Optimization Of Extraction,Purification,Identification And Biological Activities Of Flavonoids From Cappairs Himalayensis

Flavonoids in the dried fruits of Capparis himalayensis were extracted by ultrasonic assisted ethanol solution in this experiment.The extraction process was optimized by single factor experiment and response surface experiment.The optimum extraction process parameters were as follows:ethanol concentration was 60%,the extraction time was 2.5 h,the extraction temperature was 74?,the material-to-liquid ratio was 27 mL/g,the ultrasonic power was 400 W and the ultrasonic time was 15 min.Under this condition,the average yield of flavonoids was 4.86 mg/g.According to the static adsorption-desorption experiment,the resin with the best purification effect of Capparis himalayensis was selected from three different types(D101,AB-8,HPD600)macroporous resins.Results showed that AB-8 macroporous adsorption resin has a better performance.Its saturated adsorption capacity reached 31.02±0.25 mg/g,adsorption rate reached 71.04%±0.35%and desorption rate of 89.24%±0.21%.The saturated adsorption capacity of the polyamide resin was 26.39±0.47 mg/g,and the adsorption rate and the resolution were 61.85%±0.38%and 86.61%±0.32%,respectively.After being enriched and preliminarily purified by AB-8 macroporous adsorption resin,the purity of flavonoids was increased from 16.71%±0.63%to 58.59%±0.49%.Further purification by polyamide resin,the purity increased to 82.36%±0.61%.Experiments indicated that AB-8 macroporous adsorption resin and polyamide resin have good adsorption and purification effects on total flavonoids of Capparis himalayensis.Ultraviolet visible light scanning spectrum illustrated that the purified flavonoids had two absorption peaks at 372 nm and 261 nm,and the double absorption peak was the characteristic absorption peak of flavonoid.Infrared spectra showed that the extract of Capparis himalayensis contains characteristic absorption peaks of flavonoid.Combined with high performance liquid chromatography and liquid chromatography-mass spectrometry,3 kinds of flavonoids,namely kaempferol,rutin and quercetin were identified.The antioxidant activity and anti-tumor activity of flavonoids of Capparis himalayensis were determined by in vitro experiments.The antioxidant activity experiments were mainly composed of DPPH experiment,ABTS experiment and reducing power experiment.When the concentration of flavonoids was 1.0 mg/mL,the scavenging rates of DPPH and ABTS free radicals reached 84.79%and 90.08%,respectively.The concentration has a positive correlation with the ability to scavenge free radicals and the strength of reducing power.MTT assay showed that the inhibition rate increased with the increase of concentration,and the inhibition rate reached 42.56%when the concentration was 150 ?g/mL.When the growth rate of A549 cells was 50%,the average drug concentration was 193.19 ?g/mL,in other words,IC50=193.19±0.83 ?g/mL.In vitro activity experiments showed that flavonoids of Capparis himalayensis had a better antioxidant activity and a anti-tumor activity.