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To establish an HPLC-MS/MS method for simultaneous determination of 11 components（harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ）in Gengnianning.
HPLC assay was performed on C18 column（100 mm×2.1 mm，1.9 μm）with a mixture of methanol and 0.1% formic acid as the mobile phase in gradient elution at a flow rate of 0.3 mL·min-1. The column temperature was 25 ℃ and the injection volume was 1 μL. Detection was carried out on a triple quadrupole mass spectrometer in positive ion mode（harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone and harpagoside）and negative ion mode（vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ）using an electrospray ion source（ESI）. Multiple reaction monitoring（MRM）mode was employed.
The calibration curves were linear within the ranges of 1.485-29.71 μg·mL-1，1.620-32.40 μg·mL-1，7.801-156.0 μg·mL-1，0.518-10.35 μg·mL-1，0.167-3.333 μg·mL-1，0.359-7.179 μg·mL-1，1.455-29.10 μg·mL-1，1.520-30.40 μg·mL-1，0.160-3.205 μg·mL-1，0.143-2.864 μg·mL-1 and 0.157-3.136 μg·mL-1 for harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ，and atractylenolide Ⅰ，respectively. All 11 components showed good linearity（r≥0.998 0）. The average recoveries（n=6）were in the range of 95.9%-102.6% with RSDs within 0.90%-3.0%. The contents of harpagide，salidroside，nuezhenide，lobetyolin，wedelolactone，harpagoside，vaccarin，6-gingerol，atractylenolide Ⅲ，atractylenolide Ⅱ and atractylenolide Ⅰ in 10 tested samples from 5 manufactures were 14.8-104.5，37.6-288.5，335.8-1 332.8，6.2-10.1，6.6-61.8，13.7-75.1，57.4-132.8，16.9-70.6，11.8-33.9，3.4-15.4 and 6.5-12.9 μg·g-1.
The developed method is accurate and sensitive. It can be used in quality control of Gengnianning.
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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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