目的: 建立替格瑞洛原料药中的基因毒性杂质盐酸羟胺的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。方法: 采用反相色谱,以水-乙腈(0.1%甲酸)为流动相,梯度洗脱,流速0.3 mL·min-1,以ESI正离子多反应监测(MRM)模式进行检测。结果: 盐酸羟胺的检测限和定量限可分别达到0.2和0.4 ng·mL-1,在线性范围内有良好的线性关系(r=0.998 7),准确度实验低、中、高浓度回收率均在95.1%~109.2%,RSD为2.2%~6.4%,重复性实验6份加标样品溶液回收率的RSD为5.7%。在3批次替格瑞洛原料药中均未检出盐酸羟胺。结论: 该方法可用于检测替格瑞洛中的盐酸羟胺。
Objective: To establish an HPLC-MS/MS detection method of hydroxylamine hydrochloride in Ticagrelor API. Methods: Hydroxylamine hydrochloride and API sample were separated by reverse chromatography, with water-acetonitrile (0.1% formic acid) as mobile phase using gradient elution at a flow rate of 0.3 mL·min-1. Mass was performed with multi-reaction monitoring(MRM)in positive ESI mode. Results: The detection and quantitative limits of hydroxylamine hydrochloride reached 0.2, 0.4 ng·mL-1, respectively, and good linear relationship(r=0.998 7) was acquired. The recoveries of low, medium and high level in the accuracy experiment ranged from 95.1% to 109.2%, with RSD of 2.2%~6.4%. The RSD of the recovery of 6 spiked sample solution was 5.7% for the repeatability experiment. No hydroxylamine hydrochloride was detected in 3 batches of Ticagrelor API. Conclusion: The method can be used to detect hydroxylamine hydrochloride in Ticagrelor.
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