Quantitative HPLC Determination of Ftivazide. Evaluation of the Measurement Uncertainty

A novel, simple, rapid, effective, and highly sensitive HPLC method has been developed for the quantitative determination of ftivazide. Solutions of ftivazide (0.2 mg/mL) in MeCN were chromatographed over a Kinetex C18 column (250 × 4.6 mm, 5 μm) in isocratic mode using a mixture of MeCN and ammonium acetate solution (0.1 M) in a 25:75 volume ratio as a mobile phase at a flow rate of 1 mL/min. Detection was made at 330 nm using a UV diode-array detector. The retention time of the ftivazide peak was 3.9 min. The suitability of the developed method was confirmed by validation for specificity, linearity, accuracy, robustness, and precision. The developed method showed higher selectivity as compared to the certified method using UV spectrophotometry included in the regulations for ftivazide-based drugs. The measurement uncertainty of the novel method was evaluated in three ways, using bottom-up and top-down approaches and calculation of the confidence interval using the Student t-test.