Rivera Vélez Sol-Maiam, Morassi Alice, Court Michael H, Slovak Jennifer E, Villarino Nicolas F
Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
J Pharm Biomed Anal. 2016 Nov 30;131:94-102. doi: 10.1016/j.jpba.2016.08.012. Epub 2016 Aug 12.
Mycophenolic acid (MPA) is the active metabolite of the prodrug mycophenolate mofetil. In this study, we developed and validated a novel ultra-high performance liquid chromatography (UHPLC) method for the rapid quantification of MPA in plasma from dogs, cats and humans. Following the protein precipitation, calibration standards and quality controls were separated by UHPLC reversed-phase on a 1.5μm 2.1×100mmC column and quantified using UV detection at 215nm. The procedure produced a linear curve (r>0.997) over the concentration range 0.4-50μg/mL and exhibited a high degree of repeatability (CV% <11%). The limit of detection (LOD) and lower limit of quantitation (LLOQ) were 0.1 and ≤0.4μg/mL, respectively and the overall recovery was ≥87%. By combining isocratic conditions with a UHPLC column containing solid core particles, we were able to elute MPA and the internal standard (mycophenolic acid carboxybutoxy ether) within 3.0min. The short total run time makes this method ideal to study the disposition of MPA in large batches of plasma samples and/or monitor plasma drug concentrations, as recommended for patients that require optimized immunosuppression.
霉酚酸（MPA）是前体药物霉酚酸酯的活性代谢产物。在本研究中，我们开发并验证了一种新型超高效液相色谱（UHPLC）方法，用于快速定量犬、猫和人类血浆中的MPA。蛋白质沉淀后，校准标准品和质量控制品通过UHPLC反相在1.5μm 2.1×100mm C柱上分离，并使用215nm的紫外检测进行定量。该方法在0.4 - 50μg/mL的浓度范围内产生线性曲线（r>0.997），并且具有高度的重复性（CV%<11%）。检测限（LOD）和定量下限（LLOQ）分别为0.1和≤0.4μg/mL，总回收率≥87%。通过将等度条件与含有实心颗粒的UHPLC柱相结合，我们能够在3.0分钟内洗脱MPA和内标（霉酚酸羧丁氧基醚）。较短的总运行时间使该方法非常适合研究大量血浆样品中MPA的处置情况和/或监测血浆药物浓度，这对于需要优化免疫抑制的患者是推荐的。
