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To establish a liquid chromatography mass spectrometry (LC-MS) method for the detection of nepasaikosaponin K in Bupleurum marginatum var. stenophyllum, which could be used to quickly identify whether Bupleurum marginatum var. stenophyllum was substituted for Bupleurum or was mixed with Bupleurum. At the same time, nepasaikosaponin K was detected in 16 batches of Bupleurum marginatum var. stenophyllum and 157 batches of bupleurum decoction pieces.
Ultra-high performance liquid chromatography (UPLC) coupled with triple quadruple mass spectrometry (QqQ MS) was used to detect nepasaikosaponin K in Bupleurum and Bupleurum marginatum var. stenophyllum. Determination was carried out with the application of a Phenomenex Kinetex C18 (2.1 mm×100 mm, 1.7 μm) column at temperature of 25 ℃. The mobile phase was composed of acetonitrile(A) and water(B) with gradient elution(0-5 min 15% A→25% A; 5-0 min, 25% A→30% A; 30-35 min, 30% A→90% A; 35-36 min, 90% A→15% A; 36-40 min, 15% A) at a flow rate of 0.3 mL·min-1. The electrospray ionization(ESI-) source was performed in multiple reaction monitoring (MRM) mode of the transitions of m/z 943.5→797.3, m/z 943.5→635.3, and m/z 943.5→781.3.
The contents of nepasaikosaponin K detected in 16 batches of Bupleurum marginatum var. stenophyllum were much higher than minimum requirements. Nepasaikosaponin K was detected in 8 out of 157 batches of Bupleurum.
The established method is verified by methodology to be specific, so as to develop a supplementary test method for the components of Bupleurum and Bupleurum marginatum var. stenophyllum. to improve the quality control standard and authentic identification investigation, and to effectively control the quality of Bupleurum decoction pieces and guarantee its clinical efficacy.
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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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