Identification and quality evaluation of Chinese rice wine using UPLC-PDA-QTOF/MS with dual-column separation.

BACKGROUND: Chinese rice wine (CRW) is a well-known drink and functional food that is used in traditional Chinese medicine. However, there is still a lack of quality control and evaluation methods for CRWs. PURPOSE: The study aimed to establish a new method that can serve both as quality control and evaluation method and, as well as an identification method for CRWs. METHOD: Compound identification in different CRW samples and determination of uracil, xanthine, uridine, adenine, guanosine, 5-hydroxymethylfurfural, and adenosine contents from 29 CRW samples from 14 brands were performed using UPLC-PDA/TOF-MS. The dual-column chromatographic separation of CRW was performed using CORTECS T3 coupled to HSS T3. The optimal mobile phase consisted of water with 0.1% formic acid, 40 mM ammonium acetate, and methanol: acetonitrile (2:1). Furthermore, to compare the UPLC fingerprints between CRWs of different brands, a similarity analysis was performed to classify the CRW samples. Finally, network pharmacology and in vitro efficacy and toxicity tests were used to investigate the biological function of the seven components and CRWs. RESULTS: A total of 55 compounds were unambiguously or tentatively identified. Among them, nucleoside, pyrimidines and purines were reported in CRW for the first time. The seven components were successfully determined, and their contents showed large variations among different brands of CRW, which was consistent with the results of the chromatographic fingerprint similarities. The results of in vitro efficacy and toxicity tests indicated that CRWs and seven components had obvious protective effect on H9c2 cell injury induced by the H2O2 model. Network pharmacology analysis showed that these seven compounds might be the main active components of CRW that promote blood circulation and ventilation. CONCLUSION: This study revealed that dual-column chromatographic separation is an effective method for quantitative and chromatographic fingerprint analyzes of complex samples, and seven compounds can be used for the quality evaluation and control of CRWs.

Metabolic Profiling of Glabridin in Rat Plasma, Urine, Bile, and Feces After Intragastric and Intravenous Administration.

BACKGROUND AND OBJECTIVES: Glabridin is usually used to treat cardiovascular and central nervous system diseases, which is a main bioactive phytoconstituent of licorice root. In this study, our aim was to obtain metabolic profiles of glabridin in rat plasma, urine, bile, and feces after intragastric and intravenous administration. METHODS: By UPLC-QTOF-MS, the metabolites of glabridin were systematically analyzed and identified. An ACQUITY UPLC HSS T3 column (100 × 2.1 mm, 1.8 mm) and the mobile phase containing water with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B) via gradient elution was used as the chromatographic condition for separation. An extracted ion chromatogram strategy with multiple prototype/metabolite intermediate templates and 71 typical metabolic reactions was suggested to completely profile the metabolites of glabridin. RESULTS: For the first time, 13 compounds in total were recognized from urine, plasma, bile, and feces of rats with the metabolite profiling strategy. The main metabolic form of glabridin in rats consisted of the prototype, hydroxylation, glucuronide conjugation, decarbonylation, and tert-butyl to alcohol metabolites. CONCLUSIONS: The results not only indicated a comprehensive study on the probable metabolic pathways of glabridin in vivo, but also provided useful data and information for future pharmacological studies of glabridin.

An integrated network pharmacology and cell metabolomics approach to reveal the role of rhein, a novel PPARα agonist, against renal fibrosis by activating the PPARα-CPT1A axis.

BACKGROUND: Rhein, an anthraquinone compound, displays extensive antifibrotic effects; however, its potential mechanisms are not fully understood. In this study, we explored the underlying molecular mechanism of action of rhein. METHOD: An integrated network pharmacology and cell metabolomics approach was developed based on network pharmacology and bioinformatics method, and then successfully applied to speculate the potential targets of rhein and construct a rhein-target-metabolic enzyme-metabolite network. Thereafter, the antifibrotic mechanism of rhein was validated in TGF-ß- and oleic acid- induced HK-2 and NRK-52E cells in vitro as well as a unilateral ischemia-reperfusion injury Sprague-Dawley rat model. RESULTS: Based on the construction of the rhein-target-metabolic enzyme-metabolite network, we found that rhein played an antifibrotic role through the PPAR-α-CPT1A-l-palmitoyl-carnitine axis. In vitro experiments demonstrated that rhein effectively activated the expression of PPARα and its downstream proteins (CPT1A and ACOX1) to alleviate lipid accumulation and fibrosis development. In vivo experiments indicated that rhein attenuated renal fibrosis mainly by activating the fatty acid oxidation pathway and improving lipid metabolism. CONCLUSION: Taken together, our findings reveal that rhein is a novel agonist of PPARα, which contributes to its renoprotection through the regulation of the PPARα-CPT1A axis. Moreover, our study provides a novel insight into an integrated network pharmacology-metabolomics strategy for uncovering the pharmacological mechanisms of drugs from the system perspective.

The chemical and metabolite profiles of Gualou-Xiebai-Banxia decoction, a classical traditional Chinese medicine formula, by using high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and in-house software.

ETHNOPHARMACOLOGICAL RELEVANCE: Gualou-Xiebai-Banxia decoction (GXBD) was a classical traditional Chinese medicine formula for the treatment of coronary heart disease. However, the current study on the chemical and metabolite profiles of GXBD did not follow the ancient prescription and extraction method, which hindered the discovery of effective compounds and quality control. MATERIALS AND METHODS: In this study, we prepared GXBD by ancient prescription and extraction methods, and then analysed the chemical components and xenobiotics of GXBD in vivo using high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and in-house software. RESULTS: 49 chemical constituents were preliminarily identified, including 7 terpenoids, 6 flavonoids, 5 alkaloids, 17 organic acids, 8 steroids and steroidal saponins, 2 nucleosides and 4 other types of compounds, of which 10 constituents were confirmed unambiguously with authentic standards. Moreover, 129 metabolites were tentatively identified, including 83 metabolites in plasma, 39 metabolites in urine, 25 metabolites in bile and 9 metabolites in feces. Our study speculated that luteolin, adenosine, vanillic acid and curbitacin B might be possible effective components of GXBD for the treatment of coronary heart disease. Dehydration, deglycosylation, dehydrogenation, acetylation and taurine regulation were the main biotransformation reactions of GXBD. CONCLUSION: Our results provided an important basis for the discovery of effective compounds and quality control of GXBD. In addition, in-house software was an useful tool for identifcation of metabolites.