高效液相色谱法测定烧烤制品中5种杂环胺的含量

目的 建立高效液相色谱法检测烧烤制品中5种杂环胺(heterocyclic aromatic amines, HAAs)含量的方法。方法 样品经乙酸乙酯、三乙胺、氨水提取, 丙基磺酸(propanesulfonic acid, PRS)和C18固相萃取小柱净化, 采用TSK-gel ODS-80TM色谱柱分析, 以乙腈和0.05 mol/L醋酸-醋酸铵缓冲液(pH=3.4)为流动相进行梯度洗脱, 紫外-荧光检测器的串联方式检测杂环胺含量。结果 2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶(2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine, PhIP)、9H-吡啶并[4,3-b]吲哚(9H-pyrido[4,3-b]indole, Norharman)、1-甲基-9H-吡啶并[4,3-b]吲哚(1-methyl-9H-pyrido[4,3-b]indol, Harman)、2-氨基-9H-吡啶并[2,3-b]吲哚(2-amine-9H-pyrido[2,3-b]indol, AaC)在0.1~500 μg/L范围呈良好的线性关系, 2-氨基-3-甲基咪唑并[4,5-f]喹啉(2-amino-3-methylimidazo(4,5-f)quinoline, IQ)在0.1~1000 μg/L范围呈良好的线性关系, 相关系数r2均大于99.3%, 加标回收率为74.06%~93.77% (n=6), 相对标准偏差(relative standard deviation, RSD)为3.41%~5.93% (n=6)。结论 该方法简便快速、结果准确、灵敏度高, 适合于肉制品中的杂环胺含量的检测。通过检测结果可知不同原料中5种杂环胺含量不同, 相同原料烧烤条件不同, 产生的杂环胺含量也不相同。     

模型体系中PhIP、Norharman、Harman 3种杂环胺的形成与抑制

采用模型体系探讨2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶（2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine,PhIP）、9H-吡啶并[3,4-b]吲哚（9H-pyrido[3,4-b]indole,norharman）、1-甲基-9H-吡啶并[3,4-b]吲哚（1-methyl-9H-pyrido[3,4-b]indole,harman）3 种杂环胺的形成规律及抑制措施。结果表明：随着加热温度的升高与加热时间的延长,模型体系中PhIP、Norharman和Harman的形成量均不断增多,其中PhIP对加热温度较为敏感,100、125 ℃反应20 min不能形成PhIP,150 ℃加热时PhIP生成量开始逐渐增多,175～200 ℃加热时PhIP形成量急剧增加;单独加热色氨酸即能产生Norharman和Harman,加入葡萄糖时Norharman和Harman的形成量进一步提高;本研究建立的同时检测色氨酸、Norharman和Harman的色谱方法明确表明,色氨酸是Norharman和Harman的前体物;高良姜素及其类似物山柰酚和槲皮素均能在一定程度上减少PhIP的形成,但只有高良姜素能降低Harman和Norharman的形成量,而山柰酚和槲皮素不但没有抑制作用反而在一定程度上提高了Harman和Norharman的形成量,进一步证实β-咔啉类杂环胺形成的复杂性。     

前体物含量对杂环胺形成的影响

将牛、猪、羊、鸡、鸭和鹅肉分别在200℃下煎烤10 min,分析其原料肉中前体物的含量与加工肉制品中杂环胺的形成量,以探讨不同动物原料肉中杂环胺的形成量及原料肉中前体物的含量对杂环胺形成的影响。结果表明,6种原料肉中前体物含量差异较大,肌酸与葡萄糖的摩尔浓度比在0.89～9.84之间。加工肉制品中共检测出10种杂环胺,其中9H-吡啶并[3,4-b]吲哚(Norharman)与1-甲基-9H-吡啶并[3,4-b]吲哚(Harman)在加工肉品中含量最高,分别在10.90～24.16 ng/g与4.64～14.04 ng/g之间。而2-氨基-3,8-二甲基咪唑并[4,5-f]喹喔啉(MeIQx)在牛肉中形成量较高,达3.55 ng/g,2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶(PhIP)在禽肉中易于产生,其中鸡肉中含量最高,达15.26 ng/g。PhIP与肌酸和葡萄糖的摩尔浓度比存在相关性(P<0.05),随着葡萄糖浓度的增高,PhIP的形成量减少。     

