目的 建立并验证检测佐剂中QS-21含量的高效液相色谱法。方法 采用氯仿与异丙醇（体积比3∶7）对佐剂进行破乳，破乳比例为1∶1。采用C₁₈色谱柱，以体积分数0.05%三氟乙酸水溶液和0.05%三氟乙酸乙腈-水溶液梯度洗脱，流速为1.0 mL/min，进样量为10 μL，柱温为25 ℃，检测波长为210 nm。对方法进行系统适应性、线性和范围、准确度、检测限和定量限、专属性验证。结果 方法的系统适应性较好，3次实验QS-21色谱峰的保留时间和峰面积相对标准偏差均小于2.0%；标准曲线的线性良好，决定系数在0.999 6到0.999 8之间。在3个加标浓度（12.5、25.0和37.5 μg/mL）下，QS-21加标回收率在96.0%~114.4%之间，相对标准偏差为5.5%；检测限为1 μg/mL，定量限为3 μg/mL；对照溶液无QS-21峰。结论 建立的QS-21含量测定方法操作简便、线性良好、准确度高、专属性强，可有效检测佐剂中QS-21含量，保证佐剂相关质量控制。

Objective To develop and validate a high performance liquid chromatography (HPLC) method for the detection of QS-21 content in adjuvants.Methods Adjuvant was demulsified by chloroform and isopropanol mixture(3∶7, volume ratio) with a demulsification ratio of 1∶1. Chromatographic separation was performed on a C₁₈ column with gradient elution using aqueous solution containing 0.05%(volume ratio) trifluoroacetic acid (TFA) in water and 0.05% TFA in acetonitrile-water. The flow rate was 1.0 mL/min, injection volume was 10 μL, column temperature was 25 ℃, and detection wavelength was set at 210 nm. The system adaptability, linearity and range, accuracy, limits of detection and quantification, and specificity of established method were verified.Results This method demonstrated good system suitability. The relative standard deviations (RSDs) of both retention time and peak area of QS-21 chromatographic peak from 3 repeated tests were all less than 2.0%. The standard curve exhibited excellent linearity, with coefficients of determination ranging from 0.999 6 to 0.999 8. At 3 spiked concentrations levels (12.5, 25.0 and 37.5 μg/mL), the spiked recovery rates of QS-21 were between 96.0% and 114.4% with a RSD of 5.5%. The limit of detection was 1 μg/mL and the limit of quantification was 3 μg/mL.No QS-21 peak was observed in control solution. Conclusion The established QS-21 content detection method features simple operation, good linearity, high accuracy, and strong specificity, and can effectively detect the QS-21 content in adjuvants to support quality control of adjuvants.