To establish an HPLC method for simultaneous determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, forsythiaside A, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, forsythin, andrographalide and dehydroandrographalide in compound Shuanghua tablets.
The samples were extracted with 50% methanol solution under ultrasonic condition, and were performed on Agilent Zoabax SB-C18 column (250 mm×4.6 mm, 5 μm) by gradient elution of acetonitrile -0.15% phosphoric acid solution at a flow rate of 1.0 mL·min-1. The column temperature was 30 ℃, the injection volume was 10 μL, and the detection wavelength were set at 327 nm (detecting neochlorogenic acid,chlorogenic acid, cryptochlorogenic acid, forsythiaside A,3,4-O-dicaffeoylquinic acid,3,5-O-dicaffeoylquinic acid and 4,5-O-dicaffeoylquinic acid) and 226 nm (detecting forsythin, andrographalide and dehydroandrographalide).
The linear ranges of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid,forsythiaside A,3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, forsythin, andrographalide and dehydroandrographalide were 4.071-40.71 μg·mL-1(r=0.999 9), 20.16-201.6 μg·mL-1(r=0.999 9), 4.730-47.30 μg·mL-1(r=0.999 9), 4.536-45.36 μg·mL-1(r=0.999 8), 1.817-18.17 μg·mL-1(r=0.999 9), 2.266-22.66 μg·mL-1(r=0.999 7), 3.321-33.21 μg·mL-1(r=0.999 9), 3.462-34.62 μg·mL-1(r=0.999 6), 2.111-21.11 μg·mL-1(r=0.999 9) and 2.290-22.90 μg·mL-1(r=0.999 7), respectively. The average recoveries (n=6) were 101.3%(2.1%),103.0%(1.5%),100.9%(2.0%),101.1%(2.0%),98.4%(1.6%),102.2%(2.4%),98.2%(1.3%),97.8%(2.0%),99.0%(2.0%) and 96.4%(1.1%), respectively. The contents of the 10 components in 4 bacthes of Fufang Shuanghua tablets were in the range of 1.685-2.649 mg·g-1, 12.202-13.780 mg·g-1, 2.415-2.594 mg·g-1, 1.340-1.919 mg·g-1, 0.501-0.791 mg·g-1, 0.891-1.342 mg·g-1, 1.299-2.105 mg·g-1, 1.147-1.504 mg·g-1, 0.654-0.694 mg·g-1, 0.846-1.151 mg·g-1, respectively.
This simple accurate reproducible method can be used for the quality control and evaluate of compound Shuanghua tablets.
| 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 |