Study On The Chemical Ingredients And Quality Evaluation Method Of Scutellaria Barbata

Scutellaria barbata is a dry whole grass of S.barbarta D.Don.S.barbata is also known as Pythium aurantium,Hanshinia angustifolia,toothbrush grass,etc.According to the Pharmacopoeia,S.barbata is cold,has a pungent,bitter taste,act on the Lungs,Liver,and Kidney Channels.It has the functions of clearing away heat and detoxification,reducing blood stasis and diuresis,edema,jaundice and snakebite.At present,various chemical components have been isolated and purified from S.barbata,including flavonoids,flavonoid glycosides,dihydroflavonoids,dihydroflavonoid glycosides,diterpenes,polysaccharides and steroids.We studied the qualitative analysis of the chemical composition of S.barbata by UPLC/IM–QTOF–MS technology,and separate the chemical composition of the S.barbata,in order to clarify the material basis of S.barbata.It provides a basis for further research on the active substance basis of S.barbata.This paper mainly includes the following parts:1.Isolation and identification of chemical components of S.barbataUsing ODS,Silica gel,Sephadex LH–20,preparative high–performance liquid chromatography and other chromatographic methods to further isolate and purify S.barbata.We identified 15 compounds by UV,LC–MS,NMR and other spectroscopy techniques.It is epi–oleanolic acid,syringiaresinol–4–O–β–D–monodigloside,oleanolic acid,palmitic acid,naringenin,aurantiamide acetate,octadecanoic acid,teradecanoic acid,5,7,4’–trihydroxy–8–methyloxyflavone,hispidulin,apigenin,scutellarein,scutellarin,4–（4–hydroxyphenyl）–3–buten–2–one;4–allyl–2,6–dimethoxyphenyl glucoside respectively.It enriches the chemical composition of S.barbata and provides a theoretical basis for its further development.2 System characterization of S.barbata chemical composition based on LC–MS spectrometryUsing UPLC/IM–QTOF–MS technology for qualitative analysis of S.barbata,based on systematically sorting the literature of S.barbata phytochemical separation,a"self–built S.barbata database"was established,and 197 known chemical structure information（name,molecular formula,chemical structure）.The database was imported into UNIFI^TMsoftware to establish an efficient metabolite identification process based on UNIFI automatically annotated HDMSE data（including analysis method settings,data collection,data correction,data processing and identification）.Through small molecule prediction,MS1/MS2 data and database component theory mass spectrometry information matching greatly shortens the analysis time and presents reproducible identification results.By this way,a total of 93compounds were identified,mainly including 30 flavonoids,57 diterpenes,2 triterpenes and 4others compounds.It can provide a basis for clarifying the main components of S.barbata isolation and identification of chemical components.3 Quality evaluation of S.barbataIn the experiment,S.barbata chemical components were extracted and separated,and five compounds of scutellarin,scutellarein,naringin,apigenin and hispidulin were obtained.The research reported that scutellarin and apigenin have effects on human ovarian cancer.SKOV3 cells have a significant inhibitory effect;Scutellariae on human liver cancer cells（Hep3B）,human gastric cancer cells（AGS）,human lung cancer cells（A549）,mouse melanoma cells（B16）and mouse breast cancer cells（4T1）;naringenin and naringin have obvious pharmacological effects on the respiratory system,such as antitussive,expectorant and anti-inflammatory,etc.Because of their good pharmacological activity,these five compounds were determined to be indicators for the quality evaluation of S.barbata medicinal materials.The r value of the linear relationship is less than 0.9991,The RSD value of intraday precision was less than 1.49%,and the intraday precision was less than 2.28%.Stability RSD value is less than or equal to 2.09%;The RSD value of repeatability is less than or equal to 2.17%.The average recoveries were 104%,102%,96%,104%and 99%,indicating that the method had high precision,good reproducibility and high sensitivity,and could be used for the quality control study of S.barbata.The UPLC–UV technology was used to analyze 16 batches of S.barbata from different origins,and the obtained chromatographic data was imported into the traditional Chinese medicine fingerprint similarity evaluation system to establish a fingerprint.20 common peaks of 16 batches of S.barbata from different origins were obtained.SIMCA14.1 software was used for principal component analysis and cluster analysis.At the same time,the five compounds isolated in experiment 1 were used as indicator materials for statistical analysis.According to the results of cluster analysis,both methods of quality evaluation of S.barbata can divide 16 batches of medicinal materials into three categories,categories A,B and C.The total peak area of category A samples is significantly higher than Type B samples have the worst total peak area of type C samples.The statistical analysis results show that,compared with the standard medicinal materials,the total peak area of the medicinal materials of the other origins is higher than that of the standard medicinal materials in all the 16 batches of medicinal materials except type C samples.In summary,we extracts and separates the chemical components of S.barbata medicinal materials,identifies their structures,and estimates their total components.We evaluate the quality of S.barbata medicinal materials from different origins,and finally establishes 16batches of quality evaluations for S.barbata medicinal materials from different origins.The method provides a reference for multi-index evaluation of the quality of S.barbata medicinal materials,and helps to promote the further development of S.barbata medicinal materials.