Isolation, Purification And Biological Activity Of Polysaccharides Extracted From Gastrodia El Bl. And Elaeagnus Angustifolia

In recent years, with the continuous deepening of the research of polysaccharides, it is realized that Polysaccharide is not only the structure material(such as cellulose) or energy storage material(such as starch), but also a kind of important information molecules with a variety of physiological functions. Thus, polysaccharide with biological activity has become one of the hot spots in the field of domestic and abroad. Two kinds of polysaccharides are extracted and isolated in this research. Meanwhile, the physical and chemical properties and biological activity of purified homogeneous fractions were studied.This paper is divided into two parts as followed:Chapter one: the research of polysaccharide from Gastrodia el Bl.Gastrodia el Bl. is one of precious Traditional Chinese Medicines. Ancient medical records various magical effect of Gastrodia el Bl. The earliest records could track in "Shennong Materia Medica” of the Eastern Han Dynasty, Specific descriptions is listed as Kill the ghost sperm, poison smells and good to human’s health after taking a long time, which indicate that Gastrodin is able to cure dizzy, infantile convulsion and ease nerve pains. Polysaccharide is a natural macromolecular compound, and it has been involved in and maintained a variety of physiological functions of life. The purpose of this study is to isolate, purify and make a preliminary study of biological activity of polysaccharide from Gastrodia el Bl. to lay a solid foundation for the use of development of Gastrodia el Bl. polysaccharide.Taking Gastrodia el Bl. as material, the isolation, purification and biological activity of polysaccharide are studied. The results are as followed:1 In this research, the crude polysaccharide of Rhizoma is obtained using Water extraction and alcohol precipitation. Further, the refined polysaccharide is received after protein removal and dialysis of crude polysaccharide. Then, two homogeneous components polysaccharides RGP-1a and RGP-1b are gained using DEAE-52 and G-100.2 Preliminary structures of RGP-1a and RGP-1b are identified with the help of FT-IR, UV, HPLC and HPGPC.3 According to the Clearance test of DPPH·and ·OH free radical, RGP-1a and RGP-1b are of significant ability to scavenge DPPH and OH free radical. The in vitro activity test showed that RGP-1a and RGP-1b could significantly stimulate the release of NO and enhance the phagocytic ability of macrophages. Especially when the concentration of RGP-1b was 200 g/m L, the stimulating effect and the stimulation of macrophages to release NO and enhance the phagocytic ability of macrophages of RGP-1b was almost equivalent to LPS(2 g/m L), acting as the positive control.The above test data indicate that the polysaccharide has the potential to be developed into health food.Chapter two: the research of polysaccharide from Elaeagnus angustifolia LElaeagnus angustifolia L belongs to Elaeagnus genus of perennial plants and is mainly distributed in the northern mountains of Himalaya in Asia and Europe. In China, Elaeagnus angustifolia is widely used in the treatment of amebic dysentery, vomiting, jaundice, asthma, flatulence, rheumatoid arthritis disease. In this study, the extraction process of polysaccharide from Elaeagnus angustifolia L was optimized. Besides, physical and chemical properties and in vitro immune activity of Elaeagnus angustifolia L polysaccharide were tested. The results are listed as followed:1 Based on the single factor experiment, the Ultrasonic extraction process of polysaccharide from Elaeagnus angustifolia L was optimized taking advantage of Response surface optimization method. The optimal extraction parameters of Elaeagnus angustifolia polysaccharide were decided: the extraction time was 43 min, the extraction temperature was 79℃, Ultrasonic power was 282 W and ratio of material and solvent was 1:30. Under the optimal condition, the yield of Elaeagnus angustifolia polysaccharide was 9.93±0.24%.2 Two homogeneous components polysaccharides EAP-1a and EAP-1b are obtained using DEAE-52 and G-100. Preliminary structures of EAP-1a and EAP-1b are identified with the help of FT-IR, UV, HPLC and HPGPC.3 The in vitro activity test revealed that EAP, EAP-1a and EAP-1b have a significant effect on the stimulation of 264.7 macrophages, which can promote the phagocytosis of macrophages.The above research shows that Elaeagnus angustifolia polysaccharide has broad prospects in the field of medicine and functional food. However, further studies are needed on the mechanism structure and biological activity of Elaeagnus angustifolia polysaccharide.