葡萄籽提取物对烤羊肉中杂环胺形成的影响

将0.05%VE,0.1%与0.5%葡萄籽提取物,0.1%与0.5%槲皮素分别添加于羊肉表面,在200℃下煎烤10 min,利用固相萃取-配备二极管阵列检测器和荧光检测器的高效液相色谱分析烤羊肉中15种杂环胺的形成量,以探讨天然提取物对烤羊肉中杂环胺形成量的影响。结果表明:烤羊肉中共检测出10种杂环胺,其中极性杂环胺4种,非极性杂环胺6种,总量达51.68 ng/g。在极性杂环胺中2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶(PhIP)的形成量较高,达10.50 ng/g;非极性杂环胺中9H-吡啶并[3,4-b]吲哚(Norharman)与1-甲基-9H-吡啶并[3,4-b]吲哚(Harman)的形成量较高,分别达19.66 ng/g与12.23 ng/g。烤羊肉中添加天然抗氧化剂只对极性杂环胺有抑制作用,其中添加0.5%葡萄籽提取物可以显著抑制2-氨基-3甲基咪唑并[4,5-f]喹啉(IQ),2-氨基-3,8-二甲基咪唑并[4,5-f]喹喔啉(MeIQx),2-氨基-3,4,8-三甲基咪唑并[4,5-f]喹喔啉(4,8-DiMeIQx)与PhIP 4种极性杂环胺的形成(P<0.05),其生成量分别减少36.65%,67.73%,35.63%与30%。天然抗氧化剂对烤羊肉中非极性杂环胺的抑制作用不明显。     

植物多酚抑制热加工肉制品中杂环胺形成机制研究进展

杂环胺（heterocyclic amines,HAs）是肉制品热加工过程中产生的一类具有极强致畸致癌活性的有毒化合物,其中2-氨基-3-甲基咪唑并[4,5-f]喹啉（2-amino-3-methyl-imidazo[4,5-f]-quinoline,IQ）和2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉（2-amino-3,4-dimethyl-imidazo[4,5-f]-quinoline,MeIQ）,2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶（2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine,PhIP）,1-甲基-9H-吡啶并[3,4-b]吲哚（1-methyl-9Hpyrido[3,4-b]indole,Harman）和9H-吡啶并[3,4-b]吲哚（9H-pyrido[3,4-b]indole,Norharman）3类HAs在热加工肉制品中较为常见且分布广泛。本文即围绕上述3类HAs,简要分析其形成的途径及机制,并详细阐述外源添加植物多酚抑制HAs形成的作用机制,最后依据现有研究归纳总结出多酚化合物抑制HAs与其化...     

熟肉制品中杂环胺的形成与抑制研究进展

杂环胺是熟肉制品中最常见的危害物,其种类繁多,形成途径复杂,抑制机制也不尽相同。本文介绍了2 类杂环胺的结构,主要阐述喹啉和喹喔啉类、2-氨基-1-甲基-6-苯基咪唑[4,5-b]吡啶、β-咔啉类（1-甲基-9H-吡啶[4,3-b]吲哚（1-methyl-9H-pyrido[4,3-b]indole,harman）、9H-吡啶[4,3-b]吲哚（9H-pyrido[4,3-b]indole,norharman））以及结合态杂环胺的形成途径,归纳减少杂环胺形成的措施及机理（控制加工条件和前体物、添加外源物质清除自由基和中间体、抑制美拉德反应等）,并对目前的研究现状进行概括。     

陈皮对烘烤牛肉干中杂环胺含量的影响

探讨陈皮对烘烤牛肉干中杂环胺生成及牛肉干质量损失率、色差、感官指标的影响.陈皮的总酚物质含量及抗氧化能力分别通过福林-酚法及DPPH法测定,牛肉干中杂环胺含量利用高压液相色谱法测定.在没有经过陈皮水提物腌制处理牛肉干中检测出2-氨基-3,8-二甲基咪唑并[4,5-F]喹喔啉(MeIQx)、2-氨基-3,4,8-三甲基咪唑并[4,5-用喹喔啉(4,8DiMeIQx)、9H-吡啶并[3,4-B]吲哚(Norharman)、1-甲基-9H-吡啶并[3,4-B]吲哚(Harman)和2-氨基-1-甲基-6-苯基-咪唑并[4,5-B]吡啶(PhIP),总含量为44.01ng/g.160mg/mL的陈皮水提物可显著降低MeIQx(34.3％),PhIP(96.5％),但是显著增加了Norharman(32.4％);且陈皮提取液浓度越高、腌制时间越长,其对牛肉干中杂环胺含量的影响越大;陈皮腌制处理对牛肉干质量损失率没有显著影响,而色泽随着陈皮浓度的升高而明显变淡;感官评价表明经过陈皮腌制的牛肉干在颜色及风味感官品质上无不良影响.因此,陈皮提取物腌制是一种较好的抑制烘烤牛肉干中杂环胺的方法.     

