黄酮类物质的分离和纯化预案.pptVIP

;Contents;;EXPERIMENT; materials and pretreatment eight resins : D101, DM130, HPD450, LSA-21, LSA-40, 07C, LSD001 and HPD600. These resins were pretreated by soaking in ethanol for 24 h, and then washed with ethanol until there was no turbidity when a threefold volume of water was added into the eluent. The MAR was subsequently washed with distilled water until the ethanol was thoroughly replaced by the distilled water before use. ; Analysis of target polyphenols 1. UV analysis of total flavonoids 2. HPLC analysis of oleuropein Agilent 1200 HPLC system,G1315B diode-array detector,G1312A binary pump and a G1328B manual injector. C18 column (Sinochrom ODS-BP,250 ×4.6 mm, 5 μm).The column temperature was 25 ℃.The mobile phase was composed of acetic acid–water (0.2 : 99.8, v/v, A) and acetonitrile (B). The gradient elution of the mobile phase was as follows: 18–31% (B) in 0–29 min, 31–50% (B) in 29–30 min and 50% (B) in 30–34 min. The flow rate was set at 1.0 mL /min. The detection wavelength was set at 270 nm. The sampling size was 10 μL. The standard solution and different sample solutions were introduced into the HPLC system. The content of oleuropein was calculated by comparing the peak areas of standard and sample solutions. The results indicated that the working calibration curve based on oleuropein standard solutions showed good linearity over the range of 95.31–9150 μg/mL. The regression equation for oleuropein is y = 3.0625 x + 515.07(R = 0.9984), where y is the peak area of oleuropein and x is oleuropein concentration (μg/mL).;Static adsorption and desorption tests The static adsorption process:3 g hydrated test resin was placed in a 250 mL conical flask, and then 100 mL of the sample solution of 0.1 g/ mL raw leaves was added.The loaded flask was then continually shaken at 100 rpm and 30 ℃ in a shaker until adsorption equilibrium was reached. The static desorption process : After adsorption equilibrium was reac