目的 建立一种简便、灵敏的液相串联质谱（LC-MS/MS）法测定人血浆中利培酮的浓度，并应用于健康人体的药动学研究。方法 血浆样品经乙腈沉淀蛋白后，使用ZORBAX Eclipse XDB-C₁₈ （50 mm×4.6 mm，5 μm）色谱柱分离，以60%乙腈-40%甲醇、0.1%甲酸-5%乙腈-10 mmol/L乙酸铵水溶液作为流动相，梯度洗脱；在电喷雾离子化源（ESI）正离子检测条件下，采用多反应离子监测模式（MRM）对利培酮及内标利培酮-d4进行定量分析，检测离子对分别为m/z 411.3→191.2、m/z 415.3→195.2；进行专属性、系统适用性、准确度、精密度、基质效应、提取回收率、稳定性等方法学验证；8名健康成年受试者（无脱落）空腹单次口服利培酮片1 mg后，分别于给药前0 h和给药后10、20、30、45 min，1.00、1.25、1.50、2.00、3.00、4.00、5.00、6.00、8.00、12.00、16.00、24.00、36.00、48.00 h采集血样至含有肝素钠的抗凝管中，分离血浆样品，进行LC-MS/MS分析。结果 建立的LC-MS/MS法专属性良好，系统适用性良好，内标与待测物之间不存在交叉影响，人血浆中利培酮的线性范围为0.1～20.0 ng/mL，定量下限（LLOQ）为0.1 ng/mL，利培酮在空腹血浆、餐后血浆及溶血血浆中经内标归一化的基质效应分别为0.991～1.00、1.00～1.01和0.994～0.999，利培酮在人血浆中的平均提取回收率为96.8%～99.7%，准确度、精密度以及稳定性等均符合有关要求。健康受试者单次口服利培酮片1 mg后，主要药动学参数t_max、C_max、AUC_0~t、t_1/2分别为（0.969±0.248）h、（7.83±2.24）ng/mL、（25.5±12.0）h·ng/mL、（3.31±1.74）h。结论 建立的LC-MS/MS法前处理简便快速，灵敏度高，满足生物分析的法规要求，可应用于利培酮在健康人体中的药动学研究。

objective To establish a sensitive and simple high-performance liquid chromatographic tandem mass spectrometry (LCMS/MS) method for the determination of risperidone in human plasma and utilize in a pharmacokinetic study in healthy volunteers. Methods Protein was precipitated by acetonitrile in plasma samples, and the analyte and internal standard were separated on a ZORBAX Eclipse XDB-C₁₈ column (50 mm×4.6 mm, 5 μm) with a gradient procedure using 60% acetonitrile-40% methanol as the organic phase and 0.1% formic acid -5% acetonitrile-10 mmol/L ammonium acetate formate solution as the mobile phase at flow rate of 0.5 mL/min. Electrospray ionization (ESI) and multiple reaction monitoring (MRM) detection modes were used for quantitative detection of risperidone and risperidone-D4 (internal standard). In the mode of multiple reaction monitoring ofpositiveions, the monitoring ion pairs of risperidone and risperidone-d4 were m/z 411.3→191.2 and m/z 415.3→195.2, respectively. The specificity, system applicability, accuracy, precision, matrix effect, extraction recovery, stability and other methodological validation were carried out. Eight healthy adult subjects (without shedding) were given risperidone tablets 1 mg orally on an empty stomach. Blood samples were collected into anticoagulant tubes containing heparin sodium 0 h before administration and 10, 20, 30, 45 min, 1.00, 1.25, 1.50, 2.00, 3.00, 4.00, 5.00, 6.00, 8.00, 12.00, 16.00, 24.00, 36.00, 48.00 h after administration. Plasma samples were separated and analyzed by LC-MS / MS. Results The established LC-MS/MS method had good specificity and applicability, and there was no cross effect between internal standard and analyte, The linear range of risperidone in human plasma was 0.1 ~ 20.0 ng/mL, the lower limit of quantification (LLOQ) was 0.1 ng/mL, The matrix effects of risperidone normalized by internal standard in fasting plasma, postprandial plasma and hemolytic plasma were 0.991 ~ 1.000, 1.000~1.010 and 0.994 ~ 0.999, respectively. The average extraction recovery of risperidone in human plasma was 96.8% ~ 99.7%. The accuracy, precision and stability all meet the relevant requirements. The pharmacokinetic parameters, t_max、C_max、AUC_0~t and t_1/2 were (0.969±0.248) h、 (7.83±2.24) ng/mL、 (25.5±12.0) h·ng/mL and (3.31±1.74 ) h. Conclusion This LC-MS /MS method was proven simple, sensitive, rapid and suitable for pharmacokinetics study of risperidone in the healthy human.

R969.1

中国医学科学院医学与健康科技创新工程项目资助（2019-I2M-5-020）