The great prevalence of thin-layer chromatography over high-performance liquid chromatography is connected with the possibility of analyzing many samples in parallel. Therefore, the method is often used in screening and/or effect directed analysis to compare composition and chemical/biological properties of many samples in one run. It was already proved, that high performance thin-layer chromatography, in many cases, can replace high-performance liquid chromatography for quantitative analysis. The main aim of the paper is to show that simple thin-layer chromatography can also be used as a quantitative or at least as a semi-quantitative method, even when it concerns effect directed analysis e.g. direct bioautography. Chlorogenic acid content was measured in four methanol extracts of various green coffees and in one extract of black coffee using thin-layer chromatography with ultraviolet detection and thin-layer chromatography with effect directed detection. High-performance liquid chromatography was used as a reference method. Additionally, total contents of polyphenols and antioxidants were estimated using thin-layer chromatography or dot-blot on chromatography plates. These results were compared to spectrophotometric methods. It was proved that thin-layer chromatography can be used as a quantitative (using densitometry) or semi-quantitative method (using other detection methods including effect directed detection) as well as for estimating total antioxidants or polyphenols content.
K E Y W O R D Sdot-blot, effect directed analysis, high-performance liquid chromatography, quantitative analysis, thin-layer chromatography 1542
Planar chromatography is a very useful tool for analysis of wide range of different mixtures. Thanks to its possibility for rapid separation of large number of samples simultaneously, low solvent consumption and ability to analyse rough material allow to receive precise and reliable results in short time and low cost. Miniaturization of planar techniques brings a lot of advantages, such as shortening distance and time of chromatogram development, and further lowering of solvent consumption. Besides, it often allows to improve separation parameters and raise efficiency of chromatographic system. In this paper, ability of analysis of tropane alkaloids mixture from Datura Inoxia Mill. extract using conventional TLC technique with five micro TLC techniques (short distance TLC, HPTLC, UTLC, OPLC and ETLC) in maximally closed chromatographic conditions was compared in order to present abilities of micro TLC techniques in plant material analysis.
Thin layer techniques proved their usefulness in the analysis of plant extract material. Thanks to their low operation costs, high sample throughput and possibility to gather chromatographic data for the whole sample in a single act, they allowed to find, and in some cases identify, active ingredients often hidden in sophisticated plant extract matrices. It was also proved that the presence of a magnetic field changes the retention of some substances, and moreover changes the separation efficiency of chromatographic systems. In the presented experiment, retention, efficiency and separation abilities of TLC chromatographic systems for mixtures of alkaloids under the influence of magnetic field depending on inductivity of the magnetic field, the saturation of chromatographic chamber and quantity of chromatographed substances were investigated. The results obtained were tested on real plant extracts that revealed the ability of chromatography in a magnetic field for separation of ternary and quaternary alkaloids from Chelidonium majus L. Our experiments proved that the presence of magnetic field induction lines perpendicular to the chromatographic plate plane can influence the width and retention of chromatographed substances, and can be considered as a tool for separation adjustment of plant extracts containing ternary and quaternary alkaloids.
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