Formation Of Heterocyclic Amines Research Articles

Mechanism of four dihydroflavones inhibiting heterocyclic amine formation in roasted pork patties―increasing the moisture

This study explored the inhibitory effect and mechanisms of four dihydroflavones (hesperetin, hesperidin, naringenin and naringin) addition on the production of five HCAs (IQ, DMIP, MeIQx, Norharman and PhIP) in roasted pork patties. Results indicated that the moisture holding capacity of dihydroflavones played a crucial role in their inhibition on HCA formation. The importance of moisture in the HCA formation was further confirmed by adding two water-retaining edible starches (corn and sweet potato starch). Significantly, the combination of edible starch and naringenin resulted in better inhibition of HCAs. The inhibition ratio of the mixture containing corn starch and naringenin on IQ, DMIP, MeIQx, Norharman and PhIP of pork patties were even up to 76.64%, 100%, 53.29%, 91.29% and 100%, respectively. This study offers a new idea of combination of dihydroflavone and edible starch to better control the HCA formation in roasted meat products.

Inhibitory action of antimicrobial peptides against the formation of carcinogenic and mutagenic heterocyclic amines in meat

Thermal processing of meat leads to the development of Maillard's reaction intermediates, and carcinogenic toxicants. For the first time, the effectiveness of three (HX-12A, HX-12B and HX-12C) antimicrobial peptides (AMPs) against the formation of heterocyclic amines (HAs) in chemical and meat model systems. The results showed that AMPs especially 12A and 12C have strong metal chelation potential (48 and 40% at 1 mg/ml) and antioxidant activity (35 and 25% at 1 mg/ml), respectively, which were endorsed by their secondary structure (FTIR analysis) in terms of high β-sheets (1628 cm−1 and 1672 cm−1) in those AMPs. UPLC-MS analysis revealed that 12A and 12C were the most capable AMPs in MeIQx and PhIP-producing chemical models, respectively, whereas 12B promoted the HAs formation even higher than control. In particular, 12C AMP significantly (P < 0.05) decreased the most abundant carcinogenic HAs (PhIP) up to 90% at 9 mg/100 g of fresh meat, whereas 12A inhibited up to 80% of mutagenic HAs at same level compared to control and 12B. Low Field Nuclear Magnetic Resonance (LF-NMR) test showed that inhibitory effect of 12A and 12C was mediated by means of retaining water (lower T22 and T23 relaxation time) inside the macromolecules. This favorable effect was also evidenced by significantly enhanced tryptophan fluorescent intensity. Finally, based on correlation and principle component analysis, the mechanism of action has been proposed. These outcomes recommend that 12A and 12C are potential AMPs for the attenuation of HAs in thermally processed meat-based products.

Evaluation of the potential of annatto seed powder to reduce the formation of heterocyclic amines in charcoal-grilled and pan-fried beef patties

Various strategies are being explored to reduce the formation of undesirable compounds during the thermal processing of foods. This study investigates the impact of incorporating annatto seed powder (Bixa orellana L.) into beef patties to reduce the formation of heterocyclic amines (HAs) during charcoal-grilling and pan-frying. A three-level full factorial design was used to assess the effect of both annatto seed powder concentration and cooking times on HAs formation. The results showed that HA formation increased with longer cooking times and decreased with higher concentrations of annatto seed powder. A significant reduction in HA content was observed in both charcoal-grilled and pan-fried beef patties when annatto seed powder was added, with a particularly notable 91 % reduction at the 1 % addition level. These findings demonstrate that the addition of annatto seed powder is a highly effective strategy for reducing HA formation in beef patties. Chemical compounds studied in this article2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) (PubChem CID: 62275); 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) (PubChem CID: 104739); 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx) (PubChem CID: 104855); 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (PubChem CID: 1530); 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) (PubChem CID: 5284474); 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) (PubChem CID: 5284476); 2-amino-9H-pyrido[2,3-b]indole (AαC) (PubChem CID: 62805); 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAαC) (PubChem CID: 62244); Bixin (PubChem CID: 5281226).

