目的: 建立HPLC法测定枸橼酸托法替布中残留溶剂甲酸和氰乙酸的含量。方法: 采用Waters Atlantis T3（250 mm×4.6 mm,5 µm）色谱柱,以0.02 mol · L-1磷酸二氢钾溶液（用磷酸调pH 2.5）-甲醇为流动相,梯度洗脱,流速0.5 mL · min-1,柱温35 ℃,检测波长210 nm。结果: 甲酸、氰乙酸与相邻峰分离度良好（R>5）;分别在9.930~107.780 µg · mL-1、14.727~98.908 µg · mL-1浓度内线性关系良好（r = 0.999 9、0.999 8）;平均回收率（n=9）分别为100.2%、105.3%,RSD分别为2.3%、4.1%。稳定性考察（18个月）3批枸橼酸托法替布中含氰乙酸0.024%、0.026%、0.028%。结论: 经方法学验证,本法可用于枸橼酸托法替布中甲酸和氰乙酸的含量测定。
Objective: To establish an HPLC method for the determination of residual solvent formic acid and cyanoacetic acid content in tofacitinib citrate. Methods: 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. Results: 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. Conclusion: The established method is proved to be suitable for the determination of formic acid and cyanoacetic acid in tofacitinib citrate.
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