中文摘要：

目的：建立离线的二维液相色谱系统并分析珠蛋白的水解产物。方法：样品首先经第一维Hypersil SCX强阳离子交换色谱（100mm×4．6mm）分离，在pH4．0磷酸盐缓冲体系（含20％乙腈）条件下以0．5mL·min。的流速线性梯度增加盐浓度的方式洗脱。每2min切割收集一次洗脱产物再真空浓缩至0．1mL，共得到58个切割馏分。对每一部分取20止再进样到Hypersil BDS C18（3μm，50mm×2．1mm）反相色谱柱，在0．2mL·min^-1的流速下线性梯度增加乙腈浓度进行洗脱。结果：构建的离线二维液相色谱系统分辨率增加，系统峰容量将近10000，实际出峰数量也大大增加，进一步降低了样品分析的复杂程度。结论：离线二维液相色谱是复杂生物样品分离分析的强有力工具，该方法对生物活性肽的分离制备有一定的指导意义。

英文摘要：

Objective： To establish the system of off - line two - dimensional liquid chromatography and analyze the complex peptides from globin peptic hydrolysate. Method： Strong cation exchange chromatography （ 100 mm × 4. 6 mm） was used as the 1 st dimension. After the sample was loaded on SCX column, the peptides were eluted with the phosphate buffers （ pH 4. 0） which contained 20% acetonitrile by linearly increasing salt concentrations at a flow rate of 0. 5 mL · min ^-1. The elutents were cut and collected every 2 min and then were concentrated to 0. 1 mL, so we obtained a total of 58 fractions. 20 μL of the fractions was reinject into the 2nd dimension of Hypersil BDS CIs reverse phase chromatography （3 μm, 50 mm ×2. 1 mm）. The peptides were eluted with linearly increasing acetonitrile concentrations at a flow rate of 0. 2 mL · min^ -1 Result： The resolution＂ of the off - line two - dimensional system was enhanced greatly and the peak capacity of the system reached nearly 10000. The number of the peaks of the peptides from globin peptic hydrolysate increased and the analysis complexity of the sample was decreased greatly by off - line two - dimensional liquid chromatography. Conclusion： Off - line two - dimensional liquid chromatography is the powerful technology in the analysis of complex samples, and the technology can be used as the guideline to separate and extract bioactive peptides.