Simultaneous determination of the PhIP-proline adduct and related precursors by UPLC-MS/MS for confirmation of direct elimination of PhIP by proline

The effect and elimination pathway of proline on reducing PhIP and the effect of processing temperature, duration, and proline addition on the PhIP-proline adduct and its precursors were investigated. The results have demonstrated that PhIP and proline could condense to produce the adduct by direct heating, which could also be detected in the PhIP-producing model system and in beef patties with proline. The analytical method was optimized and has a good limit of detection (0.006-73 ng/mL), limit of quantification (0.021-245 ng/mL), recovery rate (about 80%-120%), and precision (below 15%). A high dose of proline (5.0%, w/w) promoted the formation of the adduct and reduction of PhIP; long heating duration and high temperature were not conducive to the formation of the adduct in beef patties. With increased addition of proline, creatine and creatinine decreased in a dose-dependent manner; phenylalanine and glucose did not show the same trend.