title: High-performance thin-layer chromatography linked with (bio)assays and mass spectrometry - a suited method for discovery and quantification of bioactive components? Exemplarily shown for turmeric and milk thistle extracts.
abstract: Extraction parameters, chemical fingerprint, and the single compounds' activity levels were considered for the selection of active botanicals. For an initial survey, the total bioactivity (i.e., total reducing capacity, total flavonoids contents and free radical scavenging capacity) of 21 aqueous and 21 ethanolic plant extracts was investigated. Ethanolic extracts showed a higher yield and were further analyzed by HPTLC in detail to obtain fingerprints of single flavonoids and further bioactive components. Exemplarily shown for turmeric (Curcuma longa) and milk thistle (Silybum marianum), effect-directed analysis (EDA) was performed using three selected (bio)assays, the Aliivibrio fischeri bioassay, the Bacillus subtilis bioassay and the 2,2-diphenyl-1-picrylhydrazyl (DPPH*) assay. As a proof of principle, the bioactive components found in the extracts were confirmed by HPTLC-MS. Bioassays in combination with planar chromatography directly linked to the known, single effective compounds like curcumin and silibinin. However, also some unknown bioactive components were discovered and exemplarily characterized, which demonstrated the strength of this kind of EDA. HPTLC-UV/Vis/FLD-EDA-MS could become a useful tool for selection of active botanicals and for the activity profiling of the active ingredients therein. The flexibility in effect-directed detections allows a comprehensive survey of effective ingredients in samples. This streamlined methodology comprised a non-targeted, effect-directed screening first, followed by a highly targeted characterization of the discovered bioactive compounds. HPTLC-EDA-MS can also be recommended for bioactivity profiling of food on the food intake side, as not only effective phytochemicals, but also unknown bioactive degradation products during food processing or contamination products or residues or metabolites can be detected. Thus, an efficient survey on potential food intake effects on wellness could be obtained. Having performed both, sum parameter assays and HPTLC analysis, a comparison of both approaches was made and discussed.
authors.affiliation: Justus Liebig University Giessen, Institute of Nutritional Science, Chair of Food Science, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany; Justus Liebig University Giessen, Institute of Nutritional Science, Wilhelmstrasse 20, 35392 Giessen, Germany.
authors.forename: Mahmoud N
authors.initials: MN
authors.lastname: Taha
authors.affiliation: Justus Liebig University Giessen, Institute of Nutritional Science, Wilhelmstrasse 20, 35392 Giessen, Germany.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Krawinkel
authors.affiliation: Justus Liebig University Giessen, Institute of Nutritional Science, Chair of Food Science, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany. Electronic address: Gertrud.Morlock@ernaehrung.uni-giessen.de.
authors.forename: Gertrud E
authors.initials: GE
authors.lastname: Morlock
mesh_terms: D048908:Aliivibrio fischeri / Q000187:drug effects; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects; D001681:Biological Assay / Q000379:methods; D001713:Biphenyl Compounds / Q000737:chemistry; D002855:Chromatography, Thin Layer / Q000379:methods; D030024:Curcuma / Q000737:chemistry*; D003474:Curcumin / Q000737:chemistry / Q000494:pharmacology; D020944:Silybum marianum / Q000737:chemistry*; D010851:Picrates / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry* / Q000494:pharmacology; D000077385:Silybin; D012838:Silymarin / Q000737:chemistry / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D000900:Anti-Bacterial Agents; D000975:Antioxidants; D001713:Biphenyl Compounds; D010851:Picrates; D010936:Plant Extracts; D012838:Silymarin; D000077385:Silybin; C004931:1,1-diphenyl-2-picrylhydrazyl; D003474:Curcumin
keywords: Bioassays; Curcumin; Effect-directed analysis (EDA); HPTLC–direct bioautography; Mass spectrometry; Silibinin