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目的建立测定BLP25脂质体疫苗中免疫佐剂MPL含量的方法。方法采用HPLC-蒸发光散射检测器(ELSD)法。色谱柱为孔径300A的WP Octadecyl C18柱;采用三元梯度洗脱;Alltech 2000型ELSD检测器的漂移管温度为80℃,载气流速为2.6L.min-1。结果所建方法能将MPL与脂质体中的其他成分完全分离;在48.6～486μg.mL-1浓度范围内,MPL校正曲线(lgA～lgC)的线性相关系数(r)为0.9998;平均加样回收率为100.0%(n=3×3);连续进样、平行配制溶液、不同操作人员、不同实验室测定结果的相对标准偏差(RSD)分别为0.8%(n=6)、1.3%(n=3)、1.7%(n=2)和2.8%(n=3)。结论该HPLC-ELSD方法的专属性、线性、准确度、精密度和重现性均符合方法学要求,适宜于BLP25脂质体疫苗中MPL的含量测定。
Objective To establish a method for determining the content of MPL in BLP25 liposome vaccine. Methods HPLC-Evaporative Light Scattering Detector (ELSD) was used. The column was a WP Octadecyl C18 column with a pore size of 300 A; a ternary gradient was used for the elution; the drift tube temperature of the Alltech 2000 ELSD detector was 80 ° C and the carrier gas flow rate was 2.6 L · min -1. Results The established method could completely separate MPL from other components in liposome. The linear correlation coefficient (r) of MPL calibration curve (lgA ~ lgC) was 0.9998 in the range of 48.6 ~ 486μg.mL-1. The relative standard deviations (RSDs) of different laboratories were 0.8% (n = 6), 1.3% (n = 6) (n = 3), 1.7% (n = 2) and 2.8% (n = 3). Conclusion The specificity, linearity, accuracy, precision and reproducibility of the HPLC-ELSD method meet the requirements of the method and are suitable for the determination of the content of MPL in the BLP25 liposomal vaccine.