Study On Extraction Process Of Active Ingredients In Chesetnut Vollen

Chestnut trees mostly grow in gentle slope and the river areas, Hebei, Shandong, and Shanxi province is the famous chestnut-producing areas. Chestnut pollen has rich nutrition and a balanced ratio of various ingredients with high medicinal value and health functions, it is called as a "complete nutrition" in the international. At present, the study of chestnut pollen is less.In this paper, the fatty acids, flavonoids, estradiol and protein of chestnut pollen are researched, according to the different nature of various components, using a suitable organic solvent to extract, finding the appropriate qualitative and quantitative methods to analyze the ingredients, and establishing the best extraction process conditions, the separation and purification conditions of flavonoids and estradiol are optimized.Extraction of fatty acids from chestnut pollen with hexane was studied, the types and percentage of fatty acids were detected by gas chromatography, the results are as follows: Extraction rate of fatty acid from chestnut pollen is3.05%, there are5sorts fatty acids in chestnut pollen:palmitic acid25.83%, stearic acid3.46%, oleic acid24.95%, linoleic acid32.89%, linolenic acid3.24%, unsaturated fatty acids accounted for61.08%of total fatty acids. Extraction rate of fatty acid from Chinese pine pollen is7.98%, there were4sorts fatty acids in Chinese pine pollen:palmitic acid26.50%, stearic acid2.60%, oleic acid36.65%, linoleic acid28.60%, unsaturated fatty acids accounted for65.24%of total fatty acids.Extraction conditions of total flavonoids from chestnut pollen are studied, the optimal extraction conditions are as follows:temperature of60℃, time of1.5h, ethanol content of80%and ratio of liquid to solid of10:1, flavonoids are extracted using the water-bath heating, In these conditions, the content of flavonoids in the extracts is1.68%, using the determination methods of "NaNO2-Al(NO3)3-NaOH".The content of flavonoids form chestnut pollen in the extracts is4.42%by HPLC quantitative determination. Compared with ultrasonic extraction, the extraction ratio of two methods is almost the same. The separation and purification of total flavonoids from Chestnut Pollen by AB-8Macroreticular Resin column chromatography, the optimal separation and purification conditions of flavonoids are as follows:effective column volume of20mL, adding6mL flavonoids extraction to the column, washing with10BV distilled water, then washing with6BV ethanol content of75%at the velocity of1mL/min, the eluent amount is6BV of resin bed.Extraction conditions of estradiol from chestnut pollen are studied, the better extraction conditions of estradiol are as follows:temperature of70℃, time of2.5h, ethanol content of80%and ratio of liquid to solid of10:1, the content of estradiol in the extracts is9.72μg/g by HPLC quantitative determination.The perfect separation and purification system of estradiol are as follows:two times column chromatography, the resin column is eluted with4BV ethanol content of60%and then4BV acetone successively.Different methods and different enzyme-assisted extraction of protein are studied, the result is the best effect of alkaline protease-assisted digestion and the protein extraction as high as5.63%.