HPLC-CAD法测定替诺福韦酯类前药ODE-TFV中ODE残留量

目的：建立测定替诺福韦十八烷氧乙基单酯衍生物（ODE-TFV）中十八烷氧乙醇（ODE）含量的高效液相色谱-电喷雾检测器（HPLC-CAD）的方法。方法：采用Agilent ZORBAX XDB-C₈柱（4.6 mm×75 mm，3.5 μm），以0.1 mol·L^-1醋酸铵缓冲液（pH 4.0）为流动相A，乙腈为流动相B，梯度洗脱，流速约为1.0 mL·min^-1，柱温40 ℃；采用电喷雾检测器（charged aerosol detector，CAD），雾化室温度30 ℃，采样频率10 Hz；进样量10 μL。结果：本法的线性范围为0.1000~10.00 μg·mL^-1（r=0.9999，n=6），最低检测限为50.00 ng·mL^-1；最低定量限为100.0 ng·mL^-1；方法平均回收率（n=9）为99.8%。结论：本文建立的方法能满足ODE-TFV原料药中ODE残留量监控的需要。

Objective: To establish the high performance liquid chromatography-charged aerosol detector (HPLC-CAD) method for the determination of octadecyloxyethyl(ODE) in the sample of octadecyloxyethyl-tenofovir(ODE-TFV). Methods: Octadecyloxyethyl-tenofovir and ODE were separated on an Agilent ZORBAX XDB-C₈ column (4.6 mm×75 mm,3.5 μm).The mobile phase was composed of 0.1 mol·L^-1 ammonium acetate buffer (pH 4.0) and acetonitrile with gradient elution mode at a flow rate of about 1.0 mL·min^-1;signals were detected with CAD;nebulization temperature was 30 ℃ and collection frequency was 10 Hz;the injection volume was 10 μL and column temperature was 40 ℃. Results: The linear range of the method was 0.1000-10.00 μg·mL^-1 (r=0.9999,n=6),and the lowest limit of detection was 50.00 ng·mL^-1;LOQ was 100.0 ng·mL^-1;the average recovery(n=9) of the method was 99.8%. Conclusion: The HPLC method established in this paper can satisfy the demand of ODE residual monitoring in ODE-TFV.

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