| The quality evaluation of traditional Chinese medicine theory is mainly based on the identification of the shape,texture,color and smell of the medicinal materials.So far,traditional macroscopic identification still occupies an important position.Traditional macroscopic identification has the advantages of quickness and simplicity,but most of it is based on empirical perception and lacks a more objective and scientific system support.The quality evaluation system of traditional Chinese medicine is constantly improving,and more and more scientific and technological achievements are applied to the quality control system of traditional Chinese medicine.High-performance liquid chromatography,liquid mass spectrometry,infrared chromatography,color detection technology,etc.are becoming the quality of traditional Chinese medicine.An important method for qualitative and quantitative analysis of chemical components in evaluation.The problems that the modern Chinese medicine quality evaluation system needs to solve are:the relationship between macroscopic characteristics and internal quality,and the combination of macroscopic characteristics and the internal components of medicinal materials to jointly evaluate the quality of Chinese medicine.1 Research purposes and methodsThis study takes,Mirabilis himalaica as the research object,using stereoscopic microscope and microscopic imaging technology to excavate the macroscopic and microscopic characteristics of different batches of Mirabilis himalaica,and establishs the extraction method of total flavonoids and extracts;SPSS software was used to calculate the correlation between the components and the macroscopic appearance;Through the liquid mass spectrometry technology,to analyze the full components of Mirabilis himalaica,and use the sequential metabolism as a guide to screen its active ingredients.Then provide new research data and methods for the quality evaluation research of Mirabilis himalaica.2 Research content(1)Establishment and quantitative analysis of shape indicatorsThe macroscopic characteristics and microscopic characteristics of Mirabilis himalaica are established by means of sight,hand touch,mouth taste,nose smell,etc.,with the help of instruments such as colorimeter,stereo microscope,optical microscope,etc.,as well as the measurement of the sample powder chromaticity index L*,a*,b*value.(2)Analysis of chemical indicatorsThe optimal extraction process of total flavonoids is optimized,and the content of total flavonoids of Mirabilis himalaica from different producing areas is quantitatively analyzed.To establish the extraction methods of extracts.The best extraction method of Mirabilis himalaica extracts was selected based on the comprehensive evaluation index.UPLC-MS/MS liquid-mass spectrometry was used to determine the chemical compositions in Mirabilis himalaica,which preliminarily analyzes the types of the compounds and main indicator components in Mirabilis himalaica.The metabolic process of Mirabilis himalaica was studied in vivo by the closed intestinal loop method-abdominal aorta blood sampling technology.The plasma samples were compared and analyzed by liquid chromatography mass spectrometry,and the active ingredients of Mirabilis himalaica were initial screened by the method of network pharmacology.(3)Quick identification of mixed products in the marketExtracts infrared fingerprints of Mirabilis himalaica powder are established through fourier infrared spectroscopy technology,to analyze its characteristic absorption peaks.The rapid identification of genuine products and Mirabilis jalapa is realized through comparing and analyzing the infrared absorption information,macroscopic and microscopic characteristics,etc.,of Mirabilis himalaica and Mirabilis jalapa.3.Research results(1)Compared with the Ministerial Standards,it is recommended to add the following contents in terms of microscopic characteristics:4～10 rounds of external ductile vascular bundles;20～30 layers of cork cells;bundles of calcium oxalate needles in parenchyma cells;The calcium oxalate are mostly distributed on the outer side of the central vascular bundle and the hetero-shaped vascular bundle,discontinuously arranged in a nearly concentric ring,and the outermost round of calcium oxalate crystals is densely arranged with obvious ring bands.The number of layers of Mirabilis himalaica ring pattern and the total number of irregular vascular bundles are positively correlated with the sample diameter.Its appearance and section color are correlated with the visible spectrum chromaticity index value.When the surface of Mirabilis himalaica sample is darker(tan),the powder color is When the index b*value is low,and the section color is lighter(quasi-white),the powder chromaticity index L*value is higher,and the powder chromaticity index a*value is lower.Mirabilis himalaica chromaticity index L*ranged from 62.43 to 78.40,a*ranged from 2.75 to 5.40,and b*ranged from 14.28 to 17.23.