| Epimedium is one of unique medicinal and edible plants in China. In order to provide a basis for scientific exploitation and comprehensive utilization of Epimedium resources in the food industry, nutritional compositions, the oxidation resistance and inhibiting myeloma cell proliferation activity of Epimedium were analyzed. A rapid chromogenic method was set up for the determination of water-soluble polysaccharides content of Epimedium by optimizating parameters of the color reaction. A rapid and nondestructive near infrared spectroscopy was established for testing the protein content of Epimedium, it had reference significance for Epimedium resources development, utilization and the medicine quality control. This paper also studied the Epimedium nutrition in different organizations and finally discussed the oxidation resistance, DNA damage protection and myeloma cell proliferation inhibitory activity analysis of Epimedium flavonoids in different organizations. This paper provided theoretical basis for the further study of Epimedium.The results of the study are as follows:(1) The aim of this study was to establish a specific, accurate, reproducible and sensitive phenol-sulfuric acid method for the quantitative assay of Epimedium polysaccharides, and to determine polysaccharides in Epimedium samples in Chinese markets. Galactose was adopted as the standard monosaccharide and486nm was chosen as detective wavelength. The optimal conditions for color reaction were obtained using single factor experiments and orthogonal test:temperature of20℃, amount of5%phenol of0.3mL, amount of concentrated sulfuric acid of3.5mL, incubation time of20min, and addition sequence of phenol-sample-sulfuric acid. And the colored sample solution after chromogenic reaction exhibited high stability within2h. Calibration curve was found to be linear with the range of5.00-60.00μg/mL and correlation coefficient of regression equation was0.999. LOD and LOQ were1.65and5.00μg/mL, respectively. Recovery, intraday precision, interday precision, and accuracy varied from97.43%to103.80%, from0.73%to3.48%, from1.21%to2.75%and from97.74%to101.62%. respectively. Polysaccharides in26samples of Epimedium collected from different provinces of China were quantified by the proposed colorimetric method. And a large variation of contents of polysaccharides was observed among these samples.(2) Near-infrared reflectance spectroscopy technique was applied to rapid and nondestructive determination of protein in Epimedium. Calibration model was developed under the following conditions:2nd derivative, standard multiplicative scatter correction, principal components analysis and modified partial least squares regression. R squared, standard error of cross-validation and1minus variance ratio were0.923,0.554and0.717, respectively. There’s no significant difference between protein contents determined by near-infrared reflectance spectroscopy and Kjedahl nitrogen determination methods, and their relationship was close (R2=0.9339). The proposed near-infrared reflectance spectroscopy method is proved to be a simple analytical technique with satisfactory accuracy and repeatability (RSD=0.937%). Contents of protein in eight species of Epimedium were determined by near-infrared reflectance spectroscopy method for the first time.(3) Different organizations of Epimedium were selected as experiment material. The results are as follows:Epimedium leaf has highest water soluble ash (2.26%). water insoluble ash(5.23%), protein content(10.46%), the essential amino acid content(43.78mg/g) and AAS value (139.32%); Stems are rich of crude fat (5.67%) and crude fiber (48.22%); Epimedium polysaccharide content (1.87%), half essential amino acids (his+Arg,16.36mg/g), E/T value(31.51%) and the flavonoids content (12.27%) in root is highest. Leucine and isoleucine become the restrictive amino acid in different organizations of Epimedium; Detection rate for macro elements(Mg, K, Ca) and trace elements(Mn, Fe, zinc) were100%, at the same time, the setection rate of As and Pb elements were100%, but Pb element in leaves exceeded industry standards specified in the detection limit. Cd and Hg element is not checked out.(4) The antioxidant capacity was obtained by removing DPPH· and ABTS-+of Epimedium flavonoids in different organizations. The sequence of remove DPPH· is: leaf>root>stem. And the sequence of remove ABTS’+is:root>leaf>stem. The ability of removing DPPH· and ABTS·+of Epimedium flavonoids in different organizations increase gradually with the increase of concentration, and the root has the strongest antioxidant capacity. The oxidative damage protection of Epimedium flavonoids in different organizations for pBR322plasmid DNA was conducted by agarose gel experiment. In this study, at the same concentrations, Epimedium flavonoids in different organizations all had the protection for the oxidative damaged pBR322plasmid DNA. And the degree of protection is:root>leaf>stem. Myeloma cell proliferation inhibition of Epimedium flavonoids in different organizations was conducted by MTT method. OD value indirectly reflects the strength of the inhibition of myeloma cell proliferation. Epimedium flavonoids (gradient concentration) in different organizations all had myeloma cell proliferation inhibition, and present a dose-dependent, by contrast, at the same concentration, the proliferation inhibition effects of root was strongest. |
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