目的 建立测定异丙托溴铵原料药中有关物质的高效液相色谱-二极管阵列检测器-质谱（HPLC-DAD-MS）法。方法 采用HPLC-MS法，Alltima C₁₈色谱柱（150 mm×4.6 mm，5 μm）；流动相：0.01 mol/L乙酸铵-乙腈，梯度洗脱；体积流量：0.25 mL/min；柱温：35℃，进样量：10 μL；检测波长：220 nm。采用离子阱（ESI）正负离子模式进行检测，源温110℃；脱溶剂温度350℃；脱溶剂气体积流量600 L/h；锥孔气体积流量50 L/h。结果 异丙托溴铵、托品酸、托品酸乙酯和杂质F在0.01~10.00、0.012~12.000、0.083~8.300、0.05~5.00 mg/mL显示良好的线性关系；异丙托溴铵、托品酸、托品酸乙酯和杂质F的最低检测限分别为1.46、0.57、4.16、5.03 ng；异丙托溴铵、杂质C、托品酸乙酯和杂质F的定量限分别为4.38、1.71、12.48、15.09 ng。结论 该方法灵敏度高、选择性高、检测限低，为异丙托溴铵的质量控制提供依据。

Objective To establish determination of the related substances in ipratropium bromide active pharmaceutical ingredients by HPLC-DAD-MS. Methods HPLC-MS method was adopted. The determination was carried out on Alltima C₁₈ column (150 mm×4.6 mm, 5 μm),which mobile phase consisted of 0.01 mol/L ammonium acetate-acetonitrile with gradient elution. The column temperature was set at 35℃ at a flow rate of 0.25 mL/min. Injection volume was 10 μL. The detective wavelength was set at 220 nm. The ion trap (ESI) positive and negative ion mode was used as detection mode. Source temperature was set at 110℃ with desolvent temperature of 350℃. The flow rate of desolvent and cone gas were 600 and 50 L/h. Results There were good linear relationships of ipratropium bromide, tropic acid, tropic acid ethyl ester, and impurity F in the concentration ranges of 0.01-10.00, 0.012-12.000, 0.083-8.300, and 0.05-5.00 mg/mL, respectively. The limit of detection was 1.46, 0.57, 4.16, and 5.03 ng with limit of quantitation 4.38, 1.71, 12.48, and 15.09 ng, respectively. Conclusion The method has high sensitivity, high selectivity, and low detection limit which can be used in quantity control for ipratropium bromide active pharmaceutical ingredients.

国家重大新药创制专项（2015ZX093001002009）