Potential of Low Cost Agro-Industrial Wastes as a Natural Antioxidant on Carcinogenic Acrylamide Formation in Potato Fried Chips.

Acrylamide is classified as a toxic and a prospective carcinogen to humans, and it is formed during thermal process via Maillard reaction. In order to find innovative ways to diminish acrylamide formation in potato chips, several extracts of agricultural wastes including potato peels, olive leaves, lemon peels and pomegranate peels extracts were examined as a soaking pre-treatment before frying step. Total phenolic, total flavonoids, antioxidant activity, and the reduction in sugar and asparagine contents were additionally performed. Proximate composition of these wastes was found to be markedly higher in fat, carbohydrate and ash contents. Lemon peels and potato peels showed almost similar phenolic content (162 ± 0.93 and 157 ± 0.88 mg GAE /g, respectively) and exhibited strong ABTS and DPPH radical scavenging activities than the other wastes. The reduction percentage of reducing sugars and asparagine after soaking treatment ranged from 28.70 to 39.57% and from 22.71 to 29.55%, respectively. HPLC results showed higher level of acrylamide formation in control sample (104.94 mg/kg) and by using the wastes extracts of lemon peels, potato peels, olive leaves, and pomegranate peels succeeded to mitigate acrylamide level by 86.11%, 69.66%, 34.03%, and 11.08%, respectively. Thus, it can be concluded that the soaking of potato slices in the tested wastes extracts as antioxidant as pre-treatment before frying reduces the formation of acrylamide and in this way, the risks connected to acrylamide consumption could be regulated and managed.

Acrylamide assessment of wheat bread incorporating chia seeds (Salvia hispanica L.) by LC-MS/MS.

We examined the acrylamide content in samples of wheat bread with chia seeds added at different concentrations (2%, 5%, 7%, 10%) and cooked at predefined conditions (20 min at 200°C) by a validated LC-MS/M method after QuEChERS extraction. The acrylamide contents of the bread samples with added chia seeds were compared with control wheat bread samples. The highest acrylamide values were found in bread with 5% chia seeds, showing a mean value of 156.5 ± 115.4 µg/kg, followed by bread with 10% chia seeds (150.2 ± 103.8 µg/kg). About 6% of the bread samples with added chia seeds reached acrylamide levels above the benchmark level set by the EU Regulation. No significant differences in acrylamide values were found between control samples and bread with different percentages of chia seeds (p > .05). The results obtained provide a first report on the possible contribution of chia to the increase of acrylamide formation in bread.

Risk and Benefit of Natural and Commercial Dark Brown Sugars as Evidenced by Phenolic and Maillard Reaction Product Contents.

Products of dark brown sugar (DBS) from different production processes and raw materials may bring different risks and benefits to human health. Therefore, this study was aimed to evaluate the quality of natural and commercial DBS products. Results showed that physicochemical properties, including pH value, turbidity, and browning degree have no significant difference between natural and commercial DBS products. Total flavonoid content of natural DBS was found to be significantly higher than that of commercial DBS (p < 0.05). Notably, the levels of harmful Maillard reaction products in natural DBS were significantly lower than that in commercial DBS as evidenced by analyses of methylglyoxal and fluorescent advanced glycation end products (p < 0.05). However, the amount of acrylamide in natural DBS was significantly higher than that in commercial DBS. In conclusion, this study provides useful information for risk-benefit assessment of DBS products, which is helpful for food safety management.

Assessment of a novel nanostructured flocculant with elevated flocculation and antimicrobial activity.

In this work, a novel process involving the preparation of nanochitosan-grafted flocculants (CPAM-g-NCS) to treat low turbid and salmonella suspensions simultaneously was introduced. Nanotechnology was employed to enhance the adsorption-adhesion and sterilization abilities of dual-functional flocculants. The monomers of chitosan, acrylamide, methacryloyl ethyl trimethyl ammonium chloride, and sodium tripolyphosphate were utilized for flocculants copolymerization. Then, using fourier-transform infrared spectroscopy, nuclear magnetic resonance hydrogen spectrum, and thermogravimetric and differential scanning calorimetry analysis, the successful synthesis of CPAM-g-NCS was verified. Scanning electron microscopy and size analysis suggested that nanostructured flocculants with irregular morphology and nanocolloids of 60.44 nm were formed. CPAM-g-NCS was applied to treat a series of simulated low turbid and salmonella suspensions. The simulation results showed that the minimum residual turbidity of 1.97 NTU and optical density of 0.16 (initial 0.89) can be achieved at dosages of 2.5 and 8.75 mg L-1, respectively, which were superior to conventional organics flocculants. Mechanistic studies suggested that the excellent adsorption property, and large numbers of quaternary ammonium and amino groups of nanoflocculants contributed to the superior flocculation and antibacterial performance of CPAM-g-NCS.

