The influence of demographic,physical, behavioral,and dietary factors on hemoglobin adduct levels of acrylamide and glycidamide in the general U.S. population

Purpose: This study aims to better understand the individual characteristics and dietary factors that affect the relationship between estimated consumption of acrylamide and measured acrylamide hemoglobin adduct levels (HbAA) and glycidamide hemoglobin adduct levels (HbGA). Methods: Acrylamide levels in individual food items, estimated by the U.S. Food and Drug Administration, were linked to data collected in the 2003–2004 National Health and Nutrition Examination Survey. Multivariable linear regression was used to evaluate the relationship between estimated consumption of acrylamide and HbAA. Results: A significant association between acrylamide intake and HbAA was observed, after adjustment for gender, race/ethnicity, smoking status, age, and BMI (R² = 0.34). Across quartiles of acrylamide consumption, HbAA and HbGA levels increased monotonically. Among nonsmokers, an evaluation of three heavily consumed, high AA concentration foods showed a positive trend between the consumed amount of fried potatoes and HbAA in children, adolescents, and adults. A significant positive trend between the consumed amount of potato chips or coffee was indicated in adolescents, adults, and seniors. Conclusions: Consumption of some individual foods affects HbAA concentrations more strongly and in an age-dependent manner. Our results suggest that effective dietary guidelines for controlling acrylamide intake should be subpopulation specific.     

Assay of advanced glycation endproducts in selected beverages and food by liquid chromatography with tandem mass spectrometric detection

Food and beverages contain protein glycation adducts--both early-stage adducts and advanced glycation endproducts. We determined the concentrations of glycation adducts in selected food and beverages by liquid chromatography with triple quadrupole mass spectrometric detection. Cola drink contained low concentrations of glycation free adducts, whereas pasteurised and sterilised milk were rich sources of heat-stable glycation adduct residues--Nepsilon-carboxymethyl-lysine and Nepsilon-carboxyethyl-lysine. Laboratory rodent food was a rich source of advanced glycation endproducts. Measurement of glycation adducts in 24 h urine samples of normal and diabetic rats indicated that < 10% of glycation adduct residue consumption was excreted. Induction of diabetes by streptozotocin led to a 2-fold increase in urinary excretion of Nepsilon-carboxymethyl-lysine and a 27-fold increase in urinary excretion of methylglyoxal-derived hydroimidazolone Ndelta-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine - the latter was decreased by high-dose thiamine therapy that also prevented the development of nephropathy. We conclude that cola drinks are a poor source of glycation adduct whereas thermally processed milk is rich in glycation adducts. Dietary glycation adducts residues probably have low bioavailability. Experimental diabetes is associated with a marked increase in exposure to endogenous formation of methylglyoxal-derived hydroimidazolone which is linked to the development of diabetic nephropathy.     

Formation of protein adducts of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine in cooked foods

Heterocyclic amines (HCAs) are mutagenic and carcinogenic compounds found in cooked meat and fish. Although HCAs are known to form adducts with protein after metabolic activation, adduct formation during cooking has not been elucidated. In this study, we showed that 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) is released from high molecular weight compounds by acid or enzymatic hydrolysis of cooked foods. Formation of free and protein adduct forms of PhIP was dependent on cooking temperature and time, and PhIP–protein adducts were estimated to form after formation of free PhIP. We also demonstrated that PhIP–protein adduct is formed by heating of PhIP and albumin as a model protein. A new adduct peak including [M+H]⁺ (m/z=225) of PhIP as a fragment ion was detected in the high molecular weight fraction of heat‐treated protein by LC–MS analysis. From model experiments by heating of PhIP and amino acids, the adduct was estimated to be produced by condensation of the amino group of PhIP and the carboxyl group of protein. PhIP–protein adducts were detected in several cooked meat and fish at ng/g food level as PhIP content. These results suggest that food‐borne protein adducts of HCAs may influence human HCA exposure and carcinogenic risk.     

