Search

Abstract: In the present studies, several hypotheses were tested to explain previously reported differential effects of soy and casein on colon cancer biomarkers like cell proliferation, fecal fat, fecal bile acid, alkaline phosphatase, and magnesium excretion in rats. In Study 1, the effect of methionine, a limiting amino acid in soy protein and an amino acid that is thought to have a marked effect on colonic cell proliferation, was tested. It was concluded that methionine up to 1% in the diet had no effect on cell proliferation, using the3H-thymidine assay. The same study revealed that fecal alkaline phosphatase excretion is a good marker for colonic epithelial damage and fecal magnesium excretion is not. In Study 2, the hypothesis was tested that soy fractions enriched with isoflavones and saponins may increase fat excretion and so influence colonic cell proliferation in rats. It was indeed shown that soy protein isolate and an ethanolic extract from soy protein isolate slightly increased fecal fat excretion (up to 1.7-fold). However, fecal water bile acid and free fatty acid concentrations were decreased after feeding soy protein-based diets compared with casein, and no difference in fecal alkaline phosphatase excretion was observed. In Study 3, the lytic potential of soy saponins and the interaction between saponins and some lytic bile acids were tested in vitro. Data suggest a protective effect from soy saponins by reducing lytic activity of cholic acid. The overall conclusion is that soy protein compared with casein influences several colon cancer risk parameters, indicating a more protective rather than a stimulating effect on colon cancer risk. [ABSTRACT FROM AUTHOR]

In a previous study, raw cashew kernels were assayed for the fungal contamination focusing on strains belonging to the genus Aspergillus and on aflatoxins producers. These samples showed high contamination with Aspergillus section Nigri species and absence of aflatoxins. To investigate the diversity of secondary metabolites, including mycotoxins, the species of A. section Nigri may produce and thus threaten to contaminate the raw cashew kernels, 150 strains were isolated from cashew samples and assayed for their production of secondary metabolites using liquid chromatography high resolution mass spectrometry (LC-HRMS). Seven species of black Aspergilli were isolated based on morphological and chemical identification: A. tubingensis (44%), A. niger (32%), A. brasiliensis (10%), A. carbonarius (8.7%), A. luchuensis (2.7%), A. aculeatus (2%) and A. aculeatinus (0.7%). From these, 45 metabolites and their isomers were identified. Aurasperone and pyranonigrin A, produced by all species excluding A. aculeatus and A. aculeatinus, were most prevalent and were encountered in 146 (97.3%) and 145 (95.7%) isolates, respectively. Three mycotoxins groups were detected: fumonisins (B2 and B4) (2.7%) ochratoxin A (13.3%), and secalonic acids (2%), indicating that these mycotoxins could occur in raw cashew nuts. Thirty strains of black Aspergilli were randomly sampled for verification of species identity based on sequences of β-tubulin and calmodulin genes. Among them, 27 isolates were positive to the primers used and 11 were identified as A. niger, 7 as A. tubingensis, 6 as A. carbonarius, 2 as A. luchuensis and 1 as A. welwitschiae confirming the species names as based on morphology and chemical features. These strains clustered in 5 clades in A. section Nigri. Chemical profile clustering also showed also 5 groups confirming the species specific metabolites production., Competing Interests: The authors have declared that no competing interests exist.

