Lipid Lowering Activity Of Total Flavonoids Fraction Of Microcos Paniculata And The Corresponding Qualitative And Quantitative Analysis Of Apigenin-C-glycosides

Microcos paniculata Linn.is a Chinese herbal medicines,"medicine and food homologous" plants,has been widely used in clinical and folk tea.Since the Qing Dynasty,was recorded in the Lingnan region of the books,followed by ?Ling nan cai yao lu?,?Lu Chuan ben cao?are also documented.After 2010,the ?People's Republic of China Pharmacopoeia? have been included,showing its medicinal value has been fully affirmed.Unfortunately,most of them are used for herbal tea production rather than medical care.In order to further develop its resources,this paper refers to other literature and products.We extracted the total flavonoids fraction and separated them to obtain MpTFF and Apigenin-C-glycoside compounds.Then,we established the quality standard of MpTFF.Finally,we investigated the antioxidant and lipid-lowering activity of MpTFF.1.In this paper,the total flavonoids were extracted by heating and condensing,and AB-8 resin was used to enrich the active constituents.Subsequently,gel column chromatography(Sephadex LH20)was used for purification.Finally,MpTFF powder was obtained by spray drying(inlet temperature 210?;Spray air flow 538L·h-1;pump output 85%;pump output 10%).Subsequently,8 flavonoid glycosides contained in MpTFF were qualitatively analyzed by LC-MS combined with precise molecular weight,relative retention time and MS and MSE cleavage patterns.They were: vicenin-2,isoshaftoside,shaftoside,vitexin,vicenin-1,isovitexin,astragalin,narcissoside.2.A colorimetric method for the determination of total flavonoid glycosides(55.73±0.65%)and a HPLC method-Kromasil C18 column(250 mm×4.6 mm,5 ?m),The mobile phase consisted of methanol(A)and water(containing 0.2% phosphoric acid,B)at a flow rate of 1 m L · min-1.Graded elution: 0 ~ 12 min,82% ? 70%(B);12 ~ 45 min,70% ? 55%(B);45 ~ 60 min,55% ? 20%(B);The detection wavelength was 276 nm and the column temperature was 35?-for simultaneous determination of 8 flavonoids were established.The combination of the two methods can provide more scientific quality control for MpTFF.The solubility of MpTFF in water,pH 6.8 phosphate buffer(PBS)and 0.1 mo L·L-1 hydrochloric acid was determined by shake flask method,which was 1.46 ± 0.02(slightly soluble),7.48 ± 0.07(dissolved)and 25.84 ± 0.16 g · 100 m L-1(soluble).Among them,the solubility of MpTFF in three media were hydrochloric acid> PBS> water.This phenomenon may be caused by the complexity of MpTFF's composition;in the acid or alkali environment,the interaction between each other to increase its solubility.Therefore,in order to more accurately determine the solubility of traditional Chinese medicine components,we need a more scientific method.3.The activity of MpTFF was studied by in vitro clearance of DPPH free radical test and lipid lowering test in HLP mice.The results showed that it had a good ability to scavenge free radicals(EC50 value of 0.038 mg · mL-1),and at a dose of 312 mg ·(Kg·d)-1,MpTFF could effectively reduce the body weight of HLP mice Increased,decreased TC,TG and LDL-C levels,increased HDL-C levels.At the same time,for the AST and ALT two transaminase activity levels,MpTFF has a reduced effect.The above experiments show that MpTFF has a certain Anti-oxidative activity,lipid-lowering activity and protection of liver activity.4.MpTFF lipid-lowering active components were analyzed by computer simulation of molecular docking.First,access to the relevant literature,download lipid-related receptors;then,according to the composition of MpTFF to establish a ligand database,CDOCKER model for molecular docking.The results showed that vitexin,isovitexin and astragalin were able to bind to the receptor through more hydrogen bonds.The results further demonstrate that isovitexin,vitexin and astragalin are the major lipid-lowering active ingredients in Mp TFF.5.The product was hydrolyzed on the basis of MpTFF and purified by gel column chromatography,reversed phase ODS column chromatography and semi-PHPLC.1H-NMR,13C-NMR and HRMS were used to characterize the structure.Finally,4 glycosides were obtained.vicenin-2,vitexin,isovitexin,isoviolanthin Among them,vicenin-2 is the first separated from the Microcos paniculata,but also for the first time separated from the Microcos plants.