New molecular and field evidences for the implication of mycotoxins but not aristolochic acid in human nephropathy and urinary tract tumor

To find out whether ochratoxin A (OTA), citrinin (CIT), aristolochic acids (AA) are etiologic agents of Balkan endemic nephropathy (BEN) or Chinese herbal nephrotoxicity, and associated urinary tract tumor (UTT), we have compared (i) in human kidney cell culture, the DNA adduct formation and persistence of OTA/CIT and AA adducts (ii) analyzed DNA adduct in several tumors from human kidney suspected to be exposed to either OTA and CIT, or AAs (iii) analyzed OTA, CIT, and AA in food. In kidney cell cultures, formation of specific OTA-DNA adduct and AA-DNA adduct were detected in the same range (around 10 adducts/10(9) nucleotides) and were time- and dose-dependent. After 2 days all disappeared. DNA adduct related to OTA and CIT are found in human kidney tissues from Balkans, France, and Belgium whereas no DNA adducts related to AA could be found in any tumors of BEN patients from Croatia, Bulgaria, or Serbia. No DNA adduct was found in kidney biopsy or necropsy of the French women suspected to be exposed to AA. OTA and CIT are more frequently found in rural area. AA was never detected. All these plead for implication of mycotoxins, especially OTA, in BEN and UTT.     

丙烯酰胺体内暴露生物标志物解析和风险评估研究进展

丙烯酰胺是一种重要的化工原料,也是美拉德反应的副产物之一,广泛存在于烟草烟雾和经高温加工且富含碳水化合物的食品中。丙烯酰胺可经过消化道、呼吸道、皮肤等多种途径被机体吸收,具有神经毒性、生殖发育毒性、基因毒性及致癌性。丙烯酰胺在体内经细胞色素P450作用转化为环氧丙酰胺,进一步形成主要的4种巯基尿酸加合物、2种血红蛋白加合物和4种DNA加合物,这3类加合物均可作为评价丙烯酰胺体内暴露水平的生物标志物,并可通过进一步测定其含量以评估和关联生物体(人体)在环境或膳食中丙烯酰胺的暴露水平。本研究对丙烯酰胺的危害作用、体内代谢途径、体内暴露生物标志物、生物标志物的检测方法、体内暴露风险评估和体内暴露控制与预防措施进行了概述,以期为丙烯酰胺的风险评估与监控提供参考。     

Internal doses of acrylamide and glycidamide in mice fed diets with low acrylamide contents

The formation of acrylamide during heating of certain foodstuffs constitutes a potential health hazard. The health risk assessment should be based on knowledge about the relation between dietary exposure to acrylamide and internal doses of acrylamide and its genotoxic metabolite glycidamide. The primary aim of this study in mice was to measure these relationships at low levels of acrylamide intake through the diet. A secondary aim was to clarify which extraction method should be used when analyzing acrylamide in food in order to obtain a correct measure of the acrylamide that is available for absorption. In the analysis procedure, alkaline extraction has earlier shown much higher measured acrylamide levels in certain foods compared to water extraction. In this subcronic study the administered diets were composed to give five levels of acrylamide intakes between 3 and 50 mug/kg body weight per day (calculated on figures obtained after water extraction). Internal doses of acrylamide and glycidamide were measured through hemoglobin (Hb)-adducts. The results showed linear relationships between the exposure of acrylamide and Hb-adduct levels from both acrylamide and glycidamide at these low exposure levels. The study also showed that the "extra" acrylamide measured with alkaline extraction does not correspond to bioavailable acrylamide.     

