HPLC-MS/MS法测定人血浆及尿液中贝他斯汀浓度及药动学研究

目的: 建立液相色谱-串联质谱法(HPLC-MS/MS法)测定人血浆及尿液中贝他斯汀的药物浓度,并用于苯磺酸贝他斯汀片临床药动学研究.方法: 血浆/尿液经固相萃取后,采用HPLC-MS/MS测定.色谱柱为Atlantis^TM HILIC Silica色谱柱(2.1 mm×150 mm,3 μm),以乙腈-水-200 mmol·L^-1甲酸铵溶液(pH 3.0)(75:20:5)为流动相,流速0.3 mL·min^-1,柱温30 ℃;质谱采用ESI⁺,多重反应监测(MRM)模式;选择检测离子反应对为m/z 388.8→201.7(贝他斯汀),m/z 299.9→226.7(甲氧氯普胺,内标).结果: 人血浆中贝他斯汀的线性范围为1~400 ng·mL^-1,定量下限为1 ng·mL^-1.批内、批间精密度分别小于7.9%和10.4%,专属性良好.在临床药动学研究中,应用此法检测了12名健康受试者口服5 mg苯磺酸贝他斯汀片后血浆及尿液中贝他斯汀的浓度.结论: 本方法简便、准确,专属性强,可满足苯磺酸贝他斯汀片的临床药动学研究.

Objective: To establish a sensitive and specific liquid chromatograph-tandem mass spectrometric(HPLC-MS/MS)method for the determination of bepotastine in human plasma and urine. Methods: The sample was prepared by solid phase extraction.The Atlantis^TM HILIC Silica column(2.1 mm×150 mm, 3 μm)was used with the mobile phase of acetonitrile-water-200 mmol·L^-1 ammonium formate(pH 3.0)(75:20:5)at the flow rate of 0.3 mL·min^-1. The column temperature was 30 ℃. The detection was performed by electrospray ionization mass spectrometry in the multiple reaction monitoring(MRM)mode.The positive ion scan mode was used to detect bepotastine and metoclopramide(internal standard, IS).The MRM transitions of m/z 388.8→201.7 and 299.9→226.7 were used to quantify bepotastine and IS, respectively. Results: The assay was linear in the concentration range of 1-400 ng·mL^-1 for bepotastine in human plasma. The limit of quantification was 1 ng·mL^-1.The intra-day and inter-day RSDs in plasma were lower than 7.9% and 10.4%, respectively.The method was successfully applied to a pharmacokinetic study of detection of bepotastine in human plasma and urine of healthy volunteers after oral administration of bepotastine besilate tablets(5 mg). Conclusion: The HPLC-MS/MS method is simple, accurate and selective for the clinical pharmacokinetic study of bepotastine besilate tablets.

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