This project aimed at investigating the impact of morphine exposure on brain endothelial cells via an in vitro monoculture of primary human brain microvascular endothelial cells (HBMECs) and state of the art mass spectrometry-based proteomics, using a data-independent acquisition (DIA) MS-based quantitative approach. HBMECS were exposed to morphine at three different concentrations (1, 10, 100 uM) and for two different time points (24h and 48h) to explore dose- and time-dependent effects.