Study On The Method Of Multi-index Quality Analysis Of Angelicae Pubescentis Radix

In this study,the Angelica Pubescentis Radix of umbelliferae was taken as the research object.Modern chromatographic and spectral techniques were used to quantitatively determine osthol,isoimperatorin,and columbianadin in the Angelica Pubescentis Radix,so as to achieve the purpose of quality control.The main research is divided into three parts:The goal of this work was firstly to establish a high-performance liquid chromatography（HPLC）method that simultaneously determined and quantified osthol,isoimperatorin,and columbianadin using a quantitative analysis of multicomponents by single marker（QAMS）method,which usesd internal reference substances and relative correction factors to perform the synchronous determination and quantitation of multiple components.Taking into account the price and availability of standard substances,osthol was selected as internal reference substance for analysis.The compatibility results showed that the relative correction factors of all compounds showed good reproducibility under different chromatographic column,different column temperature and different flow rate.The established method was validated by methodology and the results were all well.Besides the external standard method（ESM）and QAMS were used to determine the contents of three coumarins in Angelicae pubescentis Radix from nine different producing areas and its Chinese patent medicine named Duhuo Jisheng pills.It turnrd out there were little differences between the two methods.In summary,QAMS is an alternative approach for the quality control of Angelicae pubescentis Radix.The goal of this work was to establish a method to identify and quantify the content of main active components in Angelicae Pubescentis Radix quickly,simply and accurately.This paper firistly reported a novel method which can determine osthol,isoimperatorin,and columbianadin using ¹H-qNMR simultaneously and quantitatively.Dimethyl sulfoxide-d₆（DMSO-d₆）was selected as an optimal ¹H-qNMR solvent and pyrazine was used as internal standard substance（δ8.66 ppm）.The quantitative peaks of three active components were atδ7.54-7.56 ppm for osthol,δ6.83–6.85 ppm for columbianadin andδ6.31-6.32 ppm for isoimperatorin.The results showed that the method has good precision,stability,and repeatability.Furthermore,the experimental process was simple and the test time was short（1 min）.Graphene was introduced on the basis dispersive liquid–liquid microextraction method based on solidification of floating organic drop（DLLME-SFO）with graphene/1-dodecyl alcohol used as extractant,and this method combined with quantitative hydrogen nuclear magnetic resonance spectroscopy（¹H-qNMR）for the first time to simultaneously qualitative and quantitative osthole,columbianadin and isoimperatorin in Angelicae Pubescentis Radix.In this experiment,magnetic stirrer was used for extraction,all NMR analyses were performed on a 600 MHz spectrometer（Bruker Advance III）with DMSO-d₆ as deuterium substitute solvent and pyrazine as the internal standard.The influencing factors and NMR parameters in the extraction process were also investigated and optimized.The optimal extraction conditions were as follows:the volume ratio of ultrapure water to test solution was 9:1,the concentration of graphene in dodecyl alcohol was 0.5 mg/m L,the volume of extractant was 80μL,the pH of the solution was 5,the concentration of Na Cl solution was5%,the rotation speed was 800 rpm,and the extraction time was 20 min.The results showed that the method has good precision,stability,and repeatability,the established method was also used to determine osthol,columbianadin and isoimperatorin in different areas.Through the comparison and exploration of the above methods,the method of QAMS can be used for the quality detection of large quantities of Chinese medicinal materials because of its low cost.As a novel quantitative method,¹H-qNMR can be used for academic research because of its expensive equipment,and its large-scale production application still need to be popularized.The DLLME-SFO can effectively extract the trace components in traditional Chinese medicine.The methods established in this study have their own advantages and disadvantages,which can be used separately or comprehensively to evaluate the quality of traditional Chinese medicine according to the characteristics of the research object.