The Antioxidative Efficient Component Recognition for Curcuma Longa L. Based on the Quality Control Pattern of Constituent Knock-out/Knock-in
|School||Kunming University of Science and Technology|
|Keywords||Quality control for TCM Efficient component recognition Constituent knock-out/knock-in Biological activity determination Antioxidative Curcuma longa L. Pattern and method|
To seek a new breakthrough of the quality control for Traditional Chinese medicine (TCM), a tentative idea about efficient component recognition and quality control pattern for TCM based on constituent knock-out/knock-in is proposed. In this article, Curcuma longa L. was studied as a model due to its universal clinical application and relative clarity chemical components. We focus on its antioxidative activity and explore the pattern of efficient component recognition and quality control for TCM based on constituent knock-out/knock-in, which could perfect the current system of the quality control for TCM and assure its efficacy and safety.The different constituent knock-outed samples were prepared respectively using by Thin-Layer Chhromatography (TLC) and supercritical extraction technology. The Ultra Performance Liquid Chromatography (UPLC) detection method was established to initially analyze the different constituent knock-outed samples, which is base for the research of spectrum-effect relationship and further chromatographic fingerprint investigations.The microsomal lipid peroxidation assay in 96-well plate was developed suggesting as the biological antioxidative activity determination method for TCM. The central composite design and response surface methodology (CCD-RSM) was used to optimize the measuring conditions. The optimum conditions of the assay were obtained to get the appropriate maximal absorbance:the concentration of Fe(Ⅱ),150μM; the molar ratio of ascorbic acid (AsAH2)/Fe(Ⅱ),2.09/1-2.62/1; and the thiobarbituric acid (TBA) color reaction time,90 min. We also systematically characterized the influences of trichloroacetic acid (TCA), surfactant and sample solvent on the reaction system of the optimal lipid peroxidation assay. The biological antioxidative activity determination method for TCM is consistenting with the quality control pattern’s requirements of efficiency, sensitiveness and rapid.The antioxidative activities of different constituent knock-outed samples were measured by the method. Variance Analysis was used to evaluate the contributions of different constituents about the antioxidative activity. Besides, the assay of protective effect on oxidative stress-induced damage in PC 12 cells was carried out to validate the results. Finally, the purpose of antioxidative efficient component recognition for Curcuma longa L. could be realized, as Curcumin and Demethoxycurcumin are the antioxidative efficient component recognition for Curcuma longa L.. The research initially explored the feasibility, rationality and scientificity of the efficient component recognition and quality control pattern for TCM based on constituent knock-out/knock-in.Taken together, the efficient component recognition and quality control pattern for TCM based on constituent knock-out/knock-in sufficiently reflect the holistic and systematic characteristics for TCM, which illustrates the essence of TCM theory. It could be concluded that the new pattern offer a new view and technology for the quality control of TCM, and its value would be demonstrated soon.