Development and Validation of an HPLC Method Using Fluorescence Detection for the Quantitative Determination of Curcumin in PLGA and PLGA-PEG Nanoparticles

Abstract

In this paper, a rapid and effective chromatographic procedure for determining the curcumin encapsulation efficiency in poly(lactic-co-glycolic acid) (PLGA) and poly(lactic-co-glycolic acid)-polyethyleneglycol (PLGA-PEG) nanoparticles via reversed-phase high-performance liquid chromatography (RP-HPLC) using a fluorescence detector and low flow rate is described. Chromatographic runs were performed on a RP C18 column (250 mm x 4.6 mm, 5 μm) with a mixture of ethanol, water and acetonitrile (80:10:10, v/v/v) as the mobile phase and a flow rate of 0.8 mL/min in the isocratic mode. Curcumin was detected using a fluorescence detector operating at an excitation wavelength of 365 nm and an emission wavelength of 512 nm. This method was validated in terms of the selectivity, linearity, precision, accuracy, robustness, limit of detection and limit of quantitation. The analytical curve was linear over the concentration range of 1– 50 μg/mL, and the limits of detection and quantitation were 9.65 and 50 ng/ml, respectively. The mean recovery for curcumin was 101.14 ± 2.8% (n = 9). The intra- and inter-assay coefficients of variation were less than 3.73%. The method was robust for changes in the mobile phase, column temperature and flow rate. The maximum relative standard deviation was 3.08%. The method was successfully used to determine the encapsulation efficiency of curcumin in PLGA and PLGA-PEG nanoparticles. Keywords: Biodegradable polymers, Curcumin, Encapsulation efficiency, Fluorescence, HPLC, Nanoparticles, Particle size, PLGA, PLGA-PEG, Validation