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1、第 二 章 色谱分离技术的理论基础第一节 色谱法简介211 色谱法的历史将一滴含有混合色素的溶液滴在一块布或纸片上，随着溶液展开可以观察到一个个同心圆环出现，古代罗马人就是用这种简单方法来分析燃料与色素。大约在100多年前，德国化学家Runge对该方法做了重要改进，使其有更好的重现性和定量能力。这项技术后来就发展成了今天的纸上色谱技术。首先认识这种层析现象在分离分析方面有重要价值的是俄国植物学家M.C.，M.C.在1901年将植物色素石油醚提取液倒入一根装有颗粒碳酸钙的玻璃管中，以石油醚冲洗柱子，经过一段时间，在玻璃柱上出现了不同颜色的彩色环带，这些彩色环带就是各种色素分离后所显示的结果，这就
2、是最早的柱色谱分离。1903年3月21日M.C.在波兰华沙自然科学年会生物学会议上发表了“一种新型吸附现象及其在生化分析上的应用”的演讲。1906年在一篇论文中正式将这种方法命名为色谱法（Chromatography），它的意思就是彩色的环带。在1907年德国生物学会议上，M.C.第一次向人们公开展示了用色谱法提纯植物色素溶液的色谱图。最初，M.C.还不是著名的植物学家，其色谱论文又是以俄文发表的，因此色谱学并未引起人们的重视。20多年后，Kuhn和Lederer参考了M.C.的文章之后，用碳酸钙的玻璃柱成功地从蛋黄素中分离了植物叶黄素。Kuhn和Lederer研究成果证实了M.C.的方法可以
3、用来进行制备分离。但是在实验室中将色谱法真正作为一种分离分析工具始于三十年代，1938年出现了薄层色谱（Thin Layer Chromatography，TLC）。1940年 A. Tiselius 提出了吸附色谱法和电泳法，并因此获得了1948年的诺贝尔化学奖。1941年A. Tiselius又进一步提出发展了迎头、顶替、冲洗液相色谱技术。1941年英国科学家A. J. P. Martin 和R. L.M. Synge提出了分配色谱的概念，A. J. P. Martin 和R. L.M. Synge采用以水饱和的硅胶固定相，以含有乙醇的氯仿为流动相分离了乙氨基氨基酸。它们在文章中指出“采用
4、气体代替液体作为流动相的可能是存在的。”由于当时战争的原因，检测技术没有跟上，最后他们采用自动滴定仪检测了脂肪酸，创立了气-液色谱法，在1951年分析化学牛津分析化学会议上，James和Martin总结了他们的一系列研究成果。A. J. P. Martin 和R. L.M. Synge因此获得了1952年诺贝尔化学奖。A. J. P. Martin 和R. L.M. Synge所发展的气-液分配色谱中是以吸附为主的分配机理，而James和Martin发展了气-液溶解分配色谱，采用高沸点液体作为固定相，它弥补了吸附分配中吸附剂种类较少的缺陷，大大扩大了气相色谱的应用范围，因此James和Mart
5、in 1951年的文章在分析化学界产生了巨大的影响。特别是1958年Golay又提出了分离效能极高的毛细管气相色谱法。从此以后，气相色谱法得到了蓬勃的发展，导致了化学实验室技术上的一个史无前例的改革。液-固色谱是最先建立的色谱方法，但是早期的色谱柱多是用碳酸钙、硅胶、氧化铝装填的玻璃管，流动相靠重力作用自由流下，检测也是靠肉眼观察或离线分析。自创立液-固色谱以后的50多年里，它并没有实质性进展。到60年代，气-液色谱理论的完善、发展指导人们制造出了高效微径填充剂，以及新的高压泵和检测器，大大提高了液相色谱的分离检测能力。于七十年代中后期出现了离子色谱、超临界流体色谱，八十年代后期出现了高速逆流
6、色谱，现已成为现代色谱领域分离纯化生物大分子和天然产物的重要技术之一。212分类 分析化学的一个重要的组成部分是与分离科学相联系的。就一般而论，分离可以看作是一个混合物至少被分成具有不同组成的两部分的操作过程。分离通常是通过物理的途径达到，但是在该过程中有时也可能包括化学反应，可以把实现分离所依据的物理或物理化学现象分成两大类：平衡过程和速度过程。平衡过程是依据被分离的各组分之间的平衡性质的差异而实现彼此分离，如蒸馏过程、沉淀分离等。速度过程分离技术是依据组分间动力学性质的不同而实现彼此分离，如电泳、离心、热扩散、渗析、透析等。色谱法是一种以分配平衡为基础的分离方法。色谱分离体系包括两个相，一
7、个是固定的，一个是流动的，当两相作相对运动时，反复多次地利用混合物中所有组分分配平衡性质的差异使彼此得到分离。至今，色谱学已经发展成为包括有许多分支的重要的分离技术。色谱的种类很多，从不同的角度出发，有各种不同的分类方法。 (1)按流动相所处的物理状态分类，可以将色谱法分为气相色谱(GC)、液相色谱(LC)和超临界流体色谱(SFC)。气相色谱又可以分为气-固、气-液色谱，液相色谱可分为液-固和液-液色谱。 (2) 按固定相的形状分类，色谱法可分为柱色谱、纸色谱和薄层色谱。 (3) 按分离过程的物理化学现象分类，色谱法可分为吸附色谱、分配色谱、离子交换色谱和排阻色谱。213 色谱学文献 色谱工作
8、者或从事分离科学及相关科学的研究人员，应该了解色谱学的研究动态、前沿进展等。一般可以从查阅科技文献、参加学术会议等渠道解决。（1） 期刊杂志 中文色谱， 大连化物所，中国化学会； 综合性 J. Chromatography A, 新理论、方法 J. Chromatography B, 生物医学 Chromatographic Science 理论 High Resolution Chromatography 理论 Liquid Chromatography 综合 Biomedical Chromatography 生物医学 J Capillary Electophoresis 综合 Elect
9、rophoresis 综合 其他分析化学、药学、生物医学杂志均有报道。（2） 学术会议 国内国际上有各种类型的关于色谱和毛细管电泳方面的学术会议，国内的有： 北京国际分析测试及仪器展览会，每2年在北京召开一次，明年将召开第9次会议。 全国色谱学术报告会，每2年召开一次，明年将召开第十三次会议。全国毛细管电泳及相关微分离学术报告会，每3年召开一次，今年召开了第四次会议。地区色谱会议，2-3年召开一次。全国分析化学会议、有机及微量分析学术会议等在国际上有各种类型，不同规模的学术会议，下面是1989-1999年的境外会议情况，当然这还并不全面。International symposium on p
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