| Akebia trifoliate(Thunb.) Koidz. is a deciduous woody vine,which belongs to Lardizabalaceae. It distribute in many provinces in China. Its dry root and caudex known clearing fire in human heart, diuresising, regulating catamenia and hastening the exudation of latex have been widely used as Chinese medicine for centuries and recorded in many related literatures. Its value in pharmacology and economic benefit will be highly recognition.In order to find a more reasonable method of quality control,we attempted to establish its method for high performance liquid chromatographic fingerprint analysis.The sample preparation methods were screened and the chromatographic conditions including the mobile phase, detection wave, etc were optimized in this paper. This method was used to analysis the sample of different places. Moreover the main active components in Fructus Akebiae were determined by high performance liquid chromatography in order to establish its general quality control standard. The main results are listed as follows:1.The paper establishes a method for high performance liquid chromatographic (HPLC) fingerprint analysis. The sample preparation methods were screened and the chromatographic conditions were optimized in this paper. The sample preparation method is:the powdered sample was accurately weighed (0.5g) and extracted with 25mL methanol under ultrasonic for 45min, the extracts were filtered. The filtrate was evaporated to dryness in vacuo. The residues were diluted with 10mL water,20mL methanol and 1.0mL HC1 at 80℃in a water bath for 1h. The filtrate was evaporated to dryness and the residue was dissolved in methanol and diluted to 10 mL in a volumetric flask. Hibar C18 (250x4.6 mm,5μm)column was used, with mixtures of water and acetonitrile as mobile phase in a gradient mode. The flow rate was 0.8 mL/min, the temperature of column was 20℃,the wavelength of detection was 210 nm, the injection volume was 20μL.2.HPLC was used to establish fingerprint chromatograph of 26 batch of Akebia trifoliate in different provinces.The result showed that 14 peaks were common. We used the software named chromatographic fingerprints of TCMs similarity evaluation system 2004 A Edition to analyze and evaluate the obtained chromatogram.The similarity of Hubei, Hunan, Gansu is 0.90.The pink area of Shanxi's samples are different from others, so the similarity lower than others. Cluster analysis was used to cluster the similar sample. At last the software divided the 26 batch samples into 3 clusters. The method was simple,stable, reliable,and reproducible, can provide a reference for the development of Akebia trifoliata(Thunb.)Koidz quality standards.3.In this paper, we establish an HPLC method for determination of saponins PD and Oleanolic in Fructus Akebiae. The results showed different samples had obvious distinction. It valid composition total content from high to low is Zhen'an, FengYukou, Songbai, Fangxian, Shimen, Cili, Shibing, Zhenyan. The method was simple, stable, reliable and reproducible.SPSS16.0 was used to analysis the correlation between the content of saponins PD and environment factors in order to ascertain the main factors which can affect the content of saponins PD Stepwise regression analysis was used to establish an equation:Y=0.305+0.870X1+0.033X2-0.115X3(R2=0.987,P<0.01)and X1 is latitude,X2 is annual mean sunlight hours, X3 is annual average temperature. The results are the content of saponins PD showed appositively correlation to latitude and annual mean sunlight hours; the content of saponins PD showed a negative correlation to annual average temperature. |
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