Optimization Of Extraction Process And Preliminary Study On Serum Pharmacochemistry Of Puerariae Radix

Puerariae Radix is a commonly used Chinese herbal medicine in China.According to the 2000 edition of the Chinese Pharmacopoeia,the roots of Puerariae Lobatae Radix(PLR)and Puerariae Thomsonii Radix(PTR)are used as Pueraria.However,the two species were distinguished in the 2005 edition of the Chinese Pharmacopoeia.The dried root of Pueraria thomsonii Benth,a leguminous plant,is the dried root of the leguminous Pueraria lobata(Willd.)Ohwi.They have significant pharmacological activity in the treatment of cardiovascular diseases,hypertension and diabetes,and are homologous plants for food and medicine.After the two kinds of Puerariae Radix roots were crushed,they could hardly be distinguished,but the curative effect of the two kinds of pueraria roots was not exactly the same.This topic intends to use liquid-mass spectrometry and serum metabolomics theory to study the differences and their metabolic pathways between the components of PLR and PTR.To provide a new analytical method and means for the identification of similar and easily confused Chinese medicines.(1)Extraction process and in vitro antioxidant researchThe extraction process of isoflavones from Pueraria lobata was studied by ethanol reflux method and ultrasonic assisted reflux extraction.The yield of total flavonoids and the yield of dry extract were selected.The comprehensive single factor and orthogonal test results were used to determine the kudzu.The optimal extraction process is: 40% ethanol,material-liquid ratio 1:20,80 °C constant temperature water bath extraction for 2h.The best extraction process of isoflavones in the powdered pueraria is: 50% ethanol,the ratio of material to liquid is 1:10,75 °C ultrasonic for 65 min.The DPPH scavenging ability,total reducing ability and hydroxyl radical scavenging ability of PLR and PTR were determined by UV spectrophotometry.Both of them showed certain antioxidant activity and the antioxidant activity of PLR in vitro.To be higher than PTR,it may be related to the content of isoflavones higher than that of PTR.(2)In vitro material basic researchIn this experiment,24 compounds such as puerarin,3’-hydroxy puerarin and 3’-methoxy puerarin were detected from the PLR by liquid phase and chromatography.Sixteen compounds including puerarin,daidzin,puerarin xyloside and iris flavonoid glycosides we re identified from the PTR.3’-hydroxy puerarin and 3’-methoxy puerarin are two different substances,possibly chemical markers,used to distinguish between two species of Puerariae Radix.(3)Serum metabolomics researchIn this experiment,rats were used as a blood-inducing animal model,and the collected sera of PLR and PTR were analyzed by LC-MS.The endogenous metabolites and exogenous substances of two kinds of pueraria were studied by metabolomics research.For screening,data with VIP>1,P<0.05 were selected,and potential biomarkers were identified by database comparison.The results showed that they were mainly glycerophospholipids and glycerophospholipids.62 species of potential biomarkers were found in PLR,and a prototype compound,puerarin B,was found.Its metabolic pathways in the blood i nclude glycerolipid metabolism,sphingolipid metabolism and glycosylphosphatidylinositol(GPI)-anchor biosynthesis.The analysis of PTR,found 79 potential biomarkers,and a prototype compound soyasapogen A,its metabolic pathways in the blood are sphingolipid metabolism,glycerophospholipid metabolism,Glycolysis or Gluconeogenesis,Inositol phosphate metabolism and Purine metabolism.