1.Purification process of total flavonoids of Crataegus leaves with macroreticular resin
Yongxue GUO ; Nan LI ; Yanlai ZHANG ; Changquan HAO
Chinese Traditional Patent Medicine 1992;0(01):-
AIM: To study the technological parameters of the purification process of total flavonoids of Crataegus leaves with HPD-600 macroreticular resin. METHODS: Using orthogonal experiments,the adsorptive characteristics and elutive parameters of the process were studied by taking the elutive effects and purification ratio of total flavonoids of Crataegus leaves. RESULTS: Two significant factors which were the concentration of crude flavonoids solvent and elutant component were observed. CONCLUSION: The microwave-assisted extraction and purification process with macroreticular resin is employed,the content of total flavonoids is 80.75%, the rutin content is (1.38)%, The macroreticular resin can be used in the purification of flavonoids of Crataegus leaves.
2.in vitro Metabolism Pathway of Effective Component of Bletilla striata as Militarine in Liver Microsomes and Kinetics Char- acteristics of Enzyme-catalyzed Reactions
Tao XIA ; Changquan WANG ; Hao CHEN ; Lin ZHENG ; Zipeng GONG ; Yueting LI ; Yongjun LI ; Yong HUANG ; Jie PAN
China Pharmacy 2019;30(10):1316-1320
OBJECTIVE: To study in vitro metabolism pathway of effective component of Bletilla striata as Militarine in liver microsomes and kinetics characteristics of enzyme-catalyzed reactions. METHODS: The in vitro incubation system of rat and human liver microsomes was established, and incubation reaction of Militarine was performed. UPLC-QTOF-MS was used to identify the structure of its metabolites in combination with UNIFI database and references. Using puerarin as internal standard, UPLC-Triple Quad-MS was used to quantitatively analyze metabolic transformation of Militarine in rat liver microsomes. The kinetic parameters (vmax, km, CLint) of Militarine enzyme-catalyzed reactions with/without reducing coenzyme Ⅱ (NADPH) were calculated by fitting the curves with GraphPad Prism 5.0 software. RESULTS: After incubation in rat and human liver microsomes, Militarine produced a chemical formula C21H29O11, which was presumed to be a metabolite of Militarine ester bond hydrolysis. The kinetic study of enzyme-catalyzed reactions showed that vmax of Militarine enzyme-catalyzed reactions with/without NADPH were 1.955, 2.129 nmol/(h·mg); km were 8.601, 9.854 nmol/mL; CLint were 0.227 3, 0.216 1 mL/(h·mg); there was no significant difference between with NADPH and without NADPH. CONCLUSIONS: The main metabolic pathway of Militarine in liver microsomes is the hydrolysis of C1 and C4 ester bonds. Its metabolism does not depend on the pathway of cytochrome P450 enzymes initiated by NADPH.