牛磺酸对富含淀粉食品中丙烯酰胺的抑制作用

运用天冬酰胺/葡萄糖模式反应体系研究了牛磺酸对富含淀粉的食品中丙烯酰胺生成的影响及其抑制机理。试验结果显示:牛磺酸能够显著抑制天冬酰胺/葡萄糖反应体系中丙烯酰胺的生成,且抑制率与浓度呈剂量关系。运用LC-QTOF分析得到2种主要的反应产物:丙烯酰胺-牛磺酸加合物和丙烯酰胺二聚体-牛磺酸加合物。此外,牛磺酸与葡萄糖共存于同一体系中加热时,葡萄糖的含量显著下降,说明牛磺酸可与葡萄糖发生反应。通过模式反应体系证明:牛磺酸能够显著降低富含淀粉食品中的丙烯酰胺含量;抑制机理主要是牛磺酸与丙烯酰胺直接发生反应,反应产物为丙烯酰胺-牛磺酸加合物和丙烯酰胺二聚体-牛磺酸加合物,从而除去体系中已经生成的丙烯酰胺。此外,牛磺酸能够与葡萄糖发生美拉德反应,从而与天冬酰胺竞争消耗体系中的葡萄糖,减少了食品中的丙烯酰胺含量。     

Recent advances in heterocyclic aromatic amines: An update on food safety and hazardous control from food processing to dietary intake

Heterocyclic aromatic amines (HAAs) as probable carcinogenic substances are mainly generated in meat products during thermal processing. Numerous studies have contributed to the analysis, formation, and mitigation of HAAs during food processing. However, few articles have comprehensively reviewed food safety aspects from both food processing and dietary intake regarding the formation, mitigation, metabolism, biomarkers for exposure, hazard control, and risk assessment of HAAs, and related food safety researches. Several factors may influence the generation of HAAs, including processing temperature, processing time, and chemical composition of the meat. Nonetheless, these mutagenic compounds are attenuated to different levels by the addition of natural or synthetic flavorings and antioxidant‐rich marinades, as well as pretreatments using technique such as microwave heating. After dietary intake, different types of HAAs are metabolized in humans by several enzymes, including cytochrome P450s, peroxidases, N‐acetyltransferases, sulfotransferases, uridine diphosphate‐glucuronosyltransferases, and glutathione S‐transferases. Their primary metabolites are further conjugated with DNA or ultimately excreted in urine and feces. The 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine in hair as well as DNA, hemoglobin, and serum albumin adducts has been considered as biomarkers for exposure assessment. Dietary intake information obtained from questionnaires and the results of epidemiological investigations have shown a positive relationship between the intakes of red meat and processed meat and high risk of cancer incidence. As several cancers have been reported to be associated with HAAs, HAAs should be both effectively reduced during food processing and controlled from dietary intake to facilitate human health.     

Identification of glucosinolate congeners able to form DNA adducts and to induce mutations upon activation by myrosinase

Scope: Juices from Brassicales are mutagenic in Salmonella typhimurium and characteristic adducts are formed with the endogenous DNA in Brassicales homogenates. These effects require myrosinase activity, suggesting an involvement of breakdown products of glucosinolates (GLs). We aimed to identify GLs congeners producing these effects. Methods and results: We investigated twelve individual GLs for mutagenicity in S. typhimurium TA104 and TA100 and for adduct formation with herring sperm DNA using the 32P‐postlabelling/thin‐layer chromatography method. All bacteriotoxic and mutagenic effects observed required the presence of myrosinase. Neoglucobrassicin, 4‐methoxyglucobrassicin and sinalbin showed mutagenicity over wide concentration ranges, with neoglucobrassicin being the most potent congener. Six other GLs led to modest increases in the number of revertants in a small concentration range, before toxicity overshadowed this effect. The remaining three GLs showed some toxicity, but no mutagenicity. However, all twelve GLs formed DNA adducts. Clearly the highest adduct levels were detected with the indole GLs tested. They matched the major adduct spots formed in Brassicales homogenates. Conclusion: The observation that GLs are genotoxic demands follow‐up studies on possible genotoxic and carcinogenic effects of these common food compounds in animal models and humans. Our study may be used to prioritize the congeners in further studies.     

