Content of heterocyclic amines and polycyclic aromatic hydrocarbons in pork,beef and chicken barbecued at home by Danish consumers

It is a well-known fact that, when meat is barbecued, several harmful components, including heterocyclic amines (HCA) and polycyclic aromatic hydrocarbons (PAH), may be formed. The aim of this study was to determine the HCA and PAH content in meat (pork, chicken and beef) when barbecued at home by Danish consumers according to their normal practice. With regard to HCA, beef contained the highest concentrations of 9H-pyrido[3,4-b]indole (norharman) and 2-methyl-β-carboline (harman), while chicken contained more 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) than pork and beef. The analysis of PAH showed a markedly higher concentration of PAH in beef compared with pork and chicken. In general, a correlation between the HCA content and the surface colour of the meat was found, the darker the colour the higher the HCA concentrations.     

Effects of cooking methods and levels on formation of heterocyclic aromatic amines in chicken and fish with Oasis extraction method

Heterocyclic aromatic amines (HCAs) are potent mutagenic and carcinogenic compounds formed during heat processing of proteinaceous food such as beef, poultry, and fish. The objective of this study was to measure nine HCAs in chicken chops and fish fillets cooked by various methods (microwave, oven, hot plate, pan-frying, and barbecuing) to different degrees of doneness (rare, medium, well done, and very well done). Total HCA amount in chicken changed between 0.24 and 8.21 ng/g, and not only the highest total amount but also the lowest total HCA amount was found in microwave cooked chicken samples. The highest total HCA amount found in fish for microwave, oven, hot plate, pan-frying, and barbecuing were 18.09, 4.28, 3.12, 6.98, and 5.22 ng/g, respectively. The results show that microwave cooking alone is found to possess the highest total HCA amount, followed by pan-frying, and barbecuing of meat samples, and the total HCA amount in cooked samples is low if cooked to rare and medium degrees of doneness. 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP), 2-amino-9H-pyrido[2,3-b]indole (AαC), and 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAαC) were not detected in any samples.     

Effects of skin and grilling method on formation of heterocyclic amines in chicken pectoralis superficialis muscle

Mutagenic heterocyclic amines (HAs) originate in processed proteinaceous foods. The effects of the presence of skin (with vs. without) and of grilling method (two plate vs. infrared) on the content of HAs in grilled chicken pectoralis superficialis muscle (temperature, 220 °C) were investigated. HA precursors (creatine, creatinine, free amino acids and carbohydrates) and HAs of these raw and grilled breast muscles were determined. The muscles originated from 24 birds of either sex (provenance Ross; aged 40-45 days). The HA content was determined in homogenates of the upper and lower surface slices of the grilled muscles (T_i = 82 °C). A higher content of total free amino acids was seen for the muscle (27.1 mmol kg⁻¹ raw meat) than for the skin (21.7 mmol kg⁻¹ raw meat). The creatine, creatinine and carbohydrate levels in the skin were below the limits of detection. The contents of creatine (31.8-38.7 mmol kg⁻¹) and creatinine (0.24-0.33 mmol kg⁻¹) in the breast muscle were determined. Relatively high levels were seen for glucose (23 mmol kg⁻¹ raw meat) and fructose (10 mmol kg⁻¹ raw meat) in the muscle, with other sugars present at low levels (<2 mmol kg⁻¹ raw meat). For the chicken muscle grilled on a two-plate grill, the contents of total HAs (PhIP, MeIQx, DiMeIQx, Harman and Norharman) were lower with the skin in place than in the muscle grilled without the skin (3.5 μg kg⁻¹vs. 4.8 μg kg⁻¹). Also, during infrared grilling with the skin, lower amounts of HAs were formed than with grilling on the two-plate grill (2.4 μg kg⁻¹vs. 3.5 μg kg⁻¹). On average, the infrared-grilled samples with skin contained 3-fold more total HAs than similar samples without the skin (2.4 μg kg⁻¹vs. 0.8 μg kg⁻¹), with the highest levels seen for PhIP and MeIQx.     

