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A method was developed for the determination of toosendanin in Toosendan Fructus by HPLC coupled with charged aerosol detector (CAD) in the present study. Solid phase extraction (SPE) was employed for sample pretreatment to remove the impurities and enrich the target compounds. The methanal extracts of Toosendan Fructus were dissolved in 5 mL of 30% methanol and loaded to a C18 SPE column, followed by eluting with 6 mL of 30% methanol and 6 mL of methanol, respectively; and the components eluted with methanol was collected and used for preparation of the sample solution. Samples were separated on an Agilent ZOBAX SB C18 column (4.6 mm × 250 mm, 5 μm) using the mixture of acetonitrile and water (33∶67) as the mobile phase, the column temperature was 30 ℃. The nitrogen inlet pressure of the CAD detector was 55 psi and the nebulizer chamber temperature was 35 ℃. As results, the HPLC-CAD spectrum of Toosendan Fructus sample was clear with little interference, and the separation of the target compound toosendanin was good. The established method was well validated, and showed high sensitivity, precision, good repeatability, and satisfactory stability. The limit of detection of toosendanin was 24.0 ng, the RSD values for precision, repeatability, and stability tests were all less than 5.0%; the recoveries of toosendanin at high, middle, and low levels were ranged from 95.9% to 99.7%. The established HPLC-CAD method was applied for quantification of toosendanin in 10 batches of Toosendan Fructus samples and compared with those of the LC-MS method. Pearson correlation analysis showed that the contents of toosendanin determined by the two methods were very similar and was highly positively correlated (the correlation coefficient was 0.995 0, P < 0.001). In conclusion, a method for determination of toosendanin in Toosendan Fructus was established by HPLC-CAD coupled with SPE, providing references for the quality control and evaluation of Toosendan Fructus.
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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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