| Objective: Radix Codonopsis is a traditional tonic medicine commonly used in traditional chinese medicine.Radix Codonopsis is the dried root of Codonopsis pilosula(Franch.)Nan-nf.,Codonopsis pilosula Nannf.Var.modesta(Nannf.)L.T.Shen or Codonopsis tangshen Oliv.In Codonopsis genus campanulaceae family,in2015 edition of chinese pharmacopoeia,which mainly distributes in northeast,north,southwest and northwest of China.It has the effect of invigorating the spleen and benefiting the lung,nourishing the blood and giving birth to the blood.Mainly used for spleen and lung qi deficiency,cough asthmatic deficiency,qi and blood deficiency,and so on.(1)There are many basic sources of Radix Codonopsis,and the distribution field is wide.At present,there are few researches on the differences between Radix Codonopsis from different sources.In this paper,3 kinds of legal basis sources and 3non legal base sources of Codonopsis(Codonopsis tubulosa,Codonopsis clematidea and Codonopsis xizangensis)as the research object,using HPLC-PDA,LC-MS combined with other modern analytical techniques combined with statistical methods to study the 6 kinds of chemical composition of different species of Radix Codonopsis differences,and to find out the differences to distinguish between different base the source for the identification of different components of Radix Codonopsis,Radix Codonopsis source,to provide reference and basis for the study of quality classification and quality control.(2)The chemical constituents before and after fumigation of Codonopsis pilosula were compared to find the marked components before and after fumigation of Codonopsis pilosula.Methods:(1)According to the relevant requirements of the 2015 edition of the Chinese Pharmacopoeia,the contents of water,45% alcohol extracts and sulfur dioxide residues in the samples of Radix Codonopsis samples were determined,which laid a foundation for the next comparative study different sources of Radix Codonopsis and before and after fumigation of Codonopsis pilosula.(2)To establish an LC-MS method for determination of 4 kinds of compounds in Radix Codonopsis including Lobetyolin,AtractylenolideⅢ,adenosine and syringin.the content of 4 kinds of compounds in different sources Radix Codonopsis and before and after fumigation of Codonopsis pilosula were compared.(3)To establish an evaluate the detection method of HPLC fingerprint in different sources of Radix Codonopsis and before and after fumigation of Codonopsis pilosula.Its difference were compared.(4)The chemical constituents of 6 kinds of Radix Codonopsis from different sources and before and after the fumigation of Codonopsis pilosula were determined by LC-MS.and the collected data were pretreated by Xcalibur 3.0 software.The pre-processed data were imported into Simca p13.0 software for data processing and analysis,and the differences the 6 kinds of different sources of Radix Codonopsis and before and after sulfur fumigation of Codonopsis pilosula were analyzed,and the landmarks of the differences were found out.Results and conclusion :(1)The content of 45 % alcohol extract in different sources of Radix Codonopsis and before and after fumigation of Codonopsis pilosula ranged from 40.04 % to 82.30 %,among which the extract content of 3 kinds of legal basis sources Radix Codonopsis was more than 55 %,which was in accordance with the requirements of the content of the extract under the Chinese Pharmacopoeia(2015).At the same time,The content of 45% alcohol extract of Codonopsis pilosula was increased in different degree after fumigation with sulfur.The content of extract of Codonopsis pilosula was analyzed by SPSS software before and after sulfur fumigation.The results showed that there was a significant difference(P < 0.05)in the content of extract from Codonopsis pilosula before and after fumigation.(2)The differences content of lobetyolin,adenosine,syringin and atractylenolide Ⅲin the 6 kinds of Radix Codonopsis samples were obvious.The content of lobetyolin was between 119.36 μg/ml and 937.08 μg/ml.the content of lobetyolin was most abundant in Codonopsis tubulosa.The content of atractylenolide Ⅲ was between 0.19 μg/ml and 56.97 μg/ml.the content of atractylenolide Ⅲ was most abundant in Codonopsis pilosula.The content of adenosine was between 6.71μg/ml and 337.90 μg/ml.the content of adenosine was most abundant in Codonopsis tangshen.The content of syringin was between 0.61 μg/ml and 110.73 μg/ml.thecontent of syringin was most abundant in Codonopsis xizangsis.There is a different degree of reduction in the contents of lobetyolin,adenosine and syringin in Codonopsis pilosula after Sulphur Fumigation.While the increase of the content of atractylenolide Ⅲ.