肉制品中PhIP的形成及抑制机理研究进展

2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶（2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine,PhIP）是肉制品在热加工中产生的有害物——杂环胺（heterocyclic amines,HCA）的重要成分。PhIP通过肉制品介质进入人体后,有致癌和致突变的风险。为了避免或降低PhIP对人体的危害,在不影响肉制品品质的前提下合理控制PhIP的含量,是今后研究的一个重点方向。该文就PhIP的危害、形成及抑制机理进行综述,以期为有效抑制肉制品中PhIP的含量提供理论参考。     

热加工肉制品中β-咔啉类杂环胺的形成、检测及毒性研究进展

为了提高食品感官质量和保证饮食健康,人们通常会选择热加工的烹饪方式。在食品热加工过程中,除了赋予食品独特的风味与色泽外,高温还会导致一些有害物质的生成。杂环胺(heterocyclic aromatic amines,HAAs)是一类多在食品热加工过程中产生的有毒化合物,具有致癌性和致突变性,长期摄入还可能导致一些慢性病如帕金森综合征、阿尔兹海默症的发生。β-咔啉类HAAs,9H-吡啶并[4,3-b]吲哚(Norharman)和1-甲基-9H-吡啶并[3,4-b]吲哚(Harman)是热加工肉制品中两种主要的HAAs。随着消费者对食品安全关注度的增加,关于HAAs的未知风险也得到了人们较多关注。本文从β-咔啉类杂环胺Norharman和Harman的形成途径出发,总结了这类物质的抑制方法以及近年来国内外对于β-咔啉类HAAs的分析检测方法研究进展,同时对其毒性研究进展进行了概述,以期为控制热加工肉制品中的β-咔啉类杂环胺生成以及带来的食品安全问题提供一定的理论支撑和参考。     

食品中PhIP的危害和检测方法的研究进展

肉制品在高温环境下会产生一种致癌致突变物质——杂环胺（heterocyclic aromatic amines,HAAs）,2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶（2-amino-1-methyl-6-phenylmimidazole and[4,5-b]pyridine,PhIP）是肉制品中含量最高的一类杂环胺。在由于摄入杂环胺而可能的致癌风险中,有半数风险是由PhIP引起的,PhIP会导致前列腺癌、乳腺癌和结肠癌等疾病。因此,PhIP的定量检测对指导人们减少PhIP的摄入,进而降低相关疾病的患病风险有重要的意义。本文就PhIP的危害和检测方法进行综述,以期为寻找具有快速高效、高灵敏度和高选择性的PhIP含量的检测方法提供依据。     

Effects of traditional chinese cooking methods on formation of heterocyclic aromatic amines in lamb patties

Different amounts of the potent mutagenic and/or carcinogenic heterocyclic aromatic amines (HAAs) are formed in muscle-based foods under different cooking methods. HAAs (9 varieties) in lamb patties cooked using traditional Chinese cooking methods (roasting, frying, panfrying, and stewing in seasonings) were investigated. The total HAAs contents in roasted, fried, pan-fried, and stewed patties were 4.39-123.15 ng/g, 3.59-43.24 ng/g, 0.71-10.05 ng/g, and 51.07-120.32 ng/g, respectively. Amounts of HAAs increased as cooking time increased. 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP) was the dominant HAAs in deep roasted and fried samples, while 1-methyl- 9H-pyrido [3,4-b] indole (Harman) and 9H-pyrido [3,4-b] indole (Norharman) were the main HAAs in pan-fried and stewed samples. Types and contents of HAAs formed at different cooking times using different methods are unique. Stewing in seasoning generated a higher HAAs content than the high-temperature cooking methods roasting, frying, and pan-frying.     

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.     

Effects of breeds on the formation of heterocyclic aromatic amines in smoked lamb

Heterocyclic aromatic amines (HAAs) are mutagenic and carcinogenic compounds, which are commonly detectable in cooked meat products. The objective of this study was to evaluate the effect of sheep breeds on the formation of HAAs in smoking cooked lamb. The results showed that HAAs in smoked lamb meat products were generally low (2.74–5.42 ng g^?1), with most being Harman and Norharman. IQ, MeIQx, 4,8‐DiMeIQx, Trp‐p‐2, PhIP and MeAaC were not detected in smoked lamb meat products in the present study. The total content of HAAs differed between meat products of different sheep breeds, but no difference in the order of magnitude was determined. Smoking altered the content of protein, fat, moisture and free amino acids in lamb meat products, which was probably mostly contributed by the reduction in meat moisture. Free tryptophan decreased in all breeds after smoking, which was probably used to synthesise HAAs. In summary, HAAs were low in smoked lamb meat products of all sheep breeds; thus, consumption of smoked lamb meat products should contribute very limited intake of HAAs.     

