Study On The High Polar Ginsenosides In Panax Ginseng C.A. Mey

Panax ginseng C.A.Mey belongs to the Araliaceae family,and has greatly tonifying qi,tonifying spleen,benefit lung,sedative educational and other effects.The main effective components in this plant are ginsenosides which belong to triterpenoid saponins and are also the main components of Panax quinquefolium L.and Panax notoginseng?Burk.?F.H.Chen.Modern pharmacological researches have shown that ginsenosides has anti-tumor,regulating the immune system,regulating the central nervous system,regulating circulation system,anti-aging and other effects.To date,there are many reports on the chemical constituents and pharmacological effects of ginsenosides,and most of them are concentrated on the middle and low polarity of the common ginsenoside,and rare ginsenosides,such as ginsenoside Rb₁,Rb₂,Rb₃,Rc,Rd,Re,Rg₁,Rg₂,Rg₃,Rh₁,Rh₂,Rk₁,Rg₅,PPD,PPT and so on.However,studies on the high polar ginsenosides such as ginsenoside R0,Ra₁,Ra₂ and Ra₃ are rarely reported.In order to fill the blank of the content and biological activities of high polar ginsenosides in Panax plants,we studied the extraction,isolation,structural identification,content determination and biological activities of high polar ginsenosides in Panax ginseng C.A.Mey.In this paper,six high polar ginsenosides were firstly isolated from the 80% ethanol extract of the roots of ginseng.According to their physical,chemical properties and spectra characteristicts of 1H NMR,13 C NMR,MS,the chemical structures of the six compounds were identified as ginsenoside Ra₁,Ra₂,Ra₃,Rb₁,Rb₂ and R₀.A method for the content determination of ginsenoside Ra₁,Ra₂ and Ra₃ in Panax ginseng by HPLC method was also established in this paper.After investigation to extraction solvent,extraction method,extraction time and solvent volume which were factors affecting the extraction rate,we have established the optimum extraction method,and then try to study methodology.The results show that when the ginsenoside Ra₁ and Ra₂ were in the range of 0.7²2.5 ?g and ginsenoside Ra₃ in the range of 0.82⁵2.5 ?g,there were good linear relationships between the peak area?y?and the amount of sample?m?.The RSDs of intraday and interday precision,repeatability,stability and recovery experiments were less than 2%,and the results show that the established method had high sensitivity,good precision and high accuracy.After that,the contents of ginsenoside Ra₁,Ra₂ and Ra₃ in three Panax plants were determined by the above analysis method.The results showed that ginsenoside Ra₁,Ra₂,Ra₃ were determined from the roots,stems and leaves of Panax ginseng C.A.Mey,the roots and leaves of Panax quinquefolium L.,the leaves and stems of Panax notoginseng?Burk.?F.H.Chen.The content of ginsenoside Ra₁ in the roots of Panax quinquefolium L.was the highest and the content was 0.56%;The content of ginsenoside Ra₂ in the leaves of Panax notoginseng?Burk.?F.H.Chen was the highest and the content was 0.26%;The content of ginsenoside Ra₃ in the roots of Panax quinquefolium L.was the highest and the content was 1.24%.There are many studies on the anti-tumor activities of ginsenosides,especially the anti-tumor activities of the low polarity ginsenosides.However,studies on the anti-tumor activities of ginsenoside Ra₁,Ra₂ and Ra₃ are rarely reported,which may be related to the difficult separation and purification of high polarity ginsenosides.Therefore,we studied their anti-cancer activities by CCK-8 method,and the proliferation inhibition effects of ginsenoside Ra₁,Ra₂ and Ra₃ on lung cancer cells?A549?,breast cancer cells?MCF-7?,hepatocellular carcinoma?Hep G2?cells and melanoma cells?A375?were investigated.The results showed that ginsenoside Ra₁,Ra₂ and Ra₃ could inhibit the proliferation of lung cancer cells and hepatoma cells.In summary,in this paper,six high polar saponins were isolated and identified from the roots of Panax ginseng C.A.Mey.;The method for the content determination by HPLC of ginsenoside Ra₁,Ra₂ and Ra₃ in Panax ginseng was established;The anti-tumor activities in vitro of ginsenoside Ra₁,Ra₂ and Ra₃ were studied.The results of this paper provide a scientific basis for the medical application of high polar ginsenosides and the improvement of quality standards of Panax ginseng C.A.Mey.