Synthesis and assessment of phenylacrylamide derivatives as potential anti-oxidant and anti-inflammatory agents.

Oxidative stress and inflammation are major causes of numerous life-threatening human diseases. In the present study, we synthesized a series of phenylacrylamide derivatives as novel anti-oxidant and anti-inflammatory agents. Biological evaluation showed that compound 6a could more potently protect HBZY-1 mesangial cells from H2O2-caused oxidative stress than positive controls resveratrol and sulforaphane by dose- and time-dependently impairing the ROS accumulation. Preliminary anti-oxidant mechanism studies indicated that compound 6a could activate Nrf2 and increase the protein and mRNA expression of downstream anti-oxidant enzymes, ie. NQO-1, HO-1, GCLM and GCLC. Notably, 6a could inhibit the production of NO and the activity of NF-κB in LPS-stimulated HBZY-1 mesangial cells, indicating its potential anti-inflammatory activity. Interestingly, both effects could be significantly attenuated by Nrf2 inhibitor TRG, HO-1 inhibitor ZnPP or GCL inhibitor BSO at non-toxic concentrations, confirming that the anti-oxidant and anti-inflammatory activity of 6a is related to the activation of Nrf2 signaling pathway. These results, together with the relatively safety profile, indicated that compound 6a could be a promising lead to develop novel anti-oxidant and anti-inflammatory agents, thus preventing diseases induced by oxidative stress and inflammation.

Marine microbial L-asparaginase: Biochemistry, molecular approaches and applications in tumor therapy and in food industry.

The marine environment is a rich source of biological and chemical diversity. It covers more than 70% of the Earth's surface and features a wide diversity of habitats, often displaying extreme conditions, where marine organisms thrive, offering a vast pool for microorganisms and enzymes. Given the dissimilarity between marine and terrestrial habitats, enzymes and microorganisms, either novel or with different and appealing features as compared to terrestrial counterparts, may be identified and isolated. L-asparaginase (E.C. 3.5.1.1), is among the relevant enzymes that can be obtained from marine sources. This amidohydrolase acts on L-asparagine and produce L-aspartate and ammonia, accordingly it has an acknowledged chemotherapeutic application, namely in acute lymphoblastic leukemia. Moreover, L-asparaginase is also of interest in the food industry as it prevents acrylamide formation. Terrestrial organisms have been largely tapped for L-asparaginases, but most failed to comply with criteria for practical applications, whereas marine sources have only been marginally screened. This work provides an overview on the relevant features of this enzyme and the framework for its application, with a clear emphasis on the use of L-asparaginase from marine sources. The review envisages to highlight the unique properties of marine L-asparaginases that could make them good candidates for medical applications and industries, especially in food safety.

Mimicking new receptors based on molecular imprinting and their application to potentiometric assessment of 2,4-dichlorophenol as a food taint.

Innovative host-tailored polymers were prepared, characterized and used as recognition elements in potentiometric transducers for the selective quantification of 2,4-dichlorophenol (DCP).The polymer beads were synthesized using DCP as a template molecule, acrylamide (AM),methacrylic acid (MAA) and ethyl methacrylate (EMA) as functional monomers and divinylbenzene (DVB) and ethylene glycol dimethacrylate (EGDMA) as cross-linkers. The sensors were fabricated by the inclusion of MIPs in plasticized polyvinyl chloride (PVC) matrix. Response characteristics of the proposed sensors revealed anionic slopes of -59.2, -49.7 and -80.6 mV/decade with detection limits of 5.6 × 10-5,5.9 × 10-5 and 13.2 × 10-5 mol/L for MIP/AM/DVB, MIP/MAA/DVB and MIP/EMA/EGDMA membrane based sensors, respectively. Good selectivity was observed over common inorganic/organic anions. Validation of the assay method according to IUPAC recommendations was justified ensuring the synthesis of good reliable novel sensors for DCP determination. The method was successfully applied for routine analysis of food taint in fish and fish farms water samples.

Exposure assessment of process-related contaminants in food by biomarker monitoring.

