Study On Anti-gout Effective Components Of Xiaofeng Prescription

Xiaofeng prescription consists of Atractylodes Lancea,Phellodendron Chinense,Smilax Glabra and Caulis Lonicerae.Our preliminary studies have screeninged out volatile oil from Atractylodes Lancea,alkaloids from Phellodendron Chinense have the effects of anti-inflammatory,analgesic effect,lowering uric acid value.Early experiments also established a volatile oil,alkaloid and organic acid preparation method,and flavonoids,saponins content determination method.This study will respectively study the methods of extraction and purification each component of Caulis Lonicerae and Smilax Glabra.Anti gout effective part of Caulis Lonicerae and Smilax Glabra will be screened out.With the combination of volatile oil and alkaloids,anti gout effective parts of Xiaofeng prescription will be screened out.The pharmaceutical research foundation on anti-gout five categories of new drugs about Xiaofeng prescription will be provided.The main content of the paper is listed as follows:1.The study of anti-gout activity of each part of the Caulis LoniceraeOn the basis of preliminary study,we established the flavonoids determination method and iridoid glycoside HPLC content determination method.In order to determine the optimal preparation method,we compared macroporous resin,polyamide resin,solvent extraction of separation and purification of flavonoids,saponins and iridoid glycoside site.Preparation of organic acid site,flavones site and glycosides site,with inhibit xanthine oxidase activity,xylene induced mouse ear swelling anti-inflammatory,acetic acid-induced writhing analgesic,mouse hyperuricemia model uric acid as index,anti gout effective part was screeninged out.The research showed that the Caulis Lonicerae in all parts of the optimal method of preparation is:using solvent extraction and purification of flavonoids and glycosides site(flavonoids purity were 60.25%,glycosides purity were 63.99%).The pharmacodynamic data shows that organic acid,flavone and glycosides without inhibiting the activity of XOD and uric acid lowering action.The three parts have analgesic effects,organic acids and flavones have anti-inflammatory effect.2.The study of anti-gout activity of each part of Smilax GlabraThrough the inhibition of XOD activity effect,we initial screened the anti gout effective part of Smilax Glabra.Determination method of flavone content was established.Through the reflux extraction and percolation extraction comparison,the optimum extracting process of total flavones from Smilax Glabra was established.Through the comparison of solvent extraction and macroporous resin purification,the optimal purification process was established.With xylene induced mouse ear swelling anti-inflammatory,acetic acid-induced writhing analgesic,mouse hyperuricemia model uric acid as index,anti-gout effective part of Smilax Glabra was screened out.By using UPLC-MS analysis of ethyl acetate fractions of chromatographic peak,flavonoids in ethyl acetate extract was determined.The research showed that the best method of preparation flavonoids is:percolation extractio n and extracted with ethyl acetate flavonoids fraction(90.06%purity).Flavonoid fraction had sig nificant inhibition of XOD activity,anti-inflammatory and analgesic effects.The ethyl acetate fra ctions detected seven flavonoids,respectively:epicatechin,astilbin,neoastilbin,isoastilbin,neois oastilbin,quercetin and naringenin.The epicatechin,astilbin,quercetin,naringenin inhibit the act ivity of XOD.The epicatechin,astilbin,quercetin,naringenin have the function of inhibiting the activity of XOD.3.The Xiaofeng prescription of anti-gout activity evaluationPreparation of the volatile oil fraction from Atractylodes Lancea by steam distillation,alkaloid fraction from Phellodendron Chinense by macroporous resin,organic acid fraction from Caulis Lonicerae by anion exchange resin,lavonoid fraction and glycosides fraction by Solvent method,flavonoids parts of Smilax Glabra by extracting with ethyl acetate.Becouse we have confirmed anti-gout activity of the volatile oil and the alkaloid,with xylene induced mouse ear swelling anti-inflammatory,acetic acid-induced writhing analgesic,mouse hyperuricemia model uric acid and inhibition of the XOD active as the index,we evalued of the combination part of anti-gout activity and screeninged out effective parts of Xiaofeng prescription.The Xiaofeng prescription anti-gout efficacy data showed that:the parts which are consisted of the volatile oil from Atractylodes Lancea,alkaloids from Phellodendron Chinense,flavonoids from Smilax Glabra,orgnic acids and flavonoids from Caulis Lonicerae have the best anti-inflammatory,analgesic and drop uric effects.The inflammatory inhibition rate was 62.4%,pain inhibition rate was 59.2%,uric acid value inhibition rates in 2 hours and 4 hours were 42.4%and 45.3%;XOD active inhibition rate was 15.71%.This shows the volatile oil from Atractylodes Lancea,alkaloids from Phellodendron Chinense,flavonoids from Smilax Glabra,orgnic acids and flavonoids from Caulis Lonicerae are the anti-gout pharmacodynamic material basis.