Previous study on the crude leaf extract of Pandanus tectorius showed a diuretic activity using Sprague-Dawley rats. To corroborate this claim, the crude extract was fractionated and the isolation of squalene (1) was done on the chloroform fraction. A positive diuretic effect on three increasing doses of the chloroform extract in comparison to the negative control was observed. The highest diuretic effect was observed at a dose of 500 mg/kg BW and a diminished effect occurred at the highest dose of 1000 mg/kg BW which may indicate a low-ceiling loop diuretic mechanism. A positive diuretic activity on 1 was observed in test animals administered with 15 and 60 mg/kg BW doses. Although both 1 and chloroform extract exhibited positive diuretic effects, the results were not comparable to the positive control, Furosemide, which is a known potent diuretic agent.