Inhibition of heterocyclic amines formation in grilled beef patties by red pepper stalk extracts

ABSTRACT To effectively inhibit the formation of heterocyclic amines (HAs) is crucial for human health. Plant extracts with strong antioxidant activity could be applied to meat processing to prevent the formation of HAs. The results indicated that red pepper (Capsicum annuum L.) stalk extracts (RPSEs) exhibited antioxidant activity during the thermal processing of meat by decreasing protein and lipid oxidation and neutralizing free radicals. And effectively inhibited the formation of HAs in beef patties with an inhibition rate ranging from 29.5% to 89.3%. These benefits were achieved without adversely affecting the physical or chemical properties of grilled beef patties, except for a slight increase in hardness. Betaine-based deep eutectic solvent extracts resulted in a significant reduction of 65.4–89.3% and 67.5–84.7% for free and protein-bound HAs. Our research introduces a novel approach to diminish the levels of HAs in meat, enabling food producers to create more safety meat options.

Effects of potential key substances in woodchips smoldering smoke on the formation of heterocyclic amines and polycyclic aromatic hydrocarbons in Frankfurter sausages

The Frankfurter sausages smoked with beech, oak, and alder, respectively, were used to clarify the underlying impact of the smoke chemical composition on the levels of heterocyclic amines (HAs) and polycyclic aromatic hydrocarbons (PAHs). The result indicated that different wood types significantly affected the profiles of target substances in food matrices. The beech-smoked samples had lower contents of total free HAs (5.98–6.80 ng/g dry-weight-DW), PAH4 (3.31–3.83 ng/g DW), and PAH8 (10.0–10.8 ng/g DW), whereas the alder pyrolysis usually led to higher hazardous residues (8.26–9.19 ng/g DW of total free HAs, 4.24–6.60 ng/g DW of PAH4 and 14.1–23.3 ng/g DW of PAH8). In addition, the differences in smoke chemical composition were attributed to the different proportions of 15 key identified substances. Among them, two aldehydes (5-methyl-furfural & furfural) and two phenols (phenol & 5-hydroxymaltol) may have synergistic or competitive inhibitory effects on the formation of HAs and PAHs in smoked meat products.

Effects and mechanisms of using “clean” smoke particles in the smoking process on the formation of β-carboline heterocyclic amines (β-CHAs) in smoked meat patties

β-Carboline heterocyclic amines (β-CHAs), known for their synergistic neurotoxic and carcinogenic effects, are predominantly produced by humans through cigarette smoke and food and are found particularly in meats cooked at high temperatures. Few studies have explored the differences in the mechanisms of accumulation of β-CHAs in smoked meat and meat processed at high temperatures. In this research, the concentration of β-CHAs in smoked meats prepared using a variety of wood materials was measured using LCMS/MS. Additionally, key volatile organic compound markers associated with β-CHAs accumulation in smoke were identified through GCMS and multivariate statistical analysis and subsequently confirmed in a chemical simulation system. Three types of strainers, each with a distinct aperture size, were used to assess the efficacy of particle filtration in reducing β-CHAs levels in smoked meat. The findings indicated that smoke exposure indeed increases the β-CHAs content of meat. However, only the strainer capable of filtering PM2.5-sized particles reduced the amount of β-CHAs present compared to the control group. In contrast, strainers with larger pore sizes facilitated excessive accumulation of β-CHAs. The presence of aldehydes such as 1 H-pyrrole-2-carboxaldehyde, 5-methylfurfural, benzaldehyde, furfural, and nonanal exhibited a positive correlation with the accumulation of β-CHAs. Conversely, phenolic compounds, including 2-methoxy-4-vinylphenol, 2-methoxy-5-methylphenol, p-cresol, phenol, 2-methoxy-4-(1-propenyl)-, (Z)-, phenol, 3-ethyl-, and phenol, 4-ethyl-2-methoxy-, showed a negative correlation. Thus, filters made from chelated carbonyl trap materials both chemically and physically disrupt the buildup of β-CHAs in smoked meats. The use of this approach will not only improve the quality of these products but will also contribute to decreasing the amount of inhalation pollutants released into the environment.

