| Objective:Codonopsis Radix is traditional Chinese medicine for Buzhongyiqi. In the Chinese Pharmacopoeia of 2010, Dangshen come from three base sources, Codonopsis pilosula(Franch.) Nannf, Codonopsis pilosula Nannf.var.modesta(Nannf.) L.T.Shen and Codonopsis tangshen Oliv. It has long history of cultivation, and Commodity Dangshen was mostly cultivar Dangshen.And Dangshen quality standards was formulated the Character identification, microscopic identification, TLC identification and lobetyolin extract limits and so on.Because of the planting area is wide and the natural environment,cultivation and processing methods was have Significant differences, the quality of Dangshen was irregularity. Lu Dangshen come from Gu luzhou of shanxi(now City Changzhi and Jincheng). It is Shanxi famous-region drug, with high quality and long history. But at present, the existing Pharmacopoeia standards was inadequate, and lack of local quality standards. It is about character identification and microscopic identification items of Local standards for Dangshen and the literature of identify for different base sources. It is provided the method for formulate the quality standards of Lu Codonopsis,identificating different based source of Dangshen, and provided the theoretical basis for guiding the production, and assurancing the quality of Dangshen.Methods:It is extremely urgent of enhance the quality control methods and improve the quality standards. So the RP-HPLC specific chromatography was adopted to determined thecontents of lobetyolin and atractylenoide III; and study the HPLC specific chromatograms of 54 batches with three different sources of Codonopsis Radix.The difference of the HPLC specific chromatograms between the Lu Dangshen and other different Codonopsis Radix was compared, and separate and identify the chemical composition. The methanol ultrasonic extraction of Lu Dangshen, was separated and purificated through D101 macroporous resin, ODS reverse phase silica gel and so on, 8 compounds was obtained, the structure was idetificated by ESI-MS, NMR.Results:1. The determination of lobetyolin and atractylenoide III in Codonopsis RadixThe content of Lobetyolin and atractylenoide III have significantly difference.Lobetyolin and atractylenoide III in Lu Dangshen was 0.356 mg·g-1 and 0.035 mg·g-1, the content is higher and the quality is better of Lu Dangshen. But, after spraying Zhuanggenling, the content of two ingredients was significantly reduced. Lobetyolin can be detected in the Codonopsis Radix from different base sourses. The content of Codonopsis pilosula Nannf.var.modesta(Nannf.) L.T.Shen is highest, Fengdang was 2.236mg·g-1, significantly higher than Wendang. We think that the base source have the biggest effect to the content of lobetyolin.While, the content of atraxtylenoide III in Taidang is highest and was 0.174 mg·g-1,Lu Dangshen was followed and was 0.035 mg·g-1, significantly higher than Baitiaodang.We speculate that the producing area has biggest to the atraxtylenoide III and atraxtylenoide III was the Characteristic components of Lu Dangshen.2. Study on the HPLC specific chromatograms of Codonopsis Radix from different sources.2.1 the HPLC specific chromatograms of Lu DangshenThe HPLC specific chromatograms of Lu Dangshen control medicine was established and the relative retention times was determined. It is 0.195(peak 1), 0.323(peak 2),0.443(peak 3), 0.616(peak 4), 1.00(peak 5), 1.01(peak 6)、1.36(peak 7).The characteristic peaks was identified by the separated chemicalcomposition(codonopyrridilium B, tangshenoside I) and the purchased standards(syringin,lobetyolin, atractylenoide III) in the HPLC specific chromatogram: peak 1th was codonopyrridilium B, peak 2th was springin, peak 4th was tangshenoside I, peak 5th was lobetyolin, and peak 7th was atractylenoide III.2.2 the comparison of similarity of different producing area of Codonopsis RadixLu Dangshen, Baitiaodang and Taidang belong to the same base source, Codonopsis pilosula(Franch.) Nannf. The similarity of Lu Dangshen and Taidang was above 0.8, and with Baitiaodang is 0.4~0.8, but to Codonopsis tangshen Oliv. and Codonopsis pilosula Nannf.var.modesta(Nannf.) L.T.Shen. was only 0.1~0.4. It is show that the difference of base sources was bigger than producing area.2.3 the comparison of HPLC specific chromatograms of Codonopsis RadixIn the Baitiaodang HPLC specific chromatograms, the peak area was less than Ludangshen, in especial, 1th(codonopyrrolidium B), 4th(dangshenoside I),6th and7th(atractylenoide III), and the relatively peak area was significantly different with Lu Dangshen. The Lu Dangshen and wild Taidang has more different. Wild sample has not detected the peak 6th and the peak area of 1th(codonopyrrolidium B), 3th and7th(atractylenoide III) was bigger than Lu Dangshen.The comparison of different sources samples. Codonopsis tangshen Oliv. was not detected the peak 6th and the 7th(atractylenoide III), and the peak area of 3th and4th(dangshenoside I) was especially less than Lu Danagshen and the ralative peak area was significantly different. Codonopsis pilosula Nannf.var.modesta(Nannf.) L.T.Shen. was not detected the peak 6th, and the meanpeak area of peak 4th(dangshenoside I) and 7th(atractylenoide III) was also lower than Lu Dangshen. and the relative peak area significantly different.2.4 the comparison of different cultivation, different processing of Codonopsis Radix.After spraying Zhuanggenling, the similarity was reduced and the HPLC specific chromatograms has significantly difference. The peak area of peak 3th, 4th(dangshenoside I)and 7th(atractylenoide III) is correlation of growth years, and the peak 6thwas not detected in the Codonopsis Radix with the longer harvest years. In addition, the similarity of thesulfured Baitiaodang was between 0.4 and 0.5, and the HPLC specific chromatograms was main detected peak 2th(springin), 4th(dangshenoside I) and 5th(lobetyolin) and the peak area was less than Ludangshen.3. The separate and identify of chemical composition in the Lu Dangshen.There ingredients was determined: codonopyrrolidium B(IV), dangshenoside I(V),lobetyolin(VI), and three Oligosaccharide(I, II, III) n-hexyl- β-sophoroside(VII) and9,10,13-trihydroxy-(E)-11-octadecenoic acid(VIII) to be determined.Conclusions:Lu Dangshen has good quality and genuineness.The vary based sources, producing area and the cultivation or processing methods of dangshen, the HPLC specific chromatograms has significant differences. The HPLC specific chromatograms has significantly difference. Lu Dangshen has more two peak of 6th and 7th(atractylenoide III)than Codonopsis tangshen Oliv. And has more one peak of 6th than Codonopsis pilosula Nannf.var.modesta(Nannf.) L.T.Shen. And the peak area was bigger than Baitiaodang, At the same time, the 4th(dangshenoside I), 6th and 7th(atractylenoide III) can be used as the characterisitics ingredients reflecting the genuineness of Lu Dangshen,It is reflect the uniqueness and the genuineness, and control the quality of Lu Dangshen, and identify the different base source, and provide a basis for the cultivation of Dangshen of the established the HPLC specific chromatograms. |
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