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

Background: Naringenin is a flavanone having strong antioxidant potential. It is an anti-hyperlipidemic, antidepressant, 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 C 18 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. Bioanalytical 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.

Nrf2 Activators as Dietary Phytochemicals Against Oxidative Stress, Inflammation, and Mitochondrial Dysfunction in Autism Spectrum Disorders: A Systematic Review

“…Naringenin exerts its antioxidant effect by inhibiting the NF-κB pathway to reduce oxidative damage to DNA by mice exposed to radiation (83). However, due to the low solubility and high metabolism of naringenin, its bioavailability before entering the systemic circulation is poor, which hinders its clinical efficacy (84).…”

Anti-cancer effect of nano-encapsulated boswellic acids, curcumin and naringenin against HepG-2 cell line

“…The mobile phase of curcumin was acetonitrile and water (50:50 v/v) acidified with 2% acetic acid at a flow rate of 1 mL min − 1 and UV detection at 425 nm [ 68 ]. The mobile phase of naringenin was methanol: water (70:30) acidified by orthophosphoric acid with a flow rate of 1 mL min − 1 and ultraviolet detection at 289 nm [ 69 ]. The mobile phase used for boswellic acid was acetonitrile and water (85:15) acidified with (0.1% acetic acid) and flow rate of 0.5 mL min − 1 and UV detection at 254 nm [ 70 ].…”

A Novel Rp-HPLC Gradient Elution Technique for Bioanalytical Method Development and Validation for Estimating Gallic Acid in Wistar Rat Plasma

“…Before being injected into the HPLC equipment in a consecutive manner, the clear supernatant was filtered using 0.42 membrane filter paper. At concentrations of 0.5, 1, 2, 5, 10, 25, 50, and 100 µg/ml, the final linearity and quality control samples were made [18,19].…”