黄酮类物质的分离和纯化解析

Determination of the adsorption isotherms 100 mL feeding solutions with different concentration of raw leaves (0.01, 0.02, 0.04, 0.06, 0.08 and 0.1 g /mL) were placed in contact with 3 g hydrated resin LSA-21, respectively. The adsorption experiments were carried out for 3 h at temperatures of 25, 35, 45 and 55 ◦C. The concentrations of total flavonoids and oleuropein at equilibrium were determined by UV and HPLC, respectively.

Adsorption kinetics on selected resin
Eleven feeding solutions of 0.1 g /mL of raw leaves were added to eleven 250 mL conical flasks containing 3 g hydrated resin. All 11 flasks were continually shaken under the same conditions, but the adsorption time was different for each within a 350 min range. The concentrations of total flavonoids and oleuropein were monitored at different time intervals until equilibrium was reached.
Simultaneous separation and purification of flavonoids and oleuropein from Olea europaea L. (olive) leaves using macroporous resin
Contents
INTRODUCTION
2018/10/21
EXPERIMENT

Chemicals and reagents Chromatographic grade acetonitrile, analytical grade ethyl acetate and ethanol,deionized water,distilled water. Rutin and oleuropein standards . Preparation of the sample solution The air-dried powder of olive leaves (0.3 kg) was soaked with 3.6 L 80% ethanol for 1 h, and then heated and refluxed at 80℃ for 2 h. The extraction was then repeated. The two extracts were filtered and combined, and then the ethanol in the filtrate was collected under reduced pressure.The hydrated residue was diluted to 3 L with distilled water. After filtering, the transparent solution provided the sample solution, with a concentration of raw leaves of 0.1 g /mL.
Extraction Methods
Silica-gel column chromatography, liquid–liquid extraction, solidphase extraction, high-speed countercurrent chromatography ,and dynamic ultrasoundassisted extraction .
Other Methods
Biopolymers and polymeric absorbents have been used in separating and purifying target compounds from natural plant resources. Besides biopolymers, macroporous resin (MAR) is an important polymeric absorbent.

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).

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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.

Dynamic adsorption and desorption tests First dynamic adsorption and desorption Glass column ( 400 mm × 20 mm), which was wet-packed with about 40 g hydrated resin LSA-21. The packing length was 190 mm. 700 mL of sample solution (0.1 g /mL raw leaves) flowed through the glass column at a flow rate of 200 mL/ h, and concentrations of the measured compounds in the elution solutions were monitored at 10 mL intervals by UV and HPLC at ambient temperature. After reaching adsorptive equilibration, the resin column was first washed with 100 mL distilled water and then desorbed with 200 mL of 70% ethanol solution. The contents of total flavonoids and oleuropein in the desorption solutions were monitored by UV and HPLC .Finally, the crude extract was obtained by drying at 105 ℃ to achieve constant weight. The extract was made into powder for the second dynamic adsorption and desorption cycle.