A Novel and Green UFLC-MS/MS Method for Quantification of Amantadine and Levodopa in Polymeric Nanoparticles: Application to determine Drug loading (%DL), Drug entrapment (%DEE) and Drug release profile

Introduction: Levodopa, a dopamine precursor, widely prescribed drug in Parkinson’s disease management possesses a side effect of Levodopa-induced dyskinesia (LID). Amantadine hydrochloride, an NMDA receptor antagonist is co-encapsulated along with levodopa and formulates as polymeric nanoparticles (NPs) so as to overcome the side effects and act synergistically to enhance therapeutic outcome. Methodology: In current work, we developed a novel, green Ultra-Fast Liquid Chromatography-Tandem Mass Spectrometry (UFLC-MS/MS) technique for the simultaneous quantification of amantadine and levodopa in polymeric nanoparticles. A triple quadrupole analyser with a multiple reaction monitoring (MRM) scan mode and an atmospheric pressure chemical ionization (APCI) source. A Waters Symmetry C8 column (150 × 4.6 mm, 3.5 μm) maintained at 40 °C was used for the chromatographic separation. In order to ensure sensitive and specific analyte detection, the mobile phase consisted of 0.1 % formic acid in water and methanol (40:60) with a total run time of 5 min. Excellent linearity, recovery, accuracy, and sensitivity were validated by method validation. Greenness assessment was done by AGREE, GAPI and AES metrics. Results: Proposed green UFLC-MS/MS method effectively quantifies Levodopa and Amantadine in polymeric nanoparticles, followed by accurate evaluation of %DL, %DEE, and drug release profiles. %DEE values were observed for Levodopa (89.6%) and Amantadine (90.16 %), with corresponding %DL values of 20.5% and 24.10%, indicating substantial drug loading capacity. The in vitro drug release profiles demonstrated sustained release behaviour, with 91.89% ± 0.362 of Levodopa and 92% ± 0.362 of Amantadine released over the study period. Conclusion: Using GAPI, AGREE, and AES criteria, the created analytical method's greenness was carefully assessed. The outcomes were then compared to previously published methods in the literature. For the simultaneous measurement of amantadine and levodopa in nanoparticles, our innovative UFLC-MS/MS technology offered a dependable and extremely sensitive method.