目的 建立并验证人生长激素（human growth hormone，hGH）注射液中壬基酚浸出物超高效液相色谱-串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)检测方法。方法 参照人用药品注册技术要求国际协调会杂质相关指导原则，计算壬基酚安全限度值；采用UPLC-MS/MS法检测hGH注射液中壬基酚浸出物含量，并对该方法的线性、专属性、定量限、准确度、精密度、稳定性和耐用性进行验证。用建立的方法对3批hGH注射液进行检测。结果 壬基酚的安全限度值为4.5 μg/瓶。壬基酚浓度在8.2~65.5 μg/L范围内时，峰面积与浓度线性关系良好；空白溶液中未检测到壬基酚；定量限为1.2 μg/瓶；低、中、高水平加标样品的平均回收率均在70%~125%之间，相对标准偏差（relative standard deviationg, RSD）均＜15%；重复性和中间精密度RSD均＜15%；供试品溶液中壬基酚在室温下20 h内稳定；色谱条件中的流速设为0.27和0.33 mL/min时，6次检测壬基酚峰面积RSD＜10%；3批样品的检测结果远低于4.5 μg/瓶的限值。结论 成功建立了hGH注射液中壬基酚浸出物的UPLC-MS/MS检测方法，该方法的线性、专属性、定量限、准确度、精密度、稳定性和耐用性良好。

Objective To establish and validate an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of nonyl phenol leachable in human growth hormone (hGH) injection.Methods Referring to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines related to impurities, the safety limit of nonyl phenol was calculated. The UPLC-MS/MS method was used to determine the content of nonyl phenol leachate in hGH injection, and the linearity, specificity, limit of quantitation (LOQ), accuracy, precision, stability and robustness of the method were validated. The established method was used to detect 3 batches of hGH injection.Results The safety limit of nonyl phenol was 4.5 μg/vial. The method had a good linear relationship between peak area and concentration in the nonyl phenol concentration range of 8.2-65.5 μg/L.No nonyl phenol was detected in the blank solution.The LOQ was 1.2 μg/vial.The average recoveries of spiked samples at low, medium and high levels were all between 70%-125%, with relative standard deviations (RSDs) all ＜15%.The RSDs of repeatability and intermediate precision were both ＜15%.Nonyl phenol in the test solution was stable at room temperature for 20 h; when the flow rate in the chromatographic conditions was set to 0.27 and 0.33 mL/min, the RSD of nonyl phenol peak area in 6 detections was ＜10%.The detection results of 3 batches of samples were much lower than the limit of 4.5 μg/vial.Conclusion UPLC-MS/MS method for the determination of nonyl phenol leachate in hGH injection is successfully established, and the linearity, specificity, LOQ, accuracy, precision, stability and robustness of this method are good.