Tanathitiphuwarat Napatsanan, Tanudechpong Pannee, Chariyavilaskul Pajaree, Prompila Nantaporn, Tansrisawad Nat, Tubtimrattana Apinya, Wittayalertpanya Supeecha
Interdisciplinary Program in Pharmacology, Graduate School, Chulalongkorn University, Bangkok, Thailand.
Department of Pharmacology, Clinical Pharmacokinetics and Pharmacogenomics Research Unit, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
J Adv Pharm Technol Res. 2020 Oct-Dec;11(4):207-212. doi: 10.4103/japtr.JAPTR_40_20. Epub 2020 Oct 10.
A reliable method has been validated using ultra-performance liquid chromatography mass spectrometry (MS)/MS for simultaneous evaluation of human plasma concentration of mycophenolic acid (MPA) and its major metabolites both total and free form. All analytes were extracted from plasma by simple protein precipitation procedure with methanol. Samples for determination of their free form concentration require a preanalytic spin through an ultrafiltration system. The chromatographic separation was completed using Ccolumn at 0.3 ml/min with a gradient condition. Method validation was performed as the United State Food and Drug Administration guidelines for bio-analytical methods concerning precision, accuracy, linearity, selectivity, recovery, and matrix effect. Linearity was obtained over concentration of 0.05-4, 0.5-60, and 0.025-3 μg/ml for total MPA, mycophenolic acid glucuronide (MPAG) and mycophenolic acid acyl-glucuronide (AcMPAG), respectively. The linearity of the method for free form of analytes was confirmed in the range of 10-500, 125-10,000, and 0.5-300 ng/ml for MPA, MPAG, and AcMPAG, respectively. The intra- and interday accuracy ranged from 85.73%-102.01% for total form, and 87.23%-111.89% for free form, and the precisions of all analytes were lower than 15%. The mean recoveries of the analytes ranged from 85.54% to 94.76% and the matrix factor ranged from 0.88-1.06. The developed method is rapid, sensitive and convenient for pharmacokinetic study or therapeutic drug monitoring in patients after oral administration of enteric-coated mycophenolate sodium or mycophenolate mofetil.
已验证一种可靠的方法，该方法使用超高效液相色谱质谱联用仪（MS）/MS同时评估人血浆中霉酚酸（MPA）及其主要代谢物（总形式和游离形式）的浓度。所有分析物通过甲醇简单蛋白沉淀法从血浆中提取。用于测定其游离形式浓度的样品需要通过超滤系统进行分析前离心。使用C柱在0.3 ml/min的流速和梯度条件下完成色谱分离。按照美国食品药品监督管理局生物分析方法指南进行方法验证，涉及精密度、准确度、线性、选择性、回收率和基质效应。总MPA、霉酚酸葡萄糖醛酸苷（MPAG）和霉酚酸酰基葡萄糖醛酸苷（AcMPAG）的线性范围分别为0.05 - 4、0.5 - 60和0.025 - 3 μg/ml。分析物游离形式的方法线性分别在MPA、MPAG和AcMPAG的10 - 500、125 - 10,000和0.5 - 300 ng/ml范围内得到确认。总形式的日内和日间准确度范围为85.73% - 102.01%，游离形式为87.23% - 111.89%，所有分析物的精密度均低于15%。分析物的平均回收率范围为85.54%至94.76%，基质因子范围为0.88 - 1.06。所开发的方法快速、灵敏且方便用于口服肠溶霉酚酸钠或霉酚酸酯后患者的药代动力学研究或治疗药物监测。