Effects of ageing and low internal temperature of grilling on the formation of heterocyclic amines in beef Longissimus dorsi muscle

The mutagenic heterocyclic amines (HAs) originate in processed proteinaceous food. The effects of ageing (non-aged - i.e. 24 h post mortem vs. 14 and 28 days post mortem kept at 1 °C) and final internal temperature on cooking (T_i, 65 and 80 °C) on the content of HAs in grilled steaks (two-plated grill, temperature of 220 °C) were studied. HA precursors (creatine, creatinine, and free amino acids) and ageing indicators, such as instrumentally measured colour values, pH_ultimate values and length of myofibrilar fragments on raw and cutting values on grilled beef Longissimus dorsi muscles were determined. The muscles originated from eight commercially slaughtered Simmental bulls, 19-20 months old. The content of HAs was determined by a solid-phase extraction procedure. Meat ageing is accompanied by large changes in the chemical composition and structure of muscle tissues. In general, all the ageing indicators and precursors of HAs were influenced by ageing time at the 5% level or less. Creatine content declined significantly (non-aged: 6.00 mg g⁻¹, 14 days: 5.82 mg g⁻¹, and 28 days: 5.55 mg g⁻¹) and creatinine increased with days of ageing (non-aged: 0.19 mg g⁻¹, 14 days: 0.24 mg g⁻¹, and 28 days: 0.26 mg g⁻¹). Higher contents of total free amino acids were determined after 14 and 28 days of storage (28.18 μmol g⁻¹ and 37.59 μmol g⁻¹) than in non-aged beef (19.00 μmol g⁻¹). In this study, two HAs were determined: MeIQx (2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline) and PhIP (2—amino-1-methyl-6-phenylimidazo-[4,5-b]pyridine). The content of HAs increases with ageing. At lower T_i, more MeIQx was formed; at higher T_i, more PhIP was formed. MeIQx was present in all samples while PhIP was found only in samples grilled to higher T_i. Samples treated to T_i = 80 °C generally contained less HAs (non-aged meat: 0.20 ng g⁻¹, 14 days: 0.26 ng g⁻¹, and 28 days: 0.28 ng g⁻¹) than samples treated to T_i = 65 °C (non-aged meat: 0.19 ng g⁻¹, 14 days: 0.36 ng g⁻¹, and 28 days: 0.39 ng g⁻¹) on account of MeIQx thermolability.     

Effects of high pressure/thermal treatment on lipid oxidation in beef and chicken muscle

Lipid oxidation was studied in beef and chicken muscle after high pressure treatment (0.1–800 MPa) at different temperatures (20–70 °C) for 20 min, prior to storage at 4 °C for 7 days. Pressure treatment of beef samples at room temperature led to increases in TBARS values after 7 days storage at 4 °C; however, the increases were more marked after treatment at pressures ?400 MPa (at least fivefold) than after treatment at lower pressures (less than threefold). Similar results were found in those samples treated at 40 °C, but at 60 °C and 70 °C pressure had little additional effect on the oxidative stability of the muscle. Pressure treatments of 600 MPa and 800 MPa, at all temperatures, induced increased rates of lipid oxidation in chicken muscle, but, in general, chicken muscle was more stable than beef to pressure, and the catalytic effect of pressure was still seen at the higher temperatures of 50 °C, 60 °C and 70 °C. The addition of 1% Na₂EDTA decreased TBARS values of the beef muscle during storage and inhibited the increased rates of lipid oxidation induced by pressure. The inhibition by vitamin E (0.05% w/w) and BHT (0.02% w/w), either alone or in combination, were less marked than seen with Na₂EDTA, suggesting that transition metal ions released from insoluble complexes are of major importance in catalysing lipid oxidation in pressure-treated muscle foods.     