Health biomarkers in a rat model after intake of organically grown carrots

BACKGROUND: Organic food is perceived as being of better quality and healthier than conventional foods although the scientific research on organic foodstuffs is highly contradictory. The aim of the present study was to investigate if intake of carrots from four different cultivation systems grown in two consecutive years would influence various biomarkers of health in a rat model. All rats were fed a diet with 40% carrot content. The carrots were grown under conventional (C), ‘minimalistic’ organic (O1), organic (O2), or ‘very’ organic cultivation systems (O3). A control group (CO) being fed standard rat chow was included. RESULTS: The plasma α‐tocopherol concentration was higher in the O2 carrot‐based diet group than in the C carrot based‐diet group in one year, while all other health biomarkers or nutrient content differences were observed between the CO diet and the carrot‐based diets. CONCLUSION: This well‐controlled field study demonstrated no clear influence of cultivation methods or harvest year on the nutritional quality of carrots or effect of cultivation methods on health‐related biomarkers in a sensitive rat model. However, the experimental set‐up and selected biomarkers could be used as a framework for further studies of health in relation to organic foodstuff. Copyright © 2012 Society of Chemical Industry     

Acrylamide–taurine adducts formation as a key mechanism for taurine’s inhibitory effect on acrylamide formation

Amino acids have attracted increasing attention in developing effective inhibitors on acrylamide (AA) formation during food processing. The main purpose of this study was to evaluate the inhibitory effect and mechanism of taurine (Tau) on AA formation. In the presence of Tau, AA formation was effectively inhibited in asparagine/glucose (Asn/Glc) model system. Results also showed that Glc content was decreased as Glc–Tau solution was heated, which indicated that Tau could react directly with Glc in the Maillard reaction. LC‐QTOF MS revealed two peaks in extracted ion chromatograms [M + H] ⁺ at m/z 197 and m/z 268 during the thermal treatment, which were identified as AA–Tau and AA dimmer–Tau adducts, respectively. Our results demonstrated that inhibitory effect of Tau on AA formation was not only mediated through contesting consumption of Glc with Asn, but also directly reacting with AA to promote formation of AA–Tau and AA dimer–Tau adducts.     

Determination of acrylamide content of food products in Korea

Acrylamide (AA) contents of commercial market‐purchased foods and of traditional home‐cooked Korean foods were investigated. The effect of cooking method on AA amount in potatoes was also studied. AA contents of roasted barley and corn ranged from 116 to 449 µg kg^?1 and of chips ranged from 12 to 3241 µg kg^?1, representing a very high variation in AA contents among tested samples. Lower levels of AA were found in wheat flour chips compared to potato chips. AA contents of ramen noodles were below 37 µg kg^?1, whether they contained potato starch or not. The AA contents of Korean home‐cooked oil fried foods increased in the following order: yakwa < whole deep‐fried sea tangle < ground and deep‐fried lotus root < ground and pan‐fried lotus root < French fries < ground and pan‐fried potato. Lotus root and garlic also had the potential to produce AA during cooking. Pre‐treatment such as grinding, boiling, freezing and thawing increased the AA formation in oil‐fried potato. Temperature was more influential than time on AA formation during deep‐oil frying. Removing water‐soluble fractions reduced AA content of cooked potatoes. Copyright © 2006 Society of Chemical Industry     

Development of a Highly Sensitive Competitive Indirect Enzyme-Linked Immunosorbent Assay for Detection of Acrylamide in Foods and Water

Acrylamide (AA) has been classified as a probable human carcinogen and forms in certain foods, particularly plant-based foods that are rich in carbohydrates and low in proteins, during processing or cooking at high temperatures. In this study, polyclonal antibodies were raised against a hapten derived from acrylamide and 3-mercaptobenzoic acid (3-MBA). An indirect competitive enzyme-linked immunosorbent assay was developed to rapidly quantify AA in complex food matrices and water. The assay was very specific to the AA-3-MBA derivative and showed no cross-reactivity to asparagine, the main precursor of AA formation in foods, aspartic acid, AA, or 3-MBA. The assay was very sensitive with a limit of detection of 5.0 ng/g in model for food matrices to 0.1 μg/L in water. Good recoveries for AA were observed in all matrices tested, and the results using this method were comparable to those obtained from mass spectrometry methods including Food Analysis Performance Assessment Scheme control samples and results for different food products.     

