Kerstin Skog

Carcinogenic Heterocyclic Amines in Model Systems and Cooked Foods: A Review on Formation, Occurrence and Intake

Frying or grilling of meat and fish products may generate low ppb levels of mutagenic/carcinogenic heterocyclic amines (HAs). Many heterocyclic amines are formed via the Maillard reaction from creatine, free amino acids and monosaccharides; compounds naturally occurring in protein-rich foods of animal origin. The formation and yield of HAs are dependent on physical parameters, such as cooking temperature and time, cooking technique and equipment, heat and mass transport, and on chemical parameters, especially the precursors to HAs. This paper reviews the current knowledge on the formation of HAs in cooked foods and model systems, and summarizes data on the content of HAs in various cooked foods, and estimates of the dietary intake of HAs. It should be noted that the presence of carcinogens of other types in food (e.g. nitrosamines, aromatic amines, cholesterol oxide products) and that their generation during frying and grilling are outside the scope of this review.

Dietary heterocyclic amines and cancer of the colon, rectum, bladder, and kidney: a population-based study

BACKGROUND Heterocyclic amines formed in cooked meat and fish are carcinogenic in animal models and form DNA adducts in human beings. We undertook a study to assess whether these substances are related to the risks of cancer in the large bowel and urinary tract. METHODS In a population-based case-control study, cases were identified from the Swedish cancer registry. Controls were randomly selected from the population register. Information on intake of various foods and nutrients was assessed by questionnaire, with photographs of foods cooked at various temperatures. We measured the content of heterocyclic amines in foods cooked under these conditions. FINDINGS Information was retrieved from 553 controls, 352 cases of colon cancer, 249 cases of rectal cancer, 273 cases of bladder cancer, and 138 cases of kidney cancer. The response rate was 80% for controls and 70% for cases. The estimated daily median intake of heterocyclic amines was 77 ng for controls, and 66 ng, 63 ng, 96 ng, and 84 ng for cases with cancer of the colon, rectum, bladder, and kidney, respectively. The relative risk for the intake of heterocyclic amines (highest vs lowest quintile) was 0.6 (95% CI 0.4-1.0) for colon cancer, 0.7 (0.4-1.1) for rectal cancer, 1.2 (0.7-2.1) for bladder cancer, and 1.0 (0.5-1.9) for kidney cancer. Seven cases, but no controls, had an estimated daily intake of heterocyclic amines above 1900 ng. INTERPRETATION Intake of heterocyclic amines, within the usual dietary range in this study population, is unlikely to increase the incidence of cancer in the colon, rectum, bladder, or kidney. For daily intakes above 1900 ng, our data are consistent with human carcinogenicity, but the precision was extremely low.

Cooking procedures and food mutagens: A literature review

Commonly eaten meat products prepared from beef, pork, mutton and chicken show some level of mutagenic activity following normal frying. Food preparation methods have a significant influence on the formation of the mutagenic activity. The main food mutagens found in cooked meat products are heterocyclic amines. Several of them have been tested in long-term animal studies and shown to be carcinogenic in rodents. From a health point of view, it is desirable to reduce or prevent the formation of food mutagens. Therefore, a deeper understanding of the precursors and reaction conditions for mutagen formation during normal domestic cooking is very important. Modelling experiments are useful tools for studying the influence of different physical parameters and various precursors on the mutagenic activity. The identification of several thermic mutagens from the modelling experiments support the theory that creatine or creatinine, amino acids and sugars are precursors in the formation of thermic mutagens. Creatine is generally accepted to be a precursor of the mutagens and, interestingly, the conversion of creatine to creatinine has been shown to be blocked by an excess of sugars, which also caused the mutagenic activity to decrease. The mutagenic activity differed for different amino acids used in the model systems, and various thermic mutagens were produced from the amino acids. The incorporation of carbon atoms originating from glucose into food mutagen molecules has shown glucose to be a precursor. Sugar has also been shown to either enhance or inhibit the yield of mutagenic activity, depending on its molar ratio versus the other reactants, which suggests that the Maillard reaction may be used to control the formation of mutagens.

Formation of heterocyclic amines using model systems.

