Identification Of 200 Kinds Of Traditional Chinese Medicine By Three Dimensional Fluorescence Spectroscopy And Fluorescence Quantitative Analysis Of Forsythia Suspensa

Objective:Traditional Chinese medicine is the treasure of ancient Chinese science.It plays an important role in the prevention and treatment of diseases and epidemic prevention.It is an important guarantee for the continuous development of the Chinese nation.At present,there are many kinds of traditional Chinese medicine circulating in the market,with complex components and confused authenticity.It is of great significance to identify and control the authenticity and quality of traditional Chinese medicine with modern advanced instruments and analytical methods.In this paper,traditional Chinese medicine fluorescence identification method and traditional Chinese medicine three-dimensional fluorescence spectrum method are used to identify the authenticity of traditional Chinese medicine,and BP neural network algorithm is used to establish the chemical component classification recognition model of traditional Chinese medicine three-dimensional fluorescence spectrum,which provides a new method for the authenticity identification and quality evaluation of traditional Chinese medicine.At the same time,taking Fructus Forsythiae as an example,a fluorescence method for the determination of total phenylethanoid glycosides in Fructus Forsythiae was established,which provided a basis for the scientific and comprehensive evaluation of the quality of Fructus Forsythiae and laid a foundation for the future application of fluorescence method in the quantitative analysis of traditional Chinese medicine.Methods:1.The traditional fluorescence identification method of traditional Chinese medicine was used to observe the fluorescence color of herbal pieces,herbal powder,water extract and methanol extract under the ultraviolet lamp,take photos and record.2.The methodology of three-dimensional fluorescence spectrum identification of traditional Chinese medicine was studied.The effects of extraction solvent,extraction method,extraction time,concentration,p H value and ionic strength were investigated to determine the general method of three-dimensional fluorescence spectrum sample treatment of traditional Chinese medicine.At the same time,the scanning range,slit width,voltage,scanning speed and sampling interval were investigated to determine the best instrument parameters.Three dimensional fluorescence spectrum identification belongs to qualitative identification,scanning the three-dimensional fluorescence spectrum of 200 kinds of standard Chinese medicine,and selecting some counterfeit medicine,easily confused medicine and medicine from different places for identification,and comparing with the standard medicine spectrum to judge the authenticity of the test sample.At the same time,the three-dimensional fluorescence spectra of 200 kinds of traditional Chinese medicine were summarized.3.Based on the correlation between three-dimensional fluorescence spectrum and chemical components,the BP neural network algorithm is used to establish the classification and recognition model of chemical components in three-dimensional fluorescence spectrum of traditional Chinese medicine.4.Establish a fluorescence quantitative method for the determination of total phenylethanoid glycosides in Forsythia suspensa.The excitation wavelength was 330 nm and the emission wavelength was 430 nm.The content of total phenylethanoid glycosides in Forsythia suspensa was determined by fluorescence quantitative method.Results:1.The traditional fluorescence identification of traditional Chinese Medicine found that the fluorescence phenomenon of root,rhizome,stem wood and bark was obvious under the ultraviolet lamp with wavelength of 365 nm;There was no obvious fluorescence phenomenon in leaves,flowers,fruits,seeds and grasses.The fluorescence color of the herbs with the same chemical composition was similar.Most of the herbs with quinones showed yellow fluorescence,and a few showed blue fluorescence;Most of the herbs containing alkaloids show blue or yellow fluorescence,which may be related to their conjugated state;Most of the medicinal materials containing phenylpropanoids,coumarins,lignans,terpenoids and saponins showed blue fluorescence,and the blue fluorescence of the medicinal materials containing coumarins and lignans was particularly obvious,while the blue fluorescence of the medicinal materials containing phenylpropanoids,terpenoids and saponins was slightly weak;Most of the herbs containing flavonoids showed blue,yellow,green and red fluorescence.2.The basic method of three-dimensional fluorescence spectrum identification is:water as solvent,ultrasonic extraction,ultrasonic time 30 min,sample concentration between 0.1～1 mg/ml.The instrument parameters are: scanning range,excitation wavelength 200～700 nm,emission wavelength 220～700 nm,slit width 5 nm,voltage 400 V,scanning speed 60000 nm/min,sampling interval(ex/EM)10 nm/10 nm.The fluorescence spectra of 200 kinds of standard medicinal materials were measured,and the standard three-dimensional fluorescence spectra of traditional Chinese medicine were established.The applicability of three-dimensional fluorescence spectrum was verified by qualitative identification of different medicinal parts,different origins and some counterfeit drugs.It is found that there is a certain correlation between the three-dimensional fluorescence spectrum of traditional Chinese medicine and the chemical components of traditional Chinese medicine.The three-dimensional fluorescence spectrum of traditional Chinese medicine containing the same chemical components has similar characteristic peaks.3.Based on the correlation between chemical components of traditional Chinese medicine and three-dimensional fluorescence spectrum,the BP neural network was used to establish the classification and recognition model of chemical components of three-dimensional fluorescence spectrum.According to this model,the main chemical components of traditional Chinese medicine can be predicted,and some counterfeit products which are different from the authentic chemical components can be identified,with an accuracy of 99 %.4.A fluorescence quantitative method for the determination of total phenylethanoid glycosides in Forsythia suspensa was established μ The regression equation was y=15.085x+20.9(r=0.9993).The average recovery was 99 %(RSD = 1.51 %).The method is simple,sensitive,reproducible and can be used for the determination of total phenylethanoid glycosides in Forsythia suspensa.Conclusions:The traditional Chinese medicine fluorescence identification method is simple,easy to use,and can be used for rapid identification.The method is simple,sensitive and reproducible.BP neural network was used to establish the classification and recognition model of chemical components of traditional Chinese medicine,which can predict the chemical components of unknown traditional Chinese medicine.At the same time,a fluorescence quantitative method for the determination of total phenylethanoid glycosides in Forsythia suspensa was established,which provided a new idea and method for the comprehensive control of the quality of traditional Chinese medicine.