Analytical profiling of Lentinan in hybrid nanostructures for intranasal Delivery: Method development and Validation

Abstract

Lentinan (LNT) a polysaccharide obtained from mushroom Lentinus edodes, possesses potent anticancer and antitumor activity. To improve the bioavailability, encapsulation of LNT in polymer -lipid hybrid nanostructures (PLHNs) is done and administration via nasal pathway will bypass hepatic first-pass metabolism, which possibly will give high prospect for treatment of brain tumors. The estimation of entrapped LNT within Hybrid Nanostructures is one of the important parameters for the characterization of nanoparticles. Therefore, an analytical method was developed and validated for estimating plain LNT and LNT-PLHNs in Nasal Simulated Fluid (NSF) by conjugating with Congo Red (CR) in dilute alkaline solutions by employing high-performance liquid chromatography (HPLC). The developed method was validated for selectivity, linearity, precision, accuracy, robustness. The linearity was checked over the concertation range of 2–10 μg/mL the and limit of detection (LOD) and limit of quantification (LOQ) were 38.02 ± 0.21 ng/mL and 86.71 ± 0.82 ng/mL respectively. The mean percent recovery of LNT was 99.18 %; relative standard deviation (RSD) of 0.386 % and intra- and interday coefficient of variation was <2 %. The developed HPLC method was found suitable for assessing percent entrapment efficiency (%EE), in-vitro drug release, intracellular uptake study and stability profile of LNT-PLHNs obtained by solvent injection technique.