中文摘要：

目的 建立芬太尼在国人体外肝微粒体代谢的方法.方法 择期肝脏手术患者30例,术中取边缘正常的肝组织制备肝微粒体,建立芬太尼的人肝微粒体孵育体系,采用高效液相色谱法测定孵育液中0、5、10、15、20、30 min的芬太尼浓度,以舒芬太尼为内标,正己烷-无水乙醇进行萃取,采用Grace C18色谱柱（4.6 mm×250.0 mm,5 μm）,以乙腈-KH2PO4缓冲液为流动相,流速1.0 ml/min,检测波长205 nm,进样量20 m.采用最小二乘法进行线性回归分析.取0.6、2.4、10.0μ g/ml终浓度芬太尼的空白孵育体系样品,测定孵育体系芬太尼浓度的回收率、精密度和稳定性.计算肝微粒体中芬太尼的代谢速率.结果 芬太尼与内标舒芬太尼分离完全,不受微粒体内源性物质的干扰,保留时间分别为5.730 min和9.336 min.回归方程C=0.945 8A-0.1404,R2 =0.999 2,其中C为芬太尼浓度,A为芬太尼与内标物舒芬太尼的峰面积比值.低、中、高浓度的芬太尼孵育体系测定的回收率85％-115％,相对标准偏差＜10％,日内、日间精密度和稳定性相对标准偏差＜10％,符合生物样品测定要求,30例人肝微粒体中平均每毫克蛋白的芬太尼代谢速率（1.6±0.8） nmol/min.结论 国人体外肝微粒体代谢芬太尼的方法简便,检测灵敏度高,可用于芬太尼的体外代谢研究.

英文摘要：

Objective To establish in vitro metabolism of fentanyl by human liver microsomes in Chinese population.Methods Thirty patients undergoing elective operation on liver were enrolled in the study.Normal liver specimens were obtained during removal of liver and gall for preparation of liver microsomes （by calcium precipitation） which were used for establishment of the liver microsomal incubation system for fentanyl.Fentanyl served as the metabolic substrate in the incubation reaction.The concentration of fentanyl in the incubation medium was detected at 0,5,10,15,20 and 30 min of incubation using HPLC-UV.Sufentanil served as the interior label element.The n-hexane-ethanol absolute was used to extract the sample.The chromatographic column used in this method was Grace C18 （4.6 mm × 250.0 mm,5 μm）.The mobile phase was methyl cyanide-KH2PO4 buffer solution with the flow rate of 1.0 ml/min,detection wavelength of 205 nm and sample size of 20 μl.Linear regression analysis was performed by using the least-squares method.The specimens of the blank incubation system with the final concentration of fentanyl 0.6,2.4 and 10.0 μg/ml were obtained to determine the recovery,precision and stability.The metabolic rate of fentanyl in human hepatic microsomes was calculated.Results Fentanyl and the interior label element sufentanil were separated completely,and the retention time were 5.730 and 9.336 min,respectively.Endogenous matrix of microsomes did not interfere with the analysis.Regression equation was C =0.945 8A-0.140 4,R2 =0.999 2.C was the concentration of fentanyl,and A was the peak area ratio of fentanyl versus sufentanil.The recovery of incubation system with low,medium and high concentrations of fentanyl was 85％-115％,and relative standard deviation （RSD） was less than 10％.The RSD of intra-and inter-day precision and stability was less than 10％.The method was proved to meet the requirements of biological sample analysis.The metabolic rate of fentanyl was （1.6 ＋ 0.8） nmol/min per milligram prot