A method for the direct determination of levoglucosan at the picogram per milliliter level in less than 1 mL of Antarctic ice has been developed. Chemical analysis is performed by high-performance liquid chromatography with triple quadrupole tandem mass spectrometric detection. Levoglucosan, a specific molecular marker for biomass burning, is identified by negative ion electrospray mass spectrometry using m/z 161/113, 161/101, 161/85, and 161/71 as monitoring ion transitions. Contamination problems were carefully taken into account by adopting ultraclean procedures during sampling and sample pretreatment phases. The limit of detection is 3 pg mL(-1) (0.3 pg absolute amount injected); the repeatability ranges between 20% and 50% at a concentration of 20 and 9 pg mL(-1), respectively. This methodology allowed the direct determination of levoglucosan in a 1 mL sample of Antarctic ice with concentration ranges between 4 and 30 pg mL(-1). To our knowledge these are the first levoglucosan concentrations reported for Antarctic ice.