| Herba Epimedii is barren wort genus, alias "Xianlingpi" in Chinese. It is one of the most common in traditional Chinese medicine. The study found that Epimedii contain many kinds of effective components, including flavonoids, alkaloids, polysaccharide, polyphenols, lignans and so on. At present, investigators have found many kinds of flavonoid components from Epimedium koreanum Nakai. (E. koreanum). Modern pharmacology study showed that Epimedii have many kinds of pharmacological activity, including anti-osteoporosis, resistance to disease of heart head blood-vessel, anticancer, anti-depression, anti-inflammatory, antibacterial, antioxidant activity, anti-aging activity, enhance sexual function and so on.In the paper, we study on extraction, separation polysaccharides and flavonoids effective ingredients from E. koreanum. According to E. koreanum pharmacological activity, developed health wine, and solved the problems existing in the health wine storage. Finally, determining antioxidant activity of extract of E. koreanum. The research on E. koreanum provides basis for the further development and utilization on E. koreanum. Specific research work including:1. Study on extraction technology of polysaccharides from E. koreanum.Determination the effect of the ethanol concentration, extraction pressure, extraction time and solvent content on high-pressure extraction technology of polysaccharides. The result showed that the optimal ultra-high pressure extraction process of polysaccharides was: extraction solvent,30% ethanol solvent, extraction pressure,300 MPa, extraction time,30 s, solid/liquid ratio (w/v),1:50 (w/v, g·mL-1).Determination extraction technology of polysaccharides by others methods, ultrasonic, extraction solvent,30% ethanol solvent, extraction power,60 w, and extraction time,90 min, solid/liquid ratio (w/v),1:50 (w/v, g·mL-1); hot reflux, extraction solvent,30% ethanol solution, extraction time,120 min, solid/liquid,1:50 (w/v, g·mL-1); microwave-assisted, extraction solvent,30% ethanol solution, extraction time,60 s, solid/liquid,1:50 (w/v, g·mL-1), respectively. Compared on different extraction methods, we can conclud that the extraction yield of total sugar and reducing sugar extracted by the hot reflux extraction and microwave extraction were higher than the high pressure extraction and ultrasonic extraction. The contents of polysaccharides extracted by four extraction method were no significant difference. And from HPLC chromatogram can be seen that extract composition of four extraction method have no essential difference nearly, just contents distribution of each ingredient have slightly difference. But high-pressure extraction was more appropriate than other extraction methods in extraction time, the energy consumption.2. Separation and purification flavonoid components of E. koreanum by HSCCC.Determination two-phases solvent systems was chloroform-methanol-water (8:7:5). In addition to remaining ratio of stationary phase as an index, inspected Effect of revolution, mobile phase flow rate and testing temperature on the remaining ratio of stationary phase. The result showed that the optimal operating condition of separation and purification flavonoid components of E. koreanum by HSCCC was:revolution 850 r·min-1, mobile phase flow rate,1.5 mL·min-1, testing temperature,25?.According to two phase solvent systems chloroform-methanol-water (8:7:5) and the optimal operating conditions, seven compounds were isolated from E. koreanum Determine the structure of the compounds by ESI-MS. Compounds 2,3 and compounds 5,6 were isomers, respectively. Because we can not judge the concrete structure according to MS and HPLC, therefore can only be sure that compound 2,3 is Caohuoside A, B or Korepimedoside A. Compounds 5,6 is Caohuoside E or Epimedokoreanoside?. Compound 1 is Icariside?. Compound 4 is Sagittatoside B. Compounds 7 is Icariin.3. Antioxidant activity of effective ingredients.Determination antioxidant activity of effective ingredients using DPPH scavenging activity, hydroxyl radicals scavenging activity and reducing power. The experimental research shows that crude extract, crude flavonoids and effective components by different solvent extraction all have antioxidant activity, along with increasing levels of antioxidant activity enhanced. Determine correlation of different antioxidant activity determination methods by SPSS statistics software, the results showed that antioxidant activity results determined with different methods is consistent.The test result found that epimedii main antioxidant effect components are mainly concentrated in the effective components of ethyl acetate and chloroform extraction. When the content of extracts to 0.3 mg·mL-1, its antioxidant activity was quite to TBHQ, and when contents reached 0.5 mg·mL-1, its antioxidant activity was higher than TBHQ.4. Establish quality standards of epimedii health wine.Refer to the People's Republic of China national standards for liquor, wine, blending and brewing wine quality standards regulations, sampled inspection and investigation chemical composition, such as flavonoid and polysaccharide, physiochemical index, such as Alcoholicity, total sour, pH value, total solid of health wine, respectively. Establish one quality standards of epimedii health wine for industrial production.5. Investigation on the turbidity factors of health wine.Determination the main ingredients of epimedum health wine precipitation, composition containing polyphenol, flavones, total sugar and protein. Contents of composition are polyphenol,24%, flavones,9%, total sugar,7%, protein,6%, respectively. We can speculate that the reasons of the formation of wine precipitation were the total polyphenol, protein and polysaccharide. In addition,54% unknown components of wine precipitation were not detected in literature, which was probable pectin, esters and fiber vegetables that easily form the muddy waters of the beverage component, these components may also the factors of wine turbidity forming. Compared to the resultant precipitates quantity and wine clarification, it can be concluded that the ester and metal ions in the sorghum white wine has certain influence to the formation of health wine turbidity.6. Epimedii health wine clarification experiment.Application clarifying agent and ultra-filtration membrane clarification on health wine, determine the concents of flavonoid and polysaccharide. Effective component content of health wine were changed after treatment, which less than 10% of flavonoids loss. It does not affect the effect of concrete. But the polysaccharide adsorption of ZTC-III, gelatin clarify agent is quite obvious, loss of 50% will degrade the efficacy of concrete. And the contents of polysaccharide after membrane treatment changed little, remaining radio above 90%. Therefore, the membrane treatment was better than clarify agent in effective components remain.Investigation on color and transmittance after clarification treatment. Found that stability of health wine is obviously improved after clarification treatment. Color and transmittance very little change under room temperature for four months. The stability and effect of clarification treatment with membrane filtration and ZTC-?clarify agent were better than gelatin clarification. Investigation on pH change whether join clarify agent in health wine. It can see that the pH value of health wine lower than before health wine clarification, but no significant difference. Contrasting HPLC chromatogram in 254 nm and 270 nm of health wine before and after processing, it can see that the ingredients that the composition of health wine nearly change after treatment.Through a series of checking, we can draw the conclusion. Compared with other clarify agent, no matter from the effective component reserved or clarification effects perspective, membrane processing would have better effect. Ultra-filtration membrane has advantages of energy saving, simple etc. It will have a broad application prospect in dealing with health wine and beverage. |
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