Exposure assessment is a fundamental part of the risk assessment paradigm, but can often present a number of challenges and uncertainties. This is especially the case for process contaminants formed during the processing, e.g. heating of food, since they are in part highly reactive and/or volatile, thus making exposure assessment by analysing contents in food unreliable. New approaches are therefore required to accurately assess consumer exposure and thus better inform the risk assessment. Such novel approaches may include the use of biomarkers, physiologically based kinetic (PBK) modelling-facilitated reverse dosimetry, and/or duplicate diet studies. This review focuses on the state of the art with respect to the use of biomarkers of exposure for the process contaminants acrylamide, 3-MCPD esters, glycidyl esters, furan and acrolein. From the overview presented, it becomes clear that the field of assessing human exposure to process-related contaminants in food by biomarker monitoring is promising and strongly developing. The current state of the art as well as the existing data gaps and challenges for the future were defined. They include (1) using PBK modelling and duplicate diet studies to establish, preferably in humans, correlations between external exposure and biomarkers; (2) elucidation of the possible endogenous formation of the process-related contaminants and the resulting biomarker levels; (3) the influence of inter-individual variations and how to include that in the biomarker-based exposure predictions; (4) the correction for confounding factors; (5) the value of the different biomarkers in relation to exposure scenario's and risk assessment, and (6) the possibilities of novel methodologies. In spite of these challenges it can be concluded that biomarker-based exposure assessment provides a unique opportunity to more accurately assess consumer exposure to process-related contaminants in food and thus to better inform risk assessment.

Determination of acrylamide levels in potato crisps and other snacks and exposure risk assessment through a Margin of Exposure approach.

Potato crisps, corn-based extruded snacks and other savoury snacks are very popular products especially among younger generations. These products could be a potential source of acrylamide (AA), a toxic compound which could develop during frying and baking processes. The purpose of this study was the assessment of the dietary intake to AA across six groups of consumers divided according to age through the consumption of potato crisps and other snacks, in order to eventually evaluate the margin of exposure (MOE) related to neurotoxic and carcinogenic critical endpoints. Different brands of potato crisps and other popular snacks were analyzed through a matrix solid-phase dispersion method followed by a bromination step and GC-MS quantification. The concentration of detected AA ranged from 21 to 3444 ng g-1 and the highest level occurred in potato crisps samples which showed a median value of 968 ng g-1. The risk characterization through MOE assessment revealed that five out of six consumers groups showed higher exposure values associated with an augmented carcinogenic risk.

Assessment of novel maleic anhydride copolymers prepared via nitroxide-mediated radical polymerization as CaSO4 crystal growth inhibitors.

Calcium sulfate is one of the dominant scales which, unlike carbonate scale, are not easily removable by acid. To inhibit CaSO4 scale formation in artificial cooling water systems, well-defined low molecular weight maleic anhydride and n-alkylacrylamide copolymers (YMR-S series) were synthesized via nitroxide-mediated radical polymerization initiated by benzoyl peroxide in the presence of 2,2,6,6-tetramethyl-1-piperidinyloxy at varying concentrations. These polymerizations exhibit living polymerization characteristics; that is, they show linear growth in chain length as a function of monomer conversion, and have narrow molecular weight distributions. Resultant polymers were characterized by means of 1H-NMR and 13C-NMR. The inhibition behavior of these YMR-S series polymers against CaSO4 was evaluated using the static scale inhibition method and a dynamic tube block test. The inhibition ability on the CaSO4 scale is 99.5% with 9 ppm dosage level at pH 10.45 and temperature 70°C. Scanning electronic microscope analysis proved the morphological changes of the CaSO4 scales due to the strong inhibition action of YMR-S polymers. It is also observed that the antiscaling effect of the copolymers greatly depends on the molecular weight, and the optimum range is below 20,000 and approximately in the range 500-2000.

Reduction in Dietary Acrylamide Exposure-Impact of Potatoes with Low Acrylamide Potential.

Acrylamide forms primarily from a reaction between reducing sugars (e.g., glucose and fructose) and an amino acid (asparagine, Asn) formed naturally in foods, including potatoes. This reaction occurs when carbohydrate-rich foods are heated at temperatures above 120 °C. Multiple potato varieties were transformed with potato genomic DNA that results in down-regulation of the expression of the asparagine synthetase-1 gene (Asn1), significantly reducing synthesis of free Asn, and consequently lowering the potential to form acrylamide during cooking. These potatoes with low acrylamide potential (LAP) were tested in agronomic trials, and processed into French fries and potato chips. Decreased levels of acrylamide were measured in these cooked food products when derived from LAP potatoes compared with those derived from conventional potatoes. These reductions can be directly attributed to reduction in Asn levels in the LAP potatoes. The corresponding average reduction in exposure to acrylamide from French fry and potato chip consumption is estimated to be 65%, which would amount to approximately a 25% reduction in overall dietary exposure. Considering that children consume nearly three times more acrylamide than adults on a per kg body weight basis, they would experience the most impact from the reduced acrylamide associated with LAP potatoes. The potential public health impacts, in context of dietary acrylamide exposure reduction, are discussed in this study.

