使用同位素稀释-HPLC-MS/MS建立了检测游离和蛋白质结合的晚期糖基化终产物（AGEs）和4-甲基咪唑（4-MI）的定性和定量方法，并成功应用于cookie和模型系统。讨论了不同温度（160-220°C）对饼干中游离和蛋白质结合的AGEs和4-MI形成的影响，并验证了蛋白质结合AGEs和4-MI的可能模型系统（使用小麦面筋蛋白葡萄糖蔗糖的美拉德反应途径1；使用小麦面筋蛋白CML/CEL/4-MI的直接添加途径1）。结果表明,蛋白质结合的CML,CEL,和4-MI高于游离含量，具有随温度先增加后降低的趋势，在饼干中达到200°C的最大值。在模型系统中，蛋白质结合的CML水平，CEL,4-MI高于自由CML，CEL,4-MI蛋白质结合的CML，CEL,和4-MI在模型系统1中分别占总量的90.73、87.64和97.56%，而在模型系统2中分别占总量的68.19、59.00和50.96%。相比之下,蛋白质结合的CML，CEL,4-MI可以很容易地通过美拉德反应直接生成。
A qualitative and quantitative method for detecting free and protein-bound advanced glycation end products (AGEs) and 4-methylimidazole (4-MI) was established using isotope dilution-HPLC-MS/MS, and successfully applied in cookies and model systems. The effects of different temperatures (160-220 °C) on the formation of free and protein-bound AGEs and 4-MI in cookies were discussed, and the possible model systems (Maillard reaction pathway 1 using wheat gluten protein + glucose + sucrose; direct addition pathway 1 using wheat gluten protein + CML/CEL/4-MI) of protein-bound AGEs and 4-MI were verified. The results showed that the contents of protein-bound CML, CEL, and 4-MI were higher than free content with a tendency of increasing first and subsequently decreasing with temperature, reaching a maximum at 200 °C in cookies. In the model systems, the levels of protein-bound CML, CEL, and 4-MI are higher than those of free CML, CEL, and 4-MI. The protein-bound CML, CEL, and 4-MI accounted for 90.73, 87.64, and 97.56% of the total amount in the model system 1, while accounting for 68.19, 59.00, and 50.96% in the model system 2, respectively. In comparison, protein-bound CML, CEL, and 4-MI could be easily generated directly by Maillard reaction.
