HS-SPME coupled with GC-FID was applied for the quantification of 2-acetyl-1-pyrroline (2AP) in Pandanus amaryllifolius, the best available natural source of (2AP). Conditions for qualitative and quantitative analysis of aroma volatiles with special reference to 2AP from P. amaryllifolius have been optimized using HS-SPME/GC-FID. About 10 mg powdered leaf material at 80 ° C temperature for 35 min was found to be the most effective combination for maximum release of 2AP. Volatile analyses led to the identification of 31 compounds, of which 21 are reported for the first time in P. amaryllifolius. 2AP contents and other volatiles were found to vary in small and large growth forms.