A rapid micro-scale solid-phase micro-extraction (SPME) procedure coupled with gas-chromatography with flame ionized detector (GC-FID) was used to extract parts per billion levels of a principle basmati aroma compound 2-acetyl-1-pyrroline (2-AP) from bacterial samples. In present investigation, optimization parameters of bacterial incubation period, sample weight, pre-incubation time, adsorption time, and temperature, precursors and their concentrations has been studied. In the optimized conditions, detection of 2-AP produced by Bacillus cereus ATCC10702 using only 0.5 g of sample volume was 85 g/kg. Along with 2-AP, 15 other compounds produced by B. cereus were also reported out of which 14 were reported for the first time consisting mainly of (E)-2-hexenal, pentadecanal, 4-hydroxy-2-butanone, n-hexanal, 2-6-nonadienal, 3-methoxy-2(5H) furanone and 2-acetyl-1-pyridine and octanal. High recovery of 2-AP (87 %) from very less amount of B. cereus samples was observed. The method is reproducible fast and can be used for detection of 2-AP production by B. cereus. (c) 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1356-1363, 2014