To the best of our knowledge, this is the first report describing optimization of extraction process of phytochemical compounds and XO inhibitory activity from E. hirta. Therefore, this study aimed at optimizing the extraction temperature, time, and solid-to-liquid ratio for the extraction of total flavonoid content, total phenolic content, and anti-gout activity from E. hirta whole plant excluding roots using RSM. The gene discussed is XDH; the disease is gout.