To establish an ultra-performance liquid chromatography-diode array detector (UPLC-DAD) method for the simultaneous determination of 5-hydroxymethylfurfural, morroniside, chlorogenic acid, loganin, paeoniflorin, 4, 5-O-dicaffeoylquinic acid, benzoylpaeoniflorin, and paeonol in Qiju Dihuang condensed pills.
Separation was performed on a Waters ACQUITY HSS T3 C18 column (2.1 mm × 100 mm, 1.8 μm) using a gradient elution program consisting of methanol, acetonitrile, and 0.3% phosphoric acid. Detection wavelengths were set at 230 nm (paeoniflorin, benzoylpaeoniflorin), 240 nm (morroniside, loganin), 274 nm (paeonol), 284 nm (5-hydroxymethylfurfural), 325 nm (chlorogenic acid), and 330 nm (4, 5-O-dicaffeoylquinic acid). The column temperature was maintained at 35 ℃, with a flow rate of 0.3 mL·min -1 and an injection volume of 1 μL.
The calibration curves for 5-hydroxymethylfurfural, morroniside, chlorogenic acid, loganin, paeoniflorin, 4, 5-O-dicaffeoylquinic acid, benzoylpaeoniflorin, and paeonol exhibited good linearity within concentration ranges of 4.69-234.53 (r = 0.999 3), 2.43-121.52 (r=0.999 8), 1.12-56.09 (r = 0.999 9), 2.81-140.33 (r = 0.999 9), 2.55-127.64 (r = 0.999 8), 1.17-58.60 (r = 0.999 8), 2.22-110.75 (r=0.999 5), and 4.93-246.25 μg·mL-1 (r = 0.999 9), respectively. The average recoveries were 99.67%, 99.30%, 97.64%, 99.88%, 99.90%, 102.60%, 99.65%, 100.59% with corresponding RSDs of 0.97%, 1.36%, 0.75%, 1.03%, 1.72%, 0.64%, 1.01% and 1.00%.
The developed method is rapid, accurate, sensitive, efficient, and reproducible, making it suitable for the determination of multi-index components, quality control and quality evaluation of Qiju Dihuang condensed pill.
| 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 |