Effects of Phenols from Olive Vegetation Water on Mutagenicity and Genotoxicity of Stored-Cooked Beef Patties.

This explorative study aimed to assess the mutagenicity and genotoxicity of stored-cooked beef patties formulated with and without phenols (7.00 mg of phenols/80-g patty) extracted from olive vegetation water (OVW), as related to the formation of cholesterol oxidation products (COPs) and heterocyclic amines (HCAs). The patties were packaged in a modified atmosphere, sampled during cold storage (4 °C) for 9 days, and grilled at 200 °C. The genotoxicity was evaluated by the Comet assay. The patty extract was found to be genotoxic on primary peripheral blood mononuclear cells (PBMCs), while no mutagenicity was detected. The addition of OVW phenols significantly decreased the genotoxicity of the patty extract and reduced the total COPs content in stored-cooked patties (4.59 times lower than control); however, it did not affect the content of total HCAs (31.51-36.31 ng/patty) and the revertants' number. Therefore, these results demonstrate that the OVW phenols were able to counteract the formation of genotoxic compounds in stored-cooked beef patties.

Effects of basic amino acids on heterocyclic amines and quality characteristics of fried beef patties at low NaCl level

The impact of basic amino acids (Lysine, Arginine, Histidine) on the formation of total heterocyclic amines (HAs) was investigated in fried beef patties at 1% NaCl level. Different levels of basic amino acids (0.1%, 0.5%, 1%) significantly inhibited the formation of the total and individual HAs at 1% NaCl, and the inhibitory effect was more effective than 3% NaCl (6.19 ng/g, 26.93% inhibition) (P < 0.05). Lys at 1% reduced total HAs the most (2.46 ng/g, 70.88% inhibition), followed by 1% His (2.79 ng/g, 67.03% inhibition) and 1% Arg (3.43 ng/g, 59.51% inhibition). Compared to the 3% NaCl, the quality characteristics (moisture content, frying loss, texture profile, and color) of the fried beef patties were significantly improved when basic amino acids were added at 1% NaCl (P < 0.05). The lipid oxidation of fried beef patties was significantly inhibited by 1% Arg and 1% Lys at 1% NaCl level (P < 0.05). The results indicated that basic amino acids could inhibit the formation of total HAs while maintaining the quality of meat products at low NaCl condition.

Effect of Chinese quince proanthocyanidins on the inhibition of heterocyclic amines and quality of fried chicken meatballs and tofu.

Heterocyclic amines (HCAs) have potential carcinogenic and mutagenic activity and are generated in cooked protein-rich foods. Adding proanthocyanidins (PAs) to these foods before frying is an effective way to reduce HCAs. In this study, polymeric PAs (PPA) and ultrasound-assisted acid-catalyzed/catechin nucleophilic depolymerized PAs (UAPA, a type of oligomeric PA) were prepared from Chinese quince fruits (CQF). Different levels of PPA and UAPA (0.05%, 0.1%, and 0.15%) were added to chicken meatballs and tofu; then these foods were fried, and the content of HCAs in them after frying was investigated. The results showed that PPA and, particularly, UAPA significantly inhibited the formation of HCAs in fried meatballs and tofu, and this inhibition was dose-dependent. The inhibition of HCAs by both PPA and UAPA was stronger in the chicken meatballs than in fried tofu. The level of total HCAs was significantly reduced by 57.84% (from 11.93 to 5.03ng/g) after treatment of meatballs with 0.15% UAPA, with inhibition rates of 78.94%, 50.37%, and 17.81% for norharman, harman, and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), respectively. Of note, there was a negative correlation between water, lipid, protein, creatine, and glucose content and HCA content in the crust, interior, and whole (crust-plus-interior) measurements of all fried samples. Interestingly, PPA and UAPA were found more effective in inhibiting HCAs in the exterior crust than in the interior of the fried chicken meatballs. These results provide evidence that further studies on the reduction of the formation of harmful HCAs in fried foods by adding CQF PAs could be valuable to the fried food industry. PRACTICAL APPLICATION: Chinese quince proanthocyanidins treatments significantly inhibited the generation of heterocyclic amines (HCAs) in chicken meatballs and tofu when deep-fried. These results suggest that Chinese quince proanthocyanidins can be used as natural food additive for reducing HCAs in fried foods, laying the foundation for using Chinese quince fruit proanthocyanidins for HCA inhibition in the food industry.

