UPLC-MS/MS与化学发光微粒子免疫分析法测定甲氨蝶呤血药浓度的比较分析

doi:10.5246/jcps.2025.05.032

中国药学(英文版) ›› 2025, Vol. 34 ›› Issue (5): 423-431.DOI: 10.5246/jcps.2025.05.032

UPLC-MS/MS与化学发光微粒子免疫分析法测定甲氨蝶呤血药浓度的比较分析

时正媛¹^,²^,^*(), 李京峰¹^,², 杨春静¹^,², 续茜桥¹^,², 姜德春¹^,²

^1. 首都医科大学附属北京世纪坛医院药学部, 北京 100038
^2. 临床合理用药生物特征谱学评价北京市重点实验室, 北京 100038

收稿日期:2024-11-24 修回日期:2025-01-20 接受日期:2025-02-07 出版日期:2025-06-02 发布日期:2025-06-01
通讯作者: 时正媛

Comparative analysis of UPLC-MS/MS and chemiluminescence microparticle immunoassay for serum methotrexate measurement in pediatric intracranial tumor patients

Zhengyuan Shi¹^,²^,^*(), Jingfeng Li¹^,², Chunjing Yang¹^,², Xiqiao Xu¹^,², Dechun Jiang¹^,²

¹ Department of Pharmacy, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China
² Beijing Key Laboratory of Bio-Characteristic Profling for Evaluation of Clinical Rational Drug Use, Beijing 100038, China

Received:2024-11-24 Revised:2025-01-20 Accepted:2025-02-07 Online:2025-06-02 Published:2025-06-01
Contact: Zhengyuan Shi
Supported by:
The Science and Technology Fund of Beijing Shijitan Hospital (Grant No. 2022-C06), the National Key Research and Development Program of China (Grant No. 2020YFF01014606).

摘要/Abstract

摘要：

本研究旨在建立一种基于超高效液相色谱-串联质谱(UPLC-MS/MS)的高准确度的检测方法, 用于测定血清中甲氨蝶呤(MTX)的浓度, 并将该方法与化学发光微粒子免疫分析法(CMIA)进行比较。收集接受大剂量MTX治疗的211例颅内肿瘤患儿血清样本, 分别用UPLC-MS/MS和CMIA法进行测定, 对测定结果进行相关性分析和一致性分析。结果表明, 两种方法测得的MTX血清浓度在低浓度区间内(0.04–1.5 μmol/L)有统计学差异, 而在高浓度区间内(1.5–400 μmol/L)无统计学差异, 在以上两个浓度区间, 两组之间均有较好的相关性(P < 0.05), 相关系数分别为0.9913和0.9893。Bland-Altman分析结果表明, 在0.04–1.5 μmol/L的浓度范围内(107例), 两种方法的偏差为0.018。研究表明, 两种方法测定的甲氨蝶呤血清浓度不能直接替代, 可通过方程式进行结果互算, 为临床个体化用药方案的制订提供参考。

关键词: 甲氨蝶呤, 治疗药物监测, 超高效液相色谱-串联质谱法, 化学发光微粒子免疫分析法

Abstract:

This study aimed to establish a highly accurate method for quantifying methotrexate (MTX) concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system (UPLC-MS/MS) and to compare its performance with the chemiluminescence microparticle immunoassay (CMIA). A total of 211 serum samples from pediatric patients with intracranial tumors undergoing high-dose MTX treatment were analyzed using both techniques. Correlation and agreement analyses were performed to assess the level of concordance between these methods. The results demonstrated a significant correlation (P < 0.05) between the two methods, with correlation coefficients of 0.9913 and 0.9893, respectively. However, a statistical difference was noted in MTX serum concentrations at lower levels (0.04–1.5 μmol/L), while no significant difference was observed at higher concentrations (1.5–400 μmol/L). Specifically, in the 0.04–1.5 μmol/L range (107 cases), Bland-Altman analysis indicated a bias of 0.018 between the two methods. Given the observed discrepancies, particularly at lower concentrations, it is advised that the UPLC-MS/MS method should not be used interchangeably with the CMIA assay for therapeutic drug monitoring in patients receiving high-dose MTX treatment.

Key words: Methotrexate, Therapeutic drug monitoring, UPLC-MS/MS, Chemiluminescence microparticle immunoassay

Supporting:

时正媛, 李京峰, 杨春静, 续茜桥, 姜德春. UPLC-MS/MS与化学发光微粒子免疫分析法测定甲氨蝶呤血药浓度的比较分析[J]. 中国药学(英文版), 2025, 34(5): 423-431.

Zhengyuan Shi, Jingfeng Li, Chunjing Yang, Xiqiao Xu, Dechun Jiang. Comparative analysis of UPLC-MS/MS and chemiluminescence microparticle immunoassay for serum methotrexate measurement in pediatric intracranial tumor patients[J]. Journal of Chinese Pharmaceutical Sciences, 2025, 34(5): 423-431.

图/表 4

Figure 1. Typical chromatograms of MTX and MTX-D3 as IS determined by UPLC-MS/MS method (A. blank serum; B. blank serum spiked with IS; C. LLOQ; D. patient sample).

Table 1. Accuracy and precision of MTX in human serum determined by UPLC-MS / MS method (n = 5).

Table 2. Recovery and matrix effect of MTX in human serum determined by UPLC-MS/MS method (n = 5).

Figure 2. Passing-Bablok regression analysis and Bland-Altman plots of differences over mean values of paired MTX concentrations obtained by CMIA and UPLC-MS/MS, analyzers on 195 serum samples. Passing-Bablok regression analysis (A. 0.01–1.5 μmol/L; B. 1.5–400 μmol/L); and Bland-Altman plots (C. 0.01–1.5 μmol/L; D. 1.5–400 μmol/L).

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UPLC-MS/MS与化学发光微粒子免疫分析法测定甲氨蝶呤血药浓度的比较分析

Comparative analysis of UPLC-MS/MS and chemiluminescence microparticle immunoassay for serum methotrexate measurement in pediatric intracranial tumor patients

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