Andrographis paniculata Burm. F Nees used to treat COVID-19 in several Asian countries because it has metabolites with pharmacological activity that may decrease the pathogenesis features of COVID-19. The extraction solvent significantly affected the composition and concentration of the metabolites. Therefore, the inhibitory effects of nitric oxide of A. paniculata extract based on differences in extraction solvent concentrations were investigare and also identified its metabolite profile. The leaves of A. paniculata were extracted with three different solvents like water solvent, 50% ethanol and pure ethanol. Nitrite oxide (NO) inhibitory activity on LPS-activated RAW 264.7 macrophage cells was used to evaluate the anti-inflammatory activity of the extract, the identification of metabolite profile using LC-MS/MS. Extract pure ethanol had the highest inhibition potential against NO at 91.02%, whereas water and 50% ethanol solvents had inhibitory potentials of 21.35% and -8.13%, respectively. The metabolites 14-deoxyandrographoside and 14-deoxy-17-hydroxyandrographolid were only identified in the pure ethanol extract. This study shows that differences in solvent concentrations play an essential role in enhancing the pharmacological potential of A. paniculata as an anti-inflammatory candidate for COVID-19 treatment. © 2024 Asian Publication Corporation. All rights reserved.