| The development of traditional Chinese medicine is one of the important national development strategies.The “Opinions on Promoting Inheritance,Innovation and Development of Traditional Chinese Medicine” issued by the State Council in 2019 clearly stated that "vigorously promote the quality improvement of Chinese medicine and the high-quality development of the industry",requiring the strengthening of the quality control of traditional Chinese medicine and the quality promotion of Chinese patent medicines and Chinese herbal medicines.Fingered Citron contains a wide range of biochemical activities with antioxidative,anti-inflammatory,hypoglycemic,hypolipidemic and antitussive effects.It is of great significance for the treatment of glycolipid metabolism disorder and the development of new drugs.In recent years,the research on Fingered Citron has made significant progress,mainly in the extraction and separation of Fingered Citron volatile oil,flavonoids,and polysaccharides,determination of chemical components,the establishment of fingerprints,and the study of pharmacological effects.There are many domestic and foreign researches about the identification of Fingered Citron ingredients,but the reflection of the composition differences between Fingered Citron and processed Fingered Citron,the types and quantities of chemical components in Fingered Citron,the quality of traditional Chinese medicine and the overall curative effect of traditional Chinese medicine isn’t shown.With the development of modern analysis technology,the use of new technologies to comprehensively identify the chemical components of Fingered Citron and construct the fingerprint of Fingered Citron is of great significance not only for the quality control and efficacy evaluation of traditional Chinese medicine Fingered Citron,but also for research on the pharmacological effects of Fingered Citron.First of all,Ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF MS)technology was used to analyze and identify the chemical components of processed Fingered Citron.The sample was extracted 1 hour by methanol and the chromatographic separation was performed on a Poroshell Infinity Lab 120 EC-C18 column(3.0×150 mm×2.7 μm),mobile phases consisted of eluent A(0.1% formic acid and 5 mmol/L ammonium acetate in water)and eluent B(acetonitrile)with a flow rate of 0.35 m L/min.The main chromatographic peaks in the total ion chromatogram(TIC)were characterized in positive and negative ion modes.Based on the accurate mass,fragment ions,retention behaviors,net-work database and the fragmentation pathways,a total of 67 components were identified from processed Fingered Citron including 22 coumarins,12 flavonoids and flavonoid glycosides,15 organic acids,6 glycosides,5 limonin,and 7 other compounds,24 compounds of them were first time discovered.This method is suitable for the detection of material basis of processed Fingered Citron,with many peaks and high sensitivity.Secondly,based on the above study,the UPLC-Q-TOF fingerprints in the negative model of raw Fingered Citron and its processed were established respectively,which was applied in the establishment of fingerprints for Fingered Citron first time.The results show that a total of 24 common peaks were identified in the UPLC-Q-TOF fingerprint of raw Fingered Citron,and 19 components were identified through confirmation;totals of 22 common peaks were identified in the UPLC-Q-TOF fingerprint of bergamot,of which 14 components were identified.The established UPLC-Q-TOF fingerprinting method of raw and processed Fingered Citron is stable,has many common peak components,and can fully characterize the quality of Fingered Citron.This method can be used to identify the authenticity of Fingered Citron,quality assessment and herbs traceability.Finally,this study built a method to analyze the difference in ingredients of processed Fingered Citron and its steamed homologous one.This method uses Agilent Profinder10 software to process its collected data,and Simca14.1 software is used to perform principal component analysis(PCA)and Orthogonal Partial Least Squares Discrimination Analysis(OPLS-DA)on the data.After screening the components with VIP value>1 and p(corr)value>0.58,the significant difference components were obtained pass independent sample T test.Through component identification,a total of6 components were found and identified in the positive mode and the negative mode:Plumbagin-5-O-glycoside,Nomilin acid,7-methoxy-5-prenyloxycoumarin,Palmitic acid,Oleic acid and Oxypeucedanin hydrate.This method analyzes all the collected data,and comprehensively filters the significant difference components that can most affect the raw Fingered Citron and processed Fingered Citron,ensuring the integrity and reliability of the results.At the same time,the method is scientific and rigorous,and it can be used as a new way to study the material difference. |
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