Biopharmaceutical assessment of naloxone permeation through human respiratory epithelial tissues: A chromatographic-mass spectrometric approach with cloud based aerosol dosing and delivery.

Abstract

Understanding naloxone permeation is important for optimizing nasal delivery and supporting comparative assessment of nasal drug products. In this study, a stability-indicating LC-MS/MS method was developed and validated for the simultaneous quantification of naloxone and its related impurities, naloxone N-oxide and noroxymorphone, in in vitro permeation test receptor media. The validated method was applied to characterize naloxone permeation following cloud-based aerosol dosing across a synthetic Nuclepore Track-Etched membrane and a differentiated human EpiAirway™ mucociliary tissue model under finite-dose conditions. The analytical procedure demonstrated linearity over 0.25-20.0 ng/mL in Dulbecco's Phosphate-Buffered Saline and Krebs-Ringer Bicarbonate Buffer, with acceptable accuracy and precision. No degradation products or additional impurities were detected in permeation samples, confirming the stability-indicating capability of the method. Naloxone exhibited rapid early-time permeation across the synthetic membrane, whereas transport across the epithelial tissue model was attenuated and plateaued, reflecting physiological barrier function. Integration of cloud-based aerosol delivery with a validated LC-MS/MS platform enables mechanistic evaluation of nasal naloxone permeation and provides a supportive in vitro framework for formulation characterization and comparative assessments.