Abstract
A simple, fast, and selective HPLC method is presented to quantified amphotericin B (AmB) in a nose-to-brain nanoliposomal pharmaceutical formulation. The development was based on the design of experiments (DoE) approach. The chromatographic analysis was validated on a C18 Zorbax reversed-phase column (250 mm x 4,6 mm I.D.) with 5 µm of particle size using mobile phase, consisting of a binary mixture of ultra-purified water and an organic composition of acetonitrile, methanol, and tetrahydrofuran (75:17:8, v/v). The isocratic flow rate was 1.0 mL.min-1, and the detection at 383 nm. The 12 minutes running time being selective between the main peak from possible degradation products, linear and accurate for the concentration range of 0.5 to 7.0 μg.mL-1, and precise demonstrating a relative standard deviation of 0.01 % (n = 6). Application of this method to assay and stability studies of AmB in nanoliposomal lipid-based is provided.
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