Pharmacokinetics Of Four Homoisoflavanones From Polygonati Odorati Rhizoma By LC-MS

LC-MS is one of the most commonly used techniques for the determination of trace components.It has the advantages of fast analysis speed,high sensitivity and high accuracy,so it is commonly used for rapid qualitative and quantitative analysis.This technique has significant advantages in the study of pharmacokinetics of drug components in vivo.Polygonati Odorati Rhizoma is the dried rhizome of Polygonatum odoratum.It is a traditional Chinese medicine for both medicine and food.It has the functions of lowering blood sugar,blood lipid and anti-tumor.It is widely used in medicine,health food and cosmetics industries.The chemical constituents of Polygonatum odoratum mainly include polysaccharides,saponins,flavonoids and amino acids.The ethanol extract of Polygonatum odoratum has good pharmacological activity,rich in more than 20 kinds of homoisoflavanone components.10 kg Polygonati Odorati Rhizoma was extracted by 80%ethanol,separated by silica gel column chromatography,and purified rapidly by semi-preparative high performance liquid chromatography（HPLC）and preparative thin layer chromatography（TLC）.Four components with high content were obtained.The structures of these four compounds were identified by mass spectrometry,nuclear magnetic spectroscopy and other analytical methods,and their micro infrared spectra and micro Raman spectra were collected,micro Raman spectra was collected for the first time.The differences of the four homoisoflavanones in infrared and Raman spectra were discusessed as while.The contents of four homoisoflavanones（Ⅰ,Ⅲ,Ⅳ,Ⅴ）were higher in Polygonatum odoratum,It was found that the four homoisoflavanones（Ⅰ,Ⅲ,Ⅳ,Ⅴ）had strong antibacterial,hypoglycemic and anticancer activities,but no pharmacokinetic study was reported in vivo.In this experiment,A UPLC-MS/MS method for the simultaneous determination of four homoisoflavanones in the blood of Polygonatum odoratum was established for the first time,and the method was applied to the determination of the blood drug concentration in rats for the first time.1.Four homoisoflavanones compounds were successfully isolated from Polygonatum odoratum,the mass of which were 13.52 mg,63.83 mg,105.95 mg and 54.15 mg respectively,and identified as 6-methyl-8,4?-dimethoxy-5,7-dihydroxy-homoisoflavanone（Ⅰ）,6,8-dimethyl-5,7,4?-trihydroxy-homoisoflavanone（Ⅲ）,6-methyl-8-methoxy-5,7,4?-trihydroxy-homoisoflavanone（Ⅳ）,6-methyl-5,7,4?-trihydroxy homoisoflavanone（Ⅴ）.2.A UPLC-MS/MS method was established for the determination of four homoisoflavanone in blood.The mobile phase was 0.1%formic water-0.1%formic acetonitrile.The four components and internal standard were completely separated in 8minutes.The chromatographic and mass spectrometric conditions of the method were investigated,and then the specificity,linearity,precision,accuracy,recovery,matrix effect and stability of the method were investigated.It was found that the linear range of homoisoflavanoneⅢ,Ⅳ,Ⅴ,Ⅰwas 0.2-32,0.2-26,0.0875-14,0.0875-14,respectively.The results showed that the relative coefficient r was more than 0.9990,the detection limit of quantification was 0.2,0.2,0.0875,0.0875 ng/m L,the total precision RSD was 1.91%～6.23%,3.95%～6.23%,the recovery was more than 90%,the matrix effect and stability met the requirements of biological sample analysis,which provided the basis for the pharmacokinetic study of these four homoisoflavanones.3.The established UPLC-MS/MS method was used to detect the plasma samples of SD rats fed with the mixture of homoisoflavanoneⅢ,Ⅳ,Ⅴ,Ⅰ,the plasma concentration time curve was established,and the Cmax,Tmax,T_1/2Z,AUC_（0-t）,CL_Z/F,V_Z/F and other parameters of homoisoflavanoneⅢ,Ⅳ,Ⅴ,Ⅰin vivo was obtained.This method can be used to study the pharmacokinetics of these homoisoflavanones in rats.