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To establish an ultra-high performance liquid chromatography-tandem mass spectrometry method (UPLC-MS/MS) for the determination of the contents of cholic acid, imperatorin, quercetin, isoquercetin, ferulic acid, chlorogenic acid, rutin, and cryptochlorogenic acid in the Huoxuexiaozhong capsule.
The analysis was performed by UPLC-MS/MS. Chromatographic conditions: separation was achieved on an Agilent Poroshell 120 EC-C18 column (2.1×150 mm, 1.9 μm). The mobile phase consisted of acetonitrile (A) and 0.1% formic acid in water (B) with gradient elution, at a flow rate of 0.3 mL·min-1 and a column temperature of 35 ℃. Mass spectrometry conditions: multiple reaction monitoring (MRM) with positive/negative ion switching mode was used electrospray ionization source. The ion source voltage was set at +5.5 kV and -4.5 kV, with the ion source temperature maintained at 550 ℃. The gas settings were as follows: GS1 344.74 kPa, GS2 344.74 kPa, and CUR 241.32 kPa. The specificity, standard curves and limits of detection and quantification, precision and recovery, and stability were investigated.
The analytical method demonstrated good linearities for cholic acid, quercetin, isoquercitrin, ferulic acid, chlorogenic acid, cryptochlorogenic acid, rutin and imperatorin over the concentration ranges of (0.26-25.70), (0.03-1.68), (0.03-1.51), (0.03-1.47), (0.40-10.34), (0.56-27.05), (0.03-1.37), and (2.50-759.65) μg·mL-1, respectively (R2 ≥ 0.9911). The relative standard deviations (RSD) of peak areas for reference standards and test samples were ≤ 2.44% and ≤ 2.87%, respectively. Recoveries for all components ranged from 96.54% to 107.86%, with RSD values between 0.47% and 5.41%, indicating good precision and accuracy. The stability of the method showed an RSD of ≤ 2.77%.
The UPLC-MS/MS method was established to determine the concentration of eight components in Huoxuexiaozhong capsule simultaneously. This method has high sensitivity, specificity and stability, which can be used as a reliable reference for the quality control of the capsule.
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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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