The Chemical Composition And Pharmacokonetic Studies Of Iridoids On Picrorrhiza Scrophularaeflora Pennell

Picrorrhiza scrophularaeflora Pennell is a perennial herb of the PicrorrhizaRoylegenus from Scrophulariaceae.The dry roots and rhizomescan abate vacuity heat,chancroid fever and damp-clearing heat.It usually used for bone steaming hot flashes,infantile chancre fever,damp-heat diarrhea,jaundice and urine redness,sores and sores and so on.The components of P.scrophularaefloracontains iridoids,phenylethanosides,phenolic glycosides.Iridoids are the main active and distinctive constituents.Modern pharmacological studies have shown that the compounds contained in P.scrophularaeflora have remarkable effects in protecting liver and gallbladder,anti-bacterial and anti-inflammatory,protecting damaged nerve cells and apoptotic cardiomyocytes,etc.In order to study the iridoids of P.scrophularaeflora,the dried roots and rhizomes were extracted under reflux with ethanol-water(75:25,v/v),and then filtrated.The concentrated extract was mixed with water,and then the ethyl acetate part of P.scrophularaeflora was extracted.Using silica gel column chromatography and ODS column chromatography,the ethyl acetate fraction of P.scrophularaeflora was systematically separated and purified by HPLC preparation chromatography.As a result,9 compounds were separated and the chemical structures were fully determined as:6"-O-trans-caffeoyl catalpol(1)、Picroside Ⅱ(2)、5"-methoxy-ampicoside(3)、Minecoside(4)、Picroside Ⅰ(5)、Picroside Ⅲ(6)、Sweroside(7)、Vanillic acid(8)、2-O-glucoside of deacetoxycucurbitacin B(9),among which compounds 1-7 were iridoids,and compounds 1 and 3 were obtained for the first time fromPicrorrhizaRoyle genus.In order to determine the content of 5 iridoids in P.scrophularaefloraand evaluate the quality of different batches of P.scrophularaeflora from different habitats,an UPLC-ESI-MS/MS method for the simultaneous determination of the contents of Picroside Ⅰ-Ⅲ,Sweroside and Minecosidein different batches was established,The chromatographic column was ACTUITY UPLC? BEH Amide Column(2.1 mm × 100 mm,1.7 μm),the mobile phase was eluted by 0.1%formic acid aqueous solution-0.1%acetonitrile formate.The flow rate was 0.4 mL/min,and the negative ion mode was used to monitor the scanning mode of MRM.The linear range of the method and the precision of the chromatographic system were investigated.The results showed that the method was rapid,sensitive,reliable and accurate,which could be used to evaluate the quality of P.scrophularaeflora.The 5 iridoids showed a good linear relationship within the limit range.The method was used to determine the content of 5 components in 25 different batches of P.scrophularaeflora.The results showed that the contents of 5 iridoids in different batches of P.scrophularaeflora were quite different,and the quality of some batches of online shopping was not up to standard.The quality of medicinal materials is on the market is intermingled.In this paper,the pharmacokinetics of 5 iridoid compounds from P.scrophularaeflora in SD rats was studied,and the simultaneous determination of Picroside Ⅰ-Ⅲ,Sweroside and Minecoside in plasma of SD rats was establishedby UPLC-ESI-MS/MS.Paeoniflorin was chosen as internal standard,plasma sample was pretreated by protein precipitation method.The chromatographic column was ACTUITY UPLC? BEH Amide Column(2.1 mm ×100 mm,1.7μm),column temperature was 40℃,the mobile phase was eluted by 0.1%formic acid aqueous solution-0.1%acetonitrile formate.The flow rate was 0.4 mL/min,and the negative ion mode was used to monitor the scanning mode of MRM.The optimized ion-pairs of Picroside Ⅰ-Ⅲ,Sweroside,Minecoside and internal standard were m/z 491.2/147.0,m/z 511.3/234.8,m/z 537.4/261.0,m/z 537.5/281.4,m/z 403.2/195.0,m/z 525.2/449.1respectively.The LLOQ of the 5 analytes was 0.687 ng/mL、0.519 ng/mL、0.504 ng/mL、0.384 ng/mL、0.561 ng/mL,and Tmax was 1.1±0.2 h、1.1±0.1 h、0.8±0.1 h、1.0±0.2h、2.1±0.1 h。In this paper,a systematic study on the iridoids of P.scrophularaeflora was carried out,which was of great significance in clarifying the substance basis of its pharmacodynamics,the process of absorption and metabolism in vivo,and the establishment of quality control standards.It also provides theoretical and scientific basis for the development and utilization of natural plant resources and clinical drug use of P.scrophularaeflora.