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To establish an HPLC fingerprint of Zuoyu’an San and conduct quality control by combining chemometrics and quantitative analysis of multi-components with a single-marker（QAMS）.
HPLC was used to establish the fingerprint spectra of 12 batches of Zuoyu’an San and perform similarity evaluation. Cluster analysis (CA)，principal component analysis (PCA)，and orthogonal partial least squares discriminant analysis (OPLS-DA) were combined to perform chemical pattern recognition analysis on different batches of Zuoyu’an San. At the same time，QAMS method was established using rhein as the internal standard reference. The relative correction factors between the internal reference and jatrorrhizine，palmatine，berberine，aloe-emodin，emodin，chrysophanol，and physcion were established. The external standard method (ESM) and QAMS method were used to determine jatrorrhizine，palmatine，berberine，aloe-emodin，rhein，emodin，chrysophanol，and physcion in Zuoyu’an San，respectively. To verify the reliability of QAMS metod results，the contents of 8 components of physcion were determined.
A total of 23 common peaks were calibrated in the fingerprint，and 8 components were identified by comparing with the mixed reference solution. The similarity between the samples and the control spectrum was between 0.952 and 0.994；CA divided 12 batches of Zuoyu’an San samples into 3 categories；PCA showed that there were certain differences in the chemical composition of different batches of Zuoyu’an San，and extracted 5 principal components that affected the quality evaluation of Zuoyu’an San；OPLS-DA screened out 13 potential marker components (VIP>1) among 23 common peaks that caused quality differences between different batches of Zuoyu’an San samples；There was no significant difference between the QAMS method calculated value and the ESM measured value.
The established HPLC fingerprint and multi-index component determination method was accurate and simple，and combined with chemometric methods，could provide reference for the quality control and evaluation of Zuoyu’an San.
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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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