Simultaneous Identification and Quantification of Phenylalanine, Glycine and Lysine amino acids and conjugated heterocyclic amines in Maillard chemical model system using RP-HPLC-DAD

Abstract

In this study, we developed a simple reversed-phase high-performance liquid chromatography (RP-HPLC) methodology for simultaneous identification and quantification of three types of mix: (1) phenylalanine and PHIP; (2) glycine and MeIQx; (3) lysine and 4,8 DiMeIQx in Maillard chemical model systems. In addition, we investigated the effects of different conditions on decreasing amino acids and heterocyclic amines (HCA) formation in chemical model systems. The results show that the developed methods used to separate the proposed mixes gave a relative standard deviation (RSD)% of less than 2% of peak area and retention time, thus proving the robustness and high reproducibility of the proposed method. The relationship between concentration and peak areas is linear with high statistical significance (p<0.001). The residual and slope of linear regression give a limit of detection LoD) and (LoQ) of phenylalanine and PHIP were (2.92- 0.029 ppm),(9.73- 0.098 ppm),respectively, which indicate the high sensitivity of the developed method. Moreover, the findings illustrate that a water ratio of 20% leads to a decrease in phenylalanine versus the largest amount of PHIP, and phenylalanine is the most active amino acid with a decreasing percentage up to 99%.