Effects of grilling conditions on heterocyclic amines and quality attributes of grilled grass carp

SummaryThe effects of different grilling temperatures (220 °C, 250 °C, 280 °C, 310 °C) reaching the central temperature of 70 °C on heterocyclic amines (HAs) formation, moisture content, cooking loss, texture, colour, and sensory properties of grilled grass carp (Ctenopharyngodon idella) were investigated. The grilling condition significantly (P &lt; 0.05) affected cooking loss, moisture content, heterocyclic amines (HAs) formation, texture, colour and sensory properties. The results showed that 5 kinds of free and 9 kinds of protein‐bound heterocyclic amines were detected. The total amount of protein‐bound heterocyclic amines was much higher than that of the free state. Harman and Norharman, belonging to β‐carboline, constituted over 80% of the total content of heterocyclic amines. Considering the HAs formation and quality attributes, when the grilling condition was 280 °C for 10 min, the total content of HAs was the lowest, and the fish had moderate moisture content, firm texture, bright colour, and high sensory evaluation score.

A Disconnection for Rapid Access to Heterocyclic Benzylic Amines with Fully Substituted α-Carbons.

2- or 4-Pyridyl benzylic amines represent a privileged motif in drug discovery. However, the formation of heterocyclic benzylic amines with fully substituted α-carbons can require the execution of lengthy synthetic routes, which limit their application. Addition of various nucleophilic agents to Ellman's imines has been well established; however, there is no precedented literature reported for pyridyl-type nucleophiles, which are very important for medicinal chemistry. In this letter, we disclose the development of a one-step synthesis of heterocyclic benzylic amines with fully substituted α-carbons from heteroaryl halides and sulfinyl imines. Starting from 2,4-dibromopyridine, regioselective synthesis of 2- or 4-pyridyl benzylic amines could be achieved by choosing toluene or MTBE as a solvent.

Effects of seasoning addition and cooking conditions on the formation of free and protein-bound heterocyclic amines and advanced glycation end products in braised lamb

The accumulation of heterocyclic amines (HAs) and advanced glycation end products (AGEs) in thermally processed meats has been arising safety concerns. The effects of cooking conditions and seasoning addition on the formation of HAs and AGEs in Chinese traditional braised lamb were investigated by UPLC-MS/MS analysis. Soy sauce significantly increased the formation of HAs and AGEs, among which light soy sauce had the greatest promoting effect (69.45–15300.62 %). Conversely, spices inhibited HAs and AGEs formation, the inhibition rate of free HAs and AGEs reached 22.06–34.72 % when using 70 % ethanol extract. Hot blanching treatment and adding soy sauce and spices at a later stage could significantly suppress HAs and AGEs production. Flavonoids, including galangin, hesperidin, narirutin, etc., were identified as key effectors in spices. These findings help to promote awareness of the formation of HAs and AGEs in braised lamb and provide valuable insights for optimizing processing techniques to minimize their production.

