| Ding-Zhi-Xiao-Wan(DZXW) is a form of traditional Chinese medicine that was initially recorded in the Essential Recipes for Emergent Use Worth A Thousand Gold(Beiji Qianjin Yao Fang). It is commonly used in the treatment of emotional disease characterized by feelings of sadness and anhedonia, which are associated with depressive disorders. DZXW is composed of four component herbs, namely Ginseng Radix, Poria, Polygalae Radix, and Acori Tatarinowii Rhizoma, in a ratio of 3:3:2:2. Due to the structural complexity and low natural abundance of Traditional Chinese Medicine(TCM), makes the phytochemical research becomes quite complex and difficult. UPLC-Q-TOF-MSE provided a sensitive, selective, and rapid analytical method for characterizing the chemical constituents of TCM.Objective:In this study, LC-MS and GC-MS methods were established to simultaneously determine the the chemical constituents of DZXW andits absorption in rats’ plasma and brain. In combination with pharmacological information, we predicted the bioactive constituents of DZXW for treatment of depression. Our results provide useful chemical information for further study of the pharmacodynamic material basis of DZXWantidepressant effects.Methods : In this study, a new rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MSE) method was established to analysis the constituents of DZXW, Chromatographic separation was carried out at on Waters CORTECTS UPLC C18 column(2.1×50mm, 1.6μm) with mobile phase A(0.1% formic acid) and mobile phase B(acetonitrile). We summarized the mass spectrum pathway of five standards through both MS full scan and MSE high energy information, and the data was controlled using Masslynx 4.1 software. In addition, we analyzed the volatile constituents in DZXW by GC-MS. Then, we established a depressive model rats by chronic unpredictable mild stress(CUMS). Following a three-day oral administration of DZXW, we collected the plasma, brain, bile, urine, feces from the rats, these samples were analyzed by UPLC-Q-TOF-MSE and processed using Metabolynx. Finally, 10 compounds were predicted as the bioactive constituents of DZXW for treatment of depression.Results:A total of 97 compounds were identified from DZXW, including 50 from Polygala and 41 from Ginseng, besides, six identified compounds were not observed in the individual drugand three potentially novel compounds were reported for the first time. After three-day oral administration of DZXW, 60 and 28 compounds were observed in the plasma of normal and depressive rats, respectively. At the same time, ten volatile compounds were identified from DZXW and three of them were detected in depressive rat brain. We summarized ten compounds(Re, Rg1, Rb1, Rc, Ro, tenuifolin, 3,6-disinapoylsucrose, Tenuifoliside C, Tenuifoliose K, polygalaxanthone) as bioactive constituents of DZXW, and analyzed the invivo metabolites and metabolic pathway in depressive rats.Conclusion : A sensitive and selective UPLC-Q-TOF-MSE method was eatablished for the simultaneous determination of DZXW and its invivo metabolites. Besides, the GC-MS detected the volatile compounds from DZXW. We can get the metabolites and types of target compounds by Metabolynx software quickly and exactly. In addition, the use of the CUMS model may increase the clinical significance of our study, which could be employed for further study of the pharmacodynamic material basis of DZXW’santidepressant effects. |
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