Development and Validation of a Fast RP-HPLC Method for Determination of Methotrexate Entrapment Efficiency in Polymeric Nanocapsules

• Published in: Journal of chromatographic science 2008-07, Vol.46 (6), p.505-509
• Main Authors: Sartori, Tatiane, Murakami, Fabio Seigi, Cruz, Ariane Pinheiro, de Campos, Angela Machado
• Format: Article
• Language: English
• Subjects: Analysis; Antineoplastic Agents - chemistry; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; General pharmacology; Medical sciences; Methotrexate - chemistry; Nanocapsules; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Polymers - chemistry; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet
• ISSN: 0021-9665; 1945-239X
• DOI: 10.1093/chromsci/46.6.505

A rapid and effective isocratic chromatographic procedure is successfully developed to determinate methotrexate (MTX) entrapment efficiency (EE) in polymeric nanocapsules using reversed-phase high-performance liquid chromatography. The method employed a RP-C18 Shimadzu Shim-pack CLC-ODS (150 mm × 4.6 mm, 5 µm) column with mobile phase constituted by a mixture of water-acetonitrile-tetrahydrofuran (65:30:5 v/v/v; pH 3.0) at a flow rate of 0.8 mL/min. The eluate is monitored with a UV detector set at 313 nm. The parameters used in the validation process are: linearity, specificity, precision, accuracy, and limit of quantitation (LOQ). The linearity is evaluated by a calibration curve in the concentration range of 10–50 µg/mL and presented a correlation coefficient of 0.9998. The polymers (PLA or PLA-PEG), oil, and surfactants used in the nanocapsule formulation did not interfere with analysis and the recovery was quantitative. The intra and inter-day assay relative standard deviation were less than 0.72%. Results are satisfactory, and the method proved to be adequate for the determination of methotrexate in nanocpsules formulations.