Abstract
The richness of bioactive compounds in plant materials encourages continuous development of separation methods and bioassays for their isolation and identification. Thin-layer chromatography-direct bioautography links separation on the adsorbent layer with biological tests performed directly on it. Therefore, the method is very convenient for searching plant constituents with biological activity, such as antibiotics. Test bacteria grow directly on a plate surface excluding places where antibacterials are located. They can be detected with reagents converted by living bacteria. TLC-DB is a high throughput method enabling analyses of many samples in parallel and the comparison of their activity. Both screening and semi-quantitative analysis is possible. The targeted compounds can be identified using spectroscopic methods, mostly mass spectrometry, that can be performed directly on a TLC plate. This paper discusses all above mentioned aspects of TLC-DB, illustrating them with literature, schemes and our own results.
Highlights
The world of plants constitutes an almost unlimited source of biologically active substances which are or could be used in the treatment of many human, animal and plant diseases [1,2]
thin-layer chromatography (TLC) linked to biological assay or to chemical detection indicating biological properties (e.g., TLC-DPPH for testing radical scavenging properties) is generally call TLC-bioautography (TLC-B)
Thin-layer chromatography-direct bioautography (TLC-DB) is one of three variants of a TLC-B method in which separation, a given assay and visualization are performed directly on a TLC layer. This is used for testing antimicrobial properties directly on a TLC plate covered with a thin layer of bacteria or fungi, as opposed to a thick layer of inoculated agar in agar overlay bioautography [6,10,11]
Summary
The world of plants constitutes an almost unlimited source of biologically active substances which are or could be used in the treatment of many human, animal and plant diseases [1,2]. It is difficult to link this information with the information on biological (e.g., antibacterial) properties This problem can be solved by effect-directed analysis (EDA) which provides information on biologically relevant substances even in very complex matrices, such as plant tissue extracts [6]. Thin-layer chromatography-direct bioautography (TLC-DB) is one of three variants of a TLC-B method (the others are contact and agar overlay bioautography) in which separation, a given assay and visualization are performed directly on a TLC layer This is used for testing antimicrobial properties directly on a TLC plate covered with a thin layer of bacteria or fungi, as opposed to a thick layer of inoculated agar in agar overlay bioautography [6,10,11]. The image conversion and spreadsheet programs enable data interpretation [31] Another luminescent test, using a genetically modified plant, pathogenic bacterium Pseudomonas syringae pv. There are no such tests available for other, non-luminescent bacteria
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