Effect of temperature, pH, metal ions, and oil type on Chinese quince proanthocyanidins' thermal stability, antioxidant activity, and inhibition of heterocyclic amine formation during simulated frying

Heterocyclic amines (HCAs) are potent carcinogenic and mutagenic compounds generated during the frying of meat. The addition of Chinese quince proanthocyanidins (PAs) to fried food can reduce HCA formation. However, differences in food parameters (such as temperature, pH, presence of metal ions, and oil type) affect the efficacy of PA-mediated inhibition of HCA formation. In this study, two PAs (P1 and P2) of different molecular weights were prepared from Chinese quince. The effects of pH, metal ions, and oil type on the thermal stabilities, antioxidant capacities, and HCA-inhibitory abilities of P1 and P2 were studied during simulated frying. After simulated frying, the degree of polymerization (DP) and molecular weights of P1 decreased dramatically from 15224 Da to 2280 Da, while they decreased slightly in P2. Fe3+ decreased the thermal stability of P1 and P2 more than high pH; however, both factors reduced the HCA formation-inhibitory activity. Notably, the thermal stabilities of P1 and P2 in sunflower oil were lower than those in other oils; however, the inhibition of HCAs by P1 and P2 in sunflower oil (69.31% and 72.11%, respectively, for norharman) differed only slightly from that in the other oils. Multivariate analysis revealed positive correlations among thermal stabilities, antioxidant capacities, and HCA-inhibitory activities. Thus, Chinese quince PAs may be potentially used as food additives to mitigate the production of harmful HCAs in fried food.

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

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&lt;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%.

Formation and prediction of heterocyclic amines and N-nitrosamines in smoked sausages using back propagation artificial neural network.

Heterocyclic amines (HAs) and N-nitrosamines (NAs) are formed easily during the thermal processing of food, and epidemiological studies have demonstrated that consuming HAs and NAs increases the risk of cancer. However, there are few studies on the application of back propagation artificial neural network (BP-ANN) models to simultaneously predict the content of HAs and NAs in sausages. This study aimed to investigate the effects of cooking time and temperature, smoking time and temperature, and fat-to-lean ratio on the formation of HAs and NAs in smoked sausages, and to predict their total content based on the BP-ANN model. With an increase in processing time, processing temperature and fat ratio, the content of HAs and NAs in smoked sausages increased significantly, while the content of HA precursors and nitrite residues decreased significantly. The optimal network topology of the BP-ANN model was 5-11-2, the correlation coefficient values for training, validation, testing and all datasets were 0.99228, 0.99785, 0.99520 and 0.99369, respectively, and the mean squared error value of the best validation performance was 0.11326. The bias factor and the accuracy factor were within acceptable limits, and the predicted values approximated the true values, indicating that the model has good predictive performance. The contents of HAs and NAs in smoked sausages were significantly influenced by the cooking conditions, smoking conditions and fat ratio. The BP-ANN model has high application value in predicting the contents of HAs and NAs in sausages, which provides a theoretical basis for the suppression of carcinogen formation. © 2024 Society of Chemical Industry.

Modified Chinese quince oligomeric proanthocyanidin protects deep-frying oil quality by inhibiting oxidation

Chinese quince (Chaenomeles sinensis) fruit is an underutilized resource, rich in proanthocyanidins with antioxidant ability but poor lipid solubility. In this study, a novel modified oligomeric proanthocyanidin (MOPA) was prepared, which exhibited favorable lipid solubility (354.52 mg/100 g). It showed higher radical scavenging abilities than commercial antioxidant-BHA (butylated hydroxyanisole), both at 0.4–0.5 mg/mL. The addition of MOPA (0.04 %wt.) significantly increased the oxidative stability index of the soybean oil from 5.52 to 8.03 h, which was slightly lower than that of BHA (8.35 h). Analysis of the physicochemical properties and composition of oil during deep-frying showed that MOPA demonstrated significant antioxidant effects and effectively restricted the oil oxidation. This inhibition also delays the formation of heterocyclic amines (HAs) in fried food, thereby reducing the migration of HAs from food to deep-frying oil. Therefore, MOPA is a promising novel liposoluble antioxidant for protecting the quality of deep-frying oil.