Current issues in dietary acrylamide: formation,mitigation and risk assessment

Acrylamide (AA) is known as a neurotoxin in humans and it is classified as a probable human carcinogen by the International Agency of Research on Cancer. AA is produced as by‐product of the Maillard reaction in starchy foods processed at high temperatures (>120 °C). This review includes the investigation of AA precursors, mechanisms of AA formation and AA mitigation technologies in potato, cereal and coffee products. Additionally, most relevant issues of AA risk assessment are discussed. New technologies tested from laboratory to industrial scale face, as a major challenge, the reduction of AA content of browned food, while still maintaining its attractive organoleptic properties. Reducing sugars such as glucose and fructose are the major contributors to AA in potato‐based products. On the other hand, the limiting substrate of AA formation in cereals and coffee is the free amino acid asparagine. For some products the addition of glycine or asparaginase reduces AA formation during baking. Since, for potatoes, the limiting substrate is reducing sugars, increases in sugar content in potatoes during storage then introduce some difficulties and potentially quite large variations in the AA content of the final product. Sugars in potatoes may be reduced by blanching. Levels of AA in different foods show large variations and no general upper limit is easily applicable, since some formation will always occur. Current policy is that practical measures should be taken voluntarily to reduce AA formation in vulnerable foods since AA is considered a health risk at the concentrations found in foods. © 2013 Society of Chemical Industry     

Aromatic DNA adducts in relation to dietary and other lifestyle factors in Spanish adults

The aim of this study was to assess the relationship between bulky DNA adducts in white blood cells (WBC) and lifestyle factors in a sample from the Spanish cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC). 296 subjects aged between 35 and 64 years, from five regions, were included. Food intake was estimated with a computerized version of dietary history questionnaire. Daily intake of polycyclic aromatic hydrocarbons (PAH) was estimated using a database with information on food content of potential carcinogens. Data on lifestyle and health factors were collected and DNA adducts measured using the nuclease P1 ³²P-postlabelling technique. Geometric means of adducts were similar for men and women (4.11/10⁹ and 3.94/10⁹ nucleotides, respectively). Highest levels of adduct were observed in non-smokers and non-occupationally exposed. Meat intake, oils and fats were associated with higher levels of adducts, but all non-statistically significant. Higher intakes of calcium, sodium and phosphorus were associated with lower adducts levels. Summarising, our study shows that bulky adducts measured by ³²P-postlabelling in DNA from WBC do not correlate with the usual diet of healthy Spanish adults. Although it has been proposed that diet be the main source of PAH in nonsmokers without occupational exposure, DNA adducts do not seem to be suitable biomarkers of dietary PAH in general population.     

Enhancing Nutraceutical Performance Using Excipient Foods: Designing Food Structures and Compositions to Increase Bioavailability

The oral bioavailability of many bioactives (pharmaceuticals, dietary supplements, nutrients, and nutraceuticals) is limited because of physicochemical and physiological events that occur within the gastrointestinal tract (GIT) after their ingestion. These events include: (i) restricted liberation from drugs, supplements, or foods; (ii) extensive metabolism or chemical transformation during passage through the GIT; (iii) low solubility in intestinal fluids; (iv) low permeation through the intestinal cell monolayer; and (v) efflux from epithelium cells. Bioactive bioavailability can often be improved by designing the composition and structure of food matrices to control their liberation, transformation, solubilization, transport, absorption, and efflux in the GIT. This article reviews the potential impact of food composition and structure on the oral bioavailability of bioactives, and then shows how this knowledge can be used to design excipient foods that can improve the bioavailability profile of bioactives. The bioactive may be incorporated within an excipient food or co‐ingested with an excipient food. The suitability of oil‐in‐water emulsions as excipient foods is highlighted. The utilization of excipient foods may provide a new strategy for improving the efficacy of nutraceuticals, supplements, and pharmaceuticals.     

