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To establish an HPLC method for the determination of residual solvent formic acid and cyanoacetic acid content in tofacitinib citrate.
The separation was performed on a Waters Atlantis T3(250 mm×4.6 mm, 5 μm) column with a gradient elution of 0.02 mol·L-1 potassium dihydrogen phosphate (pH was adjusted to 2.5 with phosphoric acid) (A) - methanol (B) as the mobile phases. The flow rate was 0.5 mL·min-1 and the column temperature was 35 ℃. Detection wavelength was 210 nm.
Formic acid and cyanoacetic acid were well separated from the adjacent peaks (R>5). The linearity was good in the concentration ranges of 9.930-107.780 μg·mL-1 and 14.727-98.908 μg·mL-1 (r = 0.999 9, 0.999 8), respectively. The average recoveries (n=9) were 100.2% and 105.3%, with RSDs of 2.3% and 4.1%, respectively. The determination results of three batches of samples were 0.024%, 0.026%, 0.028% (cyanoacetic acid), respectively.
The established method is proved to be suitable for the determination of formic acid and cyanoacetic acid in tofacitinib citrate.
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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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