| Objective:Codonopsis Radix is the dried root of Codonopsis pilosula(Franch.)Nannf.,a famous medicinal material.As a special commodity,Chinese medicinal materials are an important indicator for evaluating their quality.However,the current medicinal materials market of Codonopsis Radix is still based on the 1984 "76 kinds of medicinal commodity specifications",which can no longer meet the market demand.This paper conducts a grade study on Lu Dangshen.Established the evaluation standard of the medicinal grades of Lu Dangshen;and through the comparative study on the different quality grades of medicinal materials of Lu Dangshen,the quality standards of Lu Dangshen medicinal materials were formulated.Methods:1.In this study,Investigate the standards and related books such as "76 kinds of medicinal products specifications","Several varieties of electronic trading specifications","Chinese medicine adjustment and identification color illustration","Chinese medicine commodity science" and "Chinese medicine identification" and Shanxi Lu Dangshen main producing area and the Codonopsis Radix production area of Gansu and other places,as well as Anguo,Bozhou and other medicinal materials markets and the distribution of Codonopsis Radix,planting processing and grade division,etc.and the collection of different grades of Codonopsis Radix Medicinal materials were used as experimental samples.The collection of Codonopsis Radix samples was quantified according to traditional empirical indicators such as “diameter,length,impurity content,mildew,oil,color”,and analyzed by statistical methods such as variability analysis,correlationanalysis and one-way analysis of variance.The correlation between the product grade and appearance traits of Codonopsis Radix was screened out,and the appearance traits of the Codonopsis Radix medicinal materials were screened out,and the grading standards of Codonopsis Radix medicinal materials were established.2.This topic uses the commonly used biopharmaceutical identification method,and consults the related books such as "Chinese Flora" and "Chinese Higher Plants Illustration",and refers to the 2015 edition of the "Chinese Pharmacopoeia" to identify the source and identification of the collected Codonopsis Radix,microscopic identification and thin-layer identification,then the qualitative identification method and standard of Lu Dangshen are established.Refer to the 2015 edition of the Chinese Pharmacopoeia(Appendix 1)and the routine inspection of Codonopsis Radix,including water,total ash and alcohol soluble extractives,and statistical software for the different grades of Codonopsis Radix on the aspect of water、total ash and alcohol soluble extractives,and comparison of the differences between Lu Dangshen、IOUs Dangshenan and Wen Codonopsis Radix,and make the limits for water,ash,and soluble extractives of Lu Dangshen.Based on the previous research of the research group,UV-visible spectrophotometry was used to determine the polysaccharide content of Codonopsis Radix at 490 nm,and the methodological verification was carried out,including standard curve and linear range investigation,stability,precision and repeatability.The sample recovery rate test was carried out;the polysaccharide content of different grades of Codonopsis Radix was determined,and the difference of the content of polysaccharides in Lu Dangshen、IOUs Dangshenan and Wen Codonopsis Radix was compared,and the limit of polysaccharide content of different grades of Codonopsis Radix was established and then quantitative research methods for Lu Dangshen was established.The HPLC method was used to study the characteristic map of Codonopsis Radix,and the methodological verification was carried out,including stability,precision,repeatability and durability.The HPLC characteristics of 87 batches of different grades,different origins and different varieties of Codonopsis Radix sampleswere compared.The characteristic map of Lu Dangshen was identified,the HPLC characteristic map of Lu Dangshen was improved,and the quality standard of Lu Dangshen medicinal materials was established.Results:1.Through literature and field research,it was found that Codonopsis Radix was first published in the Qing Dynasty’s "Materia Medica Renewal",and in the Qing Dynasty,"Materia Medica" recorded the Tao Real Estate Area of Shanxi Lu Dangshen,mainly distributed in Shanxi Changzhi Pingshun,Huguan,Jincheng linchuan and other surrounding counties.The research results of Lu Dangshen medicinal materials grade literature are as follows: the three grades are divided into the diameter,which are first class(1.0cm or more),second class(0.8cm or more),third class(0.4cm or more).The results of the field investigation are as follows: divided into multiple grades based on the diameter,which are large strips(0.8cm or more),medium strips(0.6cm or more),small strips(0.4cm or more),and unified goods(not screened).The research group collected a total of 87 batches(49 batches of Lu Dangshen,34 batches of IOUs Dangshenan,4 batches of Wen Codonopsis Radix),and 261 samples of different grades,different origins and different varieties of Codonopsis Radix.This experiment refers to the "76 kinds of medicinal products commodity specifications" and other standards and related literature,and combined with the market situation,selects the measurable traits of diameter,length and impurity content(observing the mildew and oil spill of each batch of samples).Through variability analysis,correlation analysis and one-way analysis of variance,the diameter and length as the traits of the Lu Dangshen grade were selected.According to the mean value,the grading standard of the Lu Dangshen medicinal materials was formulated as follows:First-class products: the diameter is more than 0.8cm and the length is more than23 cm,no