(2)The best extraction process for total flavonoids in Mirabilis himalaica is as follows:ethanol concentration is 61%,material-to-liquid ratio(g:ml)is 40,and extraction time is 65 minutes.Hot dip method,50%ethanol as solvent is the best extraction method for extracts.The extracts content from different batches of Mirabilis himalaica ranged from 17.48%to 28.16%.The mass fraction of total flavonoids in extracts ranges from 0.899%to 1.834%.The sample diameter is negatively correlated with the total flavonoid content.The extracts content increases with the increase of the chromaticity index L*value,and decreases with the increase of the a*value.The chromaticity can be used as a preliminary evaluation index for the content of extracts.(3)Using liquid chromatography-mass spectrometry technology to analyze the water extract of Mirabilis himalaica to obtain 31 components,and analyze the ethanol extract to obtain 59 components,including Choline,DL-Arginine,Oleamide,Salsolinol,(-)-catechin,4-Hydroxybenzoic acid,Adenosine,ferulic acid,Linolenic acid are 9 common components,and a total of 81 compounds were analyzed.Through the closed intestinal ring method,a total of 21 components were detected as prototype components,including L-Glutamic acid,DL-Arginine,DL-Tryptophan 3 amino acid components,Trigonelline,Salsolinol 2 alkaloid components,Biochanin A and Naringenin 2 flavonoids,Scoparone,Coumarin 2 coumarins,and Adenine,Adenosine,Creatinine,N-Acetylserotonin,4-Methoxycinnamic acid,Oleamide,5-Methylcytosine,6-Methylquinoline,Isoquinoline,Methyl palmitate,Chrysogine,Tolycaine 12 other ingredients.(4)The main difference between Mirabilis himalaica and Mirabilis jalapa in shape characteristics is:Mirabilis himalaica is cylindrical,Mirabilis jalapa is long conical;Mirabilis himalaica has rough surface,gray-brown or tan,and Mirabilis jalapa is relatively smooth and light,white or gray yellow,keratinous cross-section;Compared with Mirabilis himalaica,Mirabilis jalapa is often gelatinized due to boiling process,and does not have brown cork cells.The infrared spectrum analysis results of Mirabilis himalaica powder and extracts samples:3487 cm-1,3421 cm-1,3340 cm-1,3063 cm-1,1607 cm-1,1318 cm-1,1007 cm-1,782cm-1,521 cm-1 is the main characteristic fingerprint peak of Mirabilis himalaica.3284cm-1,2927 cm-1,2855 cm-1,1728 cm-1,1600 cm-1,1390 cm-1,1323cm-1,1280 cm-1,1202 cm-1,1102 cm-1,1044 cm-1,990 cm-1,924 cm-1,854 cm-1,667 cm-1 are the main characteristic fingerprint peaks of Mirabilis himalaica extracts.In the range of 3670～2160 cm-1,Mirabilis himalaica and Mirabilis jalapa have similar infrared spectrum peaks,but the latter has higher absorption intensity.In the wavelength range of 1600～1000 cm-1,the peak shape and absorption intensity of the two are different.The absorption intensity of starch in Mirabilis jalapa is higher than that in Mirabilis himalaica.The common pattern map in the range of Mirabilis himalaica 1600～1000cm-1 was used as the control map to calculate the similarity between Mirabilis himalaica and Mirabilis jalapa.The average similarity of Mirabilis himalaica in different batches was 0.997,and the average similarity of Mirabilis jalapa was 0.950.The statistical software SAS was used to perform clustering analysis on the similarity of 28 batches of Mirabilis himalaica and Mirabilis jalapa.Mirabilis himalaica was grouped into one category,and Mirabilis jalapa was another category,indicating that there is a significant difference between Mirabilis himalaica and Mirabilis jalapa infrared spectrum.4.Innovation(1)The visible spectrum method was used for the first time to detect the chromaticity value of Mirabilis himalaica powder and extract,and it was found to be connected with the color results of traditional Chinese medicinal materials and decoction pieces identified by traditional experience,providing a certain basis for the quantitative detection of color judgment.(2)The content of extracts determination method was established,under the condition of comprehensive reference to 4 indicators.This provides a reference for the formulation of the test standard for extract content.(3)For the first time,liquid-mass spectrometry was used to analyze and identify the chemical components in Mirabilis himalaica water extract and alcohol extract and their original blood components. |
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