新型P2Y12受体拮抗剂TSD-1的有关物质研究

Study on related substances of TSD-1——a novel P2Y12 receptor antagonist

为了对TSD-1原料药中有关物质进行定量分析,本文采用核磁共振波谱仪和超高效液相-质谱联用仪对制备的杂质进行结构确证,并建立一种高效液相色谱法测定TSD-1中有关物质。采用Agilent ZORBAX Eclipe XDB-C8(250 mm×4.6 mm,5µm)色谱柱,以50 mmol·L^(-1)乙酸铵溶液(乙酸调节至pH 5.8)为流动相A,乙腈为流动相B,梯度洗脱,流速1.0 mL·min^(-1),检测波长240 nm,柱温30℃。结果表明,TSD-1与杂质A、杂质B、TSD-D和TSD-F的色谱峰均能良好分离,并分别在0.242~48.4µg·mL^(-1)(r=1.0000)、0.244~9.75µg·mL^(-1)(r=0.9999)、0.244~4.80µg·mL^(-1)(r=0.9999)、0.254~1.02µg·mL^(-1)(r=0.9999)、0.247~0.987µg·mL^(-1)(r=0.9999)范围内线性关系良好(n=7)。杂质A、杂质B、TSD-D和TSD-F的定量下限分别为0.244、0.244、0.254、0.247μg·mL^(-1);各杂质的平均回收率在99.08%~103.00%之间,准确度高。3批TSD-1原料药中TSD-D、TSD-F均未检出,杂质A、杂质B检出量均未超限。建立的HPLC方法简便、准确,适合TSD-1的有关物质测定,可为TSD-1的后续研究与开发提供有价值的参考。

For quantitative analysis of related substances in TSD-1 active pharmaceutical ingredient,struc‐tures of prepared impurities were confirmed by NMR and UHPLC-MS,and a high performance liquid chromato‐graphic method was established to determine the related substances in TSD-1.The analytical column was an Agilent ZORBAX Eclipe XDB-C8(250 mm×4.6 mm,5µm).The mobile phase A was 50 mmol·L^(-1) ammonium acetate solution(adjusted pH to 5.8 with acetic acid)and the mobile phase B was acetonitrile.The whole run was carried out by gradient elution at a flow rate of 1.0 mL·min^(-1).The detection wavelength was set at 240 nm and the column temperature was 30℃.The resolutions among peaks of TSD-1,impurity A,impurity B,TSD-D,and TSD-F were good.The calibration curves(n=7)of TSD-1,impurity A,impurity B,TSD-D and TSD-F were linear in their respective weight ranges of 0.242-48.4µg·mL^(-1)(r=1.0000),0.244-9.75µg·mL^(-1)(r=0.9999),0.244-4.80µg·mL^(-1)(r=0.9999),0.254-1.02µg·mL^(-1)(r=0.9999),and 0.247-0.987µg·mL^(-1)(r=0.9999).The lower limits of quantitation were 0.244,0.244,0.254,and 0.247µg·mL^(-1) for impurity A,impurity B,TSD-D,and TSD-F,respectively,and the average recovery of each impurity ranged from 99.08%to 103.00%with high accuracy.TSD-D and TSD-F were not detected in the three batches of TSD-1 active pharmaceutical ingredients,and impurity A and impurity B were not detected beyond the limit.The established HPLC method is simple,accurate,and suitable for determination of related substances of TSD-1,which can provide a valuable reference for the subse-quent development of TSD-1.