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A new RP-HPLC method was developed for the simultaneous determination of 5 active constituents, including loganic acid, swertiamarin, gentiopicroside, sweroside, isoorientin in aerial part of Gentiana straminea. Analysis was achieved on a Hypersil ODS analytical column (4.6 mm x 250 mm, 5 microm) eluted with methanol and water containing 0.02% phosphoric acid in gradient elution. The flow rate was 1 mL x min(-1), the column temperature was set at 35 degrees C and detection wavelength was set at 254 nm. The results showed that 5 active components were separated well and showed good linearity. The corelation coefficints and concentration ranges of the calibration curves were as follow: r = 0.999 9, 0.364-2.18 microg for loganic acid. r = 0.999 9, 0.275-1.65 (microg for swertiamarin. r = 0.999 9, 0.614-3.68 microg for gentiopicroside. r = 0.999 9, 0.065 6-0.394 microg for sweroside. r = 0.999 9, 0.089 9-0.539 microg for isoorientin. The contents of loganic acid and gentiopicroside far exceed that of the other 3 active components, and made up 60% in the total contents. The developed method was proved to be rapid, sensitive, accurate, credible and repeatable. It can be applied to quality control of aerial part of Tibetan medicine Gentiana straminea.
High-speed counter-current chromatography (HSCCC) was successfully applied for the first time to isolate and purify four cis-trans isomers of coumaroylspermidine analogs from Safflower. HSCCC separation was achieved with a two-phase solvent system composed of chloroform-methanol-water (1:1:1, v/v/v) with the upper phase as the mobile phase. In a single run, a total of 1.3mg of N(1), N(5), N(10)-(E)-tri-p-coumaroylspermidine (EEE), 4.4mg of N(1)(E)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (EZE), 7.2mg of N(1)(Z)-N(5)-(Z)-N(10)-(E)-tri-p-coumaroylspermidine (ZZE), and 11.5mg of N(1),N(5),N(10)-(Z)-tri-p-coumaroylspermidine (ZZZ) were obtained from 100mg of crude sample. High Performance Liquid Chromatography (HPLC) analysis showed that the purities of these four components are 95.5%, 98.1%, 97.5% and 96.2%, respectively. The chemical structures were identified by ESI-MS, (1)H NMR and (13)C NMR.