Study On The Extraction And Separation Technology Of Active Substances From Pomegranate Flowers

There are many compounds in pomegranate flowers, like polyphenol , flavones,triterpene and so on.These compounds have all kinds of functions,such as softening vessels,digesting food,relieving diarrhea and itching,removing toxicity,making hypothermia and resisting too much gastric acid.In recent years,antioxidation,curing diabetes and protecting liver can also be founded in pomegranate flowers,and the pomegranate flowers is high in research and development.Pomegranate flowers are used as raw material in this dissertation.After drying, crushing,grinding,we use the enzyme and ultrasonic extraction technology and the method of response surface optimization to determine the best parameters,so as to extract polyphenol,flavones,triterpene largely.Then we measure its content by the TLC and ultraviolet spectrophotometry,and separate and purify these active substances in the last.The final results are as follows:1.Based on a single factor experiment, we study the effects of the quantity of enzyme,the time of enzymolysis,the concentration of ethanol,the frequency of ultrasonic and the time of ultrasonic extraction in enzymolysis process.Using the influence of Chinese software Minitab15 response surface analysis,we determine the best technological conditions:the quantity of enzyme is 0.05g,the time of enzymolysis is 60min,the concentration of ethanol is 80%,the frequency of ultrasonic is 36 kHz,and the time of ultrasonic extraction is 20min.2.We use the TLC and ultraviolet spectrophotometry to determine the content of three active substances in pomegranate flowers.After separating the sample and reference substance by the TLC and ultraviolet spectrophotometry,we determine and calculate the absorbancy of polyphenol,flavones and triterpene under various conditions,and then ascertain the content of each compound.Through the TLC separation,the volume ratio of chloroform,ethyl acetate,methanol,formic acid is 6:8:4:2,which is determined as the optimal spreading reagent;Based on the orthogonal experiment results and the ultraviolet spectrophotometry,we determine the content and extract polyphenol,flavones,triterpene,the respective content is 58.6mg/g,68.1mg/g and 11.0mg/g in the best experiment conditions.3.Through the static adsorption test from twelve kinds of macroporous adsorption resin,HPD-600 macroporous adsorptive resin,AB-8 macroporous adsorption resin,D-101 macroporous adsorption resin were selected to purify polyphenol,flavones, triterpene.These three technologies have high absorbance and desorption.Through researching on single factor experiment and orthogonal experiment to study the pH of sample liquid,the concentration of eluting solvent,the velocity of adding sample and the concentration of adding sample and the dosage of eluting solvent.Then we purify the three active substances through gradual separation technology.The results are as follows: The pH of sample liquid is 6.0, the concentration of ethanol is 90%, the velocity of adding sample is 2.0BV/h, the concentration of polyphenol is 2.02mg/mL, the dosage of eluting solvent is 3.0BV,in this conditions we may obtain the maximum adsorption of polyphenol, it is 93.85 mg/g, thus the recovery is 80.1%; The pH of sample liquid is 6.0, the concentration of ethanol is 90%, the velocity of adding sample is 1.0BV/h, the concentration of flavones is 1.88 mg/mL, the dosage of eluting solvent is 2.0BV,in this conditions we may obtain the maximum adsorption of flavones,it is 69.93mg/g,thus the recovery is 92.5%; The pH of sample liquid is 6.0, the concentration of ethanol is 90%, the velocity of adding sample is 3.0BV/h,the concentration of triterpene is 0.80 mg/mL, the dosage of eluting solvent is 3.0BV,in this conditions we may obtain the maximum adsorption of triterpene,it is 61.57mg/g,thus the recovery is 84.2%.