Heating of protein- and sugar-containing materials is considered the primary factor affecting the formation of advanced glycation end products (AGEs). This study aimed to investigate the influence of heating conditions, digestion, and aggregation on the binding capacity of AGEs to the soluble AGE receptor (sRAGE). Samples consisting of mixtures of whey protein and lactose were heated at 130 °C. An in vitro infant digestion model was used to study the influence of heat treatment on the digestibility of whey proteins. The amount of sRAGE-binding ligands before and after digestion was measured by an ELISA-based sRAGE-binding assay. Water activity did not significantly affect the extent of digestibility of whey proteins dry heated at pH 5 (ranging from 3.3 ± 0.2 to 3.6 ± 0.1% for gastric digestion and from 53.5 ± 1.5 to 64.7 ± 1.1% for duodenal digestion), but there were differences in cleavage patterns of peptides among the samples heated at different pH values. Formation of sRAGE-binding ligands depended on the formation of aggregates and was limited in the samples heated at pH 5. Moreover, the sRAGE-binding activity of digested sample was changed by protease degradation and correlated with the digestibility of samples. In conclusion, generation of sRAGE-binding ligands during extensive heat treatment of whey protein/lactose mixtures is limited in acidic heating condition and dependent on glycation and aggregation.

Pulsed electrical field (PEF) technology offers an alternative to thermal pasteurisation of high-acid fruit juices, by extending the shelf life of food products, while retaining its fresh taste and nutritional value. Substantial research has been performed on the effect of electrical field strength on the inactivation kinetics of spoilage and pathogenic micro-organisms and on the outgrowth of spoilage micro-organisms during shelf life. However, studies on the effect of electrical field strength on the inactivation and outgrowth of surviving populations during shelf life are missing. In this study, we assessed the influence of electrical field strength applied by PEF processing and storage temperature on the outgrowth of surviving yeast and mould populations naturally present in fresh fruit smoothie in time. Therefore, an apple-strawberry-banana smoothie was treated in a continuous-flow PEF system (130L/h), using similar inlet and outlet conditions (preheating temperature 41°C, maximum temperature 58°C) to assure that the amount of energy across the different conditions was kept constant. Smoothies treated with variable electrical field strengths (13.5, 17.0, 20.0 and 24.0kV/cm) were compared to smoothies without treatment for outgrowth of yeasts and moulds. Outgrowth of yeasts and moulds stored at 4°C and 7°C was analysed by plating and visual observation and yeast growth was modelled using the modified logistic growth model (Zwietering model). Results showed that the intensity of the electrical field strength had an influence on the degree of inactivation of yeast cells, resulting in a faster outgrowth over time at lower electrical field strength. Outgrowth of moulds over time was not affected by the intensity of the electrical field strength used. Application of PEF introduces a trade-off between type of spoilage: in untreated smoothie yeasts lead to spoilage after 8days when stored at 4 or 7°C, whereas in PEF treated smoothie yeasts were (partly) inactivated and provided outgrowth opportunities for moulds, which led to spoilage by moulds after 14days (7°C) or 18days (4°C)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Lupinus mutabilis has protein (32.0-52.6 g/100 g dry weight) and lipid (13.0-24.6 g/100 g dry weight) contents similar to soya bean (Glycine max). The Ω3, Ω6, and Ω9 contents are 1.9-3.0, 26.5-39.6, and 41.2-56.2 g/100 g lipid, respectively. Lupins can be used to fortify the protein content of pasta, bread, biscuits, salads, hamburgers, sausages, and can substitute milk and soya bean. Specific lupin protein concentrates or isolates display protein solubility (>90%), water-absorption capacity (4.5 g/g dry weight), oil-absorption capacity (3.98 g/g), emulsifying capacity (2000 mL of oil/g), emulsifying stability (100%, 60 hours), foaming capacity (2083%), foaming stability (78.8%, 36 hours), and least gelation concentration (6%), which are of industrial interest. Lupins contain bitter alkaloids. Preliminary studies on their toxicity suggest as lethal acute dose for infants and children 10 mg/kg bw and for adults 25 mg/kg bw. However, alkaloids can also have medical use for their hypocholesterolemic, antiarrhythmic, and immunosuppressive activity. Bitter lupins can be detoxified by biological, chemical, or aqueous processes. The shortest debittering process requires one hour. This review presents the nutritional composition of lupins, their uses (as food, medicine, and functional protein isolates), toxicology, and debittering process scenarios. It critically evaluates the data, infers conclusions, and makes suggestions for future research.

