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The Dioscorea nipponia quality control and evaluation

Author: ZhangZuo
Tutor: KangTingGuo
School: Liaoning University of Traditional Chinese Medicine
Course: Pharmacognosy
Keywords: Dioscorea nipponica Maknio chemical constituent determination Polysaccharides inorganic element HPLC fingerprint X-ray Diffraction fingerprint
CLC: S567.239
Type: Master's thesis
Year: 2011
Downloads: 33
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Abstract

Purpose: Dioscorea nipponica M. have apparent efficacy and widely been used in clinical practice. The saponins and sapogenins in it have hypolipidemic effect, anti-platelet aggregation effect, coronary heart disease treatment effect, anti-tumor effect, etc. Dioscorea nipponica M. with its medical value and enormous perspective had been used as important industrial raw material and widely been cultivated. The chemical constituents of Dioscorea nipponica M., the quantitative analysis, the Inorganic elements of the drug, HPLC fingerprint and XRD fingerprint will be studied systematically. The results of these studies will provide an academic reference for the research of the reference substances, the quality control and cultivation of Dioscorea nipponica M..Materials and methods:1. The compounds were isolated and purified from the EtOAc and n-BuOH extracts of Dioscorea nipponica Maknio by repeated Silica-gel column chromatography, SephadexLH-20 column chromatography, and re-crystallization. Their structures were identified on the basis of their chemical and physical properties as well as the spectra data of 1H-NMR, 13C-NMR etc, compared with data reported in literatures.2. A HPLC method for the determination of Protodioscin and Dioscin had been developed, Correlate analysis and principal component analysis were applied to study a new aggregative indicator to evaluate the quality of the crude drug. There were 35 samples from various origins had been tested and evaluated.3. The independent variables were extraction temperature, extraction time and liquid-to-solid ratio, the dependent variable was extraction rate of polysaccharides in Dioscorea nipponica M. Linear or no-linear mathematic models were used to estimate the relationship between independent and dependent variables. Central composite design and Response surface methodology (CCD-RSM) were used to optimize the process of extraction. The prediction was carried out through comparing the observed and predicted values.4. The content of 13 elements in ten Dioscorea nipponica M. samples and ten soil samples were determined by means of ICP/MS, and Zn/Cu had been calculated. The principal component analysis of SPSS and Q-type cluster analysis were applied for the study of characteristic elements in Dioscorea nipponica M.. And calculating the correlations among 13 elements in crude drugs to research the cooperation of these elements. Meanwhile, calculating the correlations between the crude drugs and relevant soil samples, and the constants of element accumulation had been calculated also. The results provided a academic reference for cultivation of Dioscorea nipponica M..5. There were 36 samples of wild and cultivated from various origins were tested by the means of HPLC. And the data of chromatography of 28 samples had been analyzed by the means of statistical analysis such as principal component analysis (PCA) and Cluster analysis. Finally, 11 samples were selected to apply to establish the common pattern of the HPLC fingerprint.6. 10 sources of Dioscorea nipponica had been tested by X ray Diffraction, located by the 2θ, calculated the similarity by d/I/I0, and the record had been analyzed to obtain a standard XRD Fourier fingerprint of Dioscorea nipponica M..Results and Conclusion:1. 6 compounds had been isolated from the extracts, 4 compounds were identified as:β-sitosterol (Ⅰ), Dioscin(Ⅱ), Methylprotodiosgenin-26-O-Glc(Ⅲ), Protodioscin(Ⅳ).2. The determination of HPLC method is simple,accurate and reproducible.The two indexes were moderate negative correlated. The principal component score is proposed to be used for the quality control of Dioscorea nipponica M..3. The optimum technological parameters were as follows: extracting temperature 91.5℃, extracting time 172.9 min, solid -to- liquid ratio 1:25.8 and 2 times for extraction. Bias between observed and predicted of rates was 1.3%. Extraction technology of Polysaccharides optimized by CCD-RSM is simple, reliable and highly predictive.4. The analysis results show that Cr,Mn,Fe,Ni,Cd,Hg may be the characteristic elements in Dioscorea nipponica M.. The results of Q-type cluster analysis show that the samples could be clustered reasonably into two groups, and the elemental distribution characteristics are related to the ecology and origin of Dioscorea nipponica M.. The Zn/Cu of Dioscorea nipponica M. is apparently lowwer than the average value of Dicotyledons which can partly explain the mechanism of the anti-coronary artery heart disease. There are multi-correlations among some elements in the crude drug,in the contrast, there are no apparent correlations between the crude drug and its relevant soil. The pathway of inorganic element absorption is selective absorption in Dioscorea nipponica M..5. The data of HPLC had been analyzed by PCA and Cluster analysis. The results of the statistical analysis had been the base of the selection of samples which can be applied to establish the common pattern of the HPLC fingerprint.6. XRD fingerprint can be applied to identify Dioscorea nipponica M., and the similarity analysis can be used to evaluate the quality of this crude drug. A new identified method of D. nipponica by X-ray Diffraction fingerprint has been established.

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