Studies On Extraction, Purification And Efficacy Of Total Flavonoids From Exocarpium Citri Grandis

At present, traditional Chinese medicine has attracted lots of attention and researchersshow great interest about its physiological effects，which has the high value and potentialmarket prospect. Exocarpium citri Grandis is a kind of traditional Chinese herbal medicine.According to ancient record and modern research, Exocarpium citri Grandis contains avariety of active ingredients, with phlegm and liqi, invigorating spleen, Improve digestion,relieve coldness, suitable for Phlegm stagnation, cough due to wind-cold evil chest coldcough, throat itching asthma, food stagnanted, vomiting new frowsty etc. Exocarpium citriGrandis flavonoids has became a research focus. Therefore, it’s essential to studyExocarpium citri Grandis flavonoids content and a hige efficient method for Exocarpiumcitri Grandis flavonoids extraction, separation and purification,thire structure as well aspotential pharmacological function.Exocarpium citri Grandis form Xiushui as the test material, we researched theextraction method of Exocarpium citri Grandis flavonoids, then discussed thire separationand purification.Preliminary do the research of the structure in Exocarpium citri Grandisflavonoids. A variety of methods was used to detect the antioxidant activity in Exocarpiumcitri Grandis flavonoids, and we made a study on the cough expectorant and antiphlogisticanalgesic effects.Based on the experiment of single factor to optimize extraction yield of totalflavonoids. The results showed that the optimal extraction conditions of total flavonoidsfrom Exocarpium citri Gandis were ethanol concentration30%, ratio of the liquid tosolid1:100g/mL, the ultrasonic power150W, extraction temperature80℃.Under theseconditions, the predicted value of extraction yield of total flavonoids was0.884%.Response surface method design is no significant difference with the predicted value (p>0.05), and verify the feasibility of the experimental model.Taking the absorbability and desorption properties as parameters, we investigated thepurifying conditions of total flavonoids from Exocarpium citri Grandis by macroporousresins, compared with five types of macroporous resins. The result showed that D-312macroporous resins was the best one. The optimal adsorption process conditions were asfollows: pH4,1.0mL/min adsorption velocity. Besides, the best desorption conditions: pH7,1.0mL/min desorption velocity of flow, concentration of desorption soluntion80%. Thepurity of general flavonoids extracted from Exocarpium citri Grandis was raised from14.16%to53.5%. After polyamide resin purification of Exocarpium citri Grandisflavonoids, purity reached81.37%, is1.52times of the macroporous adsorption resinpurification. HPLC gradient elution conditions was determined by the test.The analysis wasperformed on column with mobile phase contained acetic acid (0.1%)-acetonitrile.Theflow rate was0.8mL/min and the temperature of column was30℃,and the detectionwavelength was set at280nm.The injected volume was10μL. Using HPLC-MS method,preliminary analysised the contents of total flavonoids, The result is probablyIsorhamnetin-3-O-neohespeidoside, quercetin-3-O-β-D-galactopyranoside, NaringinDihydrochalcone.Meanwhile, the antioxidant activity of Exocarpium citri Grandis was evaluated byreducing poweron, hydroxyl free radical, superoxide anion and DPPH radical scavengingsystems, The total reducing poweron and DPPH radical scavenging ability compared withVc was no significant difference (p>0.05). Hydroxyl free radical scavenging ability wassignificant difference compared with Vc (p<0.05).And the superoxide anion removal forcewas extremely significant different form Vc (p<0.01).Exocarpium citri Grandis total flavonoids’ effect on anti-inflammatory, expectorant,relieve a cough was studied. Effect on inflamed mouse auricle caused by dimethybenzeneshowed that all dose group and positive control group were extremely significantdifference, compared with blank control group (p<0.01). Ammonia cough test, low doseand blank control group were no significant differences, while low dose group and highdose group had a extremely difference (p<0.01), and middle and high dosage groupcomparing with positive control group there stood a extremely difference (p<0.01).Tracheal phenol red method, phlegm experiment showed that pummelo Exocarpium citriGrandis all dosage group and positive control group were extremely significant comparedwith blank control group (p<0.01), and there were no significant differences between dosegroups (p>0.05). Effect of total flavonoids Exocarpium citri Grandis groups phlegm andthe positive control group were no significant difference (p>0.05). All those experimentscould illustrate Exocarpium citri Grandis flavonoids can diminish inflammation, inhibitcough and reduce phlegm.