ScienceAsia 19 (1993): 073-086 |doi: 10.2306/scienceasia1513-1874.1993.19.073

SOME EMPIRICAL ASPECTS ON THE PRACTICAL USE OF FLASH CHROMATOGRAPHY AND MEDIUM PRESSURE UQUID CHROMATOGRAPHY FOR THE ISOLATION OF BIOLOGICALLY ACTIVE COMPOUNDS FROM PLANTS.

PER CLAESON,^1,2 PATOOMRATANA TUCHINDA¹ AND VICHAI REUTRAKUV³

ABSTRACT: The applications of flash chromatography and medium pressure liquid chromatography (MPLC) for the isolation of pure compounds from plants are presented with the emphasis on the practical aspects of both techniques. The pressure for flash chromatography, which operates at about 1-2 bars, is provided by an aquarium pump or several pumps in parallel. The simple necessary glass apparatus, which can be constructed in a glass blowing workshop, are described diagrammatically. The practical operation of the technique is included. Up to 200 grams of crude plant extract can be readily separated in a single run. The approximate relationship among sizes of samples, columns and fractions collected is given. MPLC can be used as a complement to flash chromatography. The pressure required is in the range of 5-20 bars and it can be provided by a piston pump with a variable flow rate. Crude plant extract fractionated by flash chromatography can be further purified by MPLC to give pure materials. Up to 50 grams of semipurified fraction can be conveniently chromatographed in a single run. The use of gradient elution is essential for the success of both flash chromatography and MPLC. A gradient can be constructed between any two miscible organic solvents. Initially, the polarity is increased slowly and more steeply toward the end of the separation. A brief guideline for the use of various normal phase column packing materials is given. An example illustrating the utilization of the two techniques in a multi gram scale isolation of pure compounds from the plant Curcuma xanthorrhiza Roxb. (Zingiberaceae) is described.

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Received May 15, 1993