目的 建立高效液相色譜法（HPLC）-二極管陣列檢測器（DAD）法測定2-氨基吡啶原料藥中的有關(guān)物質(zhì)。方法 使用Waters XBridge C₁₈色譜柱（250mm×4.6mm，5μm），流動相為10mmol/L磷酸氫二鉀（磷酸調(diào)pH值至7.5）–甲醇，梯度洗脫；檢測波長為225 nm（2,4-二氨基吡啶、2,6-二氨基吡啶）、260 nm（4-氨基吡啶）；體積流量1.0mL/min；柱溫為35℃；進樣量：10μL。結(jié)果 4-氨基吡啶、2,4-二氨基吡啶、2,6-二氨基吡啶在0.01～2.00、0.01～2.00、0.06～2.00μg/mL線性關(guān)系良好。4-氨基吡啶、2,4-二氨基吡啶、2,6-二氨基吡啶的平均回收率分別為99.6%、97.5%、102.7%，RSD值分別為4.6%、2.6%、3.5%。結(jié)論 建立的方法操作簡便、準(zhǔn)確、靈敏度高、專屬性強，可用于2-氨基吡啶原料藥的質(zhì)量控制。

Objective To establish an HPLC-DAD method for the determination of related substances in 2-aminopyridine active pharmaceutical ingredients. Methods The analysis was performed on an Waters XBridge C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase was 10 mmol/L dipotassium phosphate (pH value was adjusted to 7.5 with phosphoric acid) - methanol with gradient elution. The detection wavelength was 225 nm (2,4-diaminopyridine and 2,6-diaminopyridine) and 260 nm (4- aminopyridine), the flow rate was 1.0 mL/min, column temperature was 35 ℃, and injection volume was 10 μL. Results The calibration curves of 4-aminopyridine, 2,4-diaminopyridine, and 2,6-diaminopyridine were linear within the concentration ranges of 0.01 — 2.00, 0.01 — 2.00, and 0.06 — 2.00 μg/mL, respectively. The average recoveries for 4-aminopyridine, 2,4-diaminopyridine, and 2,6-diaminopyridine were 99.6%, 97.5%, and 102.7% with RSD values of 4.6%, 2.6%, and 3.5%, respectively. Conclusion The method is simple, accurate, sensitive, and specific, and can be used for the quality control for 2-aminopyridine active pharmaceutical ingredients.

R927.2

淮安市科技項目（HAP201901）