Shanbhag, C.C.Salimuddin, R.M.H.Iyyaswami, R.Belur, P.D.2026-02-032025Preparative Biochemistry and Biotechnology, 2025, 55, 9, pp. 1118-113410826068https://doi.org/10.1080/10826068.2025.2483240https://idr.nitk.ac.in/handle/123456789/20732Simultaneous extraction and purification of principal bioactive compounds, anthocyanins (ACNs), garcinol (GL), and hydroxycitric acid (HCA) from the rinds of Garcinia indica (Kokum) fruits in a single-step using Three Liquid Phase Systems (TLPS) were investigated. Among the various phase-forming components studied, TLPS formed by n-hexane-ethanol-(NH<inf>4</inf>)<inf>2</inf>SO<inf>4</inf>-water system was considered for partitioning GL into the n-hexane-rich top phase, ACNs into the ethanol-rich middle phase, and HCA into the aqueous salt-rich bottom phase. The present system was even able to separate carbohydrates into the bottom phase, which can be detrimental to the stability of ACNs. The effect of n-hexane, ethanol, and (NH<inf>4</inf>)<inf>2</inf>SO<inf>4</inf> concentration on the partitioning behavior of biomolecules was analyzed. The TLPS composed of water-n-hexane-ethanol-(NH<inf>4</inf>)<inf>2</inf>SO<inf>4</inf> could purify and extract 95.08% of ACNs, 95.33% of GL, and 67.98% of HCA in a single-step extraction process while the other extraction methods require multi-step extraction process to separate these three compounds. The effect of pH studies on the partitioning characteristics of biomolecules revealed that pH 4 is optimum and more efficient than the native pH of the system to achieve maximum yield of all the bioactive compounds. © 2025 Taylor & Francis Group, LLC.alcoholanthocyanincitric acidgarcinolhexanehydroxycitric acidn-hexaneplant extractterpenechemistryfruitGarciniaisolation and purificationliquid liquid extractionpHproceduresAnthocyaninsCitratesEthanolFruitHexanesHydrogen-Ion ConcentrationLiquid-Liquid ExtractionPlant ExtractsTerpenes