Research On Rapid Identification Technology Of Rhizoma Coptidis Based On High Resolution Mass Spectromtryand Isotope Mass Spectromtry

Rhizoma Coptidis is one of the precious traditional Chinese medicine with thousands of years of medicinal history.The 2015 edition of the Chinese Pharmacopoeia records that it has the effects of clearing away heat and dampness,clearing fire and detoxifying.Studies have reported that the effective ingredient content of Rhizoma Coptidis from different places has significant differences,and the pharmacological effects are also significantly different.Therefore,it is of great significance to establish a simple,rapid and efficient method for tracing origin of Rhizoma Coptidis.Ultra-high performance liquid chromatography-high resolution mass spectrometry is considered to be an effective analytical method and has been widely used for traceability analysis of the origin of various agricultural products and foods.The method can provide excelent sensitivity,accuracy and precision,and can assay multiple chemical components in various substances with high throughput.The chemical composition of plants growing in different environments has obvious differences in quantity and content,such as the growth altitude,annual average temperature,air humidity and fertilization methods,etc.will lead to the difference in the quantity or content of the same plant chemical composition.In addition,stable isotope mass spectrometry analysis has also been proven to be an effective identification technique of origin.The stable isotope ratio can directly or indirectly reflect the growth environment of the plant.The carbon isotope ratio is mainly related to the light intensity of the area where the plant grows;the nitrogen isotope ratio is mainly related to the fertilization conditions for plant growth;the hydrogen and oxygen isotope ratio is mainly related to the water source required for plant growth.The growth conditions of Rhizoma Coptidis in different regions are different,and the isotope ratios are significantly different.Therefore,this paper adopts high-resolution mass spectrometry to analyze the various chemical components in Rhizoma Coptidis to provide a strong basis for the origin of Rhizoma Coptidis;at the same time,an isotope ratio analysis method has been established to explore the origin of Rhizoma Coptidis.Part?Based on high-resolution mass spectrometry combined with chemometric analysis for discriminating geographical origins of Rhizoma CoptidisObjective:Novel analysis approach based on UPLC-HRMS combined with chemometric,was established for determining origins of Rhizoma Coptidis and screening the differential markers.Methods:After extracting Rhizoma Coptidis with methanol-water,various chemical components in the sample were analyzed by high-performance liquid chromatography-high resolution mass spectrometry combined with chemometrics techniques.Results:A total of 11 compounds including 8known compounds and 3 unknown compounds have been found and confirmed as unambiguous and reliable markers for discriminating origins of Rhizoma Coptidis.Among them,Scorpinone,Bicuculline,Berberine,compound 1 and compound 2 were used for distinguishing the samples from Chongqing?CQ?and Sichuan?SC?.Hydroprotopine,Chelidonine and Pinoresinol were used for distinguishing the samples from Chongqing?CQ?and Hubei?HB?.Flakealkali,Palmatine and compound 3 were used for distinguishing the samples from HB and SC.Meanwhile,the reliable orthogonal partial least squares discriminant analysis?OPLS-DA?models were established based on these markers and the samples from different origins were successfully discriminated,as well as the accuracy was up to 100%.It showed that the established models could be used for predicting the geographic origins of unknown Rhizoma Coptidis,and those markers could provide a guidance for discriminating origins of Rhizoma Coptidis.Conclusion:In conclusion,high-resolution mass spectrometry combined with chemometric could serve as a valid strategy to analyze the origins,such as Rhizoma Coptidis.Part?Combination of stable isotopes and multi-elements analysis with chemometric for determining the geographical origins of Rhizoma CoptidisObjective:Rhizoma Coptidis is a valuable medicine food homology plant,whose geographic origin is topic of interest for both consumers and producers since the geographic origin determines its price and efficacy.A novel method for discriminating the geographic origins of Rhizoma Coptidis was established based on stable isotopes and multi-elements analysis combined with chemometric.Methods:Combining two analytical approaches,4 light stable isotope ratios and 32 inorganic elements of 103 samples from six regions were determined.Results:It was shown that the isotopes ratios and multi-elements contents were significantly different among Rhizoma Coptidis from six regions.Al,Sn,Mg,?¹⁵N and?¹³C were considered as the most powerful differentiation markers and used for discriminating the geographical origins.Meanwhile,the built and validated predictive model,based on linear discriminant analysis?LDA?and orthogonal partial least squares discriminant analysis?OPLS-DA?,successfully classified samples according to their regional origins with 100%accuracy and successfully predicted unknown samples with 100%accuracy.Conclusion:Therefore,the stable isotopes and multi-elements analysis combined with chemometric tools can be successfully used to discriminate the geographic origins of Rhizoma Coptidis.