fritters,impurities,insects,mildew.Second-class products: the diameter is more than 0.6cm and the length is more than 19 cm,no fritters,impurities,insects,mildew.Third-class products: the diameter is more than 0.5cm and the length is more than 18 cm,no fritters,impurities,insects,mildew.General goods: not screened,no impurities,insects,mildew.2.The 49 batches of Lu Dangshen collected by our group are the dried roots of Codonopsis pilosula(Franch.)Nannf.The medicinal material has a cylindrical shape and is 13 to 43 cm in length and 0.4 to 2.6 cm in diameter.The surface is yellowish brown to grayish brown,and there are often dense annular stripes under the roots;all of them have longitudinal and sparsely long lenticels,and dark brown gums can be seen at the roots and roots.The texture is slightly hard or soft,with radial texture,cracks on the outside,yellow-white or yellow-brown skin,yellowish wood;special aroma,slightly sweet,chewy slag,its traits.Stone microscopic,milk tube,inulin,cork cells,trapezoidal or reticulated catheters are the main microscopic identification features.A thin layer of silica gel G was used,and the reference ginsenoside was used as a reference substance.N-butanol-glacial acetic acid-water(7:1:0.5)was used as a developing agent,and 10% sulfuric acid ethanol solution was used as a color developing agent,and heated at 90 °C until The spots were clearly colored and were examined by daylight and ultraviolet light(365 nm).The basic source identification,trait identification,microscopic identification and thin-layer identification of the quality standard of Lu Dangshen were established.The methodological study on the determination of polysaccharide content of Lu Dangshen showed that the polysaccharide of Codonopsis pilosula showed a good linear relationship in the concentration range of 5.15μg/ml ~ 51.5μg/ml.The regression equation was Y=0.0134x+0.017(r=0.9996,n=6).The average sample recovery rate was 99.40%.According to the appendix of the 2015 edition of the Chinese Pharmacopoeia,the moisture content of 49 batches of Lu Dangshen was 5.61%-1.26%,and the total ash content was 2.88%-7.43%.In terms of dry product,the content of soluble extractives in Lu Dangshen was 54.82%~ 75.93%,polysaccharide content 6.60% ~32.90%.According to the mean value of ±20%,combined with the characteristics of themoisture absorption of the Codonopsis Radix,it is proposed that the water quality of the Codonopsis Radix should not exceed 12.0%,and the total ash content should not exceed 5.0%,the soluble extractives content should not be less than 57.0%,and the polysaccharide content should not be less than 13.0%.The inspection items and content determination items of the quality standards of Lu Dangshen were established.A characteristic chromatogram of Lu dangshen by HPLC was established.The relative retention time of seven characteristic peaks was determined by taking peak 5as reference peak,which were 0.26(peak 1),0.39(peak 2),0.51(peak 3),0.66(peak4),1.00(peak 5),1.02(peak 6)and 1.30(peak 7),respectively.The characteristic peaks were identified by reference addition method,and the results were as follows:1.Peak codonopyrrolidium B,peak 3-geniposide,peak 4-codonopyrrolidin I,peak5-codonopyrrolidin and peak 7-atractylolide III identified five characteristic peaks,of which peak 3-geniposide was first identified.Comparing the characteristic chromatograms of different grades of Lu dangshen by HPLC,the similarity is between 0.46-0.92;there are significant differences between the relative peak areas of large strips and medium strips,unified goods and medium strips No.1,between large strips and small strips No.3,and between medium strips and unified goods No.7.For IOUs Dangshenan,Compared with the characteristic chromatograms of Lu Dangshen by HPLC,the similarity was low,ranging from 0.38 to 0.70.The relative peak areas of peaks 2,6 and 7 of IOUs Dangshenan were significantly smaller than those of Lu Dangshen.The similarity of the characteristic maps of Wen Codonopsis Radix was about 0.4.The relative peak area of No.4 and No.6 peaks was significantly smaller than that of Lu Dangshen.A grade standard for the medicinal materials of Lu Dangshen was established and submitted.Conclusion:1.On the basis of a large number of market and production research,with reference to the "76 kinds of medicinal product specifications and standards" andother literatures,statistical software was used to screen out the main indicators of the diameter and length of Lu Dangshen medicinal materials,and initially proposed the grade standard of Lu Dangshen medicine products.The formulation of this standard is guided by market research and has certain rationality,reality and scientificity.It has certain practical significance for the evaluation of the quality of Lu Dangshen.It provides scientific research for the formulation of the commodity grade standard of Lu Dangshen and the market price of high quality and good price.2.Through the research and comparison of 49 batches of different origins and different grades of Lu Dangshen,such as base identification,trait identification,microscopic identification,thin-layer identification,routine inspection,polysaccharide content determination and chemical characteristic map,Medicinal grade quality standards was established.The method established in this study has good accuracy,specificity and feasibility.Based on the 2015 edition of the Chinese Pharmacopoeia,the content of polysaccharides and the characteristic maps have been added to reflect the quality of Lu Dangshen and to formulate the party.The quality standard of medicinal materials provides a theoretical basis and provides a scientific basis for the quality control and rational use of Lu Dangshen medicinal materials. |
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