Abstract: To accurately determine the content of pyrazine compounds in the Maillard reaction, this study took the L-lysine hydrochloride/glucose model system as the research object, and conducted methodological evaluation on four analytical methods: liquid-liquid extraction (LLE), headspace-solid-phase microextraction (HS-SPME), stir bar sorption extraction (SBSE) coupled with gas chromatography-mass spectrometry (GC-MS), and direct injection coupled with high-performance liquid chromatography-mass spectrometry (HPLC-MS). To assess the performance of four methods in terms of linearity of fit, method detection limit (MDL), method quantification limit (MQL), spiked recovery, and relative standard deviation (RSD). The optimally performing LLE-GC-MS method was further applied to model reaction products such as glutamic acid/glucose and tyrosine/glucose, as well as real food samples including coffee extract, to validate the universality of this analytical approach. The results showed that both LLE-GC-MS and HPLC-MS exhibited excellent performance in terms of linearity (R² ≥ 0.999), spiked recovery rates (87.38%-124.26%, 85.80%-123.32%), repeatability (RSD < 10%), method detection limits (MDL ≤ 0.026 8 μg/mL), and method quantification limits (MQL ≤ 0.081 3 μg/mL). Among these methods, the extraction process of HS-SPME-GC-MS and SBSE-GC-MS were susceptible to external influences, resulting in slightly inferior repeatability compared with other two methods. LLE-GC-MS, featuring simple operation, efficient analytical workflow, and low instrument maintenance costs, is more suitable for high-throughput and precise quantification of samples from different systems, and has been successfully applied to the quantification of pyrazine in samples from other systems. This study provides a methodological basis for the quantitative analysis of flavor substances in Maillard reaction spices, and holds significant practical value for food flavor regulation and the development of thermally reactive spices.