Development and validation of a fast RP-HPLC method to determine the analogue of the thyroid hormone, 3,5,3′-triiodothyroacetic acid (TRIAC), in polymeric nanoparticles

Abstract

The objective of this work was to develop and validate a rapid Reversed-Phase High-Performance Liquid Chromatography method for the quantification of 3,5,3′-triiodothyroacetic acid (TRIAC) in nanoparticles delivery system prepared in different polymeric matrices. Special attention was given to developing a reliable reproductive technique for the pretreatment of the samples. Chromatographic runs were performed on an Agilent 1200 Series HPLC with a RP Phenomenex^® Gemini C18 (150 × 4, 6 mm i.d., 5 μm) column using acetonitrile and triethylamine buffer 0.1% (TEA) (40 : 60 v/v) as a mobile phase in an isocratic elution, pH 5.6 at a flow rate of 1 ml min⁻¹. TRIAC was detected at a wavelength of 220 nm. The injection volume was 20 μl and the column temperature was maintained at 35 °C. The validation characteristics included accuracy, precision, specificity, linearity, recovery, and robustness. The standard curve was found to have a linear relationship (r² = 0.9996) over the analytical range of 5–100 μg ml⁻¹. The detection and quantitation limits were 1.3 and 3.8 μg ml⁻¹, respectively. The recovery and loaded TRIAC in colloidal system delivery was nearly 100% and 98%, respectively. The method was successfully applied in polycaprolactone, polyhydroxybutyrate, and polymethylmethacrylate nanoparticles.