| Part 1 Being one of the oldest and most popular herbal medicines, Glycyrrhiza uralensis has been widely used in traditional medicines, foods, beverages, and tobacco industries. However, the main medicinal part of Glycyrrhiza uralensis were the roots and rhizomes and the plant aerial part was usually neglected. In this study, an ultra-high-performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-QQQMS) method was developed to simultaneously quantify 8 constituents including (1) 22-hydroxyglycyrrhetinic acid, (2) 3-epi-glycyrrhetinic acid, (3) glycyrrhetinic acid, (4) glycyrrhizic acid, (5) liquiritin, (6) formononetin, (7) isoglycyrol and (8) glycycoumarinin in the root, rhizome and the aerial part (stems, leaves, seeds and seed coats) of Glycyrrhiza uralensis. As a result, the method was turned out to be sensitive, simple, fast, and accurate, and could be used for the quality control of different parts of Glycyrrhiza uralensis and related food products. On the other hand, we investigated the bioactive constituents of the plant aerial part for fully use of Glycyrrhiza uralensis resources. Seven phenolic compounds including a new one (15) α,α’-dihydro-3,5,3’,4’-tetrahydroxy-2,5’-diisopentenylstilbene and six prenylated ones (9) quercetin-3-Me ether, (10) 5’-prenylquercetin, (11) 8-[(Z)-3-hydroxymethyl-2-butenyl]-eriodictyol, (12) 6-prenyleriodictyol, (13) 5’-prenyleriodictyol, (14) 6-prenylquercetin-3-Me ether were isolated and purified from the leaves of Glycyrrhiza uralensis by various chromatography methods. Compound 9-15 together with another eight compounds 16-23, which were possibly existed in the leaves of Glycyrrhiza uralensis, were selected to do the quantitative and bioassay experiments. As a result, compound 9-15 were mainly distributed in 40-80%ethanol elution parts of the Glycyrrhiza uralensis leaves extracts. Bioassay experiments showed that compounds 9-15 have strong radical scavenging activity on DPPH · and most of these compounds also demonstrated inhibitory activity on a-glucosidase and maltase. These results provided scientific evidences for the further development of Glycyrrhiza uralensis leaves or its bioactive fractions as drugs or functional foods for treating obesity and diabetes.Part 2 Bile acids (BAs), are the primary components of animal biles. Existed in the enterohepatic circulation system, BAs play an important role in fat metabolism. An ultra-high-performance liquid chromatograph-triple quadrupole mass spectrometry has been established and validated for simultaneous quantification of 15 bile acids in four traditional animal medicines and their preparations. The separation of bile acids were performed on an Agilent ZORBAX Eclipse XDB-C18 column (50 mmx2.1 mm; 1.8 μm) with methanol-0.1%formic acid as mobile phase. Glycyrrhetinic acid was added as internal standard owing to its similar physiochemical properties with the bile acids. Using this condition, detected in multiple reaction monitoring mode, the 15 bile acids, including three groups of isomers were well quantified individually. Method validation showed that the linear regression relationship (R2, 0.9993-0.9999), precisions (intra-day RSD,0.96-4.31%; inter-day, 1.73-4.43%), and recovery (95.3-120.9%) were all satisfactory. Analysis results showed that bear bile and bezoar (Niu Huang) as well as their formulations contained large amounts of most of the 15 bile acids. In addition, this research revealed for the first time the presences of bile acids in the two clinic used animal waste medication in traditional medicine, Trogopterus Dung and Hei-Bing-Pian (a traditional Mongolian medicine). The established method can be used for the quantification of other bile-or animal waste-based crude drugs and their formulated products. |
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