Amino acids effects on heterocyclic amines formation and physicochemical properties in pan-fried beef patties.

The effects of surface application of amino acids on the formation of heterocyclic amines (HCAs) and meat quality properties were evaluated in pan-fried beef patties (230 °C/15 min). Tryptophan, lysine, leucine, and proline at three concentrations, 0.05%, 0.20%, and 0.50% (w/w), were tested. The meat crusts were analyzed for HCA content using liquid chromatography-tandem mass spectrometry. Results showed that surface application of all tested amino acids significantly reduced total HCA content (P < 0.05), and the interaction of amino acid type and concentration significantly affected (P < 0.05) both individual and total HCA formation. Tryptophan at 0.50% reduced total HCAs the most (0.92 ng/g, 93% inhibition), followed by 0.50% lysine (1.94 ng/g, 84% inhibition), while leucine (3.95 ng/g, 64% inhibition) and proline (4.71 ng/g, 56% inhibition) were less effective at 0.50%. In addition, applying amino acids to meat surface significantly influenced (P < 0.05) pH and surface color change of beef crusts; particularly, lysine at 0.20% and 0.50% increased pH and a* (redness) but reduced b* (yellowness), while tryptophan and leucine at 0.50% increased L* (whiteness). No significant effect was observed on cooking loss. Adding amino acids at 0.50% affected (P < 0.05) formation of aldehydes and pyrazines (as the key flavor compounds of fried beef). Overall, the results of this study suggested that adding amino acids to ground beef patties could effectively mitigate mutagenic HCA formation during cooking.

Determination of Heterocyclic Amines in Meat Matrices Using Enhanced Matrix Removal-Lipid Extraction and Liquid Chromatography-Tandem Mass Spectrometry.

A simple, fast, and efficient method, "enhanced matrix removal of lipids" (EMR-lipid), was proposed, optimized, and validated for identifying five polar heterocyclic amines (HCAs) in meat samples that ranged from high-protein (beef and chicken) to high-fat (pork bacon) matrices. The protocol involves an initial solid-liquid phase extraction followed by a rapid dispersive solid-phase extraction using EMR-lipid sorbents and salting-out partitioning. Acetonitrile containing formic acid at two levels (1% and 2%) efficiently extracted HCAs from different meat matrices. Liquid chromatography-tandem mass spectrometry (MS/MS) with selective reaction monitoring mode was developed for qualitative and quantitative analysis. The highest MS/MS responses and better peak separation of analytes were achieved by adjusting mobile phases to pH 3.0 with instrumental detection limits between 0.01 and 0.05 ng/mL. Good linearity of standard curves was obtained in both pure solvents and postspiked meat extracts between 0.5 and 50.0 ng/mL. The validation results showed good precision, accuracy, and sensitivity for detecting HCAs in spiked meat samples. Satisfactory recoveries of four HCAs were achieved: 65% to 111% in beef, 71% to 106% in bacon, and 42% to 77% in chicken. Matrix effects were also assessed and showed less than -20% of ion suppression in bacon extract, while a medium to high signal suppression was observed in beef (-37% to -55%) and chicken (-28% to -52%). This optimized EMR-lipid method provides acceptable results and advantages for determining trace level HCAs in complex meat matrices.

Amino Acids Inhibitory Effects and Mechanism on 2-Amino-1-Methyl-6-Phenylimidazo [4,5-b]Pyridine (PhIP) Formation in the Maillard Reaction Model Systems.

This study was to investigate the inhibitory effects of amino acids (AAs) on the formation of 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) and to evaluate the inhibition mechanism of PhIP in Maillard model systems. Different AAs were individually added into model systems heat-treated at 180 °C/1 h. The PhIP, phenylacetaldehyde (PheAce), and pyrazines derivatives were determined using HPLC and GC-MS. AAs significantly reduced (P < 0.05) PhIP levels in a dose-dependent response, ranking as: Trp = Lys > Pro > Leu > Met > Val > Ile > Thr > Phe > Asp, at the highest molar ratio. The PheAce content was gradually reduced with increasing AAs levels, suggesting that AAs may inhibit PhIP formation through scavenging the available PheAce. A correlation between PhIP inhibition and PheAce-scavenging activity of AAs was observed when PheAce and AAs were heated. The variety and quantity of pyrazines formed are highly depending on the type of AAs.