重组人生长激素胰蛋白酶肽图谱的微径LCMS研究  

Tryptic Mapping of Reco m binant Hu m an Growth Horm one by Microbore Liquid Chro m atography Electrospray Ionization Mass Spectro m etry

作　　者：韩俊[1] 盛龙生[1] 杨仲元[2] 相秉仁[1] 安登魁[1] 

机构地区：[1]中国药科大学分析计算中心,南京210009 [2]广州市药品检验所,广州510160

出　　处：《中国药科大学学报》1999年第4期269-274,共6页Journal of China Pharmaceutical University

摘　　要：采用电喷雾离子化为接口的液相色谱 质谱联用技术测定了重组人生长激素（ｒｈ Ｇ Ｈ） 胰蛋白酶水解的肽图谱。以此为例，讨论了微径柱对质谱检测灵敏度的影响，运用柱后“ Ｔ Ｆ Ａｆｉｘ”修饰技术大大提高了质谱检测灵敏度。根据肽图谱的分析结果，验证了ｒｈ Ｇ Ｈ 的一级结构，并区分了ｒｈ Ｇ Ｈ 及其甲硫氨酰化产物，综合肽段检出率１００ ％ 。This paper dealt with the methodology of tryptic peptide mapping of recombinant human growth hormone(rh G H) by microbore liquid chromatographyelectrosprayionization mass spectrometry . The rh G Hsample was digestedusing bovine T P C K treated trypsin and isolated on two microbore C18 columns with internal diameter of 1 .0 m m and 2 .1 mm respectively . Using binary gradient elution mode ,the tryptic peptide maps were obtained with tandem U Vand E S I M Sdetection . Apostcolumn addition technique ,named as “ T F A fix”modification was used to improve greatly theratio of signal to noise of M Ssignals . The composition of “ T F A fix”was optimized as propanoic acid isopropanol(20∶80) . The mechanism of “ T F A fix”modification was explained . The influence of column internal diameter on the M Sdetection sensitivity was discussed based on the results . According to the C I D mass spectra extracted from M S T I Cchromatograms,the tryptic peptide fragments of rh G H were assigned and 100 % of the expected peptides detected .rh G H was definitely differentiated from its methionylated analog based on their peptide maps and mass spectra .

关 键 词：液相色谱 ESI 重组人生长激素 肽图谱 

分 类 号：R977.13[医药卫生—药品]