Cassia Active Ingredient Extraction And Separation Of Structure By Electrospray Ionization Tandem Mass Spectrometry (esi-ms ~ N)

Cassia obtusifolia have been widely used in traditional Chinesemedicine for the treatments of hypertension activity, higher cholesterin,astriction, antimicrobial, etc. It is highly valuable to study the chemicalconstituents of Cassia obtusifolia. The extraction and separation of themain functional components from Cassia obtusifolia is necessary. The maincontents are belows.1. The extraction of effective components from Cassia obtusifolia atroom temperature was carried out by High Pressure HomogenizationExtraction (HPHE) method. Compared to heat reflux extraction method andultrasonic extraction method, the best way for the extraction of effectivecomponents from Cassia obtusifolia was HPHE method. The optimumextraction conditions of HPHE method were obtained by orthogonal test.Under the optimal conditions of HPHE method, the yield of flavonoids,anthraquinones and polysaccharide were up to 1.233%, 0.089%, 10.55%,which were at least 20 to 30% higher than the figures obtained from thereflux extraction and ultrasonic extraction. The HPHE was proved to be anew process for extraction of the effective components of traditionalChinese medicine.2. HPLC fingerprint of Cassia obtusifolia was established by Highpressure liqid chromatography. The chemical components were analyzed byHPLC-electrosprary ionization-mass spectrometry (HPLC/ESI-MS~n) and thestructure were elucidated according to mass-to-charge ratio (m/z) ofmolecule ionic peak, the fragmental and reference. There were more than 20chemical components isolated, but only 10 chemical components weredetermined. The data obtained for chemical component using this methodprovide a Cassia obtusifolia profile database useful for the rapididentification of Cassia obtusifolia. The method of HPLC/ESI-MS~n is rapidand sensitive. It is suitable to application in the field of natural productsanalysis.3. The separation of Cassia obtusifolia was carried out by High-speed countercurrent chromatography (HSCCC), with the solvent systemcomposed of n-hexane-ethyl acetate-methanol-water (4:1:3:2, v/v) anda total of five well-separated peaks were obtained in the HSCCCchromatogram and their purities were determined by HPLC-UV absorptionspectrometry, the purities of five peaks were 98%, 95%, 96%, 95%, 96%,respectively, which further verified HSCCC was a versatile separationsystem for active ingredient of plant. These peaks were characterized byESI-MS~n and the data compared with the reference standards. Five peakswere identified as 1, 2, 6-trihydroxy-7, 8-dimethoxy-3-methylanthraquinone1, 2, 6, 8-tetrahydroxy-7-methoxy-3-methylanthraquinone, 2-hydroxy-1, 6,7, 8-teramethoxy-3-methylanthraquinone, 2, 6-dihydroxy-1, 7, 8-trimethoxy-3-methylanthraquinone and 1, 2-dihydroxy-6, 7, 8-trimethoxy-3-methyl-anthraquinone, respectively.