RPLC method development and validation for the quantitative determination of phenytoin and valproic acid in tablet formulations
DOI:
https://doi.org/10.1590/s2175-97902024e24638Keywords:
Antiepileptic drugs, Binary mixtures, RPLC, Method validation, Optimization, QuantificationAbstract
In this study, the reverse phase liquid chromatography (RPLC) method was preferred to determine the chromatographic behavior of phenytoin and valproic acid, the active ingredients of antiepileptic drugs. The optimization of the developed method was based on the capacity factor values of studied compounds in water-organic solvent mixtures varying in constant mobile phase pH value and the organic solvent concentration in the mobile phase where the compounds were analyzed. At constant pH, phenytoin was analyzed in acetonitrile-water binary mixtures containing 30-40% (v/v) acetonitrile, and valproic acid was analyzed in methanol-water binary mixtures containing 10-20% (v/v) methanol at 37oC. Kinetex EVO C18 Core-Shell (250x4,6mm I.D., 5µm) and Pinnacle DB Cyano (250 x 4,6 mm I.D., 5µm) columns were preferred for the quantitative determination of the compounds. Under all conditions mentioned in the method section, excellent linearity (r>0.99) was obtained in the concentration range of 10-40 μg/mL for phenytoin and 200-600 μg/mL for valproic acid. The recovery value of the method was calculated as 98.48% and 99.40% for phenytoin and valproic acid, respectively. In this study, an analytical procedure suitable for routine use was developed, and the method was validated for the determination of the amount of studied compounds in pharmaceutical dosage forms.
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