Abstract : Andrographis paniculata (AP) is a medicinal herb claimed to possess various pharmacological activities. As the AP extracts were widely used for medicinal purposes, the present study was embarked to evaluate the best extraction methods among AP ethanol, methanol and aqueous extracts, to determine its phytochemical compound and cytotoxicity effect. The toxicity effect of AP extract was evaluated using brine shrimp lethality assay. Finally, the chemical compounds of AP extracts were also identified using Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). AP ethanol extraction resulted in the highest yield, demonstrated the superiority of the method. The AP ethanol extract was subjected to a subsequent study of brine shrimp cytotoxicity assay and demonstrated the non-toxic effect of this extract. The chemical evaluation of AP extracts showed the presence of andrographolide and neo-andrographolide in methanol and ethanol extract, while AP aqueous extract only contained andrographolide. Therefore, the findings in this study support the safety and potential use of AP ethanol extract as a novel agent for medicinal purposes. Further laboratory and clinical trial studies are needed to establish the use of AP ethanol extract as an alternative medicine