固相萃取-液相色谱-串联质谱法测定市售卤肉制品中5种杂环胺含量

建立了LC-MS/MS快速检测卤肉制品中MeIQ、MeIQx、4,8-DiMeIQx、7,8-DiMeIQx和PhIP 5种杂环胺的方法。样品经40 g/L NaOH-甲醇溶液超声提取,用聚苯乙烯二乙烯苯固相萃取小柱(Poly-Sery PSD)净化,在电喷雾离子源正离子(ESI⁺)多反应监测(MRM)模式下进行测定。5种杂环胺化合物在0.1~10 μg/L范围内线性关系良好,决定系数R²在0.9976~0.9997之间,在空白样品中的平均添加回收率为76.9%~127.7%,日内相对标准偏差(RSD)为1.0%~9.5%,日间相对标准偏差(RSD)为2.1%~8.7%。在信噪比(S/N)分别为3和10时各化合物的LODs在0.01~0.1 μg/kg,LOQs在0.025~0.25 μg/kg。该方法分析时间短,准确度高,精密度好,可满足卤肉中多种杂环胺同时检测的要求。     

Inhibitory activities of dietary phenolic compounds on heterocyclic amine formation in both chemical model system and beef patties

The correlation between radical scavenging capacity and inhibitory activity in PhIP formation of 12 food-derived antioxidative phenolic compounds was investigated. Very poor correlation was found between their radical scavenging activity assessed by Trolox equivalent antioxidant capacity assay and PhIP formation inhibition in a model system. The effects of several of these polyphenols were further evaluated using beef patties. Remarkably, theaflavin 3,3'-digallate, epicatechin gallate, rosmarinic acid, and naringenin were capable of simultaneously reducing the levels of PhIP, MeIQx, and 4,8-DiMeIQx. Moreover, the inhibition of the formation of one of these HAs was not compromised by the inhibition of the formation of another HA. Naringenin, a flavonoid found in many citrus fruits, was found to be the most promising inhibitor in both chemical model system and beef patties, suggesting its great potential for practical application in daily cuisine.     

HPLC法检测市售传统肉制品中的杂环胺含量

本文研究了市售传统肉制品中杂环胺的含量,根据加工工艺不同,对肉制品中进行分类研究,应用高效液相色谱法(high performance liquid chromatography)检测市售传统肉制品中杂环胺(Heterocyclic Amines)的种类和含量,从购买来源(个人摊点、超市和专售店)、种类及加工工序(酱卤、油炸-酱卤、酱卤-烤制以及烤制)角度分析传统肉制品中HAs的特点,并分析不同种肉制品基本化学成分。研究表明:不同肉制品中基本成分含量各不相同;个人摊点中HAs含量比超市和专售相对较高;不同加工工序中酱卤-油炸和烤制中HAs的含量相对较高,含量高达13.40 ng/g,并且所有工序中都检测到Norharman和Harman;鸡皮比鸡肉(不带皮)中HAs含量高,酱卤鸡腿、酱卤烧鸡和叫花鸡皮中HAs的含量分别是鸡肉(不带皮)HAs含量的2.8、1.5和1.5倍。     

Differentiation in improvements of gel strength in chicken and beef sausages induced by transglutaminase

This research investigated the improvement in the texture of chicken and beef sausages induced by using microbial transglutaminase (MTG). The ε-(γ-glutamyl)lysine (G-L) content and the extractability of myofibrillar proteins from these sausages were also investigated. Treatment with MTG significantly affected the breaking strength score in both meat types, especially for beef cooked at 80°C (p<0.001). The protein concentration of both meat types treated with MTG and extracted in water-soluble protein solution (WSP) was slightly decreased; compared with a significant decrease (p<0.003) in samples extracted in Guba-Straub-ATP solution (GS-ATP). The variation in protein extractability of both meat types could lead to some considerations of the mechanisms and the high affinity reaction between MTG and myosin heavy chain (MHC). SDS-PAGE analysis revealed significant changes in the density of the bands after adding MTG, especially for the beef samples. The G-L content in the presence of MTG was double that in control samples of both meat types. The amount of crosslinking in chicken and beef meat was different and found to be reasonable. Collectively, this suggests that the binding ability of myofibrillar proteins with MTG is strong and dominated by MHC. There was a unique reaction among MHC proteins with MTG molecules considered as a very advantageous reaction. This leads us to suggest that the functional properties of MTG make it a beneficial protein-binding agent, positively helping the functionality of proteins to improve the texture and gelation of meat products that are treated mechanically, such as sausages. Some variation in gel improvement level between chicken and beef sausages was observed; this resulted from the variation in meat proteins in response to MTG, as well as to the original glutamyl and lysine contents.     

