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A fast two-dimensional gas chromatography (GC-MS) method that uses heart-cutting and thermal extraction (TE) and requires no chemical derivatization was developed for the determination of anhydro-sugars in fine aerosols. Evaluation of the TE-GC-GC-MS method shows high average relative accuracy (≥90%), reproducibility (≤10% relative standard deviation), detection limits of less than 3 ng/μL, and negligible carryover for levoglucosan, mannosan, and galactosan markers. TE-GC-GC-MS- and solvent extraction (SE)-GC-MS-measured levoglucosan concentrations correlate across several diverse types of biomass burning aerosols. Because the SE-GC-MS measurements were taken 8 years prior to the TE-GC-GC-MS ones, the stability of levoglucosan is established for quartz filter-collected biomass burning aerosol samples stored at ultra-low temperature (−50 °C). Levoglucosan concentrations (w/w) in aerosols collected following atmospheric dilution near open fires of varying intensity are similar to those in biomass burning aerosols produced in a laboratory enclosure. An average levoglucosan-mannosan-galactosan ratio of 15:2:1 is observed for these two aerosol sets. TE-GC-GC-MS analysis of atmospheric aerosols from the US and Africa produced levoglucosan concentrations (0.01–1.6 μg/m³) well within those reported for aerosols collected globally and examined using different analytical techniques (0.004–7.6 μg/m³). Further comparisons among techniques suggest that fast TE-GC-GC-MS is among the most sensitive, accurate, and precise methods for compound-specific quantification of anhydro-sugars. In addition, an approximately twofold increase in anhydro-sugar determination may be realized when combining TE with fast chromatography