This study evaluated the stability and pharmacokinetics of asiaticoside (AS) dry nanoemulsion (AS-DNE) and developed a validated LC–MS/MS method for AS quantification in rat plasma. AS-DNE remained stable over 24 weeks, with minimal changes in moisture content, particle size, and zeta potential. AS degradation was temperature dependent, with the highest loss (6.73%) observed at 40 °C. Plasma samples were prepared by protein precipitation using methanol, with valproic acid used as an internal standard. The method demonstrated excellent linearity (25–2000 ng/mL, R2 = 0.9999), precision (< 7.39%), accuracy (< 11.19%), and recovery (80.95-91.19%), and AS was stable under various tested conditions. Pharmacokinetic studies revealed dose-independent behavior and significantly enhanced oral absorption: AS-DNE reached a peak plasma concentration of 211.32 ng/mL at 15 min, compared with 71.21 ng/mL at 14.4 h for pure AS. These results demonstrate improved bioavailability of AS through dry nanoemulsion formulation. © Copyright Wardiyah et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC-BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.