Optimized high performance liquid chromatography?ultraviolet detection method using core-shell particles for the therapeutic monitoring of methotrexate

MILAGROS MONTEMURRO; MARÍA MERCEDES DE ZAN; JUAN CARLOS ROBLES

Journal of Pharmaceutical Analysis

Elsevier

Año: 2016 vol. 6 p. 103 - 111

Methotrexate (MTX) is an antineoplastic drug, and due to its high toxicity, the therapeutic drug monitoringis strictly conducted in the clinical practice. The chemometric optimization and validation of ahigh performance liquid chromatography (HPLC) method using core?shell particles is presented for thedetermination of MTX in plasma during therapeutic monitoring. Experimental design and responsesurface methodology (RSM) were applied for the optimization of the chromatographic system and theanalyte extraction step. A Poroshell 120 EC-C18 (3.0 mm75 mm, 2.7 μm) column was used to obtain afast and efficient separation in a complete run time of 4 min. The optimum conditions for the chromatographicsystem resulted in a mobile phase consisting of acetic acid/sodium acetate buffer solution(85.0 mM, pH¼4.00) and 11.2% of acetonitrile at a flow rate of 0.4 mL/min. Selectivity, linearity, accuracyand precision were demonstrated in a range of 0.10?6.0 mM of MTX. The application of the optimizedmethod required only 150 mL of patient plasma and a low consumption of solvent to provide rapid results.