This study aimed to investigate the effects of sugar type on the reaction mechanism for formation of acrylamide and 5-hydroxymethylfurfural (HMF) during the baking of biscuits at 200°C using multiresponse modelling. Four types of biscuits were prepared: (1) with sucrose, (2) with glucose and fructose, (3) with fructose only and (4) with glucose only. Experimental data showed that HMF concentration was highest in biscuits with glucose and fructose, whereas acrylamide concentration was highest in biscuits with glucose, also having the highest asparagine concentration. Proposed mechanistic models suggested that HMF is formed via caramelisation and that acrylamide formation follows the specific amino acid route, i.e., reducing sugars react with asparagine to form the Schiff base before decarboxylation, to generate acrylamide without the Amadori rearrangement product and sugar fragmentation. Study results contribute to understanding chemical reaction pathways in real food products., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This study investigated the formation of N(ε)-carboxymethyllysine (CML) in two caseinate solutions containing: (1) glucose, (2) lactose, each heated at 120 °C and 130 °C. At both heating temperatures, CML concentration in lactose-caseinate solution was higher than in glucose-caseinate solution. In both solutions, more CML was formed at 130 °C than at 120 °C. Using multiresponse modelling, two degradation routes for the sugars were confirmed: (1) isomerisation of glucose or lactose and subsequent degradation via Lobry de Bruyn-Alberda van Ekenstein (LA) arrangement; (2) the Maillard reaction between the reducing sugar and lysine residues. Modelling results suggested that CML was not formed from oxidation of the reducing sugars, but from the Maillard reaction via the Amadori rearrangement product. Since CML appeared to be thermally unstable under the current study conditions, it may not be a perfect indicator for heat damage of processed foods. This is the first study in which CML formation was linked to available information on the Maillard reaction via multiresponse modelling., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Exposure to food odours increases the appetite for congruent foods and decreases the appetite for incongruent foods. However, the effect of exposure to a variety of food odours, as often occurs in daily life, is unknown., Objective: Investigate how switching between sweet and savoury odours affects the appetite for sweet and savoury products., Design: Thirty women (age: 18-45y; BMI: 18.5-25kg/m2) intensely smelled the contents of cups filled with banana, meat or water (no-odour) in a within-subject design with four combinations: no-odour/banana, no-odour/meat, meat/banana and banana/meat. Participants received one combination per test day. In each combination, two cups with different fillings were smelled for five minutes after each other. Treatment order was balanced as much as possible. The effects of previous exposure and current odour on the appetite for (in)congruent sweet and savoury products, and odour pleasantness were analysed. A change from meat to banana odour or banana to meat odour was referred to as switch, whereas a change from no-odour to meat odour or no-odour to banana odour was no-switch., Results: The current odour (P<0.001), as opposed to the previous exposure (P = 0.71), determined the appetite for (in)congruent sweet and savoury products, already one minute after a switch between sweet and savoury odours. The pleasantness of the odour decreased during odour exposure (P = 0.005)., Conclusions: After a switch, the appetite for specific products quickly adjusted to the new odour and followed the typical pattern as found during odour exposure in previous studies. Interestingly, the appetite for the smelled food remained elevated during odour exposure, known as sensory-specific appetite, whereas the pleasantness of the odour decreased over time, previously termed olfactory sensory-specific satiety. This seeming contradiction may result from different mechanisms underlying the odour-induced anticipation of food intake versus the decrease in hedonic value during prolonged sensory stimulation.

