Anxiolytic activity and phytochemical content of the methanol extract of Blumea balsamifera (L.) DC. in ICR mice

Anxiety disorders significantly impact quality of life worldwide. While anxiolytic drugs like benzodiazepines are effective, they pose risk of adverse effects. This has driven interest in plant-derived alternatives with safer profiles. Blumea balsamifera (L.) DC. is traditionally used in Southeast Asia for medicinal purposes, yet its anxiolytic activity has not been scientifically evaluated. This study investigated the anxiolytic potential of the methanol extract of B. balsamifera leaves in male Institute of Cancer Research mice using the elevated plus maze test. This was an experimental, oral dose-response study employing a randomized controlled design to compare the effects of B. balsamifera against control groups. The extract underwent photochemical screening and an acute oral toxicity test following OECD Guideline 423. Subsequently, anxiolytic activity was evaluated using the elevated plus maze test at oral doses of 250, 500, or 1000 mg/kg. Diazepam (1 mg/kg) and 1% Tween 80 (10 mL/kg) served as the positive and negative control, respectively. A power analysis determined a minimum sample size of 25 animals (five groups) to achieve ≥ 0.95 power (α = 0.05; f = 1). Phytochemical screening identified the presence of flavonoids, tannins, terpenoids, and phenols. Acute oral toxicity test indicated an estimated LD₅₀ of 5000 mg/kg. In the elevated plus maze test, ICR mice treated with 500 and 1000 mg/kg showed a statistically significant increase compared to the negative control. These findings demonstrate a favorable safety profile and dose-dependent anxiolytic-like effects, providing the like effects, providing the first scientific evidence supporting B. balsamifera leaves as a potential natural anxiolytic agent.