Studies On Extraction Purification And Bioactivities Of Flavonoids In Black Currant Leaves

Black currant commonly known as black bean fruit,is a special kind of small berries in cold area Heilongjiang,the leaves are rich in flavonoids.The study choose the leaves of black currant as the the experimental materials, In the research of flavonoids based on the best extraction process,systematically research flavonoids separation,purification,identification and biological activity.expect to provide support for black currant industrial production.In extraction process using response surface method optimized ultrasonic extraction conditions:determine the optimum theoretical conditions:ultrasonic time30min,ultrasonic power517W,liquid to material ratio15.2:1mL/g,flavonoids extracted under this conditions theoretical predicted value is4.6916mg/g,actual operating conditions:ultrasonic time30min,ultrasonic power520W,liquid to material ratio15.2:1mL/g, flavonoids extracted under these conditions the amount of up to4.5891mg/g,is very close to the predicted value4.6916mg/g.Study purification through macroporous resin as adsorption material,through the static experiment compare six macroporous resin effect of adsorption and desorption flavonoids of the black currant leaves,chose three kinds of macroporous resin:NKA-9%D4020、D101-1,through the adsorption and desorption dynamic experiment,determine NKA-9as experimental resin purified the flavonoids of black currant leaves.The optimum conditions of NKA-9dynamic adsorption and desorption the flavonoids of black currant leaves is:at25℃,the adsorption rate1mL/min (concentration0.9311mg/mL),adsorption time1h, the sample volume60mL, anhydrous ethanol as eluant,elution flow rate1mL/min,elution time100min,elution volume100mL,under these conditions,the purity of black currant leaves flavonoids is up to40.15%,compared the former(without purification)(9.53%),the purity enhanced more than4times. These show that NKA-9macroporous resin purifies well.In the separation process,the Sephadex LH-20column was applied to separation of the purified flavonoids,anhydrous methanol was chosen as elution,solvent elution flow rate was1.0mL/min,through the HPLC analysis (chromatography conditions were as:column: Agilent-1100C18column; mobile phase:methanol:water (50:50);flow rate:1.0mL/min;detection:Agilent-G1314AUVdetector;detectionwavelength:360nm;temperature:room temperature),merge the same components,gain extractive flavonoids,the separation was detective by HPLC.The result show:the main of flavonoids of black currant leaves is rutin,the purity is 86.37%.The flavonoids activity experiments investigated the inhibitory effect of the protein non-enzymatic glycation in vitro; determination of hydroxyl radical scavenging.The results show:inhibition action increased with increased of the concentration of flavonoids,along with the extend of time,inhibition rate first increased then decreased,the maximum inhibition rate emerge on the third day,61.89%.The effect of hydroxyl radical scavenging is increased with the increased of the concentration of flavonoids,when the concentration of flavonoids is up to1.18mg/mL,the rate of clearance is65.4%,IC50is0.875mg/mL.