Heat treatment is the most common way of milk processing, inducing structural changes as well as chemical modifications in milk proteins. These modifications influence the immune-reactivity and allergenicity of milk proteins. This study shows the influence of dry heating on the solubility, particle size, loss of accessible thiol and amino groups, degree of Maillard reaction, IgG-binding capacity and binding to the receptor for advanced glycation end products (RAGE) of thermally treated and glycated whey proteins. A mixture of whey proteins and lactose was dry heated at 130 °C up to 20 min to mimic the baking process in two different water activities, 0.23 to mimic the heating in the dry state and 0.59 for the semi-dry state. The dry heating was accompanied by a loss of soluble proteins and an increase in the size of dissolved aggregates. Most of the Maillard reaction sites were found to be located in the reported conformational epitope area on whey proteins. Therefore the structural changes, including exposure of the SH group, SH-SS exchange, covalent cross-links and the loss of available lysine, subsequently resulted in a decreased IgG-binding capacity (up to 33%). The binding of glycation products to RAGE increased with the heating time, which was correlated with the stage of the Maillard reaction and the decrease in the IgG-binding capacity. The RAGE-binding capacity was higher in samples with a lower water activity (0.23). These results indicate that the intensive dry heating of whey proteins as it occurs during baking may be of importance to the immunological properties of allergens in cow's milk, both due to chemical modifications of the allergens and formation of AGEs.

Mung bean (Vigna radiata (L.) R. Wilczek) has been intensively researched; scattered data are available on various properties. Data on physical, chemical, food processing, and nutritional properties were collected for whole mung bean grains and reviewed to assess the crop's potential as food and to set research priorities. Results show that mung bean is a rich source of protein (14.6-33.0 g/100 g) and iron (5.9-7.6 mg/100 g). Grain color is correlated with compounds like polyphenols and carotenoids, while grain hardness is associated with fiber content. Physical properties like grain dimensions, sphericity, porosity, bulk, and true density are related to moisture content. Anti-nutrients are phytic acid, tannins, hemagglutinins, and polyphenols. Reported nutrient contents vary greatly, the causes of which are not well understood. Grain size and color have been associated with different regions and were used by plant breeders for selection purposes. Analytical methods require more accuracy and precision to distinguish biological variation from analytical variation. Research on nutrient digestibility, food processing properties, and bioavailability is needed. Furthermore, the effects of storage and processing on nutrients and food processing properties are required to enable optimization of processing steps, for better mung bean food quality and process efficiency.

Broccoli belongs to the Brassicaceae plant family consisting of widely eaten vegetables containing high concentrations of glucosinolates. Enzymatic hydrolysis of glucosinolates by endogenous myrosinase (MYR) can form isothiocyanates with health-promoting activities. The effect of water content (WC) and temperature on MYR inactivation in broccoli was investigated. Broccoli was freeze dried obtaining batches with WC between 10% and 90% (aw from 0.10 to 0.96). These samples were incubated for various times at different temperatures (40-70°C) and MYR activity was measured. The initial MYR inactivation rates were estimated by the first-order reaction kinetic model. MYR inactivation rate constants were lower in the driest samples (10% WC) at all studied temperatures. Samples with 67% and 90% WC showed initial inactivation rate constants all in the same order of magnitude. Samples with 31% WC showed intermediate initial inactivation rate constants. These results are useful to optimise the conditions of drying processes to produce dried broccoli with optimal MYR retention for human health., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Understanding overconsumption starts with knowledge of how separate factors influence our eating behavior. Food cues such as food odors are known for their effect on general appetite and sensory-specific appetite (SSA). Active sniffing rather than passive exposure may induce satiation over time. The objective of this study was to investigate how actively sniffing banana odors affects general appetite, SSA, and subsequent food intake. In a crossover study, 61 women actively smelled cups containing natural banana, artificial banana odor, or water (no odor) for 10 min. Treatment order was randomly assigned as much as possible. General appetite and SSA were monitored by using 100-mm visual analog scales during the 10 min of active sniffing, followed by ad libitum intake of banana milkshake. Results showed that SSA was consistently high (+12 mm) during actively sniffing natural or artificial banana odors, with no decrease in SSA over time. Sniffing both banana odors increased the appetite for banana (+11 mm) and other sweet products (+4 mm), whereas the appetite for savory products decreased by 7 mm (all P < 0.01) compared with no odor. Actively sniffing banana odor did not significantly influence food intake (P = 0.68) or general appetite scores (P = 0.06). In conclusion, SSA scores during active sniffing were identical to the SSA found in a similar study that used passive smelling, suggesting that SSA is independent of the manner of sniffing and exposure time. Moreover, sweet/savory categorization may suggest that food odors communicate information about the nutrient composition of their associated foods. These data clearly show the appetizing effects of food odors., (© 2014 American Society for Nutrition.)