Effect of innovative pre-treatments on the mitigation of acrylamide formation in potato chips

The aim of this research was to evaluate the reduction of acrylamide (AA) formation in potato chips applying innovative pre-treatments. Raw potato slices were subjected for 5 and 15 min to: dipping in water; dipping in Aureobasidium pullulans L1 yeast water suspension; dipping in water or in yeast water suspension after pulsed electric fields (PEF) in order to investigate a possible synergistic effect of pre-treatments. The raw potato samples were analysed for AA precursors and, after frying, for AA by using HPLC-MS/MS. In addition, the final potato chips main quality parameters were evaluated.Compared to untreated potato chips, yeast treatment promoted a reduction of AA formation mainly at the longest dipping time (15 min). PEF treatment followed by water dipping was the most effective in reducing AA for both the studied treatment times. The combination of PEF and yeast treatments led only a slight reduction of AA formation.Industrial relevanceThe Commission Regulation (EU) 2017/2158 has established new acrylamide (AA) benchmark levels in different foods due to its negative classification as “probably carcinogenic to human”. For this reason, food industries are interested in developing different processing methods to reduce the AA formation and at the same time to maintain an acceptable quality of final products. Fried potatoes (French fries and chips) are the most vulnerable foods in terms of high content of AA formation, being rich in the main Maillard reaction substrates, such as asparagine and reducing sugars, and characterized by a high surface to volume ratio. Among the strategies recently suggested for the reduction of AA in potato chips, pulsed electric fields (PEF) and yeast pre-treatments are very promising, having the potentiality to reduce AA precursors in raw potato tissues. In this study the possibility to apply yeast and PEF pre-treatments on raw potato slices with suitable processing times for a possible industrial application were evaluated.     

Acrylamide degradation by filamentous fungi used in food and beverage industries

Filamentous fungi (24 strains) used in food and beverage industries were investigated for acrylamide-degradation ability: Aspergillus oryzae KBN1010 showed the highest ability. Little acrylic acid was produced but no glycidamide was detected during AA degradation in roasted green tea; therefore, A. oryzae could be used for reducing the AA concentration.     

Phase solubility studies and stability of cholesterol/β-cyclodextrin inclusion complexes

BACKGROUND: Cyclodextrins (CDs) are able to enhance the solubility, stability and bioavailability of several bioactive hydrophobic compounds by complex formation. They can also be used for removal of undesired components (such as cholesterol, off‐flavors or bitter components) present in foods. Although many patents account for the use of cyclodextrins for removal of cholesterol from dairy foods, there is no available information on the effect of water on encapsulation efficiency and on the stability of sterols in CDs. The aim of this work was to study the inclusion properties and the factors affecting the encapsulation and stability of cholesterol in β‐cyclodextrin (BCD). The optimum encapsulation conditions (ligand–CD molar ratio, stirring time and temperature), and stability of the complexes as a function of storage time and water content were analyzed. RESULTS: Phase solubility study pointed out the formation of 1:1 stoichiometric complexes between cholesterol and β‐cyclodextrin, which was influenced by temperature variations. The process was shown to be exothermic and energetically favored. The presence of cholesterol greatly modified the BCD water sorption curves, being the amount of adsorbed water smaller in the combined systems. The principal ‘driving force’ for complex formation is the substitution of the high‐enthalpy water molecules by an appropriate hydrophobic ligand. The freeze‐dried complexes probed to be stable at different storage conditions. CONCLUSION: The phase solubility and stability data obtained could be essential for selecting the most suitable conditions when CDs are employed either for removing cholesterol or to incorporate functional ingredients (i.e. sitosterol) in the development of innovative food products. Copyright © 2011 Society of Chemical Industry