Determination of meat-cooking carcinogens from grilled pork by HPLC: Mitigation by beer marinades

Abstract

Recently the International Agency for Research on Cancer (IARC), the cancer agency of the World Health Organization, classified the consumption of red meat and processed meat as probably carcinogenic to humans (Group 2A) and carcinogenic to humans (Group 1) respectively. Grilling meat is a process that involves high temperatures that lead to production of cooking toxicants, such as heterocyclic aromatic amines (HAs) and polycyclic aromatic hydrocarbons (PAHs). To obtain realistic exposure levels of these two groups of mutagens analyses of the same samples using similar separation/detection techniques should be performed. The present study aims to determine HAs and PAHs formation in charcoal-grilled pork by high performance liquid chromatography (HPLC); and to evaluate the effect of beer marinades illustrating their impact on exposure to these carcinogens.Pork loin steaks (control) and pork loin steaks prior marinated in beer were charcoal grilled. Three beers (Pilsner beer (PB), non-alcoholic PB (P0B), and Black beer (BB)) were used as marinades. Antiradical activities of beers were assayed by DPPH. After solid-phase extractions, HAs were analyzed by HPLC with diode array and fluorescence (FLD) detections and PAHs were analyzed by HPLC-FLD. In charcoal grilled pork (control) eight PAHs, the PAH8 named by EFSA [1], were quantified, and their sum was around 21 ng/g. Chrysene> benz[a]anthracene> benzo[b]fluoranthene> benzo[a]pyrene contributed for 80% of PAH8. The sum of these compounds comprises the PAH4 [1]. Concerning HAs, PhIP > Trp-P-1 > 4,8-DiMeIQx > AC > MeAC were formed, totaling 20 ng/g of grilled pork.Free radical pathways are involved in the formation of these hazards, the addition of antioxidants may scavenge them. BB exhibited the strongest radical scavenging activity (68.0%), followed by P0B (36.5%) and PB (29.5%). Concerning PAHs formation, BB showed the highest inhibitory effect (53%), followed by P0B (25%) and PB (13%). The inhibitory effect of beer marinades on PAH8 increased with the increase of beer antioxidant activity. Concerning HAs, it was observed that all beers used as marinade significantly reduced PhIP, Trp-P-1 and AC (p < 0.05). 4,8- DiMeIQx was inhibited only by BB marinade. The inhibitory effect was higher using BB (90%), followed by PB (70%) and P0B (58%). A strong positive correlation was achieved between the inhibitory effects on total HAs and beer scavenging activity. Considering the consumption of 100 g of charcoal-grilled pork, that are easily attained in a single meal, PAH8 will exceed 2050 ng and similar amount of HAs. If meat was prior marinated in beer, the exposure to PAH8 decreases to 1782, 1550 and 974 ng/100 g for PB, P0B and BB respectively, and decreases HAs to 636, 869 and 197/100g for PB, P0B and BB respectively.The consumption of charcoal grilled pork entails a high exposure to PAHs exciding 2000 ng/100 g, and similar exposure to HAs. However beer marinades proved to be efficient mitigation strategies, especially black beer inhibiting 50% and 90% of formation of PAHs and HAs respectively.