| Purpose: 1.Based on the observation of the characteristic substance in the powder of Trichosanthis Pericarpium,the stone cells in the Trichosanthis Pericarpium were selected as the microscopic characteristic cells,and the microscopic quantitative research method of Trichosanthis Pericarpium was established,and the microscopic characteristic constants of Trichosanthis Pericarpium cells in 10 different regions were determined.2.By optimizing the ultrasonic extraction process of total flavonoids from Trichosanthis Pericarpium,the optimal ultrasonic extraction process of total flavonoids was optimized,and the content of total flavonoids in Trichosanthis Pericarpium was determined in 10 different regions.3.To establish a method for the content determination of five flavonoids in Trichosanthis Pericarpium,including rutin,isoquercitrin,quercitrin,quercetin and kaempferol,and to determine the content of five flavonoids in Trichosanthis Pericarpium in 10 different regions,so as to provide data reference for the quality control of Trichosanthis Pericarpium.4.The HPLC fingerprints of fructus Trichosanthis Pericarpium in 10 different regions were established,which provide research support for the qualitative identification of Trichosanthis Pericarpium.5.The correlation between the microscopic characteristic constants and the content of total flavonoids and the content of flavonoids monomers in Trichosanthis Pericarpium were established,and the results were analyzed to provide a reference for the evaluation of the quality of Trichosanthis Pericarpium in multiple dimensions.Material and method: 1.The uniform design method is adopted to investigate Trichosanthis Pericarpium medicinal material conditions for the determination of microscopic characteristic constants of stone cells,by investigating the samples quantity,granularity and adding amount of glycerol,ultimately determine the U6(63)design,optimization results through uniform design software,according to the optimum technological conditions for determination of 10 different areas of Trichosanthis Pericarpium microscopic characteristic constants of the cell.2.Based on the single factor test,the optimal extraction process conditions were optimized by L9(34)orthogonal test design.The influence of solution proportioning,hyperacoustic temperature,hyperacoustic schedule and ultrasonic rate of wark on the separation process of Trichosanthis Pericarpium were investigated.Through experimental research,it is expected to optimize the extraction process of total flavonoids from Trichosanthis Pericarpium,and provide material basis for later experiments.Ultraviolet spectrophotometry was used to determine the contents of total flavonoids in Trichosanthis Pericarpium.3.In this experiment,high-performance liquid chromatography was used to determine the contents of rutin,isoquercitrin,quercetin,quercetin and kaempferol in different collected Trichosanthis Pericarpium medicinal materials,and each component was taken as the indes for principal component analysis and cluster analysis.The quality of fructus trichosanthis peel was evaluated by the weight method of coefficient of variation.4.at the same time establish the fingerprint of Trichosanthis Pericarpium,to lay a theoretical foundation for the quality standards of Trichosanthis Pericarpium.The chromatographic column was: agilent Eclipse XDB-C18(parameter :4.6mm×250mm,5μm).The mobile phase was A: methanol-acetonitrile(1:10),B: 0.1% phosphoric acid solution.Flow rate: 1.0 m L·min-1;Detection wavelength: 360 nm;The column temperature was 30 ℃,and the amount of sampling was 20 u L.Meanwhile,cluster analysis was conducted on the samples of Trichosanthis Pericarpium from different collection fields 5.By using spss19.0 statistical software,the correlation coefficient between the microscopic characteristic constant of melon smear cells,the flavonoids in Trichosanthis Pericarpium,and the contents of total flavonoids in Trichosanthis Pericarpium was studied,which provided a multi-dimensional evaluation of the quality of melon skin.in accordance with.Results: 1.The result of the uniform design software is that the optimal condition is about 100 mg of the sample,the 120 mesh sieve is crushed,and all chioraldehyde hudrate is added to establish the correlation between the microscopic characteristic constant of the Trichosanthis Pericarpium powder and the content of each flavonoid.Sexual analysis showed that the microscopic characteristic constant