Study on Extraction, Isolation and Antioxidative Activity of the Saponins from Perennial Herbal Plants in Panax Arallaceae Family
|School||Changchun Teachers College|
|Keywords||Panax Ginseng C.A.Mey Panax quinquefolium L. Panax notoginseng (Burk) F. H.Chen saponins HPCPC-ELSD HPLC-ELSD antioxidative activity|
Panax Ginseng C.A.Mey, Panax quinquefolium L. and Panax notoginseng (Burk) F. H.Chen are three major Perennial herbal Plants in Panax Arallaceae family,. Comparison of genetic relationship among the three parties, they are similar, and both with the dammarane ginsenosides as the main active ingredient. Ginsenosides have been found to be provided with strong biological activity, therefore, it is significant to research and develop of ginsenosides.First,the present article describes several extraction processes were used respectively to extracte the active constituents from P. Ginseng, P. Quinquefoljus, P. Notoginseng, such as the warmly extraction method, heat re?ux and ultrasound-assisted extraction method, and the high-performance liquid chromatographic(HPLC) was undertaken in order to determine the compounds content. Results indicated that the ultrasound-assisted extraction method was the optimum extraction. This method is rapid, stable, efficient and low cost and without destroying components. Then Ultrasonic extraction method was used to large-scale extract the active constituents from three kinds of medicinal herbs, and the extracted crudes were separated and purified by high performance centrifugal partition chromatography.Second , the present article developed a High performance centrifugal partition chromatography(HPCPC) coupled with evaporative light scattering detector (ELSD) method for the separation and purification of saponins from P. Ginseng, P. Quinquefoljus, P. Notoginseng in one-step run. To separate notoginsenoside R1, ginsenoside Rg1, Re, Rb1 extracted from n-butanol extract of P. notoginseng using a solvent system composed of ethyl acetate–n-butanol–water(1:1:2, v/v/v), to separate ginsenoside Rc, Re, Rb1 extracted from n-butanol extract of P. Quinquefoljus using a solvent system composed of ethyl acetate–n-butanol–water(1:1:2, v/v/v) for the first time, to separate ginsenoside Rb1, ginsenosideRb2 extracted from n-butanol extract of P. ginseng using a solvent system composed of ethyl acetate–n-butanol–water(0.8:1.2:2, v/v/v) for the first time. The isolated compounds were identified by their retention time with the authentic standards, and the purities of these saponins were assessed to be over 95% by High performance liquid chromatography (HPLC) coupled with evaporative light scattering detection (ELSD). Electrospray ionization tandem mass spectrometry (ESI-MSn) data provided highly useful structural information for the saponins. The results demonstrate that HPCPC coupled with ELSD is a feasible and efficient technique for systematic isolation of non-chromophoric components from traditional medicinal herbs. HPLC-ELSD and MSn provide comfortable analysis methods for purity testing and identification of saponins.Last,to explore the antioxidative activity of the saponins in Panax Ginseng, Panax Quinquefoljus and Panax Notoginseng. Photochemi-luminescence (PCL) was first applied to determine the antioxidative activity of the saponins for the first time, and DPPH method was developed for evaluating the scavenging activity of the extracts. Results indicate that the saponins of ethanol extracts from Panax ginseng, Panax quinquefoljus, Panax notoginseng owned obvious antiradical capacity, the efficiency increases with the content of major saponins extracted from plants. Panax Notoginseng saponins, ultrasonic extracts show the strongest antioxidant capacity, in PCL is equivalent to 2.53 nmol Vc, in DPPH, the scavenging effect is 32.57%, with a total saponins 209.62 mg/g, followed by reflux extracts(1.71 nmol, 30.82%, 128.97 mg/g) and warmly extracts(1.07 nmol, 18.60%,100.44 mg/g), the efficiency increases with the content of major saponins extracted from plants. For American ginseng and ginseng total saponins, ultrasonic extracts obtain the highest saponin content(73.27 mg/g and 17.48 mg/g), followed by reflux extracts(52.26 mg/g and 10.42 mg/g) and warmly extracts(48.62 mg/g and 9.89 mg/g), however, the free radical scavenging effect is that the reflux extracts is higher (in PCL, 2.28 nmol and in DPPH, 23.62%; 0.63 nmol and 22.98%) than ultrasonic extracts(2.18 nmol and in 23.51%; 0.61 nmol and 21.20%), but both more or less, the warmly extracts show the lowest antioxidant capacity(1.07 nmol and 18.60%;0.42 nmol and 14.37%), there are some differences between antioxidangt capacity and the saponin content.