目的 建立一種簡(jiǎn)便、靈敏的液相串聯(lián)質(zhì)譜（LC-MS/MS）法測(cè)定人血漿中利培酮的濃度，并應(yīng)用于健康人體的藥動(dòng)學(xué)研究。方法 血漿樣品經(jīng)乙腈沉淀蛋白后，使用ZORBAX Eclipse XDB-C₁₈ （50 mm×4.6 mm，5 μm）色譜柱分離，以60%乙腈-40%甲醇、0.1%甲酸-5%乙腈-10 mmol/L乙酸銨水溶液作為流動(dòng)相，梯度洗脫；在電噴霧離子化源（ESI）正離子檢測(cè)條件下，采用多反應(yīng)離子監(jiān)測(cè)模式（MRM）對(duì)利培酮及內(nèi)標(biāo)利培酮-d4進(jìn)行定量分析，檢測(cè)離子對(duì)分別為m/z 411.3→191.2、m/z 415.3→195.2；進(jìn)行專屬性、系統(tǒng)適用性、準(zhǔn)確度、精密度、基質(zhì)效應(yīng)、提取回收率、穩(wěn)定性等方法學(xué)驗(yàn)證；8名健康成年受試者（無(wú)脫落）空腹單次口服利培酮片1 mg后，分別于給藥前0 h和給藥后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采集血樣至含有肝素鈉的抗凝管中，分離血漿樣品，進(jìn)行LC-MS/MS分析。結(jié)果 建立的LC-MS/MS法專屬性良好，系統(tǒng)適用性良好，內(nèi)標(biāo)與待測(cè)物之間不存在交叉影響，人血漿中利培酮的線性范圍為0.1～20.0 ng/mL，定量下限（LLOQ）為0.1 ng/mL，利培酮在空腹血漿、餐后血漿及溶血血漿中經(jīng)內(nèi)標(biāo)歸一化的基質(zhì)效應(yīng)分別為0.991～1.00、1.00～1.01和0.994～0.999，利培酮在人血漿中的平均提取回收率為96.8%～99.7%，準(zhǔn)確度、精密度以及穩(wěn)定性等均符合有關(guān)要求。健康受試者單次口服利培酮片1 mg后，主要藥動(dòng)學(xué)參數(shù)t_max、C_max、AUC_0~t、t_1/2分別為（0.969±0.248）h、（7.83±2.24）ng/mL、（25.5±12.0）h·ng/mL、（3.31±1.74）h。結(jié)論 建立的LC-MS/MS法前處理簡(jiǎn)便快速，靈敏度高，滿足生物分析的法規(guī)要求，可應(yīng)用于利培酮在健康人體中的藥動(dòng)學(xué)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

中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項(xiàng)目資助（2019-I2M-5-020）