Background: Exposure to palatable food odors influences appetite responses, either promoting or inhibiting food intake. Possibly, food odors are appetizing after a short exposure (of circa 1-3 min), but become satiating over time (circa 10-20 min)., Objective: To investigate the effect of odor exposure on general appetite and sensory-specific appetite (SSA) over time., Design: In a cross-over study, 21 unrestrained women (age: 18-45 years; BMI: 18.5-25 kg m(-2)) were exposed for 20 min to eight different odor types: five food odors, two nonfood odors and no-odor. All odors were distributed in a test room at suprathreshold levels. General appetite, SSA and salivation were measured over time., Results: All food odors significantly increased general appetite and SSA, compared with the no-odor condition. The nonfood odors decreased general appetite. All effects did not change over time during odor exposure. Savory odors increased the appetite for savory foods, but decreased appetite for sweet foods, and vice versa after exposure to sweet odors. Neither food odors nor nonfood odors affected salivation., Conclusions: Palatable food odors were appetizing during and after odor exposure and did not become satiating over a 20-min period. Food odors had a large impact on SSA and a small impact on general appetite. Moreover, exposure to food odors increased the appetite for congruent foods, but decreased the appetite for incongruent foods. It may be hypothesized that, once the body is prepared for intake of a certain food with a particular macronutrient composition, it is unfavorable to consume foods that are very different from the cued food.

Pulsed electrical field (PEF) technology can be used for the inactivation of micro-organisms and therefore for preservation of food products. It is a mild technology compared to thermal pasteurization because a lower temperature is used during processing, leading to a better retention of the quality. In this study, pathogenic and spoilage micro-organisms relevant in refrigerated fruit juices were studied to determine the impact of process parameters and juice composition on the effectiveness of the PEF process to inactivate the micro-organisms. Experiments were performed using a continuous-flow PEF system at an electrical field strength of 20 kV/cm with variable frequencies to evaluate the inactivation of Salmonella Panama, Escherichia coli, Listeria monocytogenes and Saccharomyces cerevisiae in apple, orange and watermelon juices. Kinetic data showed that under the same conditions, S. cerevisiae was the most sensitive micro-organism, followed by S. Panama and E. coli, which displayed comparable inactivation kinetics. L. monocytogenes was the most resistant micro-organism towards the treatment conditions tested. A synergistic effect between temperature and electric pulses was observed at inlet temperatures above 35 °C, hence less energy for inactivation was required at higher temperatures. Different juice matrices resulted in a different degree of inactivation, predominantly determined by pH. The survival curves were nonlinear and could satisfactorily be modeled with the Weibull model., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The present study investigated the effect of aroma exposure time and aroma concentration on ad libitum intake and subjective satiation. In a within-subject study, thirty-eight unrestrained, healthy female participants (age: 18-39 years; BMI: 18·5-26·0 kg/m²) were asked to consume tomato soup during lunchtime, until they felt comfortably full. Every 30 s, the participants consumed 10 g of a bland soup base while tomato soup aroma was delivered separately through the nose via a retronasal tube that was attached to an olfactometer. This gave the impression of consuming real tomato soup. For each sip, the aroma varied in exposure time (3 and 18 s) and concentration (5 × ), resulting in four different test conditions. Ad libitum food intake and appetite profile parameters were measured. A 9% lower food intake was observed when the participants were exposed to the condition with 18 s exposure time and a high concentration than when exposed to the other three conditions. These results indicate that changing the retronasal aroma release by aroma concentration and aroma exposure time affects food intake.

