ELSD和CAD测定多烯磷脂酰胆碱注射液中磷脂酰胆碱含量的方法比较

目的：建立HPLC-电喷雾检测器（CAD）检测多烯磷脂酰胆碱注射液中磷脂酰胆碱含量的方法，同时对蒸发光检测器（ELSD）和CAD测定磷脂酰胆碱含量的方法进行比较。方法：采用Luna silica色谱柱（250 mm×4.6 mm，5 μm），流动相A为甲醇-水-冰醋酸-三乙胺（85：15：0.45：0.05），流动相B为正己烷-异丙醇-流动相A（20：48：32），梯度洗脱，流速1.0 mL·min^-1，柱温40 ℃。ELSD漂移管温度80 ℃，载气流速2.0 L·min^-1；CAD雾化温度30 ℃，气压0.24 MPa。结果：2种检测方法在选定条件下，磷脂酰胆碱与样品中其他组分分离良好，ELSD测定磷脂酰胆碱检测限为0.75 μg，在7.5~75.7 μg的线性范围内峰面积的对数与质量浓度的对数呈良好的线性关系（r=0.9955），平均回收率（n=9）为100.3%；CAD测定的检测限为0.18 μg，在1.5~75.7 μg的线性范围内峰面积与质量浓度呈良好的线性关系（r=0.9994），平均回收率（n=9）为100.8%。结论：HPLC-CAD高效、准确，较ELSD线性关系更好，为多烯磷脂酰胆碱注射液提供新的质量控制方法。

Objective: To establish an HPLC-CAD method for the determination of phosphatidylcholine content in polyene phosphatidycholine injection,and to compare HPLC-ELSD and HPLC-CAD methods for the determination of phosphatidylcholine in polyene phosphatidycholine injection. Methods: The Luna silica column(250 mm×4.6 mm,5 μm)was used.The mobile phase for gradient elution consisted of two eluents: solvent A was methanol-water-acetic acid-triethylamine(85:15:0.45:0.05)and solvent B was n-hexane-2-propanol-solvent A(20:48:32).The flow rate was 1.0 mL·min^-1and the column temperature was 40 ℃.The detector was evaporative light-scattering detector(ELSD)with the temperature of drift tube for measurement at 80 ℃,the flow rate of carrier gas 2.0 L·min^-1.The nebulization temperature for the charged aerosol detector(CAD)was maintained at 30 ℃ and the gas pressure was 0.24 MPa. Results: Under the described condition,phosphatidylcholine was completely separated from other components in polyene phosphatidycholine injection.ELSD determination showed that the LOD of phosphatidylcholine was 0.75 μg,the linear range was 7.5-75 μg(r=0.9955),and the average recovery(n=9)was 100.3%.In comparison,CAD determination showed that the LOD of phosphatidylcholine was 0.18 μg,the linear range was 1.5-75.7 μg(r=0.9994),and the average recovery(n=9)was 100.8%. Conclusion: HPLC-CAD,a new method for phosphatidylcholine analysis,provides high sensitivity and reproducibility.Compared with the HPLC-ELSD method,the CAD method has better linearity,and provides a method for quality control of this kind of injection.