| The main research contents are as follows: Optimizing the conditions for the extraction of tectoridin, which was characterized by spectroscopic instrument; The inhibition of hyaluronidase by tectoridin, and its antioxidative ability; The interaction mechanism between tectoridin and hyaluronidase by fluorescence spectrum, cirular dichroism, etc.Results show that the optimum extraction conditions are as follows: ethanol concentration is 70%, the ratio of solid to liquid is 1:8(w/v), extracting 30 min each time, and under this condition the purity of tectoridin is 70.1%, the extraction rate is 1.333%. Through AB-8 macroporous resin and silica gel separation, the purity of tectoridin is 96.8%,and the extraction rate is 0.653%. Extract was tested by m.p,IR, UV,LC-MS and 1H-NMR,and compared with the standard, we ascertain that the extract is tectoridin.The isolated tectoridin has an active effect on the inhibition of hyaluronidase. Its inhibiton rate are able to reach 81.2% with the increase of tectoridin's intensity. At the same time, by the eliminate experiment of H2O2, OH·and DPPH·, the extract displays reliable ability in antioxidant. The maximum clearance rate against the 3 kinds of free radical are 68.5%, 80.7%, 39.6%, respectively. The research shows that tectoridin has good anti-allergenic.The result of Fluorescence spectrumshows us that Hyaluronidase appeared the strongest emission peak and the fluoresecence intensity decreased as tectoridin's concentration increased.The way of Hyaluronidase is static quenching,binding constants is 0.613×103L·mol-1(296K),1.643×103L·mol-1(303K),2.441×103L·mol-1and binding number≈1.At the same time, the procedure of tectoridin's effect on Hyaluronidase happens spontaneously.The bonding construction between tectoridin and Hyaluronidase is lyophobic. The synchronous fluorescence spectra (△λ=60nm) and circular dichroism spectra analysis results shows that tectoridin cause the reduction of theα-helix content of hyaluronidase. |
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