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<p>Spent hops after supercritical CO<sub>2</sub> extraction contain valuable bioactive compounds such as xanthohumol (XN). Due to its low water solubility and high thermal and light sensitivity, traditional purification remains challenging. This study investigates a sequential purification strategy using a three-liquid-phase extraction system followed by macroporous resin chromatography. A three-liquid-phase extraction system consisting of n-hexane-ethanol-NaH<sub>2</sub>PO<sub>4</sub>-water was employed to fractionate the crude extract. XN partitioned into the middle ethanol-rich phase, effectively separating it from non-polar lipids and polar impurities. Further purification was conducted using Diaion HP-20 resin. Adsorption data followed the Langmuir isotherm (<i>R</i><sup>2</sup> = 0.9989) and pseudo-second-order kinetics (<i>R</i><sup>2</sup> = 0.9999), indicating a monolayer chemisorption mechanism. The process significantly improved the XN purity, representing a 5.6-fold enrichment. UV–VIS and HPLC analysis confirmed that the resin-purified fraction closely matched the XN standard profile. Simultaneously performing anti-solvent precipitation also resulted in XN-enriched extracts; however, with slightly lower purity. These results demonstrate that combining a three-liquid-phase extraction system with macroporous resins provides a scalable, efficient, and reproducible method for high-purity XN recovery from industrial residues, contributing to the sustainable valorisation of brewery waste.</p>

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Valorisation of Spent Hops: Sequential Purification of Xanthohumol from Spent Hops Using a Three-Liquid-Phase n-Hexane-Ethanol-NaH2PO4 System and Macroporous Resin Adsorption

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月29日

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  • Aleksandra Modzelewska,
  • Anna Trusek

摘要

Spent hops after supercritical CO2 extraction contain valuable bioactive compounds such as xanthohumol (XN). Due to its low water solubility and high thermal and light sensitivity, traditional purification remains challenging. This study investigates a sequential purification strategy using a three-liquid-phase extraction system followed by macroporous resin chromatography. A three-liquid-phase extraction system consisting of n-hexane-ethanol-NaH2PO4-water was employed to fractionate the crude extract. XN partitioned into the middle ethanol-rich phase, effectively separating it from non-polar lipids and polar impurities. Further purification was conducted using Diaion HP-20 resin. Adsorption data followed the Langmuir isotherm (R2 = 0.9989) and pseudo-second-order kinetics (R2 = 0.9999), indicating a monolayer chemisorption mechanism. The process significantly improved the XN purity, representing a 5.6-fold enrichment. UV–VIS and HPLC analysis confirmed that the resin-purified fraction closely matched the XN standard profile. Simultaneously performing anti-solvent precipitation also resulted in XN-enriched extracts; however, with slightly lower purity. These results demonstrate that combining a three-liquid-phase extraction system with macroporous resins provides a scalable, efficient, and reproducible method for high-purity XN recovery from industrial residues, contributing to the sustainable valorisation of brewery waste.