摘要
目的建立UPLC-MS/MS法对富马酸丙酚替诺福韦原料药及其片剂中基因毒性杂质9-丙烯基腺嘌呤进行定量测定。方法选用ZORBAX Eclipse Plus C_(18)色谱柱(2.1 mm×50 mm,1.8μm),以0.1%甲酸水溶液为流动相A,乙腈为流动相B,梯度洗脱〔0 min(95%A-5%B)→3 min(40%A-60%B)→3.5 min(40%A-60%B)→3.6 min(95%A-5%B)→5 min(95%A-5%B)〕,流速为0.4 mL·min^(-1),进样量为2μL。ESI正离子化MRM模式下对9-丙烯基腺嘌呤进行检测,定量离子对为176.1→136.0,定性离子对为176.1→119.0。结果检测限为0.06ppm,定量限为0.13ppm;浓度在0.061~317.14ng·mL^(-1)范围内,线性方程为y=3156.2x+21549,r=0.9971;精密度试验杂质峰面积RSD为1.2%(n=6);重复性试验富马酸丙酚替诺福韦和富马酸丙酚替诺福韦片中杂质含量RSD分别为1.9%和2.1%(n=6);稳定性试验表明供试品溶液和对照品溶液在室温下至少28 h内稳定;耐用性试验富马酸丙酚替诺福韦和富马酸丙酚替诺福韦片中杂质含量绝对差值均小于5 ppm;富马酸丙酚替诺福韦和富马酸丙酚替诺福韦片中平均回收率分别为95.5%和91.0%,RSD分别为5.4%和4.5%(n=9)。结论该法操作简便,重复性好,结果准确可靠,可用于富马酸丙酚替诺福韦原料药及其片剂中9-丙烯基腺嘌呤的测定,为其质量控制提供参考。
OBJECTIVE To establish an UPLC-MS/MS method for the quantitative determination of a genotoxic impurity of 9-propenyladenine in tenofovir alafenamide fumarate and its tablets.METHODS The determination was performed on an ZORBAX Eclipse Plus C_(18)column(2.1 mm×50 mm,1.8μm),with mobile phase A was 0.1%formic acid aqueous solution and mobile phase B was acetonitrile by gradient elution〔0 min(95%A-5%B)→3 min(40%A-60%B)→3.5 min(40%A-60%B)→3.6 min(95%A-5%B)→5 min(95%A-5%B)〕.The flow rate was 0.4 mL·min^(-1)and the injection quantity was 2μL.9-propenyladenine was detected by ESI+ionized MRM mode,the quantitative ion pair was 176.1→136.0,and the qualitative ion pair was 176.1→119.0.RESULTS The limit of detection was 0.06 ppm,and the limit of quantitation was 0.13 ppm.The calibration curves for 9-propenyladenine revealed good linearities over the ranges of 0.061-317.14ng·mL^(-1),the linear equation was y=3156.2x+21549(r=0.9971);the precision test RSD of impurity peak area was 1.2%(n=6).The repeatability test RSD of impurities in tenofovir alafenamide fumarate and its tablets was 1.9%and 2.1%,respectively(n=6).The stability test showed that the test solution and the control solution were stable at room temperature for at least 28 h.The Durability test of the absolute difference of impurity content in tenofovir alafenamide fumarate and its tablets was less than 5ppm.The recoveries(n=9)of 9-propenyladenine in tenofovir alafenamide fumarate and its tablets were 95.5%(RSD=5.4%)and 91.0%(RSD=4.5%),respectively.CONCLUSION The method is simple,reproducible,accurate and reliable,and can be used for the determination of 9-propenyladenine in enofovir alafenamide fumarate and its tablets,providing reference for its quality control.
作者
王玲兰
宿亮
章海云
朱婧
徐彬滨
朱颖熹
WANG Ling-lan;SU Liang;ZHANG Hai-yun;ZHU Jing;XU Bin-bin;ZHU Ying-xi(Zhuzhou Qianjin Pharmaceutical Co.LTD.,Zhuzhou 412000,China;Hunan Qianjin Xiangjiang Pharmaceutical Co.LTD.,Zhuzhou 412000,China)
出处
《海峡药学》
2021年第11期85-88,共4页
Strait Pharmaceutical Journal
作者简介
王玲兰,女,硕士,中级工程师,从事药物分析相关研究;通讯作者:宿亮,男,硕士,高级工程师,从事药物制剂、药物合成相关研究。
