| Baohuoside 1 is a monosomic composition belonged to More hydroxy flavone, and is from berberidaceae epimedium plant Epimedium brevicornum Maxim. According to the literature reported at home an abroad, the resist lung cancer activity and biopharmaceutical characteristics is superior to the other flavonoid glycosides in Epimedium.The pharmacological studies show Baohuoside I has antitumor, anti-osteoporosis and other effects,and it has strong inhibition effects on myeloma and prostate cancer.In terms of pharmacokinetic study, before Icraiin is absorbed by the body,it is already metabolized to icaritin and Baohuoside I. The secondary glycoside of Icraiin,Baohuoside I,is easier absorbed into blood by body.Our studying team has already done previous studies on Baohuoside I isolated from herba epimedii. But the content of Baohuoside I in herba epimedii is very low. And compared to Baohuoside I, there are many compositions with similar polarity in herba epimedii. So,isolation and purification of Baohuoside I is very difficult. For this reason,we want to have further study to explore easier way to get Baohuoside I.This paper is consist of 3 parts, and the main content is taking Epimedium extract as the research object, expanding around the enzymatic hydrolytic technology, preliminarily discussing the suitability and applicability of enzymatic hydrolytic technology applied in Chinese herb extracts.In the first part,enzymatic hydrolytic technology in vitro combined with single factor experiment, Plackett-Burman design,central composite design-response surface methodology and other methods is adopted to hydrolyze Epimedium extract with enzyme, and the corresponding process which is optimized is obtained. Finally, the optimum process is suitable for the different content of Icraiin.and the verification tests is made. The final process could make almost all Icraiin in Epimedium extract transform into Baohuoside I. During the research, a certain content of some other component in the extract also transform into Baohuoside I.On this basis, using 20% Epimedium extract do verification tests. It confirmed that there are indeed other components transformed into Baohuoside I in enzymatic hydrolytic process. Through a series of process optimization, compared with the existing literature reporting process, the optimum process of Epimedium extract hydrolyzed to Baohuoside I by enzyme is improved whether in the aspects of the amount of raw materials or the end product of the conversion. At last, the adaptability of optimum process is verified by the other different content of Epimedium extract. It is proved that the two technological processes are respectively suitable for low content and high content of Epimedium extract hydrolyzed to prepare Baohuoside I by enzyme.In the second part, enzymolysis products are disposed and isolated with some methods,and then isolate and purify reaction products after disposing with some purification technologies,such as adsorption by coarse pored resin,polyamide resin adsorption method,recrystallization method,extraction and so on. Finally, high purity Baohuoside I is obtained. Determine the content of final product by HPLC, and chromatographic conditions were as follows.Column:Inertsil ODS-3(250 mm × 4.6 mm,5 μm); acetonitrile-0.1% phosphoric acid solution as mobile phase,gradient elution; column temperature:30℃;flow rate:1.0 mL/min;detection wavelength:270 nm. In purified enzymolysis products, Baohuoside I in chromatograms is completely separated from other peaks. This method is simple, accurate,reproducible,that could be as the content determination of Baohuoside I in end products.In the last part,through research and analysis of literature at home and abroad,research and prospect on biological enzymolysis technology applied in Epimedium is reviewed. |
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