Methotrexate Polyglutamates Analysis by Chromatography Methods in Biological Matrices: A Review

Area of Science:

• Pharmacology
• Analytical Chemistry
• Biochemistry

Background:

• Methotrexate (MTX) is a vital immunosuppressant and antineoplastic agent.
• MTX exerts its biological activity through conversion to MTX polyglutamates (MTXPGs).
• MTXPG levels correlate with patient response and toxicity, necessitating therapeutic drug monitoring for personalized treatment.

Purpose of the Study:

• To review and summarize chromatographic methods for quantifying total and individual MTXPGs.
• To provide insights into key method development and validation steps for MTXPG analysis.
• To assist researchers in establishing robust analytical protocols for MTXPG measurement.

Main Methods:

• Systematic literature search of Embase, Science Direct, and PubMed databases.
• Inclusion criteria focused on articles detailing "methotrexate polyglutamate" and "chromatography".
• Analysis and summarization of 14 selected studies on MTXPG chromatographic analysis.

Main Results:

• Various chromatographic techniques (UV-Vis, fluorescence, MS) are employed for MTXPG detection.
• Key method development aspects include sample pretreatment, LC-MS parameter optimization, and internal standard selection.
• Method validation parameters such as LOQ, accuracy, precision, recovery, matrix effect, and stability are critical.

Conclusions:

• Chromatographic methods are essential for accurate MTXPG quantification.
• Standardized approaches to method development and validation are crucial for reliable MTXPG analysis.
• This review offers a valuable resource for researchers developing and validating MTXPG assays for clinical application.