A validated chemiluminescence immunoassay for methotrexate (MTX) and its application in a pharmacokinetic study

Song, Zhaorui; Wang, Yufen; Dong, Yaqing; Xu, Kun; Liu, Hao; Chuan, Deng; Yin, Yongmei; Eremin, Sergei A.; Meng, Meng; Xi, Rimo

doi:10.1039/c5ay02270c

Cited by 11 publications

(6 citation statements)

References 23 publications

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“…is not meet the satisfaction in clinic [14,21] . HPLC [22][23] and LC-MS/MS have strong speci city to differentiate target drugs and metabolites, while LC-MS/MS can work at a higher sensitivity than HPLC.…”

Section: Discussionmentioning

confidence: 94%

An optimized UHPLC-MS/MS method for simultaneous determination of erythrocyte methotrexate and polyglutamylated metabolites: Application in pediatric patients with acute lymphoblastic leukemia

Wang

Guan

Qian

et al. 2023

Preprint

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Methotrexate (MTX), a widely used chemotherapeutic drug, is critical for achieving long-term complete remission in contemporary maintenance therapy of acute lymphoblastic leukemia (ALL). MTX is intracellularly converted into methotrexate polyglutamates (MTXPGn) by folylpolyglutamate synthase after transporting into the cells. The intracellular levels of active MTXPGn regulate the clinical efficacy and drug-related side effects, but it is still challenged by large inter-individual differences and narrow therapeutic index. Therefore, it is of great significance to develop a sensitive and stable method to monitor MTXPGs concentrations in the erythrocyte. To facilitate clinical application, an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantification of MTX and MTXPGn in erythrocyte. Solid phase extraction was applied to sample clean-up followed by a gradient chromatographic separation. The elution consisting of methanol/water/2 mM ammonium acetate (flow rate: 0.4 mL/min). Linearity of the assay was assured in the range of 1-500 nM (R2 > 0.99). The co-efficient of variation ranged from 2–11% for intraday precision and 0.4–8% for inter day precision. Recovery ranged from 62.51–99.75%, and the matrix-effect varied from 63.24–96.33%. Blood samples from 89 pediatric ALL patients were detected. The concentrations and distribution of MTXPGn in these samples were basically consistent with the current literatures, which confirmed that the optimized method for quantitative detection of MTXPGn in red blood cells had high sensitivity and accuracy. The sensitive method has been fully validated and successfully applied to determine the erythrocyte MTXPGn in pediatric ALL patients during continuation therapy.

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Section: Discussionmentioning

confidence: 94%

An optimized UHPLC-MS/MS method for simultaneous determination of erythrocyte methotrexate and polyglutamylated metabolites: Application in pediatric patients with acute lymphoblastic leukemia

Wang

Guan

Qian

et al. 2023

Preprint

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“…For the cytotoxicity assay, Caco-2 and L-929 cells were seeded in 96-well plates at 1 × 10 4 viable cells per mL and incubated overnight. The cells were then treated with a culture medium containing different concentrations of Ph-Cys-Au and Ph-Cys-Au+MTX (10,20,30,40,50,60,70,80,90, and 100 µM). The non-treated cells were taken as the control.…”

Section: Methodsmentioning

confidence: 99%

“…The cytotoxicity of Ph-Cys-Au and Ph-Cys-Au+MTX on Caco-2 and L-929 cell lines was investigated by treating the cells with increasing concentrations (10,20,30,40,50,60,70,80,90, and 100 µM) for different periods, i.e., 24, 48, and 72 hours, respectively, as shown in Fig. 7 and 8.…”

Section: Cytotoxicity Screeningmentioning

confidence: 99%

“…There are several analytical methods for detecting MTX, such as surface plasmon resonance, surface-enhanced Raman scattering, high-performance liquid chromatography, electrolysis, immunoassays, radioimmunoassay, enzyme-multiplied immunoassay, protein binding, enzyme inhibition, microbiological assays, and electrochemical assays. 1,3,[7][8][9][10] In general, the high-pressure liquid chromatography (HPLC) method has been used to measure MTX in blood samples. The main disadvantage of these techniques is they are very complex and costly.…”

Section: Introductionmentioning

confidence: 99%

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An optical sensing platform for the detection of anti-cancer drugs and their cytotoxicity screening using a highly selective phosphorene-based composite

Sultana,

Thanil Singh,

Khan

et al. 2023

Nanoscale

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“…There are several methods available for determining MTX, such as surface-enhanced Raman scattering (SPRS) [ 1 ], surface plasmon resonance (SPR) [ 2 ], high-performance liquid chromatography (HPLC) [ 7 , 8 , 9 ], electroanalysis [ 10 , 11 , 12 ], and immunoassays [ 13 ]. In comparison to the methods mentioned above, fluorescence has gained attention due to its remarkable sensitivity, quick response, easy operation, good biocompatibility, minimal toxicity, and the ability to detect non-destructively [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

One-Step Preparation of S, N Co-Doped Carbon Quantum Dots for the Highly Sensitive and Simple Detection of Methotrexate

Wei

Han

et al. 2022

Molecules

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(1) Background: Carbon quantum dots (CQDs) are a new class of carbon nanomaterials with favorable features, such as tunable luminescence, unique optical properties, water solubility, and lack of cytotoxicity; they are readily applied in biomedicine. (2) Methods: S, N co-doped CQDs were prepared to develop a highly selective and sensitive fluorescence technique for the detection of methotrexate (MTX). For this purpose, citric acid and thiourea were used as C, N, and S sources in a single-step hydrothermal process to prepare the S, N co-doped CQDs, which displayed remarkable fluorescence properties. (3) Results: Two optimal emissions were observed at the excitation/emission wavelengths of 320/425 nm, respectively. The two emissions were significantly quenched in the presence of MTX. Under optimal conditions, MTX was detected in the linear concentration range of 1–300 μmol/L, with the detection limit of 0.33 μmol/L. The sensing mechanism was due to the fact that the effect of the inner filter on MTX and S, N-CQDs causes fluorescence quenching. The contents of MTX in real medicine samples were evaluated with acceptable recoveries of 98–101%. (4) Conclusions: This approach has great potential for detecting MTX in pharmaceutical analysis.

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A validated chemiluminescence immunoassay for methotrexate (MTX) and its application in a pharmacokinetic study

Abstract: Polyclonal antibodies for anticancer drug methotrexate (MTX) were originally produced, and a chemiluminescence immunoassay was developed for plasma MTX determination.

Cited by 11 publications

References 23 publications

An optimized UHPLC-MS/MS method for simultaneous determination of erythrocyte methotrexate and polyglutamylated metabolites: Application in pediatric patients with acute lymphoblastic leukemia

An optimized UHPLC-MS/MS method for simultaneous determination of erythrocyte methotrexate and polyglutamylated metabolites: Application in pediatric patients with acute lymphoblastic leukemia

An optical sensing platform for the detection of anti-cancer drugs and their cytotoxicity screening using a highly selective phosphorene-based composite

One-Step Preparation of S, N Co-Doped Carbon Quantum Dots for the Highly Sensitive and Simple Detection of Methotrexate

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