全二维液相色谱的初步构建及其在山羊血清分离中的应用  

Construction of Comprehensive Two-dimensional LiquidChromatography for the Analysis of Goat Serum

作　　者：王智聪[1,2] 张庆合[2] 张维冰[2] 李彤[2] 赵中一[1] 

机构地区：[1]中国地质大学材料与化学工程学院,武汉430074 [2]中国科学院大连化学物理研究所,大连依利特分析仪器有限公司,大连116011

出　　处：《分析化学》2005年第4期437-441,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金 (No. 20375040);"十五"国家科技攻关重大项目(No. 2001BA210A01)资助课题

摘　　要：以GFC/RP模式构建全二维液相色谱系统,第一维凝胶过滤色谱柱使用ShodexProteinKW 802. 5(300mm×8mmi.d. ),以0. 2mol/LNaH2PO4 (pH7. 0)的流动相在0. 15mL/min的流速下等度洗脱,第二维反相色谱柱使用HypersilBDSC18 (35mm×4. 6mmi.d. ),在3mL/min的流速下梯度洗脱。采用平行柱交替分析的形式作切换接口, 2. 5min切换一次,两个反相柱交替富集、分析第一维洗脱产物。以5个标准蛋白混合物的分离评价该系统,在单独一维模式中不能分离的样品在全二维液相色谱中得到了较好的分离,二维系统的总峰容量为225。与一维色谱相比,系统的总峰容量、分辨率得到较大提高。并用于山羊血清的纯化分析,对一维分离中的重合谱峰进行验证,对制备纯化有一定的实际意义。The system constructed here is a comprehensive two-dimensional liquid chromatography that uses gel filtration chromatography ( GFC) followed by reversed phase liquid chromatography ( RP). Column of Shodex Protein KW-802. 5 (300 mm x 8 mm i. d.) acts as the first dimension which was eluted with 0. 2 mol/L NaH2PO4 ( pH 7. 0) at a flow rate of 0. 15 mL/min. The second dimension uses the column of Hypersil BDS C-18 (35 mm x 4. 6 mm i. d.) which was operated in gradient form. at a flow rate of 3. 0 mL/min. The interface is a 2-position-10-port valve coupled with two parallel columns which analyzed elutes from GFC alternated. The system was evaluated by 5 standard proteins that cannot be-resolved in either GFC or RP mode alone but can be resolved by the comprehensive two-dimensional liquid chromatography. The total peak capacity of this system is calculated to be 225. Compared with one-dimensional chromatography, the total peak capacity and the resolution are enhanced. The system was used for separating goat serum to validate and isolate the overlap ingredients.

关 键 词：山羊血清 分离 应用 液相色谱系统 模式构建 凝胶过滤 梯度洗脱 二维系统 实际意义 色谱柱 一维 GFC min 流动相 BDS 反相柱 混合物 维模式 分辨率 流速 切换 容量 纯化 

分 类 号：S827[农业科学—畜牧学]