Leafy Green Phenolic / Polyphenolic Extraction

Authors: HERNANDEZ, ALEXANDER - Baylor College Of Medicine; LE, ELAINE - Baylor College Of Medicine; TAK, ALVIN - Baylor College Of Medicine; Dzakovich, Michael

Submitted to: Protocols.io
Publication Type: Other
Publication Acceptance Date: 3/12/2026
Publication Date: 3/16/2026
Citation: Hernandez, A., Le, E., Tak, A., Dzakovich, M.P. 2026. Leafy Green Phenolic / Polyphenolic Extraction. Protocols.io. https://doi.org/10.17504/protocols.io.36wgqpprkvk5/v1.
DOI: https://doi.org/10.17504/protocols.io.36wgqpprkvk5/v1

Interpretive Summary: Problem: Plant foods contain many helpful compounds called phenolics and polyphenols, but measuring them can be difficult. Different labs often use different methods to extract and prepare these compounds, which can lead to inconsistent results. If the extraction or preparation is not done correctly, some compounds may be lost or measured inaccurately, making it hard to compare studies or understand the true nutritional value of plants. Accomplishment: To address this, we developed a clear and simplified protocol for extracting these compounds from plant materials and preparing them for analysis using LC-MS/MS. The method explains how to break down plant samples, choose the right solvents, and use tools like sonication to improve extraction. It also includes steps for adding internal standards and properly preparing samples—such as diluting and filtering them—before analysis. These steps help ensure that compounds are measured accurately and consistently. Contribution: This work provides a reliable and repeatable way to study phenolics and polyphenols across different plant types. With more accurate and consistent measurements, scientists can better understand the health benefits of these compounds and compare results across studies. This ultimately supports improved research in nutrition, agriculture, and food science.

Technical Abstract: This protocol describes a streamlined method for phenolic and polyphenolic extractions from plant material and subsequent preparation for analysis by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Previously optimized extraction solvents, sample disruption techniques (e.g. sonication), and the use of internal standards primarily targeted at flavonoids are covered in detail. A workflow for diluting, filtering, and otherwise preparing samples for analysis is also included. This approach enables sensitive, reproducible profiling and quantification of diverse phenolic and polyphenolic compounds across plant matrices.