ArticleTranslational and clinical pharmacology2025
Quantitation of methotrexate polyglutamates in red blood cells and application in patients with Crohn's disease.
Article in Translational and clinical pharmacology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
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1 citing paper in PubMed.
- MTX pathway gene variants, erythrocyte methotrexate polyglutamates, and treatment outcomes in rheumatoid arthritis.Biomolecules & biomedicine · 2026Article
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9 authors.
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Abstract
Methotrexate (MTX), a folate antagonist, is commonly administered at low doses for the treatment of Crohn's disease (CD). Anti-inflammatory effects of MTX are facilitated by its intracellular conversion to MTX polyglutamates (MTX-PGs). Because plasma-based monitoring of therapeutic response does not accurately reflect the therapeutic efficacy of MTX, quantifying intracellular MTX-PGs, potential biomarkers of the MTX response, is crucial. However, it is challenging to routinely monitor intracellular MTX metabolites in patients with CD due to the low concentrations of MTX-PGs. Therefore, quantitating MTX-PGs in clinical samples with a high-sensitivity method is necessary. We established a high-sensitivity method to quantify three MTX-PGs using perchloric acid deproteinization followed by high-performance liquid chromatography-tandem mass spectrometry. Calibration curves were generated using human red blood cells as biological matrix. This method was applied to analyze MTX-PGs in red blood cells (RBCs) from patients with CD undergoing MTX therapy. The method achieved a lower limit of quantification of 1 ng/mL for individual MTX-PGs. A nine-point calibration curve covering 1-400 ng/mL showed excellent linearity. Precision (relative standard deviation < 15%) and accuracy (93.41-109.37%) were satisfactory in both intra- and inter-day assays. Plasma MTX levels were not significantly correlated with any individual RBC MTX-PG level (
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