液相色谱-串联质谱法测定猴血浆对乙酰氨基酚浓度及其药代动力学研究

Determination the concentration of 4-Acetamidophenol in monkey plasma and study on its pharmacokinetics by LC-MS/MS

目的 建立一种检测猴血浆中对乙酰氨基酚浓度的液相色谱-串联质谱（LC-MS/MS）方法,并将其用于该药在猴血浆中的药代动力学（药动学）研究.方法 分别给予6只猴单剂量鼻腔灌胃给药（20 mg/kg）后,在不同时间点取血,甲醇沉淀血浆蛋白.色谱柱（Phenmenex,Synergi,50.0 mm×3.0 mm,4.0 μm）,流动相A：2 mmol/L乙酸铵混于0.1%甲酸-水溶液中;流动相B：2 mmol/L乙酸铵混于95%乙腈-水溶液中,进行色谱分离,D4-对乙酰氨基酚为内标,采用电喷雾离子源正离子多反应监测测定对乙酰氨基酚血浆浓度,计算药动学参数.结果 LC-MS/MS法在50～50 000 ng/mL时具有良好的线性关系[相关系数为0.999 7];最低定量限为50 ng/mL.给药1 h后,雌雄猴的平均最大血药浓度值分别为6 700、8 900 ng/mL;血药浓度-时间曲线下面积值分别为12.1、18.8 h/（μg·mL）;平均半衰期为1.3 h.结论 建立的测定方法准确灵敏、快速简单、结果可靠,可用于对乙酰氨基酚在猴体内的药动学研究.

Objective To establish a LC-MS/MS method for the determination of 4-Acetamidophenol in monkey plasma, and to irwestigate the pharmaeokinetics of 4-Aeetamidophenol in monkey by using this method. Methods The blood samples were collected from 6 monkeys at different time point after single dose of nasal intragastric administration of 20 mg/kg 4-Acetami- dophenol, and methanol was used to deposit the plasma albumen in the blood samples. Chromatographic separation was conducted in chromatographic column Phenmenex,Synergi with the mobile phase of acetonitrile-water-formic acid-ammonium acetate （mobile phase A ： 2 mmol/L ammonium in 0.1% formic acid-water solution ; mobile phase B ： 2 mmol/L ammonium in 95% acetonitrile-water solution） and the internal standard of 4-Aeetamidophenol. The concentration of 4-Acetamidophenol in monkey plasma was deter- mined by LC-ESI-MS/MS method with positive ion MRM, and pharmacokinetic parameters of 4-Acetamidopheuol were calculated. Results Lineafity of the LC-MS/MS method was well within the range from 50 to 50 000 ng/mL（r=0.999 7） ,with the lowest limit of quantification of 50 ng/mL. 1 hour after medication, the mean maximum plasma concentration was 6 700 ng/mL for males and 8 900 ng/mL for females ; area tinder the curve of plasma concentration-time was 12.1 h- μg/mL for females and 18.8 h.μg/mL for male with the average half period of 1.3 h. Conclusion The method of LC-MS/MS is proved to be accurate, sensitive, quick, simple and reliable. It can be used for the determination of pharmacokinetics of 4-Acetamidophenol in monkey.