Inhibitory effect of mixture herbs/spices on formation of heterocyclic amines and mutagenic activity of grilled beef

ABSTRACT

Natural antioxidants in spices and herbs have attracted considerable attention as potential inhibitors against the formation of mutagenic heterocyclic amines (HCAs) in heat-processed meat. In this study, the inhibitory activity of four spices/herbs and their mixtures on HCAs formation in grilled beef were examined. A simplex centroid mixture design with four components comprising turmeric, curry leaf, torch ginger and lemon grass in 19 different proportions were applied on beef samples before grilling at 240 ºC for 10 min. The HCAs were extracted from the samples using solid phase extraction (SPE) method and analysed using Liquid chromatography mass spectrometry LC-MS/MS. All spices/herbs in single or mixture forms were found to reduce total HCA concentrations in marinated grilled beef ranging from 21.2% for beef marinated with curry leaf to 94.7% for the combination of turmeric and lemon grass (50:50 w/w). At the optimum marinade formula (turmeric: lemon grass 52.4%: 47.6%), concentration of 2-amino-3-methylimidazo[4,5-f]quinolone (IQ), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), Harman, Norharman and AαC were 2.2, 1.4, 0.5, 2.8 and 1.2 ng/g, respectively. The results of the mutagenic activity demonstrated that this optimised marinade formula significantly (p < 0.05) diminished mutagenicity of marinated grilled beef in bacterial Ames test.     

不同食品基质中杂环胺的检测技术研究进展

杂环胺是富含蛋白质的食品原料在热加工过程中产生的一类多环芳香族化合物,具有强烈的致癌、致突变性。杂环胺在热加工食品中一般以痕量水平存在,并且食品基质复杂,因此,高效、高选择性的样品前处理方法和高灵敏度的仪器分析手段是不同食品基质中痕量杂环胺分析检测的关键。该文综述2010年～2019年不同食品基质中杂环胺的分析方法,包括固相萃取、分散固相萃取等前处理技术以及气相色谱-质谱法、高效液相色谱法和超高效液相色谱串联质谱法等仪器检测方法,并与典型检测方法的性能进行比较。最后,结合现有研究对食品中杂环胺分析的未来发展趋势进行展望。     

Effect of cooking methods on the formation of heterocyclic aromatic amines in chicken and duck breast

Heterocyclic aromatic amines (HAAs), potent mutagens/carcinogens, are pyrolysis formed during the cooking of meat and fish. In the present study, the effects of various cooking methods, pan-frying, deep-frying, charcoal grilling and roasting on the formation of HAAs in chicken breast and duck breast were studied. The various HAAs formed during cooking were isolated by solid-phase extraction and analyzed by high-performance liquid chromatography (HPLC). Results showed that chicken breast cooked by charcoal grilling contained the highest content of total HAAs, as high as 112 ng/g, followed by pan-fried duck breast (53.3 ng/g), charcoal grilled duck breast (32 ng/g), pan-fried chicken breast (27.4 ng/g), deep-fried chicken breast (21.3 ng/g), deep-fried duck breast (14 ng/g), roasted duck breast (7 ng/g) and roasted chicken breast (4 ng/g). For individual HAA, the most abundant HAA was 9H-pyrido-[4,3-b]indole (Norharman), which was detected in charcoal grilled chicken breast at content as high as 32.2 ng/g, followed by 1-methyl-9H-pyrido[4,3-b] indole (Harman) and 2-amino-1-methyl-6-phenylimidazo[4,5-f]pyridine(PhIP) at 32 and 31.1 ng/g in charcoal grilled chicken breast, respectively. The content of PhIP in pan-fried duck and chicken breast were 22 and 18.3 ng/g, respectively. Generally, the type and content of HAAs in cooked poultry meat varies with cooking method and cooking conditions.     

Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, chicken and turkey

Many meat products are composed of two or more meat species. To determine the proportion of these meat fractions, a quantitative multiplex PCR was developed for the quantification of beef, pork, chicken and turkey. This system proved its applicability, precision and accuracy in examining different meat products from the market. Thus it allows the efficient control of composed meat products in official food control and production control laboratories.     

温和条件下模型体系中烧烤风味及杂环胺形成测定

以美拉德模型体系葡萄糖-肌酸酐-氨基酸（丙氨酸、苏氨酸、甘氨酸）为研究对象,探索3 个体系在130 ℃条件下反应3 h后风味物质及杂环胺的形成。采用固相微萃取-气相色谱-质谱联用方法检测表明,3 种模型均产生烧烤风味物质,主要为烷基吡嗪类。高效液相色谱分析表明,丙氨酸模型（Ⅰ）和甘氨酸模型（Ⅲ）均检测到2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉、2-氨基-3,4,8-三甲基咪唑并[4,5-f]喹喔啉和2-氨基-3,7,8-三甲基咪唑并[4,5-f]喹喔啉3 种杂环胺,苏氨酸模型（Ⅱ）检测到了2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉和2-氨基-3,4,8-三甲基咪唑并[4,5-f]喹喔啉2 种杂环胺;3 种模型体系均未检测到2-氨基-3-甲基咪唑并[4,5-f]喹啉和2-氨基-3,8-二甲基咪唑并[4,5-f]喹喔啉。130 ℃温和条件下,1.5 h内各模型体系中杂环胺的生成量很少,1.5 h后杂环胺总量随着加热时间的延长逐渐增多,3 h时模型Ⅰ、Ⅱ、Ⅲ中杂环胺总量分别为（375.50±15.80）、（414.00±18.40）、（363.50±12.20） ng/mL。本研究为肉制品加工尤其是烤肉制品加工如何控制加工条件,从而实现美拉德反应定向控制、减少杂环胺的产生提供一定参考。     

Encapsulated phosphates reduce lipid oxidation in both ground chicken and ground beef during raw and cooked meat storage with some influence on color,pH, and cooking loss

Effects of encapsulated sodium tripolyphosphate (STP), sodium hexametaphosphate (HMP) and sodium pyrophosphate (SPP) on lipid oxidation in uncooked (0, 2, 24 h) and cooked (0, 1, 7 d) ground chicken and beef during storage were determined. Ten phosphate treatments included a control (no phosphate), three unencapsulated (u) at 0.5% and three encapsulated (e) phosphates (0.5%) each at a low (e-low) and high (e-high) coating level. Two heating rates (slow, fast) were investigated. Cooking loss (CL), pH, color, orthophosphate (OP), TBARS and lipid hydroperoxides (LPO) were determined. A fast heating and uSTP resulted in lower CL (p < 0.05). Orthophosphate increased with phosphate incorporation, slow heating and storage (p < 0.05). Encapsulated phosphates and increased coating level reduced OP (p < 0.05). Unencapsulated STP increased CIE a* and pH, whereas uSPP decreased CIE a* and pH (p < 0.05). Encapsulated phosphates and the greater coating level had no effect on the pH in cooked samples. Not increased coating level but encapsulated phosphates decreased lipid oxidation in cooked samples (p < 0.05).     