Initially, modeling was used to identify the mutagenic heterocyclic amines and their precursors. Major precursors have been shown to be single amino acids or amino acids together with creatine or creatinine. There is also evidence that Maillard reactions are involved since heating sugar and amino acids together with creatine or creatinine has been shown to produce several of the mutagenic heterocyclic amines, especially the aminoimidazoazaarenes (AIA compounds), e.g., IQ, MeIQ, MeIQx, DiMeIQx and PhIP. Due to a low yield in the model systems, the mechanisms behind the formation of the mutagenic heterocyclic amines are still unclear and need further substantiation. The fact that some AIA compounds are also produced in the absence of sugar casts some doubts on an obligatory participation of the Maillard reaction; alternative routes might exist. Further work using isotopically labeled precursors needs to be done and so far such work has only been performed for PhiP. The formation of mutagenic heterocyclic amines is dependent on time, temperature, pH, concentration of the precursors, type of amino acid, and the presence of certain divalent ions. Water may have an impact both as a temperature regulator and as a solvent medium for the reactants.

Analysis of nonpolar heterocyclic amines in cooked foods and meat extracts using gas chromatography-mass spectrometry

Heat processing of muscle foods gives rise to the formation of mutagenic and carcinogenic heterocyclic amines, often at ng/g levels. A gas chromatographic-mass spectrometric (GC-MS) technique was introduced for the analysis of nonpolar heterocyclic amines in common cooked meats, pan residues, and meat extracts after solid-phase extraction. The mutagenic heterocyclic amines 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 2-amino-9H-pyrido[2,3-b]indole (A alpha C) and 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA alpha C) were identified in several samples in amounts up to 8 ng/g. Also the comutagenic substances 1-methyl-9H-pyrido [3,4-b]indole (harman) and 9H-pyrido[3,4-b]indole (norharman) were detected in the samples in amounts up to almost 200 ng/g. The GC-MS method can be applied without derivatisation of the sample. The technique offers high chromatographic efficiency, yielding detection limits for pure references in the range 0.1-2 ng per injection.

Heterocyclic amines in poultry products: a literature review.

Health risks associated with heterocyclic amines in cooked foods have been discussed and analysed since the presence of these food mutagens was first detected. Intake, metabolism, carcinogenicity and epidemiology are important parameters in the risk assessment of heterocyclic amines. It is very difficult to determine the human intake of heterocyclic amines, as the content in cooked meat is highly dependent on the type of meat and how it has been prepared. This review summarises data on estimates of the content of heterocyclic amines in heat-treated poultry products.

Screening for heterocyclic amines in chicken cooked in various ways

Chicken cooked under well-controlled conditions and commercial chicken products were screened for heterocyclic amines (HAs). Chicken samples were boiled, deep-fried, pan-fried, oven-roasted, cooked in an unglazed clay pot or in a roasting bag in the oven, and oven broiled. 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 1-methyl-9H-pyrido[3,4-b]indole (harman) and 9H-pyrido[3,4-b]indole (norharman) were identified in several samples. Chicken cooked at low temperatures contained low amounts of HAs. In pan-fried chicken breasts, MeIQx was detected in amounts below 2 ng/g, 4,8-DiMeIQx below 0.6 ng/g, and PhIP in amounts up to 38 ng/g. Harman and norharman were detected in almost all samples (below 15 ng/g). In skin from a commercially barbecued chicken, MeIQx, 4,8-DiMeIQx and PhIP were detected, while only traces of MeIQx were detected in the meat. MeIQx was detected in a commercial chicken flavour, 0.1 ng/ml. No HAs were detected in pan-fried chicken liver. The results show that the content of HAs in chicken cooked in various ways is low if prepared at low temperatures, and increases with increasing cooking temperature. PhIP formation seems to start accelerating at cooking temperatures around or above 200 degrees C. Colour development increases with cooking temperature, but no correlation with HA content was observed.

