UPLC法测定硫酸羟氯喹原料药有关物质

Determination of related substances in hydroxychloroquine sulfate active pharmaceutical ingredient by UPLC

目的建立UPLC法有效控制硫酸羟氯喹中的有关物质。方法采用Waters ACQUITY UPLC■BEH C18(2.1 mm×150 mm,1.7μm)色谱柱,以磷酸盐缓冲液(取庚烷磺酸钠0.25 g、KH_(2)PO_(4)1.36 g,加水900 mL溶解,用三乙胺调pH值为7.0,加水稀释至1000 mL,摇匀)-甲醇(体积比90∶10)为流动相A、以磷酸盐缓冲液-甲醇(体积比15∶85)为流动相B(梯度洗脱),流速为0.25 mL/min,检测波长为254 nm,柱温为45℃,进样体积为4μL。结果硫酸羟氯喹及12个杂质均能有效分离,硫酸羟氯喹在0.05~5μg/mL质量浓度范围内与峰面积呈良好的线性关系,相关系数r为0.9999;检测限和定量限分别为0.18 ng和0.60 ng;杂质B的回收率在100.7%~104.3%范围内(RSD=1.6%),杂质C的回收率在100.1%~101.8%范围内(RSD=0.7%)。结论建立的方法能准确测定硫酸羟氯喹原料药的有关物质,专属性强、灵敏度高,为硫酸羟氯喹质量控制及生产企业改进工艺提供参考。

Objective To establish a determination method to effectively control the related substances in hydroxychloroquine sulfate by UPLC.Methods A gradient elution was performed on a Waters ACQUITY UPLC■BEH C18 column(2.1 mm×150 mm,1.7μm)with phosphate buffer solution(take sodium heptane sulfonate 0.25 g and KH_(2)PO_(4)1.36 g,dissolve in 900 mL water,adjust pH value with triethylamine to 7.0,and dilute to 1000 mL with water)-methanol(90∶10)as mobile phase A,and phosphate buffer solution-methanol(15∶85)as mobile phase B at a flow rate of 0.25 mL/min.The detection wavelength was 254 nm,the column temperature was 45℃and the injection volume was 4μL.Results In the selected chromatographic conditions,hydroxychloroquine sulfate and 12 impurities can be effectively separated.When hydroxychloroquine sulfate was in the concentration range of 0.05-5μg/mL,there was a good linear relationship between peak area and concentration,and the correlation coefficient r is 0.9999.The limit of detection was 0.18 ng,and the limit of quantitation was 0.60 ng.The recovery of impurity B and impurity C was 100.7%-104.3%(RSD=1.6%)and 100.1%-101.8%(RSD=0.7%),respectively.Conclusion The established method can determine impurities in hydroxychloroquine sulfate active pharmaceutical ingredient(API)with high specificity and accuracy,which can provide a reference for quality control and process improvement of hydroxychloroquine sulfate.