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Quantitative Determination of Methotrexate, 7-Hydroxymethotrexate, and Methotrexate Polyglutamates in Human Erythrocytes by High-Performance Liquid Chromatography with Mass Spectrometry

  • STRUCTURE OF CHEMICAL COMPOUNDS, METHODS OF ANALYSIS AND PROCESS CONTROL
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Pharmaceutical Chemistry Journal Aims and scope

Methotrexate is the drug of choice for the treatment of rheumatoid arthritis. However, some patients have no or an insufficient effect if the treatment protocol is followed. Therefore, a need exists to establish patient compliance and to develop an instrumental approach to correct drug therapy. The present work was aimed at developing quantitative determination methods for methotrexate and its metabolites such as 7-hydroxymethotrexate and polyglutamate residues in erythrocytes that possess high sensitivity, selectivity, and rapidity. The proposed quantitative determination method for methotrexate and its metabolites is based on high-performance liquid chromatography with electrospray-ionization (ESI) and mass spectrometric detection (HPLC-MS/MS). The lower limit of quantitation (LLOQ) is 0.5 nmol/L for all analytes. The calibration curve is linear in the range 0.5 – 100 nmol/L. The coefficient of approximation accuracy is no less than 0.99. Samples were prepared by precipitation of a homogenized erythrocyte mass. The method was tested in clinical study No. 0514-2018-0003 using local ethics committee protocol No. 23 of Sept. 20, 2018.

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Authors and Affiliations

  1. Scientific Center for Mental Health, 34 Kashirskoe Shosse, Moscow, 115522, Russia

    N. V. Baymeeva

  2. V. A. Nasonova Research Institute of Rheumatology, 34A Kashirskoe Shosse, Moscow, 115522, Russia

    G. I. Gridneva, E. S. Aronova, E. Yu. Samarkina & Yu. V. Murav’ev

  3. N. V. Sklifosovsky Research Institute of Emergency Medicine, 3/1 Bol’shaya Sukharevskaya Sq., Moscow, 129010, Russia

    I. A. Tyurin

  4. All-Russian Research Institute of Metrological Service, 46 Ozernaya St., Moscow, 119361, Russia

    A. A. Kirilyuk

Authors
  1. N. V. Baymeeva
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  2. G. I. Gridneva
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  3. I. A. Tyurin
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  4. E. S. Aronova
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  5. E. Yu. Samarkina
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  6. Yu. V. Murav’ev
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  7. A. A. Kirilyuk
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Corresponding author

Correspondence to N. V. Baymeeva.

Additional information

Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 56, No. 3, pp. 59 – 64, March, 2022.

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Baymeeva, N.V., Gridneva, G.I., Tyurin, I.A. et al. Quantitative Determination of Methotrexate, 7-Hydroxymethotrexate, and Methotrexate Polyglutamates in Human Erythrocytes by High-Performance Liquid Chromatography with Mass Spectrometry. Pharm Chem J 56, 411–415 (2022). https://doi.org/10.1007/s11094-022-02643-w

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  • DOI: https://doi.org/10.1007/s11094-022-02643-w

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