Optimization Of Ultrasonic Extraction Process And Quality Evaluation Of Acanthopanax Senticosus Root

Acanthopanax senticosus is a genuine traditional Chinese herb mostly distributed in Northeast China,with abundant active ingredients and extensive pharmacological effects,significantly contributing to human health over the past century.Active ingredients are the material basis of TCM effects,so their qualitative and quantitative analysis,as well as efficient extraction are crucial to TCM research and development.Additionally,studies have shown that multiple factors,including temperature,humidity,and population structure,lead to inevitable quality differences of TCM.The content of active components is the most direct reflection of these differences,and it has received widespread attention.The active components of Acanthopanax senticosus root were analyzed qualitatively and quantitatively firstly in this study,which laid a foundation for subsequent research and resource utilization.The non-targeted qualitatively analysis involved establishing an UPLC Q Exactive Focus MS/MS qualitative analysis method to collect MS data and utilizing the Compound Discoverer software to establish a natural product identification workflow for non-targeted analysis.Through these efforts,we identified 35 components from extract of Acanthopanax senticosus root.29 of these compounds have been previously reported as active components of Acanthopanax senticosus root,while another 6 active ingredients have yet to be reported,and further research is needed.To ensure the authenticity of our identification results,we further performed targeted analysis on the important active components of Acanthopanax senticosus root,including syringin,eleutheroside E,isofraxidin,sesamin,chlorogenic acid,and caffeic acid.Following that,an ultrahigh performance liquid chromatography-mass spectrometry(UPLC-MS/MS)method was established for the qualitative and quantitative analysis of the target compounds within 10 minutes.Additionally,to enrich the quality evaluation system of Acanthopanax senticosus root,an UV-visible spectrophotometry method was established to determine the content of the clinical curative effect substance basis of Acanthopanax senticosus root,including total glycosides,total phenols,total flavonoids and total saponins.Secondly,the multi-component ultrasonic extraction process of Acanthopanax senticosus root was optimized to lay a foundation for quality evaluation and pharmacodynamic research.An integrated evaluation method combining the analytic Hierarchy process(AHP)and entropy weight method(EWM),known as AHP-EWM,was established.This method allowed for the subjective experience to assign index importance and also objectively reflected information regarding the index.Using UPLC-MS/MS quantitative method and AHP-EWM method,a single factor combined with BBD-based response surface method(RSM)was used to optimize the multi-component ultrasonic extraction process of Acanthopanax senticosus root.The optimal extraction processes were as follows: ultrasonic power 780 W,ultrasonic time 17.5 min,solid-liquid ratio 1/40 g/m L,ethanol volume fraction 57%,and raw material particle size 80 mesh.The final verification results indicated that the contents of syringin,eleutheroside E,isofraxidin,sesamin,chlorogenic acid and caffeic acid in Acanthopanax senticosus root were517.96±6.09 μg/g、861.25±5.30 μg/g、66.66±1.22 μg/g、99.36±0.69 μg/g、2235.84±12.17μg/g、45.75±0.77 μg/g,respectively.The comprehensive score based on the AHP-EWM method was 95.45±0.39,which was close to the predicted theoretical value.Finally,the quality differences of Acanthopanax senticosus root from different producing areas were evaluated based on the content of index components to lay a foundation for its resources utilization.A total of 45 samples of Acanthopanax senticosus root were collected from 15 different habitats with 3 batches per habitat.The content of syringin,eleutheroside E,isofraxidin,sesamin,chlorogenic acid and caffeic acid in Acanthopanax senticosus root from different habitats were determined using UPLC-MS/MS method,while the content of total glycosides,total phenols,total flavonoids and total saponins were determined using UV-vis spectrophotometry.The quality differences of Acanthopanax senticosus root from different habitats were evaluated using stoichiometric methods,including principal component analysis(PCA),cluster analysis(CA),and orthogonal partial least squares discriminant analysis(OPLS-DA).PCA and CA revealed that quality differences in Acanthopanax senticosus root were minimal between batches from the same origin,but significant differences were observed between origins,likely related to regional variation.OPLS-DA showed that samples from the same province could be better clustered and distinguished from other provinces.The marker components indicating the quality differences of Acanthopanax senticosus root in different provinces were significantly distinguished,and made different contribution.It may be due to the different metabolic pathways of index components in different habitats,which affect their content and ultimately lead to quality differences.In this study,the active components of Acanthopanax senticosus root were analyzed qualitatively and quantitatively to lay a foundation for the subsequent research.Then the multicomponent ultrasonic extraction process of Acanthopanax senticosus root was optimized based on multi-index comprehensive evaluation,which provided theoretical and practical guidance for its quality evaluation and pharmacodynamic research.The quality differences of Acanthopanax senticosus root from different producing areas were evaluated based on the content of index components,which was of great significance for drug safety,quality control and resource utilization,and also provided ideas and practical guidance for the establishment of TCM quality evaluation system.