Water Enzymatic Extraction, Antioxidation And Anti-DNA Damage Activity Evaluation Of Epimedium Sagittatum Polysaccharide And Development Of Health Oral Liquid

Herb Epimedii is one of unique medicinal and edible plants in China and Epimedium brevicornu maxim is one of five species of Epimedium, which is recorded in "The People's Republic of China Pharmacopoeia". In order to provide a basis for scientific exploitation and comprehensive utilization of Epimedium brevicornu resources, optimal extraction method was chosen and extraction procedure has been optimized to isolate polysaccharids from Epimedium brevicornu. Then the content of polysaccharides and their antioxidant activity in different tissues of Epimedium brevicornu were studied and its structure was characterized. Furthermore, healthy oral liquid containing polysaccharides of Epimedium brevicornu was developed, which have the activity of scavenging free radicals.A protocol of enzymatic water extraction was developed to isolate antiradical and DNA damage inhibitory polysaccharides from Epimedium brevicornu using single factor experiments and orthogonal test for the first time. Then a comparative study was carried out among enzymatic water extraction, heating extraction and non-enzymatic extraction to explain their extraction mechanism. The optimal concentrations of papain, cellulase and pectinase were 2.5%,2.0% and 1.5%, respectively, the optimal condition of pH, extraction temperature and extraction duration were 6.0,50 ? and 60 min. The yield of Epimedium brevicornu polysaccharides was 0.93 mg/g. Compared to conventional heating extraction, enzymatic water extraction dramatically enhanced extraction yield, shortened extraction duration, reduced extractant consumption and decreased extraction temperature. In marked contrast to non-enzymatic extraction, enzymatic water extraction exhibited an 82.4% increase in extraction yield of Epimedium brevicornu polysaccharides. Therefore, enzymatic water extraction of polysaccharides from Epimedium brevicornu showed great potential for the development and utilization of Epimedium.On the basis of the present study, further comparative researches were carried out between different tissues of Epimedium brevicornu to chose the optimal tissue for the following study. The results showed that polysaccharide content of roots is the highest and significantly higher than that of Epimedium leaves and stems(P<0.05). Comparative analysis of antioxidant activity found antioxidant activity of polysaccharide in Epimedium leaves was significantly higher than that of polysaccharide in Epimedium roots and stems (P<0.05).Structural features of Epimedium brevicornu polysaccharides were investigated by infrared spectroscopy. And some other physicochemical properties like solubility of Epimedium brevicornu polysaccharides were identified. These results showed a high purity of Epimedium brevicornu polysaccharides, which has a significantly higher solubility in acidic solution than in alkaline solution (P<0.05). Picture of infrared spectroscopy suggested that molecules of Epimedium brevicornu polysaccharides had pyranoid ring and pyranoside structure. And also, there were chains of -(CH2) in Epimedium brevicornu polysaccharides molecules, and the number of chains was less than 4 (n<4). Epimedium polysaccharides possessed protective effect against plasmid DNA damage induced by oxidative stress. In summary, enzymatic water extraction seems to be a promising technique for separating Epimedium polysaccharides, which might act as a natural antioxidant or a chemopreventive agent due to their free radical-scavenging capacity and protective activity against DNA damage caused by oxidative stress.On the basis of the optimization of the extraction protcol of Epimedium polysaccharide, physical and chemical properties analysis and in vitro activity studies, a healthy oral liquid containing of polysaccharides from Epimedium brevicornu was developed.The optimal add doses of Epimedium Polysaccharide, ascorbic acid, citric acid and xylitol were optimized by orthogonal test and fuzzy mathematics comprehensive evaluation. The best processing was adding Epimedium Polysaccharide 0.012%, ascorbic acid 0.012%, citric acid 0.020%, xylitol 1.200%, respectively. Finally, the content of Epimedium polysaccharides of the oral liquid was 0.12 mg/mL, it prompted that the processing can save the active ingredients of Epimedium. The antioxidant study found that the IC50 values of oral liquid and Vc in the DPPH assay were 3.49 g/mL and 4.32 g/mL, respectively. What's more the IC50 values of oral liquid and Vc in the ABTS assay were 35.72 g/mL, and 36.37 g/mL respectively. The results indicated that oral liquid displayed DPPH· and ABTS+· radical-scavenging activities and the activities of Epimedium oral liquid were better than those of ascorbic acid. It shows that the functional oral liquid have free radical scavenging activity.