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[用于生物基质中化合物分析与分离的不同液相色谱技术之间流动相的转移可能性]

[Transfer possibilities of the mobile phases between different liquid chromatographic techniques for the analysis and isolation of compounds of biological matrices].

作者信息

Nyiredy S

出版信息

Acta Pharm Hung. 1999 Jan;69(1):46-56.

Abstract

After the survey and characterisation of the solid/liquid chromatographic methods, the author summarized the features of overpressure layer chromatography; the disturbing zone and the multi-front effect as well as the elimination of their influence. In light of these effects, the strategy of the mobile phase transfer possibilities is demonstrated between the various analytical and preparative liquid chromatographic methods, with the OPLC playing a central role. The main point of this strategy is that the examination of biological matrices is always begun with unsaturated TLC chamber, in which the compounds to be separated are placed between the Rf values of 0.3 and 0.8. The optimized TLC mobile phase is transferred without changes to the OPLC technique where a prerun is applied. For separation of nonpolar compounds, the prerun can be performed with hexane; for separation of polar substances the prerun can be performed with any component of the mobile phase in which the components are unable to migrate. The selection of this solvent might be considered during optimization of the mobile phase. Using HPTLC chromatoplate and analytical OPLC technique, highly effective separation can be achieved. The scaling-up for the various preparative chromatographic systems can be performed on basis of the applied chromatographic circumstances. The dry-filled preparative (FC, LPLC, MPLC) columns can be equilibrated with the solvent used for the prerun in analytical OPLC, while in case of filling with slurry technique, the slurry has to be prepared using the same solvent as was used for the prerun of OPLC. The air bubbles can be eliminated in both cases by pumping over the appropriate quantity of the solvent used for prerun, afterwards the preparative separation can be started with the optimized unsaturated TLC mobile phase. The author deals separately with the mobile phase transfer possibilities between the different analytical and preparative planar (OPLC and RPC with various chamber types) chromatographic techniques, where the saturation of the vapour phase is the determinant. The strategy is demonstrated through various plant extracts. The mobile phase transfer possibilities between the different solid-liquid chromatographic methods are summarized in comprehensive figures. Figure 13 demonstrates the direct transfer possibilities, different lines show those applicable with various methods: dotted line stands for off-line and thin line for on-line methods, while the ones marked with thick line ensure the transfer of the optimized mobile phase without change between different solid-liquid planar and column chromatographic techniques in case of both--off-line and on-line--procedures.

摘要

在对固/液色谱方法进行调查和表征之后,作者总结了超压层色谱的特点;干扰区和多前沿效应以及消除它们影响的方法。鉴于这些效应,展示了在各种分析和制备液相色谱方法之间流动相转移可能性的策略,其中超压层色谱(OPLC)起着核心作用。该策略的要点是,对生物基质的检测总是从不饱和薄层色谱(TLC)展开缸开始,将要分离的化合物置于比移值(Rf)为0.3至0.8之间。优化后的TLC流动相无需改变即可转移至采用预运行的OPLC技术中。对于非极性化合物的分离,预运行可用己烷进行;对于极性物质的分离,预运行可用流动相中的任何一种在其中组分无法迁移的成分进行。在流动相优化过程中可考虑这种溶剂的选择。使用高效薄层色谱(HPTLC)色谱板和分析型OPLC技术,可实现高效分离。各种制备色谱系统的放大可根据所应用的色谱条件进行。干填充制备柱(FC、LPLC、MPLC)可用分析型OPLC中用于预运行的溶剂进行平衡,而对于采用匀浆技术填充的情况,匀浆必须使用与OPLC预运行所用相同的溶剂来制备。在这两种情况下,通过泵送适量用于预运行的溶剂可消除气泡,之后可用优化后的不饱和TLC流动相开始制备分离。作者分别探讨了不同分析和制备平面色谱技术(不同类型展开缸的OPLC和RPC)之间的流动相转移可能性,其中气相饱和度是决定性因素。该策略通过各种植物提取物进行了展示。不同固液色谱方法之间的流动相转移可能性在综合图表中进行了总结。图13展示了直接转移可能性,不同线条表示适用于不同方法的情况:虚线代表离线方法,细线代表在线方法,而粗线标记的那些确保在离线和在线程序两种情况下,优化后的流动相可在不同固液平面色谱和柱色谱技术之间无变化地转移。

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