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ARS Home » Midwest Area » East Lansing, Michigan » Sugarbeet and Bean Research » Research » Publications at this Location » Publication #401763
Research Project: Genetic Characterization for Sugar Beet Improvement

Location: Sugarbeet and Bean Research

Title: A new method for fractionation and characterization of polyphenols and tannins from grapevine leaf tissue

Author
item SOMMER, STEPHAN - University Of Missouri
item SALIE, MARNELLE - California State University
item GARCIA, ESTEBAN - California State University
item REYES, ANTHONY - California State University
item EBERSOLE, STEVEN - California State University
item Naegele, Rachel
item VAN ZYL, SONET - California State University

Submitted to: Plants
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/19/2023
Publication Date: 4/20/2023
Citation: Sommer, S., Salie, M., Garcia, E., Reyes, A., Ebersole, S.C., Naegele, R.P., Van Zyl, S. 2023. A new method for fractionation and characterization of polyphenols and tannins from grapevine leaf tissue. Plants. 12. Article 1706. https://doi.org/10.3390/plants12081706.
DOI: https://doi.org/10.3390/plants12081706

Interpretive Summary: Extracting phenolics from grape leaves is challenging, requiring large amounts of plant material and the use toxic and hazardous chemicals. A new method was developed to analyze phenolic compounds in grape leaves. The new method minimizes hazardous waste, while using less plant material, and was able to separate large and small molecules.

Technical Abstract: Plants accumulate different types of phenolic material in their tissue as a response to biotic as well as abiotic stress. Smaller molecules can serve as protection against ultraviolet radiation or pre-vent oxidative tissue damage, while larger molecules such as tannins can be the plant’s reaction to an infection or physical damage. Therefore, characterization, profiling, and quantification of di-verse phenolics can provide valuable information about the plant and the stress status at any given time. A method was developed that allows the extraction of polyphenols and tannins from leaf tissue, followed by fractionation and quantification. Extraction was performed with liquid nitrogen and 30% acetate-buffered ethanol. The separation of tannins from smaller polyphenols was achieved by bovine serum albumin precipitation and resuspension in a urea-triethanolamine buffer. Tannins were reacted with ferric chloride and analyzed spectrophotometrically. Smaller non-protein-precipitable polyphenols were then analyzed via HPLC-DAD from the supernatant of the precipitation sample. This way, a more complete spectrum of compounds can be analyzed from the same plant tissue extract. The method was tested with a variety of grape cultivars and showed great improvements of the chromatography that would otherwise be impacted by tannins. With the fractionation suggested here, hydroxycinnamic acids and flavan-3-ols can be sep-arated and quantified with good accuracy and precision. Stress status and plant responses can be monitored using the total concentrations of polyphenols and tannins, as well as the ratio between those compound classes.