Precision Chemotherapy: Optimizing Calibration for Rapid Determination of Blood Methotrexate by Tandem Mass Spectrometry ± Liquid Chromatography

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry Pub Date : 2025-04-28 DOI:10.1002/rcm.10053

Dennis J. Dietzen, Connor J. Blair

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引用次数: 0

Abstract

Background

Rapid therapeutic monitoring of high-dose methotrexate (MTX) chemotherapy is essential to avoid toxicity. MTX concentrations are commonly monitored using immunoassays that are limited by narrow dynamic range and metabolite cross reactivity. Mass spectrometry may improve the molecular specificity of MTX analysis but is limited by slow throughput and extensive calibration. In this study, we examined the consequences of eliminating LC from MS/MS and foregoing external calibration to enable rapid determination of MTX.

Methods

MTX (m/z 455 → 308), was assessed using UPLC–MS/MS or flow-injection MS/MS using a six-point external calibration scheme with a single deuterated internal standard, 4-point internal calibration using ¹³C₅, ¹³C₆, ¹³C₁₁, and ¹³C₁₄-MTX, or by single point calibration with the single deuterated internal standard. Accuracy, precision, and resistance to hemolysis, icterus, and lipemia were compared with immunoassay.

Results

Across all six injection/calibration schemes, imprecision ranged from 2.5% to 10% (CV) from 0.05–10 μM. Regardless of calibration scheme, MS/MS ± LC was more resistant to interference from hemolysis and bilirubin than immunoassay. MS/MS ± LC determination of MTX was compared with immunoassay in 81–100 plasma specimens with MTX concentrations ranging from 0.05–81 μM. Intercept estimates all included zero with 95% confidence. Estimates of slopes versus immunoassay for each of the six analytic approaches ranged from 0.88 to 1.09.

Conclusions

Eliminating LC and external calibration enabled rapid, precise, and accurate determination of MTX concentrations in plasma. Minimalist but robust approaches such as these may enable the use of MS for routine MTX determination in time-sensitive clinical circumstances.

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精密化疗：串联质谱法±液相色谱法快速测定血液中甲氨蝶呤的优化校准

背景大剂量甲氨蝶呤（MTX）化疗的快速治疗监测是避免毒性的必要条件。MTX浓度通常使用免疫测定法监测，但受限于窄动态范围和代谢物交叉反应性。质谱法可以提高MTX分析的分子特异性，但受缓慢的通量和大量校准的限制。在本研究中，我们检查了从MS/MS中去除LC和前述外部校准以实现MTX快速测定的后果。方法MTX （m/z 455→308）采用UPLC-MS /MS或流动注射MS/MS，采用单氘化内标的6点外标方案，13C5、13C6、13C11和13C14-MTX的4点内标方案，或单氘化内标的单点校准方案。准确度、精密度以及对溶血、黄疸和血脂的耐药性与免疫分析法进行比较。结果在所有六种进样/校准方案中，不精度范围为2.5%至10% (CV)，范围为0.05-10 μM。无论采用何种校准方案，MS/MS±LC比免疫分析法更能抵抗溶血和胆红素的干扰。采用MS/MS±LC法测定81 ~ 100份MTX浓度在0.05 ~ 81 μM范围内的血浆标本中MTX的含量，并与免疫分析法进行比较。截距估计均包含零，置信度为95%。与免疫分析法相比，六种分析方法的斜率估计值从0.88到1.09不等。结论消除LC和外部校准可以快速、精确、准确地测定血浆中MTX浓度。这些极简但可靠的方法可能使MS在时间敏感的临床情况下用于常规MTX测定。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Rapid Communications in Mass Spectrometry 化学-分析化学

CiteScore

4.10

自引率

5.00%

发文量

219

审稿时长

2.6 months

期刊介绍： Rapid Communications in Mass Spectrometry is a journal whose aim is the rapid publication of original research results and ideas on all aspects of the science of gas-phase ions; it covers all the associated scientific disciplines. There is no formal limit on paper length ("rapid" is not synonymous with "brief"), but papers should be of a length that is commensurate with the importance and complexity of the results being reported. Contributions may be theoretical or practical in nature; they may deal with methods, techniques and applications, or with the interpretation of results; they may cover any area in science that depends directly on measurements made upon gaseous ions or that is associated with such measurements.