This review paper discusses opportunities for intelligent packaging for monitoring directly or indirectly quality attributes of perishable packaged foods. The possible roles of intelligent packaging as a tool in supply chain management are discussed as well as the barriers to implement this kind of technology in commercial applications. Cases on pasteurized milk and fresh cod fillets illustrate the application of different intelligent packaging concepts to monitor and estimate quality attributes. Conditions influencing quality (e.g., temperature-time) can be monitored to predict the quality of perishable products when the initial quality is known and rather constant (e.g., pasteurized milk). Products with a highly variable initial quality (e.g., fresh fish) require sensors monitoring compounds correlated with quality.

Consumption habits for mung bean foods were assessed by the free word association method and interview techniques. Four groups of closely related products and perceived quality were revealed. The largest group comprised sweets and snacks, which were associated with unhealthiness, expensiveness and sensory liking. Another group consisted of split dhals associated with convenience and healthiness. It appeared that under different circumstances food choices vary and are influenced more by socioeconomic restrictions then by consumer perception and preferences. Scenario analysis based on consumer perception, preferences, practices and nutritional value of products revealed dhals as the most promising food for innovation.

Increasing demand of shea products (kernels and butter) has led to the assessment of the state-of-the-art of these products. In this review, attention has been focused on macronutrients and micronutrients of pulp, kernels, and butter of shea tree and also the physicochemical properties of shea butter. Surveying the literature revealed that the pulp is rich in vitamin C (196.1 mg/100 g); consumption of 50 g covers 332% and 98% of the recommended daily intake (RDI) of children (4-8 years old) and pregnant women, respectively. The kernels contain a high level of fat (17.4-59.1 g/100 g dry weight). Fat extraction is mainly done by traditional methods that involve roasting and pressing of the kernels, churning the obtained liquid with water, boiling, sieving, and cooling. The fat (butter) is used in food preparation and medicinal and cosmetics industries. Its biochemical properties indicate some antioxidant and anti-inflammatory activities. Large variations are observed in the reported values for the composition of shea products. Recommendations for future research are presented to improve the quality and the shelf-life of the butter. In addition, more attention should be given to the accuracy and precision in experimental analyses to obtain more reliable information about biological variation.

Tenebrio molitor, Zophobas morio, Alphitobius diaperinus, Acheta domesticus and Blaptica dubia were evaluated for their potential as a future protein source. Crude protein content ranged from 19% to 22% (Dumas analysis). Essential amino acid levels in all insect species were comparable with soybean proteins, but lower than for casein. After aqueous extraction, next to a fat fraction, a supernatant, pellet, and residue were obtained, containing 17-23%, 33-39%, 31-47% of total protein, respectively. At 3% (w/v), supernatant fractions did not form stable foams and gels at pH 3, 5, 7, and 10, except for gelation for A. domesticus at pH 7. At 30% w/v, gels at pH 7 and pH 10 were formed, but not at pH 3 and pH 5. In conclusion, the insect species studied have potential to be used in foods due to: (1) absolute protein levels; (2) protein quality; (3) ability to form gels., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The objective of this study was to obtain insight into the effect of frying instructions on food handlers' control decisions in restaurants and to investigate the impact of control decisions on the variation and concentration of acrylamide in French fries. The concentrations of acrylamide and reducing sugars were analyzed, the frying temperature and time were measured, and thawing practices were observed. The results obtained before and after instructions were provided to the food handlers were compared for restaurants as a group and for each restaurant. Frying instructions supported food handlers' decisions to start frying when the oil temperature reached 175°C; all handlers started frying at the correct temperature. However, the effect of the instructions on the food handlers' decisions for frying time differed; most handlers increased the frying time beyond 240 s to achieve crispier French fries with a final color dictated by their preference. Providing instructions did not result in a significant difference in the mean concentration of acrylamide in French fries for the restaurants as a group. However, data analyzed for each restaurant revealed that when food handlers properly followed the instructions, the mean concentration of acrylamide was significantly lower (169 μg/kg) than that before instructions were provided (1,517 μg/kg). When food handlers did not complying with the frying instructions, mean acrylamide concentrations were even higher than those before instructions were provided. Two different strategies were developed to overcome the noncompliant behavior of food handlers: establishing requirements for the features of commercial fryers and strict monitoring of compliance with instructions.

