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To illustrate the overall differential material basis of three kinds of Ge-Gen Decoction (GGD) granules including Ge-Gen Decoction compound granules (CG), formula granules (mixed, FGm; decocted, FGd) and Kampo granules (KG) using UHPLC-Q-TOF-MS combining with multivariate statistical methods.
AcclaimTM RSLC120 C18 chromatographic column (100 mm×2.1 mm, 2.2 μm) was employed to separate chemical compositions and gradient elution was performed with 0.1% formic acid-acetonitrile as mobile phase. ESI ion source was used to collect data in positive and negative ion modes respectively. The data were imported into SIMCA-P software for principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Student's t-text was utilized to measure the significance values of each metabolite. The m/z of differential components were filtered according to P<0.05, VIP>3. Then, HMDB, PubChem, TCMSP, MassBank database, MS/MS ion fragments, and related references were utilized to analyze qualitatively the differential components. Finally, combining with semi-quantitative data, a comprehensive characterization for the differential components of the three GGD granules was described.
There were 40 differential components between CG and KG, and the contents of most chemical components were higher in CG at the same dose. A total of 49 differential components were identified between CG and FGm. Some of the main active components such as puerarin and ephedrine had higher contents in CG. There were 45 differential components between FGm and FGd and there was no new component detected. The contents of most components increased after decocting. However, some important components in Paeoniae Radix Alba formula granules decreased. Fifty-nine differential components were detected totally in three kinds of Ge-Gen Decoction, and they mainly varied in the relative content, not the types. Puerariae Radix had the largest number of differential components.
UHPLC-Q-TOF-MS method can rapidly identify different components of three Ge-Gen Decoction granules, providing data support for the preparation process of Ge-Gen Decoction granules and laying a foundation for the quality control and clinical application of formula granules.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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