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The chemical pattern recognition technology was used to analyze the HPLC characteristic fingerprints data of Poria, screen the differential quality markers and perform quantitative analysis, so as to provide scientific basis for quality evaluation of Poria.
Welch Ultimate Plus C18 chromatographic column (250 mm×4.6 mm, 5 μm) was used. Acetonitrile (containing 0.2% tetrahydrofuran)-0.1% phosphoric acid aqueous solution with gradient elution was used as mobile phase. The fingerprint of Poria was established with detection wavelength set at 222 nm, flow rate at 1 mL·min-1, column temperature at 30 ℃, injection volume at 10 μL. The differential quality markers were identified by similarity analysis, principal component analysis (PCA), cluster analysis (CA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The quantitative research was carried out in accordance with the requirements of "Guidelines for Validation of Analytical Methods" in Chinese Pharmacopoeia.
The similarity of 15 batches of Poria was between 0.935~0.998. Eight common peaks were calibrated and 6 were identified; The samples were classified according to the origins from PAC results, which is consistent with CA result. Pachymic acid was screened as the differential quality marker of Poria by OPLS-DA. The established method for the determination of pachymic acid was stable, reliable, durable, which meets the requirements of the Pharmacopoeia.
The methods of characteristic fingerprint combined with chemical pattern recognition technology can effectively screen the differential quality marker of Poria from different regions, providing a reference for the establishment of quality standards for Poria.
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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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