To meet the need of a fast, robust, and reliable—but also sufficiently flexible—method to quantify fragrance allergens, an analytical workflow based on comprehensive two-dimensional gas chromatography (GCxGC) coupled to time-of-flight mass spectrometry (TOFMS) has been developed. This approach reduces the number of required analytical runs per sample. Here, the analysis of 57 fragrance allergens and their relevant isomers is demonstrated with GCxGC-TOFMS using the LECO Pegasus BTX 4D system equipped with the QuadJet™ thermal modulator using liquid nitrogen (LN2) as coolant. Examples of calibration curves and precision and accuracy metrics are provided. In addition, a perfume sample was analyzed under the same conditions the compatibility of this method with other analytical approaches, such as a non-target screening or characterization.