Burden of disease of dietary exposure to acrylamide in Denmark.

Acrylamide (AA) is a process-contaminant that potentially increases the risk of developing cancer in humans. AA is formed during heat treatment of starchy foods and detected in a wide range of commonly consumed products. Increased focus on risk ranking and prioritization of major causes of disease makes it relevant to estimate the impact that exposure to chemical contaminants and other hazards in food have on health. In this study, we estimated the burden of disease (BoD) caused by dietary exposure to AA, using disability adjusted life years (DALY) as health metric. We applied an exposure-based approach and proposed a model of three components: an exposure, health-outcome, and DALY-module. We estimated BoD using two approaches for estimating cancer risk based on toxicological data and two approaches for estimating DALY. In Denmark, 1.8 healthy life years per 100.000 inhabitants are lost each year due to exposure to AA through foods, as estimated by the most conservative approach. This result should be used to inform risk management decisions and for comparison with BoD of other food-borne hazards for prioritizing policies. However, our study shows that careful evaluation of methodological choices and assumptions used in BoD studies is necessary before use in policy making.

A colorimetric detection of acrylamide in potato chips based on nucleophile-initiated thiol-ene Michael addition.

Acrylamide (AA), a neurotoxin and a potential carcinogen, has been found in various thermally processed foods such as potato chips, biscuits, and coffee. Simple, cost-effective, and sensitive methods for the rapid detection of AA are needed to ensure food safety. Herein, a novel colorimetric method was proposed for the visual detection of AA based on a nucleophile-initiated thiol-ene Michael addition reaction. Gold nanoparticles (AuNPs) were aggregated by glutathione (GSH) because of a ligand-replacement, accompanied by a color change from red to purple. In the presence of AA, after the thiol-ene Michael addition reaction between GSH and AA with the catalysis of a nucleophile, the sulfhydryl group of GSH was consumed by AA, which hindered the subsequent ligand-replacement and the aggregation of AuNPs. Therefore, the concentration of AA could be determined by the visible color change caused by dispersion/aggregation of AuNPs. This new method showed high sensitivity with a linear range from 0.1 µmol L(-1) to 80 µmol L(-1) and a detection limit of 28.6 nmol L(-1), and especially revealed better selectivity than the fluorescence sensing method reported previously. Moreover, this new method was used to detect AA in potato chips with a satisfactory result in comparison with the standard methods based on chromatography, which indicated that the colorimetric method can be expanded for the rapid detection of AA in thermally processed foods.

Use and impact of usual intake models on dietary exposure estimate and risk assessment of chemical substances: a practical example for cadmium, acrylamide and sulphites.

To estimate of food and nutrient intakes, 24-h recalls are frequently used in dietary assessment. However intake data collected for a short period are a limited estimator of long-term usual intake. An important limitation of such data is that the within-person variability tends to inflate the intake distribution leading to a biased estimation of extreme percentiles. Statistical models, named usual-intake models, that separate the within-person variability from the between-persons variability, have lately been implemented. The main objectives of this study were to highlight the potential impact that usual-intake models can have on exposure estimate and risk assessment and to point out which are the key aspects to be considered in order to run these models properly and be sure to interpret the output correctly. To achieve the goal we used the consumption data obtained by the French dietary survey INCA2 and the concentration data collected during the French TDS2, using Monte Carlo Risk Assessment (MCRA) software, release 8.0. For the three substances included in this study (cadmium, acrylamide and sulphites), the exposure of the upper percentiles was significantly reduced when using usual-intake models in comparison with the results obtained in the observed individual mean models, even if in terms of risk assessment the impact of using usual-intake models was limited. From the results it appears that the key aspects to consider when using usual-intake models are: (1) the normality of the log-transformed intake distribution, (2) the contribution per single food group to the total exposure, and (3) the independency of food consumption data on multiple days. In conclusion, usual-intake models may have an impact on exposure estimates although, referring to the results, it did not bring any changes in terms of risk assessment, but further investigations are needed.

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.

A statistical regression model for the estimation of acrylamide concentrations in French fries for excess lifetime cancer risk assessment.

