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To establish the HPLC fingerprint and multi-component quantitative analysis of Hanchuang Zupa granules, and evaluate the quality of multiple batches of Hanchuang Zupa granules by chemical pattern recognition technology.
The sample pretreatment conditions and chromatographic analysis conditions of Hanchuang Zupa granules were optimized, and the optimal HPLC fingerprint and multi-component quantitative analysis method were established as follows: stationary phase was YMC-Pack ODS-A column (250 mm×4.6 mm, 5 μm, 12 nm) was adopted, and the mobile phase was acetonitrile-water (containing 0.1% phosphoric acid) with gradient elution, the detection wavelength was 220 nm, the column temperature was 30 ℃, the flow rate was 1.0 mL·min-1. Hierarchical cluster analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were applied to evaluate the quality of 17 batches of Hanchuang Zupa granules.
Methodological investigation of HPLC fingerprint and content determination were well verified and met the analysis requirements. A total of 25 common peaks were obtained by full peak matching, and eight of them were identified by comparing with the retention time of mixed reference substances. The similarities of 17 batches of samples were above 0.90, which showed good consistency and stability between the samples. Seventeen samples could be classified into three clusters. Three principal components from 21 common peaks were extracted by PCA. Six quality differential compounds were presented in the fingerprints by OPLS-DA, including rutoside, gallic acid, ammonium glycyrrhizinate, chlorogenic acid and so on. The resolution and linear relationship of eight components in quantitative analysis were good. The average recovery rates were 98.0%-99.1% with RSD≤2.0%.
In this study, the qualitative analysis of HPLC fingerprint and quantitative analysis of multiple index components is specific, simple and accurate, which can provide a reference for the quality control and quality evaluation of Hanchuang Zupa 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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