Preparation Of Red Raspberry Ellagic Acid And Its Anti-lung Cancer Cells A549 Activity

Red raspberry is a plant in the genus Rubus of the Rosaceae family.Red raspberry is rich in nutrition and has the reputation of "fruit of life".It is closely related to its bioactive ellagic acid.Ellagic acid is a polyphenol lactone,which has excellent medicinal and health care value.Red raspberry is rich in elliptic acid.With the continuous expansion of the scale of raspberry cultivation,In-depth research is of great significance to the comprehensive development and utilization of raspberries.In this research,microwave-assisted extraction of raspberry ellagic acid was studied,and X-5macroporous resin adsorption method was used to separate and purify raspberry ellagic acid.High-purity red raspberry ellagic acid was identified by HPLC-MS and particle size analyzer.To further study the effect of red raspberry ellagic acid on the proliferation and apoptosis of lung cancer cells A549 and the correlation between antioxidant and anti-cancer effects.The main findings are as follows:(1)Microwave-assisted was used to extract ellagic acid from red raspberry.The optimal extraction process of ellagic acid was obtained through single-factor experiments and response surface optimization design.The extraction rate and oxidation resistance were compared with traditional extraction methods.The results showed that the optimal extraction conditions were: extraction time of 1.8 min,extraction temperature of 60 ?,ethanol concentration of 50%,liquid-to-material ratio of 19: 1,and microwave power of 411 W.Under this condition,the content of ellagic acid was 82.75 mg / 100 g.which was 20.42% higher than the traditional method.At the same time,the free radical scavenging ability was significantly higher than the ellagic acid obtained by the traditional hydrochloric acid hydrolysis method.It shows that the microwave extraction method is better than the traditional extraction method.(2)Raspberry ellagic acid was separated and purified by macroporous resin adsorption method,and high purity red raspberry ellagic acid was identified byHPLC-MS and particle size analyzer.The results showed that the optimal adsorption process parameters of X-5 resin for purification of red raspberry ellagic acid were: the loading concentration of 1.0 mg / m L,the diameter-to-height ratio of 1:14,the loading volume of 80 m L,and the sample p H = 5 Dynamic adsorption was performed,with an adsorption rate of 92.76% and an adsorption capacity of 3.37 mg / g.The optimal desorption process parameters were: 500 m L of water washing volume,200 m L of 50%ethanol solution,and elution at a flow rate of 1.5 BV / h.The rate was 88.38%,and the recovery rate was 83.56%.The analysis of purified ellagic acid by HPLC-MS and particle size analyzer showed that the chromatographic peak area of ellagic acid increased significantly after purification,and the particle size of purified ellagic acid was much smaller than that of crude ellagic acid.The resulting powder is more delicate and uniform.(3)MTT method,apoptosis detection and scratch test were used to evaluate the effect of red raspberry ellagic acid on lung cancer cell A549,and the anti-lung cancer cell A549 activity was explored.The results showed that: MTT assay showed that 200?g / m L red raspberry ellagic acid acted on lung cancer cell A549 for 48 h,and the cell inhibition rate reached 84.05%,indicating that red raspberry ellagic acid could effectively inhibit lung cancer cell A549 proliferation.Apoptosis results showed that the apoptosis rate of lung cancer cells A549 in the 200 ?g / m L ellagic acid treatment group was 37.9%,and the apoptosis rate was concentration-dependent.In summary,it can be shown that red raspberry ellagic acid can inhibit the proliferation of lung cancer cells A549 And promote its apoptosis and inhibit the activity of lung cancer cell A549.The scratch results showed that red raspberry ellagic acid could significantly reduce the migration rate of lung cancer cell A549,and the mobility was concentration dependent.The migration rate of lung cancer cell A549 in the 200 ?g / m L ellagic acid treatment group after 24 h was only 26.2%.By establishing the correlation,it can be seen thatthere is a significant positive correlation between the antioxidant activity of red raspberry ellagic acid in vitro and the anti-lung cancer cell A549 proliferation activity.