Abstract:An efficient and precise method for the determination of non-volatile flavor substances in carrots was developed to deeply analyze the material basis of carrot flavor formation. Gas chromatography-mass spectrometry （GC-MS） was combined with one-way analysis of variance （ANOVA） to systematically compare five different extraction methods. The methanol-water-chloroform （two-phase） extraction method was finally identified as the optimal solution，which showed good extraction efficiency for most substances and significantly improved the content of amino acids. In addition，35 combinations of derivatization temperatures were compared for optimization of the derivatization procedure，and the optimal derivatization conditions were finally determined as follows： methoxylation at 50 ℃ for 0.5 h and silylation at 70 ℃ for 1.5 h. Under the optimal derivatization conditions，the levels of 11 key flavor substances including D-fructose，D-glucose，sucrose，and malic acid were significantly higher than those under other conditions. The key flavor substances of four carrot varieties were determined and analyzed under the optimal derivatization conditions，which revealed that soluble sugars were the key flavor components distinguishing different carrot varieties. Finally，the reliability and stability of the method were verified by partial least squares discriminant analysis （PLS-DA）.