The Determination Of Five Flavonoids Ingredients In Buddleia And Study On Its Fingerprint And Antioxidant Activity

A HPLC method was built for content determination of five flavonoids in traditional Chinese medicine Buddleja.The HPLC fingerprint of Buddleja and the antioxidant activity of the total flavonoid were studied so as to provide a standard for the quality control and pharmacological activity of Buddleja.MethodsA HPLC method was used to determin the active ingredients of five flavonoids in Buddleia.A HPLC analysis was carried out on a Hypersil ODS(4.6 mm×200 mm,5?m),the mobile phase consisted of methyl alcohol(A)and 0.2% phosphoric acid solution(B).The flow rate was 1.0m L/min.The detection wavelength was set at 256 nm and the column temperature was30?.Five flavonoids were used as the reference to study the characteristic fingerprinting and evaluate its similarity.The antioxidant activity of the total flavonoid was evaluated by researching the total reducing power of hydroxyl radical(·OH),superoxide anion radical(·O2-)and DPPH free radicals and the Vc was taken as the positive control.ResultsThe calibration curves were linear with in the range of 1.63~75?g/m L(r=0.9995)for buddleoside,0.42~26?g/m L(r=0.9997)for rutin,0.77~152?g/m L(r=0.9994)for luteolin,1.02~108?g/m L(r=0.9996)for quercetin and 0.91~99?g/m L(r=0.9992)for apigenin.The average recovery of buddleoside,rutin,luteolin,quercetin and apigenin were 99.7%,97.9%,99.7%,99.5% and 99.1%,the RSD were 1.3%,1.7%,0.7%,1.1% and 1.0%.12 batches of Buddlejas were determined and the characteristic spectrum was established.TCM fingerprint evaluation software was used to calculate the similarity of 12 batches of Buddlejas.Ten common peaks were identified.The similarities of 11 batches were proved higher than 0.90 and 1 batches were proved less than 0.90.In this experiment,the total flavonoids of Buddleja were studied,and the antioxidant activity of total flavonoids of Buddleia was evaluated by in vitro antioxidant model.When the total flavonoid concentration was 80?g/m L,the clearance rates of DPPH radical,superoxide anion radical and hydroxyl radical were 75.32%,48.45% and 60.31%.The result showed that the total flavonoids of Buddleia have scavenging ability on DPPH,superoxide anion and hydroxyl radical,and there was a positive correlation between total flavonoid concentration and free radical scavenging rate in a certain range.ConclusionsThe HPLC fingerprint was set up and five flavonoids in Buddleia wer e determined simultaneously,the anti-oxidation activity of the polysacchari de also were studied.All those provided a more reliable and stronger evi dence for the control of the Buddleia's quality and its research of pharm acological activity.