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Development and Validation of a High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Methotrexate Quantitation in Human Serum

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Abstract

Methotrexate (MTX) is the only chemotherapy drug that is routinely monitored by therapeutic drug monitoring service in Malaysia government hospitals. The patients with MTX plasma concentration exceeding 0.4 μM for 48 h are at higher risk to develop toxicities such as nephrotoxicity, hepatotoxicity, neurotoxicity, mucositis and myelosuppression. The aim of this study was to develop and validate the ultra-high performance liquid chromatography−tandem mass spectrometry (UHPLC−MS/MS) method for MTX quantitation in human serum. Our results show that the retention times of MTX and warfarin (internal standard) were 1.82 and 2.68 min, respectively. The linearity was obtained at the concentrations from 25 to 500 ng/mL with the regression equation of y = 1025x – 11757 and r2 of 0.9901. This analyte was stable in a freezer at –80°C for up to 90 days. As a conclusion, this UHPLC−MS/MS method is an optional technique for clinicians to quantitate the patient’s MTX levels in the hospital.

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Funding

This project is funded by Universiti Teknologi MARA and conducted at Integrative Pharmacogenomics Institute. It will be used for a collaboration research project between iPROMiSE and Pediatric Institute, Hospital Kuala Lumpur (HKL) to measure the concentration of methotrexate in children with acute lymphoblastic leukemia (ALL) treated with high-dose methotrexate (HDMTX) 5000 mg/m2.

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Authors and Affiliations

  1. Integrative Pharmacogenomics Institute (iPROMISE), Universiti Teknologi MARA (UiTM), Puncak Alam Campus, Selangor, Malaysia

    Rizal Husaini Razali, Mohd Salleh Rofiee, Lay Kek Teh & Mohd Zaki Salleh

  2. Department of Pediatric, Pediatric Institute, Hospital Kuala Lumpur, Kuala Lumpur, Malaysia

    Kok Hoi Teh & Hishamshah Mohd Ibrahim

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  1. Rizal Husaini Razali
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  2. Mohd Salleh Rofiee
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  3. Lay Kek Teh
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  4. Kok Hoi Teh
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  5. Hishamshah Mohd Ibrahim
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  6. Mohd Zaki Salleh
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Correspondence to Mohd Zaki Salleh.

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Rizal Husaini Razali, Rofiee, M.S., Teh, L.K. et al. Development and Validation of a High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Methotrexate Quantitation in Human Serum . J Anal Chem 75, 1335–1339 (2020). https://doi.org/10.1134/S1061934820100111

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  • DOI: https://doi.org/10.1134/S1061934820100111

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