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Objective: To compare the chemical composition changes in Kehuang before and after fermentation using multi-component content determination combined with chemometric analysis. Methods: HPLC was employed using a ZORBAX Eclipse Plus C18 column (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile and 0.1% phosphoric acid aqueous solution, applied with gradient elution. The flow rate was set at 1.0 mL · min-1,the column temperature was maintained at 30 ℃, the detection wavelength was 203 nm, and injection volume was 10 μL. This method was applied to analyze the chemical compositions of Kehuang before and after fermentation and to establish their corresponding chemical fingerprint profiles. Quantitative determination was conducted for 11 chemical constituents, including chlorogenic acid, cimifugin, notoginsenoside R1, baicalin, berberine, quercetin,baicalein, wogonin, emodin, chrysophanol, and physcion. Furthermore, chemometric methods such as hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were employed to distinguish and compare the samples before and after fermentation, with the aim of identifying key differential compounds. Results: Chemical fingerprint profiles were established for Kehuang before and after fermentation, with 22 common peaks identified. The fingerprints had good specificity and can be used for quality evaluation of Kehuang. The results of multi-component content determination and chemometrics analysis revealed significant differences in the content of notoginsenoside R1 (1.023-2.927 mg · g-1),scutellarein (0.125-0.568 mg · g-1), and berberine hydrochloride (2.151-3.068 mg · g-1) among the crude Kehuang powder, fermented Kehuang powder, and the content of Kehuang capsules. Notoginsenoside R1, scutellarein and berberine were identified as differential markers between pre- and post-fermentation of Kehuang, which could serve as quality indicators for distinguishing and identifying Kehuang before and after fermentation. Conclusion: The chemical composition of Kehuang changes significantly after fermentation, and the differential compounds have been clearly identified, which provides an analytical basis for quality control in the production of Kehuang capsules.
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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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