Human exposure to acrylamide (AA) through consumption of French fries and other foods has been recognized as a potential health concern. Here, we used a statistical non-linear regression model, based on the two most influential factors, cooking temperature and time, to estimate AA concentrations in French fries. The R(2) of the predictive model is 0.83, suggesting the developed model was significant and valid. Based on French fry intake survey data conducted in this study and eight frying temperature-time schemes which can produce tasty and visually appealing French fries, the Monte Carlo simulation results showed that if AA concentration is higher than 168 ppb, the estimated cancer risk for adolescents aged 13-18 years in Taichung City would be already higher than the target excess lifetime cancer risk (ELCR), and that by taking into account this limited life span only. In order to reduce the cancer risk associated with AA intake, the AA levels in French fries might have to be reduced even further if the epidemiological observations are valid. Our mathematical model can serve as basis for further investigations on ELCR including different life stages and behavior and population groups.

Acrylamide as environmental noxious agent: a health risk assessment for the general population based on the internal acrylamide burden.

The present population-based cross-sectional study in 1008 volunteers was aimed at quantifying the internal burden of acrylamide (AM) in the general population using hemoglobin adduct levels as biomarker of exposure. Based on these adduct levels the risk of (AM) related hazardous health effects in the general population is assessed. Hemoglobin adducts of acrylamide were detected in 999 out of 1008 analyzed blood samples (99.1%). Smoking can be regarded as a main source of overall acrylamide intake in persons without occupational exposure to acrylamide but a second main influence factor, i.e. food intake, is of environmental importance. Due to the sizable study population the 95th percentile differentiated in smokers and non-smokers are actually the best available reference values for an internal environmental acrylamide burden. Based on our data neurotoxicity and reproductive toxicity due to acrylamide are not likely to occur in the general population except very high consumers. However, carcinogenicity and mutagenicity are possible hazardous health effects for the general population.

Gel immobilization of acrylamide-modified single-stranded DNA template for pyrosequencing.

A novel two-step process was developed to prepare ssDNA templates for pyrosequencing. First, PCR-amplified DNA templates modified with an acrylamide group and acrylamide monomers were copolymerized in 0.1 M NaOH solution to form polyacrylamide gel spots. Second, ssDNA templates for pyrosequencing were prepared by removing electrophoretically unbound complementary strands, unmodified PCR primers, inorganic pyrophosphate (PPi), and excess deoxyribonucleotides under alkali conditions. The results show that the 3-D polyacrylamide gel network has a high immobilization capacity and the modified PCR fragments are efficiently captured. After electrophoresis, gel spots copolymerized from 10 microL of the crude PCR products and the acrylamide monomers contain template molecules on the order of pmol, which generate enough light to be detected by a regular photomultiplier tube. The porous structure of gel spots facilitated the fast transportation of the enzyme, dNTPs and other reagents, and the solution-mimicking microenvironment guaranteed polymerase efficiency for pyrosequencing. Successful genotyping from the crude PCR products was demonstrated. This method can be applied in any laboratory; it is cheap, fast, simple, and has the potential to be incorporated into a DNA-chip format for high-throughput pyrosequencing analysis.

Acrylamide: considerations for risk management.

The presence of acrylamide in many carbohydrate-rich foods is due to its formation during conventional heating and preparation methods. Although acrylamide is established to be a toxic substance, the implications to public health from the amounts found in food are not clear. A better scientific understanding is required to help determine whether, and to what extent, formal risk management action might be necessary. Since acrylamide in food was highlighted in 2002, numerous investigations and initiatives have been developed, including international collaborations across governments, industry, research organizations, and consumer representations. The newly generated information is being used to help the overall understanding of this issue. In particular, new information on health aspects will be important to update the scientific risk assessment. The basis for decisions on possible risk management measures would then be clearer. If future risk assessment concludes that the amounts of acrylamide in food can pose a health threat, then options for risk management will need to be considered, such as limits, guide levels, codes of practice, guidance information, and advice to the food and catering industries and to consumers. In the meantime, it is possible to benefit from progress already made on how acrylamide is formed in food and on ways to lower the amounts present. Raising awareness to the approaches that can reduce the presence of acrylamide in food should be encouraged. Where feasible, such approaches can be assessed for practical use in production, processing, and preparation of the relevant food products.

[Gel-producing properties of exopolysaccharide acrylamide in composition with agar-agar]. / Zheliruiushchie svoistva ékzopolisakharidakriamida v kompozitsii s agar-agarom.

A possibility to use a new polymer--exopolysaccharide acrylamide (EPAA)--for partial substitution of agar-agar in solid nutrient media in microbiology has been studied. It has been established than the EPAA in concentration 6.0-12.0 g/l with agar in concentration 10.0 g/l produce gel which is so strong as agar in concentration 15.0-20.0 g/l. Quality of gels produced depends an the method of EPAA mixing with agar. It has been shown that usage of EPAA in nutrient media does not affect growth, colony and cell morphology and biochemical properties of microorganisms. The use of EPAA to prepare nutrient media permits us to reduce expenditures for agar.