Acrylamide in food: mechanisms of formation and influencing factors during heating of foods

Background: In April 2002, the Swedish National Food Administration and a scientific group at the University of Stockholm jointly announced that they had shown acrylamide to be formed during the preparation of food and found it to occur in many foodstuffs. These new findings were clearly of concern to many types of industrial food processing as well as to home cooking. The Swedish Food Federation (Li) initiated and financed the formation of an expert committee to look into the chemical mechanisms. The present review is the final report of that expert committee. Design: The study identified, examined and put together facts and present knowledge on reaction routes for acrylamide formation in food and causal connections to cooking and food processing conditions. The results are based on literature surveys, examination of the analytical data published by the Swedish National Food Administration and other follow-up studies, contacts with international scientific networ ks, and observations from food companies. Results: The exact chemical mechanism(s) for acrylamide formation in heated foods is unknown. Several plausible mechanistic routes may be suggested, involving reactions of carbohydrates, proteins/amino acids, lipids and probably also other food components as precursors. With the data and knowledge available today it is not possible to point out any specific routes, or to exclude any possibilities. It is likely that a multitude of reaction mechanisms is involved. Acrolein is one strong precursor candidate, the origin of which could be lipids, carbohydrates or proteins/amino acids. Acrylamide is a reactive molecule and it can readily react with various other components in the food. The actual acrylamide level in a specific food product, therefore, probably reflects the balance between ease of formation and potential for further reactions in that food matrix. There are indications in support of that the Maillard reaction being an import a nt reaction route for acrylamide formation, but lipid degradation pathways to the formation of acrolein should also be considered. Conclusions: Reliable analytical methods to measure acrylamide in foods are available. Model studies are needed to identify precursors and reaction route(s) based on current hypotheses and to elucidate possible further reactions between acrylamide and other food components. Studies are needed to optimize formulation and processing conditions to minimize acrylamide levels, taking other product quality properties into consideration. Keywords: Acrylamide, cooking, food processing, heated foods, Maillard reaction.

Influence of antioxidants in virgin olive oil on the formation of heterocyclic amines in fried beefburgers

An association between the intake of heterocyclic amines (HAs) and the development of cancer has been observed in some epidemiological studies, while in other studies no such correlation has been found. HAs are mutagenic/carcinogenic compounds formed at low levels via the Maillard reaction and a free radical mechanism during cooking of animal tissue. The addition of pure antioxidants or foods containing antioxidants has previously been shown to decrease the amount of HAs formed during cooking. In this study, beefburgers were fried in six different oils: refined olive oil, virgin olive oil, virgin olive oil depleted of phenols, rapeseed oil, virgin olive oil with rosemary extract and refined olive oil with rosemary extract. The content of antioxidative compounds in the virgin olive oil and the rosemary extract was determined. The beefburgers were analysed with regards to 12 different HAs by solid phase extraction and HPLC analysis. MeIQx, 4,8-DiMeIQx, PhIP, Harman and Norharman were detected in all beefburgers fried in the different oils, but the relative amounts varied. Frying in virgin olive oil reduced the formation of HAs compared with refined olive oil. This effect is probably due to the content of phenols in the virgin olive oil. The HA-reducing effect of virgin olive oil decreased during storage, but the addition of rosemary extract may prevent this decrease.

Problems associated with the determination of heterocyclic amines in cooked foods and human exposure

Epidemiological studies have shown diet to be an important factor in the global variation of human cancer rates. The presence of mutagenic/carcinogenic heterocyclic aromatic amines (HAs) in cooked foods has attracted a great deal of interest for more than 20 years. Accurate assessment of the human exposure to HAs requires food questionnaires that address cooking methods and reliable methods for the analysis of HAs in cooked foods, and of biomarkers of exposure. The complex food matrix, the low amounts of HAs present (ng/g), and the need for several isolation steps make accurate quantification difficult. Food composition, for example the concentrations and relative amounts of naturally occurring precursors, such as creatine, free amino acids and sugars and also the presence of enhancing or inhibiting compounds are known to greatly influence the formation of HAs. Cooking temperature and time are other important factors that affect the yield of HAs. One of the most abundant HAs, PhIP (2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine), is found typically in amounts up to around 35 ng/g, but there are some reports on much higher levels of PhIP. The levels of other HAs such as MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline) and IFP (2-amino-1,6-dimethylfuro[3,2-e]imidazo[4,5-b]pyridine) generally range from not detectable up to 10 ng/g, and AalphaC (2-amino-9H-pyrido[2,3-b]indole) up to 20 ng/g. Among the factors that influence human exposure to HAs are the type of food, cooking method, portion size and intake frequency. The estimated daily intake of HAs in different studies ranges from 0 to around 15 microg per person per day.