Abstract
Complex samples such as botanical extracts contain hundreds of compounds. Since we can only identify compounds that are stable, extractable, separable and detectable from complex botanical extracts, minimal sample treatment and different detection methods are essential. A combination of high-performance thin-layer chromatography (HPTLC) with non-targeted screening via bioassays (enzymes), microchemical and biological (microorganisms) detection allows for the fast and quantitative bioprofiling of complex samples. Further hyphenation of HPTLC with spectroscopic methods of identification enables targeted identification of bioactive natural products via Effect Directed Analysis (EDA).
Highlights
The plant kingdom is a vast source of secondary metabolites, with many of these compounds possessing therapeutic activity
The traditional strategy used to discover therapeutic activity has been challenged in the last few decades, from the laborious and expensive methods under which bioactivity was studied only after the isolation of the components, to an Effect Directed Analysis (EDA) approach
High-performance thin-layer chromatography (HPTLC) is an enhanced form of TLC, which consists of the automatization of instrumentation and increased chromatogram resolution
Summary
The plant kingdom is a vast source of secondary metabolites, with many of these compounds possessing therapeutic activity. EDA is a combined approach that directly links chromatographic separation with in vitro effect-directed (bio)assays and physico-chemical analysis for the bioactive characterization of detected compounds. After a specific biological effect is detected in a bioassay, chemical fractionation (i.e., chromatographic separation) is used to reduce the complexity of the sample (e.g., plant extracts), to bioactive chemical classes and, eventually, to individual compounds. EDA is an effect-directed nontargeted analysis, used in screening for natural products with bioactivity from complex samples, like botanical extracts, in drug discovery processes. High-performance thin-layer chromatography (HPTLC) is an enhanced form of TLC, which consists of the automatization of instrumentation and increased chromatogram resolution This advanced form of TLC enables the quantitative analysis of separated analyte components in sample mixtures. The correct activity of an enzyme in the suboptimal environment of HPLC analysis is questionable
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