Simultaneous partitioning of multiple bioactive compounds from <i>Garcinia indica</i> rinds in a three-liquid-phase extraction systems

Simultaneous extraction and purification of principal bioactive compounds, anthocyanins (ACNs), garcinol (GL), and hydroxycitric acid (HCA) from the rinds of <i>Garcinia indica</i> (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-(NH₄)₂SO₄-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 (NH₄)₂SO₄ concentration on the partitioning behavior of biomolecules was analyzed. The TLPS composed of water-n-hexane-ethanol-(NH₄)₂SO₄ 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.