HPLC-CAD法测定帕拉米韦注射液的有关物质及杂质谱解析  

Determination of related substances and impurity profile in peramivir injection by HPLC-CAD

作　　者：房思萌 段玺玉 隋玉荣 鲁鑫 韩晓捷 白海娇 FANG Si-meng;DUAN Xi-yu;SUI Yu-rong;LU Xin;HAN Xiao-jie;BAI Hai-jiao(Tianjin Institute for Drug Control,Tianjin 300070,China)

机构地区：[1]天津市药品检验研究院,天津300070

出　　处：《药物分析杂志》2024年第12期2105-2113,共9页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立HPLC-CAD法测定帕拉米韦注射液的有关物质,并对降解杂质进行杂质谱解析。方法:采用Welch Ultimate Polar RP柱(250 mm×4.6 mm,3.5μm),以5 mmol·L^(-1)甲酸铵溶液(用甲酸调pH至4.6)-乙腈(B)(95∶5)为流动相A,流动相A-乙腈(50∶50)为流动相B,梯度洗脱,体积流量为0.8 mL·min^(-1),柱温30℃,CAD雾化器温度50℃,采集频率5 Hz,滤光片3.6 s。通过LC-MS/MS,推测帕拉米韦的降解途径及杂质结构。结果:帕拉米韦与各杂质之间分离度良好,帕拉米韦与7个杂质的质量浓度约在0.2~12μg·mL^(-1)与峰面积呈良好的线性关系,定量限为4.32~12.8 ng,检测限为2.16~6.44 ng;杂质Ⅲ的平均回收率(n=9)为96.2%,RSD为1.8%;对照溶液和供试品溶液在室温条件下放置102 h内稳定;微调液相色谱参数后,对有关物质的检测结果无影响。根据LC-MS/MS,推测了降解杂质的结构及可能的降解途径。结论:该方法用于测定帕拉米韦注射液的有关物质准确、可靠,专属性强,为建立该品种质量标准提供研究基础。Objective:To establish an HPLC-CAD method for the determination of related substances in peramivir injection and to analyze the impurity profile.Methods:Welch Ultimate Polar RP column(250 mm×4.6 mm,3.5μm)was used with 5 mmol·L^(-1)ammonium formate solution(pH adjusted to 4.6 with formic acid)-acetonitrile(95∶5)as mobile phase A and mobile phase A-acetonitrile(50∶50)as mobile phase B under gradient elution at the flow rate of 0.8 mL·min^(-1).Column temperature was 30℃.CAD atomizer temperature was 50℃,acquisition frequency was 5 Hz and filter was 3.6 s.The degradation pathway and impurity structure of peramivir were estimated by LC-MS/MS.Results:The separation between peramivir and the impurities was good.The mass concentration of peramivir and 7 impurities showed good linear relationships with the peak area in the ranges of 0.2-12μg·mL^(-1).The limits of quantitation were 4.32-12.8 ng,and the limits of detection were 2.16-6.44 ng.The average recovery(n=9)of impurityⅢwas 96.2%,RSD was 1.8%.The control solution and the test product solution were stable for 102 hours at room temperature.After fine-tuning the parameters of liquid chromatography,there was no influence on the detection results of related substances.Based on LC-MS/MS,the structure and possible degradation pathway of the degraded related substances were deduced.Conclusion:This method is accurate,reliable and specific for the determination of related substances in peramivir injection,and provides a basis for the establishment of quality standards for this variety.

关 键 词：帕拉米韦注射液 高效液相色谱 电喷雾检测器 杂质谱解析 有关物质 

分 类 号：R917[医药卫生—药物分析学]