样品鉴别出来,可以作为细胞色素C的种属鉴别方法。Objective： To study crude cytochrome C from different species and commercial preparations of cytochrome C by peptide mapping analysis and establish a species identification method of cytochrome C. Methods： The cytochrome C was digested by trypsin and then assayed by HPLC. Grace Vydac 208 TP C8（ 5 mm,250 mm ×4. 6 mm） was used as the analysis column,water solution containing 0. 1% trifluoroacetic acid and acetonitrile solution containing 0. 1% trifluoroacetic acid were used as mobile phase A and B,respectively. The gradient elution process was performed. The flow rate was 0. 4 mL·min^-1,the detection wavelength was 214 nm. Results： The peptide maps of cytochrome C from equine heart and pigeon breast muscle were different with the peptide maps from pig heart and bovine heart. While the peptide map of cytochrome C from pig heart was similar with the peptide map from bovine heart. The peptide maps showed that the cytochrome C preparations were all from pig heart or bovine heart.Conclusion： According to the standard requirement,cytochrome C should be extracted from pig or bovine heart. The peptide mapping analysis can identify the samples from other sources except the standard requirement and is applicable for the species identification of cytochrome C.

关 键 词：细胞色素C 肽图分析 种属鉴别 胰蛋白酶 标准提高 

分 类 号：R917[医药卫生—药物分析学]