(3)HPLC fingerprint of Radix Codonopsis from different sources was compared.The results showed that the chemical components of the 3 kinds of legal basic sources of Radix Codonopsis were basically the same,while there is a big difference in chemical composition of the 3 kinds of legal basic-source Radix Codonopsis and the 3 kinds of non-legal basic-source Radix Codonopsis.Further analysis showed that the ratio of peak height of tangshenoside I and lobetyolin in Codonopsis pilosula was more than 1,but the ratio of peak height of tangshenoside I and lobetyolin was less than 1 in Codonopsis tangshen and Codonopsis pilosula var Modesta.which indicated that this ratio could be used as the characteristic value of Codonopsis pilosula is different from Codonopsis tangshen and Codonopsis pilosula var.modesta.It was found that the three kinds of legal base-source party were involved in the control fingerprint of Codonopsis pilosula in comparison with those of Codonopsis pilosula.There is one more peak between peak 5(tangshenoside I)and peak 4(tryptophan),and one peak between peak 7(lobetyolin)and peak 8.However,there were no corresponding peaks in the fingerprint of the three kinds of basic source Codonopsis radix,which can be used as the basis for the differentiation of the 3 kinds of legal base source Radix Codonopsis.(4)A method based on LC-MS combined with multivariate statistical analysis was es-tablished to analysis the difference of chemical composition from different kinds of Radix Codonopsis and Codonopsis pilosula before and after sulphur fumigation.This method can quickly identify the difference of chemical composition from different kinds of Radix Codonopsis and Codonopsis pilosula before and after sulphur fumi-gation.Through multivariate statistical analysis and single dimensional statistical analysis,76 different chemical components were selected to influence different kinds of Radix Codonopsis and Codonopsis pilosula before and after sulphur fumigation,and 40 of them were identified.3 components were selected as the mainlandmark difference components before and after fumigation of Codonopsis pilosula.The chemical constituents of in three legal sources Radix Codonopsis were compared.A characteristic component,Codonopyrrolidium A,could be used to Codonopsis tangshen is different from Codonopsis pilosula and Codonopsis pilosula var.modesta was obtained.Three characteristic components,Atractylenolide Ⅲ,Lobetyolinin and AtractylenolideⅡ,could be used to Codonopsis pilosula is different from Codonopsis tangshen was obtained.In 3 kinds of legal basic sources Radix Codonopsis,The chemical composition of Codonopsis pilosula and Codonopsis pilosula var.Modesta is similar,While Codonopsis pilosula and Codonopsis tangshen has more difference.The chemical constituents of from 3 kinds of legal sources Radix Codonopsis and three Non statutory sources Radix Codonopsis were compared.five characteristic components,Codonopyrrolidium A,Codonopyrrolidium C,Codonopyrrolidium A isomer,352.211 9(16.40 min),352.211 7(16.00 min),could be used to Codonopsis tangshen is different from Codonopsis clematidea and Codonopsis xizangensis was obtained.four characteristic components,352.211 7(16.00 min),352.211 9(16.40min),Codonopiloside A and AtractylenolideⅡ,could be used to Codonopsis pilosula is different from Codonopsis tubulosa,Codonopsis clematidea and Codonopsis xizangensis was obtained.Two characteristic components,Codonopiloside A and Codonopsinol B,could be used to Codonopsis pilosula var.Modesta is different from Codonopsis tubulosa,Codonopsis clematidea and Codonopsis xizangensis was obtained.The chemical constituents of from Codonopsis tubulosa and the other 5species sources Radix Codonopsis were compared.A characteristic component,350.195 9(18.06min),could be used to Codonopsis tubulosa is different from the other 5 species sources Radix Codonopsis was obtained.The chemical constituents of from Codonopsis clematidea and the other 5 species sources Radix Codonopsis were compared.A characteristic component,238.143 5(1.32min),could be used to Codonopsis clematidea is different from the other 5 species sources Radix Codonopsis was obtained.The chemical constituents of from Codonopsis xizangensis and the other 5 species sources Radix Codonopsis were compared.A characteristic component,327.108 9(8.692 min),could be used to Codonopsis xizangensis isdifferent from the other 5 species sources Radix Codonopsis was obtained.Compared with the chemical constituents before and after sulfur fumigation of Codonopsis Pilosula,it was found that the chemical composition of Codonopsis pilosula before and after sulfur fumigation did not significant change.Further analysis showed that the three main components,which could be used to the differential component between the before and after sulfur fumigation of Codonopsis pilosula,It is Codonopyrrolidium C,467.193 8(26.97 min)and 369.253 2(30.49 min).the contents of these components of Codonopsis pilosula were significantly reduced after sulfur fumigation. |
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