| [Objective] Japanese persimmon (Diospyros kaki Lf) persimmon leaves and persimmon leather alcohol extract ingredients were identified, establish ursolic acid, oleanolic acid, total flavonoids determination. And to carry out fresh persimmon peel take ursolic acid, sweet persimmon leaf total flavonoids extraction technology research, screening optimum parameters provide the basis for industrial production.[Method] fresh persimmon leather heated to reflux for 75% ethanol extraction, separation, purification of the compound 1, spectral data to identify its components. Fresh persimmon leaves soaked in methanol extraction, separation, and purification was 10 compounds, spectroscopic methods of its components. Determination of total flavonoids, ursolic acid, oleanolic acid content, complete method validation using high performance liquid chromatography. Orthogonal experiment investigated the proportion of solvent, extraction time, extraction times and extraction of flavonoids influence on fresh persimmon leaves; the use of MCI Gel enrichment of persimmon leaf flavonoids; crystallization purification of flavonoids, get fresh persimmon leaf extract the optimum flavonoids. Impact were investigated macroporous resin, solvent extraction, solid ursolic acid extraction method enrichment; orthogonal experiment investigated the proportion of solvent, extraction time, extraction times on fresh persimmon peel extract ursolic amount contrast crystallization, column chromatography and crystallization comprehensive impact refining, get fresh persimmon skin extract ursolic acid optimum extraction process.[Results] fresh persimmon peel compound made by a spectral analysis of ursolic acid. Alcohol extract of fresh persimmon leaves isolated 10 compounds identified five compounds which are:kaempferol -3-O-β-D- glucoside (1), kaempferol -3-O-β-D-galactopyranoside (2), quercetin -3-O-β-D- glucoside (3), eleutheroside E (4) 3,3,5-trimethyl-4-hydroxy-4-(γ-O-β-D-glucoside)-butenyl-5-ene-cyclohexanone (5). High performance liquid chromatography ursolic and oleanolic acid:sample using ultrasonic extraction, octadecylsilane bonded silica as a filler; methanol-0.2% phosphoric acid (87:13) as the mobile phase; flow rate per minute 0.5ml; detection wavelength was 210nm; column temperature 25℃. Oleanolic acid reference sample volume in the range of 4.996～124.900μg a good linear relationship (R2=0.9993362), the average recovery was 100.95% (RSD=4.49%). Ursolic acid reference substance in the range of 32.784～409.800μg good linear relationship (R2=0.9964001), the average recovery was 97.32% (RSD=2.85%). HPLC determination of total flavonoids:samples were extracted with methanol, octadecylsilane bonded silica as a filler; 0.4% phosphoric acid as mobile phase A, methanol mobility B; gradient elution; flow rate per minute 1.2ml; detection wavelength of 265nm; column temperature was 30 ℃. SZY-7 reference in the range of 0.052～0.1239μg insider sexual relations Department of good (r=0.99998), and the recovery was 95.18%(RSD=3.2%); SZY-6 reference in the range of 0.0257～0.0613μg good linear relationship (r= 0.99997), the recovery was 96.72%(RSD=3.31%); SZY-5 reference at 0.052～ 0.1239μg (r=0.99963), and the recovery was 98.87%(RSD=1.00%). Fresh persimmon peel take ursolic acid optimum extraction parameters were:5 times the amount of 95% ethanol extracted four times, each 60 minutes; by contrast, ursolic the most economical and simple method for solid extraction enrichment France; the best method for purification by column chromatography and crystallization synthesis method. Optimum extraction parameters sweet persimmon leaf total flavonoids were: 3 times the amount of 50% ethanol extracted three times, each time 90 minutes; MCI Gel adsorption enrichment methods for flavonoids best, MCI Gel resin work of flavonoids 60% of the full sample volume, ie 2.1mg/g; equivalent per gram of resin may accept 0.075g persimmon extract (specific gravity 1.35; 50℃); purification method for 3 to 5 times the amount of 80% to 90% ethanol solution to dissolve after filtration, crystallization.[Conclusion] persimmon peel contains large amounts of ursolic acid, oleanolic acid content is very low but, in order to obtain high purity ursolic provided favorable conditions. Persimmon peel ursolic and oleanolic acid content, the content of total flavonoids persimmon leaf method is accurate and reliable determination of the method as determined by high performance liquid chromatography. This study provides a persimmon peel ursolic acid extraction method has important significance for the needs of high purity ursolic acid materials needed pharmaceutical and cosmetics industries. Persimmon leaf flavonoid extraction method is simple, economical and practical, easy to industrial production, with good market prospects and competitiveness. |
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