Development of a new, sensitive, and robust analytical and bio-analytical RP-HPLC method for in-vitro and in-vivo quantification of naringenin in polymeric nanocarriers

Abstract

Abstract Background Naringenin is a flavanone having strong antioxidant potential. It is an anti-hyperlipidemic, anti-depressant, anti-cancer, and neuroprotective agent. However, its major limitation is its low oral bioavailability. Objective In order to overcome this limitation and to explore its antioxidant potential in autism spectrum disorders, we developed brain targeting PLGA nanocarriers of naringenin. Current study involves development of a sensitive and robust RP-HPLC method for detection and quantification (in vitro and in vivo) of naringenin in nanoparticles. Methods An isocratic RP-HPLC method was developed using C18 reversed-phase column (250 × 4.6 mm internal diameter and 5 μm particle size). Flow rate of mobile phase was 1 ml/min and temperature of column was 30 °C. Methanol and 0.5% ortho-phosphoric acid in MilliQ water (pH 2) (70:30) was used as mobile phase. The ultraviolet detection wavelength for quantification was at 289 nm. Results Calibration curve showed linearity within the concentration range from 0.5 to 40 μg/ml (R 2 = 1) for the analytical method and for plasma (6.3–200 ng/ml (R 2 = 0.9975)) and brain tissue samples (31.25–12,500 ng/ml (R 2 = 1)). Limit of detection (LOD) and limit of quantification (LOQ) were 0.15 μg/ml and 0.44 μg/ml for the analytical method. For bio-analytical method, LOD and LOQ were 9.71 ng/ml and 29.44 ng/ml for plasma and 9.06 ng/ml and 27.44 ng/ml for brain sample. Both the method was precise, accurate, and robust. Bio-analytical method showed good recovery from plasma and brain samples (> 95%). Conclusion This analytical and bio-analytical method was applied to detect entrapment efficiency, in-vitro release, and pharmacokinetic parameters of naringenin nanoparticles.

Keywords

• Brain
• Naringenin
• Plasma
• PLGA nanoparticles
• RP-HPLC