Chromatographic and Biological Screening of Chosen Species of Schisandraceae Family: Schisandra chinensis, S. rubriflora, S. sphenanthera, S. henryi and Kadsura japonica.

HPLC and TLC profiling was carried out for leaf and fruit extracts of five Schisandraceae species: Schisandra chinensis, S. rubriflora, S. spehenanthera, S. henryi and Kadsura japonica. HPLC measurements confirmed presence of lignans and phenolic compounds in fruits and leaves of all tested species. The most abundant in lignans was S. chinensis fruit extract in which 15 compounds were detected (e. g.: schisandrol A, schisanhenol, γ-schisandrin, gomisin N). The effect-directed detection, i. e., TLC-direct bioautography against Bacillus subtilis, showed exceptionally high activity for S. chinensis and S. rubriflora fruit extracts. On the other hand, TLC-DB enzyme tests (α-glucosidase, lipase, tyrosinase and acetylcholinesterase (AChE) inhibition assays) showed that all fruit and leaf extracts have ability to inhibit the above-mentioned enzymes (except for the K. japonica fruit). The leaf extracts showed much stronger antioxidant activity than the fruit ones, which were assessed and compared using both TLC-direct bioautography and spectrophotometric measurements based on ABTS, DPPH and FRAP tests.

Schisandra rubriflora Fruit and Leaves as Promising New Materials of High Biological Potential: Lignan Profiling and Effect-Directed Analysis.

The effect-directed detection (EDD) of Schisandra rubriflora fruit and leaves extracts was performed to assess their pharmacological properties. The EDD comprised TLC-direct bioautography against Bacillus subtilis, a DPPH assay, as well as α-glucosidase, lipase, tyrosinase, and acetylcholinesterase (AChE) inhibition assays. The leaf extracts showed stronger antioxidant activity than the fruit extract as well as inhibition of tyrosinase and lipase. The fruit extract was found to be extremely active against B. subtilis and to inhibit α-glucosidase and AChE slightly more than the leaf extracts. UHPLC-MS/MS analysis was carried out for the bioactive fractions and pointed to the possible anti-dementia properties of the dibenzocyclooctadiene lignans found in the upper TLC fractions. Gomisin N (518 mg/100 g DW), schisanhenol (454 mg/100 g DW), gomisin G (197 mg/100 g DW), schisandrin A (167 mg/100 g DW), and gomisin O (150 mg/100 g DW) were the quantitatively dominant compounds in the fruit extract. In total, twenty-one lignans were found in the bioactive fractions.

Effect-directed analysis as a method for quality and authenticity estimation of Rhodiola rosea L. preparations.

Adulterations of food and pharmaceutical preparations are the important global problem. On the one hand, fraud practices are becoming more and more sophisticated while on the other, monitoring and uncovering falsifications are insufficient. One of the most common consumer concern is the quality and authenticity of the purchased products, related to the confidence that they have composition and properties in accordance with the manufacturer's declaration on the label. This refers also to pharmaceuticals potentially delivering great health benefits such as Rhodiola rosea L. supplements. The aim of this study was defining authenticity and possible adulterations of two R. rosea preparations basing on their TLC-bioprofiles and the presence of biomarker compounds characteristic for this plant. The effect-directed analysis (EDA), i.e. TLC hyphenated with micro-chemical and biological assays performed directly on TLC plates followed by HPLC-ESI-MS was used for the bioprofiling of antioxidants, antibacterials, and inhibitors of lipase, acetylcholine, α-glucosidase and tyrosinase as well as for the identification of the biomarkers. The results pointed to the possible adulteration of one of the tested products related to the absence of two rosavins, the most important quality markers of R. rosea.

Effect directed analysis and TLC screening of Schisandra chinensis fruits.

The fruit of Schisandra chinensis (Chinese magnolia vine), the medicinal plant well-known in Traditional Chinese Medicine, gains great popularity in the modern phytopharmacology. This phenomena is related to the wide and powerful healing properties, including supporting immune, nervous and digestive systems activity. S. chinensis is also known for its adaptogenic properties which can support the treatment of neurodegenerative disorders, particularly Alzheimer's disease. The components of S. chinensis have been analyzed mostly using chromatography, including HPLC, GC and TLC. The last technique can be easily hyphenated with biological assays performed directly on a TLC plate that is using effect directed detection. TLC-direct bioautography (TLC-DB) for acetylcholinesterase (AChE) inhibition and TLC-DB against Bacillus subtilis were performed, followed by micro-preparative separation of fractions which were subsequently subjected to LC-MS tentative identification. Additionally, screening analysis was done using both biological detection and derivatization reagents (e.g. PMA, thymol, NP-PEG). Both TLC screening and effect-directed analysis (TLC-DB followed by MS analysis) of S. chinensis fruit revealed components with biological activity, especially antibacterials (e.g. citric acid) and inhibitors of AChE (mainly dibenzocyclooctadiene lignans). AChE inhibition activities were confirmed by TLC-DB for nine standards that is: 6-O-benzoylgomisin, deoxyschisandrin, gomisin A, gomisin G, schisandrin, schisandrin C, schisanhenol, schisantherin A and schisantherin B. These lignans were further identified by LC-ESI-MS in the isolated fraction revealing AChE inhibition. Moreover three other lignans: γ-schisandrin, schisandrin B and gomisin J were tentatively identified by LC-ESI-MS.

Determination of chlorogenic acid, polyphenols and antioxidants in green coffee by thin-layer chromatography, effect-directed analysis and dot blot - comparison to HPLC and spectrophotometry methods.

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.