生姜对烘烤猪肉饼品质及杂环胺形成的影响

选取猪肉与生姜为研究对象,探究不同生姜浓度（0.5%、1.0%、1.5%）对烘烤猪肉饼品质及杂环胺（heterocyclic aromatic amines,HAAs）生成的影响。结果表明,生姜能够降低猪肉饼的质量损失率并且改善色泽。生姜对8种HAAs（MeIQ、MeIQx、4,8-DiMeIQx、PhIP、IQx、IQ、AαC、Trp-P-2）均具有很好的抑制效果。浓度为 1.5%的生姜对烘烤猪肉饼的总HAAs含量抑制率为55.45%。随着生姜浓度的增加,对游离氨基酸的消耗减少。因此生姜对烘烤猪肉饼中的HAAs有很好的抑制活性。     

Effect of varying salt and fat levels on the sensory quality of beef patties

The interactive effects of varying levels of salt and fat on the sensory and physiochemical properties of beef patties were investigated. Twenty beef patties with varying levels of fat (30% 40% 50% 60% w/w) and salt (0.5%, 0.75% 1.0% 1.25% 1.5% w/w) were manufactured. All samples were assessed instrumentally for colour, moisture, fat, cooking loss and texture profile analysis. Sensory consumer evaluation was conducted using 25 consumers. The consumers rated each coded product, in duplicate, in terms of colour, texture, tenderness, juiciness, salt, taste, meat flavour, off flavour and overall acceptability. The data indicate that the most consumer acceptable beef patty was that containing 40% fat with a salt level of 1%. This is a 20% decrease in fat and a 50% decrease in salt levels when compared to commercial patties available in Ireland and the UK.     

Inhibitory effect of marinades with hibiscus extract on formation of heterocyclic aromatic amines and sensory quality of fried beef patties

Heterocyclic aromatic amines (HAA) are carcinogenic compounds found in the crust of fried meat. The objective was to examine the possibility of inhibiting HAA formation in fried beef patties by using marinades with different concentrations of hibiscus extract (Hibiscus sabdariffa) (0.2, 0.4, 0.6, 0.8 g/100 g). After frying, patties were analyzed for 15 different HAA by HPLC-analysis. Four HAA MeIQx (0.3–0.6 ng/g), PhIP (0.02–0.06 ng/g), co-mutagenic norharmane (0.4–0.7 ng/g), and harmane (0.8–1.1 ng/g) were found at low levels. The concentration of MeIQx was reduced by about 50% and 40% by applying marinades containing the highest amount of extract compared to sunflower oil and control marinade, respectively. The antioxidant capacity (TEAC-Assay/Folin–Ciocalteu-Assay) was determined as 0.9, 1.7, 2.6 and 3.5 μmol Trolox antioxidant equivalents and total phenolic compounds were 49, 97, 146 and 195 μg/g marinade. In sensory ranking tests, marinated and fried patties were not significantly different (p > 0.05) to control samples.     

Simultaneous formation of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatic amines (HCAs) in gas-grilled beef satay at different temperatures

This study investigated the simultaneous formation of polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatic amines (HCAs) in gas-grilled beef satay at different temperatures (150, 200, 250, 300, and 350°C). Solid-phase extraction (SPE) was used for sample clean-up. Fifteen PAHs were determined using high performance liquid chromatography with fluorescence detection (HPLC-FLD) and nine HCAs were quantified using liquid chromatography tandem-mass spectrometry (LC-MS/MS) with a gradient programme. The lowest significantly concentrations of PAHs and HCAs were generated at 150°C; the formation of PAHs and HCAs simultaneously increased with temperatures. Benzo[a]pyrene was detected in all samples and increased markedly at 300 and 350°C. The sums of 4 PAHs (PAH4) in marinated beef satay at 300 and 350°C exceeded the maximum level in Commission Regulation (EU) 2015/1125. Significant reductions of polar and non-polar HCAs (except PhIP) were found in marinated beef satay across all temperatures. Overall, PAHs and HCAs showed opposite trends of formation in beef satay with marination.