Isolation of Hydrolyzable Tannins from Castanea sativa Using Centrifugal Partition Chromatography.

Castanea sativa wood is a rich source of hydrolyzable tannins, known for their diverse bioactivities. To investigate these bioactive properties further, it is crucial to isolate and characterize hydrophilic compounds effectively. To address this issue, we developed a centrifugal partition chromatography (CPC) method and applied it to an aqueous C. sativa wood extract. We determined the partition coefficients (KD) of the six major compounds using four butanol-/water-based biphasic solvent systems. Initially, we utilized the n-butanol/propanol/water (3:1:4, v/v/v) systems for the first fractionation step. Subsequently, we employed the water/methyl tert-butyl ether/butanol/acetone (8:5:3:4, v/v/v/v) system to fractionate moderately and highly hydrophilic fractions. We calculated the KD values for major compounds of the most hydrophilic fractions using the butanol/ethanol/water (4:1:5, v/v/v) and butanol/isopropanol/water (2:1:3, v/v/v) systems. In total, we isolated 23 compounds through a combination of CPC, size exclusion chromatography, and preparative HPLC. Among these compounds, six have never been previously described. We characterized them by 1D and 2D NMR experiments and high-resolution mass spectroscopy acquisitions.

Antifungal Activities of a Grapevine Byproduct Extract Enriched in Complex Stilbenes and Stilbenes Metabolization by Botrytis cinerea.

Grapevine co-products, as canes, represent a source of compounds of interest to control vineyard diseases with a sustainable approach. We chose to study an extract that we produced from grapevine trunk and roots. This extract, enriched in complex stilbenes, strongly reduced mycelial growth and spore germination of Botrytis cinerea, the fungal agent causing gray mold. The most active stilbenes were resveratrol, r-viniferin, and ε-viniferin. This grapevine extract also inhibited the production of Botrytis laccases. Conversely, Botrytis secretome metabolized resveratrol into δ-viniferin and pallidol (2 dimers); and ε-viniferin, a dimer, into hopeaphenol, r-viniferin, and r2-viniferin (3 tetramers). r-Viniferin and hopeaphenol (2 tetramers) were not metabolized. The biotransformed extract maintained an effective antimycelial activity. This study provides evidence that a grapevine extract enriched in oligomerized stilbenes exerts different anti-Botrytis activities, notwithstanding the ability of the fungus to metabolize some stilbenes.

Stilbenes from Vitis vinifera L. Waste: A Sustainable Tool for Controlling Plasmopara Viticola.

Stilbene-enriched extracts from Vitis vinifera waste (cane, wood, and root) were characterized by UHPLC-MS. Eleven stilbenes were identified and quantified as follows: ampelopsin A, (E)-piceatannol, pallidol, (E)-resveratrol, hopeaphenol, isohopeaphenol, (E)-ε-viniferin, (E)-miyabenol C, (E)-ω-viniferin, r2-viniferin, and r-viniferin. The fungicide concentration inhibiting 50% of growth of Plasmopara viticola sporulation (IC50) was determined for the extracts and also for the main compounds isolated. r-Viniferin followed by hopeaphenol and r2-viniferin showed low IC50 and thus high efficacy against Plasmopara viticola. Regarding stilbene extracts, wood extract followed by root extract showed the highest antifungal activities. These data suggest that stilbene complex mixtures from Vitis vinifera waste could be used as a cheap source of bioactive stilbenes for the development of natural fungicides.