APPLICATION OF CPC AND RELATED METHODS FOR THE ISOLATION OF NATURAL SUBSTANCES

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APPLICATION OF CPC AND RELATED METHODS FOR THE ISOLATION OF NATURAL SUBSTANCES ( application-cpc-and-related-methods-forisolation-natural-sub )

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Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 71 No. 2 pp. 223ñ227, 2014 ISSN 0001-6837 Polish Pharmaceutical Society APPLICATION OF CPC AND RELATED METHODS FOR THE ISOLATION OF NATURAL SUBSTANCES ñ A REVIEW BARTOSZK DZIERSKI*,WIRGINIAKUKU£A-KOCHandKAZIMIERZG£OWNIAK Chair and Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, 1 Chodüki St., 20-093 Lublin, Poland Abstract: A review of research on the isolation of various alkaloids from plant material by centrifugal partition chromatography (CPC) and related preparative techniques was made, in order to provide various conditions for separation of these important plant derived secondary metabolites. First of all, the construction of the CPC apparatus was presented as well as the principle of isolation of natural products with its help, and then the influ- ence of operating apparatus parameters on the separation efficiency. Finally, a review of the alkaloids separa- tion conditions was made, specifying used parameters and best solvent system. Keywords: alkaloid separation, CPC separation, centrifugal partition chromatography, counter current chro- matography Centrifugal partition chromatography (CPC) is a type of counter-current chromatography (CCC), and the advanced form of liquid chromatography (LC) (1). CPC was developed by Yoichiro Ito in 1964 and since then it is being used for the separation and purification of compounds (mainly of herbal origin). It remains an analytical preparative liquid ñ liquid technique, that does not require solid sorbent, but only two solvents that are immiscible two phases ñ stationary and mobile (2). Typical CPC device is a group of cable chan- nels combined in a cascade, located in the cartridges forming circles around the rotor. When the rotor is in motion, system is subjected to continuous cen- trifugal force and (by the action of pump) the mobile phase flows through the stationary phase. Due to the state of two liquid phases, their roles can be reversed ñ ascending mode (when the mobile phase is the lighter phase) or descending mode (when the mobile phase is heavier phase) may be selected. During the flow of one phase against the other, a process of elu- tion (percolation) of the substance from one phase to another, and the process of retention of the sub- stance in the stationary phase (3) are taking place. Substances are separated between the mobile and the stationary phase based on differences in the par- tition coefficients (4). * Corresponding author: e-mail: bartosz.kedzierski1@gmail.com CPC compared to other modern chromato- graphic techniques, is a technique well suited to large scale separation (3), and capable of being used for the separation of substances with a wide range of polarities (4). Selection of isolation parameters. The choice of solvent system, flow rate and rotation speed The choice of solvents remains an important stage in the preparation of the analysis. Distribution of the substances dissolved in the sample between the two liquid phases, on the basis of the partition coefficient values, is essential for the CPC separation (2). The compounds contained in the test material should be soluble in both mobile and stationary phas- es. The most efficient solvent is a combination of such, at which the distribution ratio of a chosen com- pound will range around 1 (preferably) or within the values of 0.2ñ5. After mixing the two phases should be separated quickly ñ preferably within 30 s (5). Retention of active compounds depends large- ly on the partition coefficient and the volume of each phase. Following equation shows this relation- ship: Vr = Vm + P Vs (1) where Vr ñ retention volume, Vm ñ volume of the mobile phase, P ñ partition coefficient, Vs ñ volume of the stationary phase. 223

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