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OBJECTIVE To establish the HPLC fingerprints and content determination method of five index components of Dashanzha pills(DSZP), and evaluate the quality combined with chemical pattern recognition. METHODS The analysis was performed on an Eclipse plus C18 column (4.6 mm×250 mm, 5 μm) at 30 ℃. Acetonitrile-0.1% formic acid aqueous solution was used as the mobile phase for gradient elution at flow rate of 1.0 mL·min-1. The detection wavelength was set at 250 nm, and the injection volume was 10 μL. Based on 51 batches of DSZP samples, the fingerprint was established, and five index components were identified and quantitatively determined.Cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to comprehensively analyze 51 batches of samples. RESULTS The fingerprints of DSZP were established, sixteen peaks were identified, and five chemical components were identified. They were chlorogenic acid (peak 5), vanillic acid (peak 6), hyperoside (peak 7), isochlorogenic acid B (peak 9), and isochlorogenic acid A (peak 10).The linear relationship of the five index components was good (r>0.998), and the recovery rate was 96.27%-109.35%, and the RSD was 2.83%-4.07%. The contents of chlorogenic acid, vanillicacid, hyperoside, isochlorogenic acid B, and isochlorogenic acid A were 0.047 2-0.227 0, 0.009 7-0.036 2, 0.023 0-0.370 2, 0.006 4-0.063 2 and 0.021 7-0.064 5 mg·g-1, respectively. Except for manufacturer Q2, the contents of the products from the other manufacturers samples were maintained at (100±30)% of their respective average values, and the contents of the five components of manufacturer Q2 were mostly higher than 30% of the average value of all samples, indicating that there were great differences in product quality among different enterprises. The results of chemical pattern recognition showed that all samples could be roughly divided into two categories, indicating that the preparation process was relatively stable within the enterprise, but there were some differences between enterprises. The difference markers of the screened samples included the target components chlorogenic acid, hyperoside, and isochlorogenic acid B. CONCLUSION The quantitative method is accurate, rapid and feasible. Combined with chemical pattern recognition method, it can be used to evaluate the quality of DSZP.
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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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