人血浆中甲氨蝶呤的超高效液相色谱-串联质谱分析及其与荧光偏振免疫分析法的比较

UPLC-MS/MS Analysis of Methotrexate in Human Plasma and Comparison with the Fluorescence Polarization Immunoassay.

作者信息

Mei Shenghui, Zhu Leting, Li Xingang, Wang Jiaqing, Jiang Xueyun, Chen Haiyan, Huo Jiping, Yang Li, Lin Song, Zhao Zhigang

机构信息

Department of Pharmacy, Beijing Tiantan Hospital, Capital Medical University.

Department of Clinical Pharmacology, College of Pharmaceutical Sciences, Capital Medical University.

出版信息

Anal Sci. 2017;33(6):665-670. doi: 10.2116/analsci.33.665.

DOI:10.2116/analsci.33.665

PMID:28603183

Abstract

Methotrexate (MTX) plasma concentration is routinely monitored to guide the dosage regimen of rescue drugs. This study aims to develop and validate an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for plasma MTX analysis, and to establish its agreement with the fluorescence polarization immunoassay (FPIA) in patients with high-dose MTX therapy. Separation was achieved by gradient elution with methanol and water (0.05% formic acid) at 40°C with a run time of 3 min. The intra- and inter-day inaccuracy and imprecision of the UPLC-MS/MS method were -4.25 to 3.1 and less than 7.63%, respectively. The IS-normalized recovery and matrix effect were 87.05 to 92.81 and 124.43 to 134.57%. The correlation coefficients between UPLC-MS/MS and FPIA were greater than 0.98. The UPLC-MS/MS method was in agreement with the FPIA at high levels of MTX (1.0 - 100 μmol/L), but not at low levels (0.01 - 1.0 μmol/L). Further studies are warranted to confirm these results.

摘要

常规监测甲氨蝶呤（MTX）的血浆浓度以指导救援药物的给药方案。本研究旨在开发并验证一种用于血浆MTX分析的超高效液相色谱串联质谱（UPLC-MS/MS）方法，并确定其与高剂量MTX治疗患者的荧光偏振免疫分析法（FPIA）的一致性。在40°C下，采用甲醇和水（含0.05%甲酸）进行梯度洗脱，运行时间为3分钟，实现分离。UPLC-MS/MS方法的日内和日间误差分别为-4.25至3.1，不精密度分别小于7.63%。内标归一化回收率和基质效应分别为87.05至92.81和124.43至134.57%。UPLC-MS/MS与FPIA之间的相关系数大于0.98。UPLC-MS/MS方法在MTX高水平（1.0 - 100 μmol/L）时与FPIA一致，但在低水平（0.01 - 1.0 μmol/L）时不一致。需要进一步研究以证实这些结果。