Analytical validation and pilot clinical application of a UPLC-MS/MS method for determining intracellular mycophenolic acid and metabolites in kidney transplant recipients

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-19 DOI:10.1016/j.jpba.2025.116748

Xiaomei Chen , Xinhua Dai , Huan Xu , Chunxia Chen , Xueqaio Wang , Yuangao Zou , Hanjing Liu , Yunying Shi , Yi Li , Yangjuan Bai

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引用次数: 0

Abstract

There is no consensus on the strategy for therapeutic drug monitoring of the immunosuppressive drug mycophenolic acid (MPA) in organ transplant recipients. The present study proposes the utilization of ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for determining the concentrations of MPA and its metabolites: 7-O-mycophenolic acid glucuronide (MPAG) and acyl mycophenolic acid glucoside (AcMPAG) in peripheral blood mononuclear cells (PBMCs). We aimed to assess the potential application of monitoring MPA and its metabolite concentrations in PBMCs in the infection after transplantation in Chinese kidney transplant recipients (KTRs). The UPLC-MS/MS method we developed demonstrated good linearity in the quantitative ranges of 0.05–50.00 ng/mL for MPA, 0.50–50.00 ng/mL for MPAG, and 0.10–20.00 ng/mL for AcMPAG. AcMPAG in PBMCs was unstable, degrading significantly after 48 h of storage at −80°C or after 3 freeze-thaw cycles. MPA and MPAG concentrations in KTRs' PBMCs exhibited high inter-individual variability, and the MPA concentration in PBMCs was poorly correlated with that in plasma (rs = 0.206, p = 0.117). Compared with the stable group, the infected group had significantly higher MPA concentration in PBMCs at 2 and 4 h post-dosing and in plasma at 4 h post-dosing (p < 0.05). The receiver operating characteristic (ROC) analysis for post-transplantation infection revealed that PBMCs MPA-C4 and PBMCs-MPA-C2 possessed much better diagnostic efficiency than Plasma-MPA-C4. This method is easy-to-use and reliable, making it a promising clinical quantitative tool for MPA, MPAG, and AcMPAG in PBMCs. PBMC-MPA monitoring may be a potential biomarker for infection monitoring for KTRs.

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器官移植受者体内免疫抑制剂霉酚酸（MPA）的治疗药物监测策略尚未达成共识。本研究提出利用超高效液相色谱-串联质谱法（UPLC-MS/MS）测定外周血单核细胞（PBMCs）中 MPA 及其代谢物：7-O-霉酚酸葡萄糖醛酸苷（MPAG）和酰基霉酚酸葡萄糖苷（AcMPAG）的浓度。我们旨在评估监测 MPA 及其代谢物在中国肾移植受者（KTR）移植后感染的 PBMCs 中浓度的潜在应用。我们开发的 UPLC-MS/MS 方法在 MPA 0.05-50.00 纳克/毫升、MPAG 0.50-50.00 纳克/毫升和 AcMPAG 0.10-20.00 纳克/毫升的定量范围内表现出良好的线性。PBMC 中的 AcMPAG 不稳定，在-80°C 储存 48 小时或冻融 3 次后降解明显。KTR PBMCs 中的 MPA 和 MPAG 浓度在个体间存在很大差异，且 PBMCs 中的 MPA 浓度与血浆中的 MPA 浓度相关性很低（rs = 0.206，p = 0.117）。与稳定组相比，感染组用药后 2 和 4 小时 PBMC 中的 MPA 浓度以及用药后 4 小时血浆中的 MPA 浓度明显更高（p < 0.05）。对移植后感染的接收者操作特征（ROC）分析表明，PBMCs MPA-C4和PBMCs-MPA-C2的诊断效率远高于血浆MPA-C4。该方法操作简便、结果可靠，有望成为临床定量检测 PBMC 中 MPA、MPAG 和 AcMPAG 的工具。PBMC-MPA监测可能是监测KTR感染的潜在生物标志物。

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来源期刊

Journal of pharmaceutical and biomedical analysis 医学-分析化学

CiteScore

6.70

自引率

5.90%

发文量

588

审稿时长

37 days

期刊介绍： This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.