Quantification and Evaluation of Glycyrrhizic Acid-loaded Surface Decorated Nanoparticles by UHPLC-MS/MS and used in the Treatment of Cerebral Ischemia

Background: Glycyrrhizic Acid (GRA), a potent antioxidant triterpene saponin glycoside and neuroprotective properties exhibits an important role in the treatment of neurological disorders i.e. cerebral ischemia. GRA is water soluble, therefore it's have low bioavailability in the brain. Objective: To enhance brain bioavailability for intranasally administered Glycyrrhizic Acid-encapsulated-chitosan-coated-PCL-Nanoparticles (CS-GRA-PCL-NPs). Methods: Chitosan-coated-PCL-Nanoparticles (CS-PCL-NPs) were developed through double emulsification-solvent evaporation technique and further characterized for particle size, zeta potential, size distribution, encapsulation efficiency as well as in vitro drug release. UPLC triple quadrupole Qtrap MS/MS method was developed to evaluate brain-drug uptake for optimized CS-GRA-PCL-NPs and to determine its pharmacokinetic in rat's brain as well as plasma. Results: Mean particles size (231.47 +/- 7.82), polydispersity index (PDI) i.e. (0.216 +/- 0.030) and entrapment efficiency (65.69 +/- 5.68) was determined for developed NPs. UPLC triple quadrupole Qtrap MS/MS method study showed a significantly high mucoadhesive potential of CS-GRA-PCL-NPs and least for conventional and homogenized nanoformulation; elution time for GRA and internal standard (IS) Hydrocortisone as 0.37 and 1.94 min at m/z 821.49/113.41 and 363.45/121.40 were observed, respectively. Furthermore, intra and inter-assay (%CV) of 0.49-5.48, %accuracy (90.00-99.09%) as well as a linear dynamic range (10.00 ng/mL -2000.0 ng/mL), was observed. Pharmacokinetic studies in Wistar rat brain exhibited a high AUC(0-24) alongwith an amplified C-max (p** < 0.01) as compared to i.v. treated group. Conclusion: Intranasal administration of developed CS-coated-GRA-loaded-PCL-NPs enhanced the drug bioavailability in rat brain along with successfully UPLC-MS/MS method and thus preparation of GRA-NPs may help treat cerebral ischemia effectively. The toxicity studies performed at the end revealed safe nature of optimized nanoformulation.