Preparation Study Of Tanshinones From Salvia Miltiorrhiza Bunge By High-speed Counter-current Chromatography

Dan-shen, the dried roots of Salvia miltiorrhiza Bunge, is one of the most popular traditional Chinese medicines.The tanshinones which was separated form Dan-shen has been widely used for promoting blood circulation to remove blood stasis, clearing away heat, relieving vexation, nourishing blood and tranquilizing the mind and cooling blood to relieve carbuncles. Traditonal chromatographic methods for tanshinone purification are all highly sample pre-execution required, long-time sep.procedure, and high loss of samples.High speed counter current chromatography(HSCCC) is a unique liquid-liquid partition chromatography that uses no solid support matrix. Therefore, it eliminates irreversible adsorptive loss of samples onto the solid support matrix used in the conventional chromtographic column and the metamorphic change of samples. It is simple, economical, quick and with good repeatability. The method has been successfully applied to the analysis and separation of several natural products.Separation and preparation of tanshinones from Danshen crude ethanol extract by HSCCC was studied in this paper. The detection and pre-execution method for tanshinones was established. The solvent system Petroleum ether/ethyl acetate/ methanol/ water(8:6:7:3) was selected and the optimum running parameters for apparatus was obtained through tube experiments and solvents selection software. The maximal amount of sample for one injection and the operation process were also studied. The follwing optimized parameters was obtained: sample rate 150mg, water circulation temperature 22℃, coiled column rotation speed 800rpm, flow rate 2.0mL/min, after 250min, changed to 4.0mL/min.Four solid collections were separated and identified. Measured by HPLC, the purity of TanshinoneⅡA, Cryptotanshinone, TanshinoneⅠ, DihydrotanshinoneⅠwere all over 98%.