Content,causes and analysis of heterocyclic amines in Chinese traditional braised chicken

ABSTRACT

Braised chicken is a popular traditional chicken product in China with a special and complicated culinary method. However, there has been little research focused on the food safety problems caused by hazardous compounds in braised chicken. In this study, the contents of heterocyclic amines (HCAs) in representative braised chicken samples were determined. Total HCAs in braised chicken were relatively high (6.20–27.7 ng g^?1), and aminoimidazoazaarenes (AIAs) took a large proportion (54.0–86.4%). 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) was found in all samples with a high level (2.72–13.5 ng g^?1), whilst 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was low (0.14–0.41 ng g^?1). 9H-pyrido[3,4-b]indole (Norharman) and 1-methyl-9H-pyrido[3,4-b]indole (Harman) were main components in aminocarbolines (ACs). So the potential safety risk in braised chicken caused by high HCAs content should be paid attention. Principal component analysis (PCA) was used to analyse and visualise the differences of HCAs levels among braised chicken samples. The result indicated that the differences among these samples were significant, but there were no obvious similarities between samples produced in the same regions. Moreover, there were rare significant positive correlations between chemical components (including moisture, fat, protein, creatine and free amino acids) and hazardous compounds. Considering PCA and correlation analysis results, these parameters may play a major role in the HCAs levels in Chinese traditional braised chicken.     

Effects of precursor composition and water on the formation of heterocyclic amines in meat model systems

Model systems based on pressed meat from ox, pork and chicken were used to study the formation of carcinogenic/mutagenic heterocyclic amines (HAs). The composition of precursors (free amino acids, creatine and glucose) was examined and samples were heated in test-tubes under wet and dry conditions at 175 and 200°C for 30 min. Several HAs were detected, and the formation of DMIP (2-amino-1,6-dimethylimidazo[4,5-b]-pyridine), TMIP (2-amino-1,5,6-trimethyl-imidazo[4,5-b]-pyridine), IFP (2-amino-1,6-dimethylfuro[3,2-e]imidazo[4,5-b]-pyridine) and PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine) was found to be favoured by dry heating conditions. Highest amounts of PhIP and IFP were detected in heated meat juice from chicken breast, while more MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]-quinoxaline) was found in heated meat juices from roast beef and pork chop. Norharman (9H-pyrido[3,4-b]-indole) and Harman (1-methyl-9H-pyrido[3,4-b]-indole) were also detected at high levels.     

Formation of amino-imidazo-azaarenes and carbolines in fried beef patties and chicken breasts under different cooking conditions in Korea

The effect of cooking temperature and time on amino-imidazo-azaarenes (AIAs) and carbolines in fried ground beef patties and chicken breast under different cooking conditions in Korea was evaluated. Beef patties were fried at different temperatures (150, 180, and 230°C) for 4, 8, 12, and 16 min per each side and then the amount of AIAs and carbolines was evaluated by solid-phase extraction and HPLC-MS analysis. In fried ground beef patties, formations of 9H-pyrido [3,4-b]indole (Norharman) and 1-methyl-9H-pyrido [3,4-b]indole (Harman) were dramatically increased at 230°C for 16 min. Concentrations of Norhanrman and Harman formed at 230°C for 16 min/side were 12 and 40 times greater than level those of Norharman formed at same cooking condition. In fried chicken breasts, 2-amino-3,7,8-trimethylimidazo[4,5-f] quinoxaline (7,8-DiMeIQx) and 2-amino-3,4,7,8-tetramethylimidazo[ 4,5-f]quinoxaline (Tri-MeIQx) were not found at 150 and 180°C. Norhanrman formed at 230°C for 16 min was approximately 4 times higher than fried chicken breasts at 180°C. These results suggest that increase of cooking temperature and time was directly affected on AIAs and carbolines formation in Korean cooked meat.     

花椒叶提取物对烤牛肉饼杂环胺形成的影响

以牛肉和花椒叶为研究对象,研究花椒叶提取物（Zanthoxylum bungeanum Maxim. leaf extract,ZME）添加量（0.015%、0.030%、0.045%）对烤牛肉饼中杂环胺（heterocyclic amines,HAs）形成的影响。结果表明,与对照组相比,ZME能够显著抑制烤牛肉饼中总HAs的形成（P＜0.05）,对其最大抑制率为39.87%,但对不同种类HAs形成的影响不同。其中,添加0.045% ZME对2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶、2-氨基-3-甲基咪唑并[4,5-f]喹啉、2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉、1-甲基-9H-吡啶并[3,4-b]吲哚和2-氨基-9H-吡啶并[2,3-b]吲哚的抑制率分别达到71.76%、78.02%、49.07%、35.82%和100%;而0.045% ZME却促进2-氨基-3,8-二甲基咪唑并[4,5-f]喹喔啉和9H-吡啶并[3,4-b]吲哚的形成。进一步分析ZME对烤牛肉饼中HAs前体物的影响,结果显示前体物的消耗随着ZME添加量的增加呈降低趋势,相关性分析表明HAs的形成与多种游离氨基酸的消耗显著相关。上述结果表明ZME能显著抑制烤牛肉饼中HAs的形成,为其提高加工食品安全性及更广泛的应用提供理论依据。