Research On The Isolating Technology Of Ginsenoside In The Fruits Of Panax Ginseng C.A.Mey And Extracting HPLC Fingerprint

For exploring and utilizing the resource of the fruits of Panax ginseng C.A.Mey. , the technology process of macroporous resin and High-speed Counter-current Chromatography (HSCCC) for separating and enreaching saponins from the fruits of ginseng were studied ,and the part of saponins were isolated with the modern phytochemical methods, the structures of compounds were elucidated. The research lays a theoretical and methodological foundation for effective exploitation and sustainable utilization of the fruits of ginseng . The results were as follows:1. Using acetoacetate-n-butanol-water (1:6:7) as the solvent system, ginsenoside Re was isolated from the fruits of ginseng by HSCCC for the first time. The lower phase was the mobile phase. The flow-rate was 2.0mL/min; Apparatus rotate speed was 900r/min.The immobile phase retention was 40%;Retention time of Gentiopicrin was 200～230min. The purity of ginsenoside Re was over 98.84%.2. On selecting macroporous resin for enreaching ginsenoside from the fruits of ginseng, the comparison research was employed by HPLC using ginsenoside Re,Rg1,Rb1,Rc,Rd and Rb2 as contrast and the su-m of 6 kinds of ginsenosides as evaluation criterion f-or AB-8,D-101,HPD300,HPD400,HPD500,HPD450and HPD700-type macroporous resin.The result showed that HPD400-type was the best and the optimiz elution system of HPD400-type for enreaching ginsenoside was set up.3. Using ultrasound extract technology , macroporous resin and silica ge(l normal pressure, medium pressure) chromatographic column, ginsenoside in the fruits of ginseng were separated .5 compounds were isolated in the extract of 60% ethanol elution from macroporous resin by using normal pressure and medium pressure silica gel chromatographic column,identified as ginsenoside Rg1,Re,Rc,Rd and Rb1 by physicochemical properties and spectral analyses.4. HPLC fingerprint of the extract from the fruits of ginseng was established for the first time. The HPLC method was, chromatogram column: Shimdzu ODS 5μm (4.6 mm×250 mm), mobile phase A acetonitrile, B water, gradient lution,0～30min, A:20～20% ; 30～60min , A : 20～0%;60~90min,A:20~20%.Uvλ=203nm , flow rate,1mL/min, column temperature,25℃.According to the determining result of ten batches of samples, we demarcated 15 fingerprint peaks. The similarity of fingerprint was over 0.90. On the investigation of stability, reproducibility and precision, we drew the conclusion that this method was stable, reliable and with better maneuverability. Several innovations achieved in this research are as follows:1. For the first time.High-Speed Counter-Current Chromatography was used to separate and prepared ginsenoside Re from the fruit of ginseng.2. HPLC fingerprint of the extract from the fruits of ginseng was established for the first time.