Effect of mulberry leaf (Morus alba L.) extract on the quality and formation of heterocyclic amines in braised muscovy duck

Effect of mulberry leaf (Morus alba L.) extract on the quality and formation of heterocyclic amines in braised muscovy duck

Effect of structurally different pectin combined with polyphenol on water distribution, protein oxidation and heterocyclic amines formation in meat-based snacks

This research studied the effect of structurally different pectin combined with polyphenol (PP) on the physicochemical properties, inhibition of protein oxidation and heterocyclic amines (HAs) formation in fish meat snacks. The findings demonstrated that pectin with a high degree of esterification (DE) and average chain length (ACL) significantly reduced cooking loss, improved zeta potential values, and reduced particle size when combined with polyphenols, indicating an inhibition of protein aggregation and crosslinking. The addition of PP decreased the carbonyl formation as compared to individual polyphenol and control. Low Field Nuclear Magnetic Resonance (LF-NMR) analysis publicized that PP combination increased the water binding capacity, which resulted in less free water (T22 and T23) in those samples. Furthermore, LC/MS analysis revealed that PP combination, retained the water inside the macromolecules and prevented the relocation of water to the surface, which resulted in the less HAs formation as compared to control. The scanning electron microscopy (SEM) revealed that combination of PP samples formed a compact structure evidenced by less wide holes while control sample revealed loose structure. The findings suggest that 201 and 601 pectin combined with catechin have potential to improve the physicochemical properties and inhibition of hazardous compound formation.

Effects of Fingerroot (Boesenbergia pandurata) Oil on Microflora as an Antimicrobial Agent and on the Formation of Heterocyclic Amines in Fried Meatballs

This study aimed to determine the antibacterial activity of the essential oil of fingerroot (Boesenbergia pandurata) (EOF) as a natural preservative in ground meat and its effect on the formation of heterocyclic amines (HAs) in pan-fried meatballs. EOF was applied either by adding it to ground pork or marinating pork in it before grinding. In addition, the antibacterial activity of EOF was tested. Aerobic mesophilic total viable count (TVC), lactic acid bacteria (LAB), and Enterobacteriaceae bacteria were monitored. The results show that EOF exhibited strong antibacterial activity when added at concentrations of 1.0 and 2.5 wt%. Antimicrobial activity against TVC, LAB, and especially Enterobacteriaceae bacteria was observed at all EOF concentrations (0.25, 0.5, 1.0, and 2.5 wt%). A 2.5% concentration of EOF applied by marinating trimmings can extend the shelf-life of ground pork to 18 days, while 2.5% EOF applied via addition can extend the shelf-life to 15 days, compared with 3 days for the control sample. After frying the meatballs, the inhibitory effect on the formation of heterocyclic amines was only significant for MeIQx with the highest addition of EOF (2.5 wt%). Significant increases in the concentrations of all other HAs were determined by adding EOF (2.5 wt%).

Effects of quercetin and l-ascorbic acid on heterocyclic amines and advanced glycation end products production in roasted eel and lipid-mediated inhibition mechanism analysis

Eel is a commercially important marine fish, frequently featured as sushi or roasted preparations. This study determined the formation of heterocyclic amines (HAs) and advanced glycation end products (AGEs) in roasted eel and evaluated the inhibitory mechanism of quercetin and l-ascorbic acid on their formation. The results indicate a respective reduction of 75.07% and 84.72% in total HAs, alongside a decline of 23.03% and 39.14% in AGEs. Additionally, fundamental parameters of roasted eel, lipid oxidation indicators and precursors were measured to elucidate the mechanisms and impact of natural antioxidants on HAs and AGEs formation in roasted eel. Furthermore, endeavors were made to probe into the molecular mechanisms governing the influence of key differential lipids on the generation of HAs and AGEs through lipid-mics analysis. This research emphasizes the potential of natural antioxidants in preventing harmful substances formation during eel thermal processing, which is helpful to food manufacturers for healthier food production.