Rapid Identification And Quality Study Of Curcumae Radix From Different Botanical Sources

Objective:Curcumae radix is the dried tuberous root of Curcuma wenyujin YH Chen&C.Ling,Curcuma longa L.,Curcuma kwangsiensis SG Lee et CF Liang,Curcuma phaeocaulis Val.Relieves depression,clears the heart and cools blood,and has the effect of promoting gallbladder and yellowing.Curcuma wenyujin,Curcuma longa and Curcuma kwangsiensis are the geoherbs of Zhejiang,Sichuan and Guangxi respectively.In recent years,the cultivation of commercial Curcumae radix has expanded from traditional production areas to new production areas on a large scale,and its blind introduction and expansion of production areas have reduced the quality of Curcumae radix medicinal materials.Although all four kinds of curcuma radix can be used for medicinal purposes,the quality of medicinal materials varies greatly,and the prices are also quite different,causing considerable difficulties for enterprises to purchase raw medicinal materials and decoction pieces.Therefore,this paper intends to explore the differences in the botanical source and chemical composition of the four kinds of Curcumae radix,determine the quality marker(Q-marker)of Curcumae radix,and provide a basis for the quality control of Curcumae radix.Methods:Forty Curcumae radix samples from four sources were collected from Sichuan,Guangxi,Zhejiang,Fujian and Yunnan provinces.The combined techniques of NIR,UPLCTriple-TOF-MS and high performance liquid chromatography were used to analyze the Curcumae radix of different botanical sources.Specifically,it includes:1.Using near-infrared technology to measure the spectrum of 4 kinds of Curcumae radix samples at 10000-4000 cm1,and combining with Orthogonal Partial Least Squares Discriminant Analysis Model(OPLSDA)to analyze Curcumae radix.2.UPLC-Triple-TOF-MS technology was used to characterize the structures of the four botanical source Curcumae radix components by means of accurate mass numbers,secondary fragment characteristic ions,fragmentation rules,retention times,chemical databases and standard products.3.Based on the above qualitative results,using the network pharmacology method,upload the results to the Pubchem database to obtain the molecular formula of the labeled compound,import it into the Swiss Target Prediction database,and take the target with a prediction score greater than 0 as the compound target.Disease targets were obtained through the OMIM database,Genecards database,and TTD database.Finally,the compound-target information was imported into Cytoscape 3.7.2 software to obtain the compound-disease target network map,and the functional enrichment analysis of key target genes GO and KEGG was performed using the Bioconductor bioinformatics software package.4.Using HPLC method,Dikma Diamonsil Plus C18(5μm,250×4.6 mm)chromatographic column;mobile phase is acetonitrile-0.1%phosphoric acid phosphoric acid aqueous solution to measure the content of relevant quality markers in four kinds of Curcumae radix officinalis.Results:1.By establishing the OPLS-DA model,comparing the discriminative effects of each botanical source Curcumae radix under different spectral preprocessing,the results show that the OPLS-DA model established under the smoothing+first derivative processing is stable,has good predictive ability,and can quickly and accurately distinguish Curcuma wenyujin,Curcuma longa,Curcuma kwangsiensis,Curcuma phaeocaulis.2.A total of 46 compounds were identified from 4 kinds of Curcumae radix,which are curcuminoids and sesquiterpenoids of 10 structural types,including 39 in Curcuma wenyujin,40 in Curcuma kwangsiensis,40 in Curcuma phaeocaulis and 17 in Curcuma longa.There are 12 components in 4 kinds of Curcumae radix,namely curcumenolactone C,curdione,curcumenone,curcumenol,curcumol,dihydrocurcumin,demethoxycurcumin,curzerenone,curcumene,beta-elemene,turmeric and gingerene.The content of curcumin in Curcuma longa was significantly higher than that of the other three turmerics,such as curcumin,demethoxycurcumin,bisdemethoxycurcumin,and dihydrocurcumin.The content of gemanetype sesquiterpenes in Curcuma wenyujin is higher than that of the other three kinds of turmerics,such as curdione,neocurdione,furanodiene,furanodienone,germacrone and so on.The eucalyptane-type sesquiterpenes in Curcuma kwangsiensis are higher than that of the other three kinds of turmerics,such as curcolone,curcolonol,and zedoarofuran.3.A total of 46 chemical constituents and 12 common constituents were identified in the decoction pieces of Curcumae radix.Using common components as Q-marker candidates to do network pharmacology analysis,predict turmerone,curcumenone,curdione,curcumenol,curcumol,dihydrocurcumin,demethoxycurcumin,furanodienone and germacrone can act on targets such as serotonin receptors(HTR1A,HTR2A,HTR1D,HTR1B),opioid receptors(OPRK1,OPRM1,OPRD1,OPRL1),etc.,by regulating neuroactive ligand-receptor interactions,serotonergic synapse,calcium signaling pathway,cAMP signaling pathway,retrograde endocannabinoid signaling and other important pathways play an antidepressant role.Turmerone,curcumenone,curdione,curcumenol,curcumol,dihydrocurcumin,demethoxycurcum in,furanodienone and germacrone can be used as potential quality markers of Curcumae radix.4.The content determination method of curcuma curcumenone,curcumenol,curdione,curzerenone,furanodienone,curcumol and germacrone was successfully established by comparing the extraction conditions.The results showed that the contents of curcumenone,curcumenol,and gemmanone in Curcuma wenyujin,Curcuma kwangsiensis and Curcuma phaeocaulis were 0.02-0.55 mg/g,0.02-1.63 mg/g and 0.03-0.43 mg/g.From the analysis of the results,it was found that curcumenone,curcumenol,and gemmanone could be detected in Curcuma wenyujin,Curcuma kwangsiensis and Curcuma phaeocaulis.Therefore,curcumenone,curcumenol,and gemmanone could be used as Curcuma wenyujin,Curcuma kwangsiensis and Curcuma phaeocaulis content determination index.The contents of 7 compounds were not detected in Curcuma longa.Therefore,these 7 components cannot be used as the quality measurement standard of Curcuma longa.Conclusion:Based on NIR and UPLC-Triple-TOF-MS technology,this study established a set of analytical methods for rapid differentiation of different botanical sources of Curcumae radix,which can be well applied to the analysis of the botanical sources of Curcumae radix.At the same time,using UPLC-Triple-TOF-MS technology and network pharmacology method,the establishment of the chemical component library of each botanical source can quickly confirm its components,analyze and predict the potential Q-marker of Curcumae radix,and can comprehensively establish the quality evaluation method and the quality traceability system provides a theoretical basis.Through the HPLC content determination of potential Q-markers,curcumenone,curcumenol and gemmanone can be used as quality control indicators for Curcuma wenyujin,Curcuma kwangsiensis and Curcuma phaeocaulis.