Component Analysis And Property Study Of Flavonoids From Potentilla Fulgens

Potentilla fulgens is a kind of perennial herb. Their flavours: bitter, astringent, cold.Their curative effects: clearing away heat and toxic materials, relieving diarrhea withastringents, cooling blood and hemostasis. They grow in hillside, grassland, bushwood,edge and centre of forest. So far, P. fulgens hasn’t been recorded as traditional Chinesemedicinal material in Chinese Pharmacopoeia (2010), few of research reports of extraction,purification, chemical components and bioactivities at home and abroad are found,therefore it has the important research value.This paper includes four parts: the extraction of flavonoids from P. fulgens bymicrowave assisted extraction, the purification by macroporous resin, the qualitative andquantitative analysis by high performance liquid chromatography-electrospray ionizationtandem mass spectrometry (HPLC-ESI-MS/MS), the measurement of antioxidant activityby five chemical methods and cell model method.Flavonoids from P. fulgens were extracted by microwave method, the effects ofethanol concentration in extractant, ratio of material to liquid, extraction temperature,microwave power and extraction duration on flavonoids yields were determined. Theresults showed that the optimum conditions were: ethanol concentration in extractant50%,ratio of material to liquid1:30, extraction temperature60℃, microwave power400W, andextraction duration5min. The best yield of flavonoids was5.68%.Researching the adsorbing and desorbing experiments of seven kinds of macroporousresins, macroporous resin D101was found to show the best effect of purifying flavonoidsfrom P. fulgens. Dynamic method was carried out to acquire the best flow rate and pH valueof the sample solution, the best pH value and ethanol concentration of the desorbent. Theeffect of purification was characterized by HPLC. The result showed that the best purifyingcondition of resin D101for flavonoids from P. fulgens are as follow: the adsorbing flowrate is2BV/h, the pH value of sample solution is4, the pH value of desorbent is8, theethanol concentration of desorbent is60%. The purification by resin D101can remove apart of impurities from P. fulgens, and is suitable to purify strong polar flavonoids whosering A and B both have conjugated double bonds, increases the purity of flavonoids from P.fulgens from28.2%to46.6%.Using high performance liquid chromatography-electrospray ionization tandem massspectrometry (HPLC-ESI-MS/MS), qualitative analyses were carried out on flavonoidsfrom P. fulgens, according to retention times, molecular ions, fragment ions in MS2andpertinent literatures, quantitative analyses were carried out with multiple reaction monitoring (MRM) monitoring specific fragment ions. The main flavonoids from P. fulgensand their contents were apigenin-7-O-β-D-glucuronide6.66μg/mg, hyperoside2.39μg/mg,astragalin2.30μg/mg, scutellarin1.95μg/mg, luteoloside0.44μg/mg, potengriffioside A0.39μg/mg, isovitexin0.01μg/mg.Total antioxidant activity, hydroxyl radical scavenging activity, superoxide anionradical scavenging activity, DPPH free radical scavenging activity of the flavonoids from P.fulgens and their inhibition on peroxidation of yolk lipoprotein were determined, withascorbic acid as reference. The results showed that the flavonoids from P. fulgens were akind of excellent antioxidants, especially hydroxyl radical scavenging activity of theflavonoids from P. fulgens had about6times hydroxyl radical scavenging activity, and3times inhibition on peroxidation of yolk lipoprotein of ascorbic acid. Furthermore, theflavonoids from P. fulgens showed obvious antioxidant activity in colon cancer cell (HT-29)and mice macrophage (RAW264.7), which were concentration-dependent.