Abstract: In this study, we established a quantitative analytical method for the detection of histamine in various amino acid additives based on 6-aminoquinoline-N-hydroxysuccinimide formate (AccQ-Tag) derivatization coupled with ultra-performance liquid chromatography-tandem mass spectrometry (AccQ-UPLC-MS/MS). Samples were dissolved and diluted with 0.1% formic acid solution, derivatized according to the AccQ Fluor reagent protocol, and separated on a Waters ACQUITY UPLC BEH C₁₈ column using a gradient elution of acetonitrile/0.1% formic acid aqueous solution as the mobile phase. Detection was performed in the positive ion electrospray ionization (ESI⁺) mode with multiple reaction monitoring (MRM), and quantification was achieved using matrix-matched standard curves with internal standard calibration. Our results demonstrated that the derivatized histamine exhibited an excellent peak shape and was completely separated from the other amino acid derivatives. Excellent linearity was observed within the range of 1.22~5000 ng/mL, with coefficient of determination (R²) beyond 0.998. The limits of detection (LOD) and quantification (LOQ) were 1.22~2.44 and 2.44~4.88 mg/kg, respectively. At low-, medium-, and high-spiked concentrations, the recoveries of histamine in seven matrices ranged from 96.71% to 104.2%. The relative standard deviations (RSD, n=6) for precision, repeatability and 72-hour stability tests were within the ranges of 1.22%~3.66%, 1.86%~4.42%, and 1.20%~3.14%, respectively. This method allows for effectively overcoming the interference from high-concentration amino acid matrices in histamine trace-level detection, demonstrating high sensitivity, accuracy, and reliability. Taken together, this method is suitable for the detection of histamine contamination in amino acid-based additives.