The objective of this study was to obtain insight into the actual effectiveness of lowering reducing sugars concentration in par-fried potato strips on the concentration and variation of acrylamide in French fries prepared in real-life situations in food service establishments. Acrylamide, frying time, frying temperature, and reducing sugars were measured and characteristics of fryers were recorded. Data showed that the use of par-fried potato strips with lower concentrations of reducing sugars than the commonly used potato strips was an effective measure to reduce acrylamide concentrations in French fries prepared under standardised frying conditions. However, there was still large variation in the acrylamide concentrations in French fries, although the variation in reducing sugars concentrations in low and normal types of par-fried potato strips was very small and the frying conditions were similar. Factors that could affect the temperature-time profile of frying oil were discussed, such as setting a lower frying temperature at the end than at the start of frying, product/oil ratio and thawing practice. These need to be controlled in daily practice to reduce variation in acrylamide., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: A lyophilised reference serum from one patient with rheumatoid arthritis (RA) diluted with serum samples from healthy subjects was evaluated as a possible first international standard for anticitrullinated peptide antibodies (ACPAs)., Methods: The authors used 12 commercial ELISAs for ACPA detection in the reference serum and for testing the linearity of the assays by studying twofold serial dilutions. To test the effectiveness of the standardisation, sera from 20 RA patients with variable antibody concentrations were analysed, and the relative concentrations were calculated using both the kit's own curve and the six dilutions of the reference serum as a calibration curve. Fifty sera from normal healthy subjects were used to calculate cut-off values for the reference serum using each commercial kit., Results: The calibration curve obtained for each of the 12 methods using the reference sample dilutions as calibrator allowed harmonisation of the ACPA concentration of the 20 RA serum samples, significantly reducing the dispersion of the values. The mean coefficient of variation (CV) was reduced from 76.4% to 27.9% (p=0.018) and from 85.9% to 33.5% (p=0.028) for the medium/high and negative samples, respectively. Low positive sera CV was also reduced, but to a smaller degree, from 82.5% to 55.5% (p=0.043)., Conclusion: This first evaluation of the behaviour of the ACPA reference serum demonstrated that it tested positive in all the assays and that it may be used as a reference standard for establishing calibration curves, reducing the dispersion of antibody values and better comparing results obtained from different methods/laboratories.

Acrylamide is a probable human carcinogen, and its presence in a range of fried and oven-cooked foods has raised considerable health concern world-wide. Dietary intake studies observed significant variations in acrylamide concentrations, which complicate risk assessment and the establishment of effective control measures. The objective of this study was to obtain an insight into the actual variation in acrylamide concentrations in French fries prepared under typical conditions in a food service establishment (FSE). Besides acrylamide, frying time, frying temperature, and reducing sugars were measured and the actual practices at receiving, thawing and frying during French fries preparation were observed and recorded. The variation in the actual frying temperature contributed most to the variation in acrylamide concentrations, followed by the variation in actual frying time; no obvious effect of reducing sugars was found. The lack of standardised control of frying temperature and frying time (due to inadequate frying equipment) and the variable practices of food handlers seem to contribute most to the large variation and high acrylamide concentrations in French fries prepared in a restaurant type of FSE as compared to chain fast-food services, and institutional caterers. The obtained insights in this study can be used to develop dedicated control measures in FSE, which may contribute to a sustainable reduction in the acrylamide intake., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A survey among 246 people belonging to 14 ethnic groups and living in 5 different parklands in Benin revealed different practices to process shea kernels (namely boiling followed sun drying and smoking) and extract shea butter. A relation between parklands, gathering period, and sun-drying conditions was established. Moisture content and appearance of kernels were the selection criteria for users of shea kernels; color was the main characteristic to buy butter. Constraints to be solved are long processing times, lack of milling equipment and high water requirements. Best practices for smoking, sun drying, and roasting operations need to be established for further improvement.

