Cereal Chem. 88(3):271-277An extremely sensitive method for the analysis of 2-acetyl-1-pyrroline (2AP) in rice, employing stir bar sorptive extraction (Twister) was studied. The Twister stir bar is placed in the headspace of a 20-mL vial containing 1 g of rice kernels, 7.5 mL of 0.1M KOH, and 2.2 g of NaCl, along with a second Teflon-coated stir bar for mixing. Analytes are adsorbed onto the Twister for 4 hr at 40°C and then desorbed at 270°C into a GC column while cryofocusing at -80°C. The headspace sorptive extraction (HSSE) method was able to detect <0.1 ppb of 2AP in rice. The precision of the HSSE method (>10%) was not as good as the GC/FID method (≈6%). Using HSSE, 2AP was observed in all samples generally considered to be aromatic and was not observed in any nonaromatic samples. Additionally, a modified method for the synthesis of 2-acetyl-1-pyrroline was studied and the presence of a tautomer of 2-acetyl-1-pyrroline was confirmed.Numerous compounds can be observed in headspace of cooked rice. Lipid oxidation products (aldehydes, alcohols, etc.) dominate the chromatographic profiles irrespective of the analytical method employed. However, only the volatile compound 2-acetyl-1-pyrroline (2AP) has been related to a characteristic rice aroma. Initial reports suggested 2AP concentrations in aromatic rice were <100 ppb (Buttery et al 1982). Subsequent reports have shown 2AP concentrations range from several hundred ppb to several thousand ppb. When present at concentrations of several hundred ppb, 2AP gives the rice a nutty or popcorn-like aroma, and the rice is referred to as aromatic. 2AP has been reported in nonaromatic rice at concentrations of only a few ppb (Buttery et al 1988;Grimm et al 2001). However, it is unclear whether 2AP is actually present in nonaromatic rice and, if so, does it contribute to the overall aroma of rice.2AP is generated during the growing season in the plant and can be found in the leaves and stems as well as in the rice kernel (Yoshhashi 2002). The maximum 2AP concentration is varietydependent (Guofo et al 2010) and occurs four or five weeks after heading (WAH) and decreases to 20% of maximum at seven or eight WAH (Itani et al 2004). Furthermore, 2AP concentration decreases with storage time and temperature (Tananuwong and Lertsiri 2010). Consequently, even rice of the same variety grown under the same conditions may show a significant difference in 2AP concentration due to differences in harvest date and postharvest handling.Several methods have been developed for the isolation and concentration of 2AP in aromatic rice samples for subsequent analysis by gas chromatography with nitrogen-phosphorus, flame ionization, or mass spectrometry as a detector. These methods include concentration of 2AP in rice by purge and trap (Buttery et al 1988), steam distillation/solvent extraction (Lin et al 1990), solvent extraction followed by direct injection (Bergman et al 2000), solid-phase microextraction (SPME) (Grimm et al 2001) and headspace analysis (Sriseadka et al 2006). Recently, SPME ...