had a good linear relationship with the quercetin content,which was positively correlated and highly correlated.2.In this experiment,methanol was used as the extraction solvent for the process investigation.On the basis of single factor study,the optimal process for extracting total flavonoids from Trichosanthis Pericarpium by orthogonal test is as follows: material to liquid ratio 1:8(g/m L),ultrasonic time 40 min,ultrasonic temperature 60°C,ultrasonic power 180 W.Under the optimal process,the total flavonoid content of No.9 producing area(Anhui 1)was 8.22 mg/g.On the basis of the best technology,the content of total flavonoids in 10 different collections of Trichosanthis Pericarpium medicinal materials was determined,and the content of total flavonoids in different producing areas was found to be different.The No.9 producing area(Anhui 1)had the highest content of 7.68 mg/g,while the No.1 producing area(Hebei 1)and No.5 producing area(Hebei 5)had the lowest contents of 4.32 mg/g and 4.85 mg/g,respectively.3.The results of the content determination showed that 10 different collections of Trichosanthis Pericarpium medicinal materials contained rutin,isoquercitrin,quercetin,quercetin and kaempferol.The content ranges from 20.35 to 406.9,41.38 to 198.8,3.324 to 14.82,14.21 to 60.86,and 11.96 to 49.47 μg·g-1,respectively.The content of flavonoids in different collections of Trichosanthis Pericarpium cultivar was different,and the cumulative contribution rate of isoquercitrin was 95.51%.The results of cluster analysis showed that the quality of medicinal materials of Anhui 1 was the best.The fingerprint of the Trichosanthis Pericarpium was established,and 10 different batches of Trichosanthis Pericarpium were taken.4.According to the stability of chromatographic peak,representative.The fingerprints of 10 batches of melon medicinal materials and 8 batches of medicinal materials all met above 0.90,indicating that the fingerprint analysis was carried out according to the method,and the similarity was good.5.The multivariate correlation analysis between microscopic characteristic constants of cucurbitacin and quercetin content and total flavonoids was carried out by spss19.0 software.The quercetin content was independent variable X1,the microscopic characteristic constant was independent variable X2,and the total flavonoid content was determined.As the dependent variable,the output result: correlation coefficient r=0.765,determinationcoefficient.R2=0.775,F=12.025,P=0.002,Y=81.997X1+0.011X2+0.941.The results were statistically significant.It is convenient to determine the content of quercetin and total flavonoids in melon skin by microscopic quantitative method.This paper provides a new and rapid method for the determination of Trichosanthis Pericarpium.Conclusion: 1.The microscopic characteristics of stone cell of Trichosanthis Pericarpium from different collection sites were different.This can be used as the basis for distinguishing the quality of crude drugs of Trichosanthis Pericarpium of different production.The method is stable and reliable,which provides the basis for the identification of Trichosanthis Pericarpium in different regions and the establishment of microscopic quantitative standards for stone cells.2.The content of total flavonoids in Trichosanthis Pericarpium of different producing areas varied greatly.The content of total flavonoids in different Trichosanthis Pericarpium.is different,which may be caused by the different geographical environment and climate.Therefore,the use of trichosanthis peel needs to distinguish the origin.3.Established by high performance liquid chromatography determination of five kinds of flavonoids content in Trichosanthis Pericarpium method,the result is reliable,easy to operate,with different quercetin as the public factor variables,can be more comprehensive of Trichosanthis Pericarpium medicinal materials quality control,weighted variation coefficient method,No.9 collection area(An Hui 1)Trichosanthis Pericarpium medicine comprehensive evaluation score the highest.4.The established methodology can be used for zhe qualitative identification of fructus Trichosanthis Pericarpium.5.The correlation between the microscopic characteristic constants of stone cells and the content of flavonoids was established.The quality of medicinal materials was given a new reference from the perspective of microscopic identification. |
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