In Africa, nutrient intake deficiencies are widespread. We, therefore, investigated the potential contribution of cowpea dishes to the ingestion of several macro- and micronutrients. Processors and consumers were interviewed and cowpea dishes analyzed. Energy, protein, iron, zinc, and calcium contents ranged from 1647 to 2570 kJ, 10 to 25 g, 1 to 35 mg, 1.5 to 3.0 mg, 38 to 380 mg per 100 g d.w., respectively. The iron and calcium contents were highest in dishes containing leaves. The consumption of these dishes should be promoted along with research on how to further decrease the associated antinutritional factors of traditional cowpea dishes.

Forest food resources contribute significantly to food supply in areas where they grow. Three fermented baobab foods were studied: Dikouanyouri (from seeds, pH = 6.5); Tayohounta (from seed kernels, pH = 7), and Mutchayan (from baobab pulp and sorghum, pH = 4.2). Bacillus spp. (8.5 and 9.5 Log cfu /g) and lactic acid bacteria (8.9 and 8.4 Log cfu /g,) dominate in Dikouanyouri and Tayohounta, respectively. In Mutchayan, lactic acid bacteria (8.1 Log cfu/g) and yeasts (7.2 Log cfu/g) predominated. The arbitrary index of protein cleavage increases from 2.3% (unfermented products) to 13.7% in Dikouanyouri and 21.3% in Tayohounta, indicating significant protein degradation. Mutchayan is the most frequently consumed product.

The mean spherical approximation (MSA) theory is used to explain the impact of sugars on ion properties in milk-based systems by taking into account electrostatic interactions and volume exclusion effects. This study first focuses on the changes in Ca(2+) activity and pH in a solution consisting of CaCl(2), KCl, and K(3)citrate, as a function of sucrose concentration. MSA model calculations were compared with experimental results, and the model satisfactorily describes the ion properties. The excluded volume effects appear to account for a considerable increase in activity coefficient of the ions. This offers a sufficient explanation for the increase in Ca(2+) activity and the decrease in pH in milk-based systems with added disaccharides. In addition, hydration of milk proteins seems to enhance ion pair formation in milk. All disaccharides lead to similar modification of the thermodynamic properties of milklike systems, confirming that the observed effects are primarily due to volume exclusion effects.

Several authors have published about baobab food products. Data on macronutrients, micronutrients, amino acids, and fatty acids were collected from literature for pulp, leaves, seeds, and kernels of the baobab tree. The results show that baobab pulp is particularly rich in vitamin C; consumption of 40 g covers 84 to more than 100% of the Recommended Daily Intake (RDI) of pregnant women (19-30 years). The leaves are particularly rich in calcium (307 to 2640 mg/100 g dw), and they are known to contain good quality proteins with a chemical score of 0.81. The whole seeds and the kernels have a relatively high lipid content, 11.6 to 33.3 g/100 g dw and 18.9 to 34.7 g/100 g dw, respectively. The pulp and leaves exhibit antioxidant properties with a higher activity in the pulp than in the leaves. Reported nutrient contents of different baobab parts show a large variation, which may have arisen from various factors. Three recommendations are given for future research: 1. More attention should be given to accuracy and precision of analytical methods, 2. Research about digestibility and bioavailability of baobab products is needed, 3. The effect of storage and processing on the nutritional value of baobab products needs to be assessed.

The effect of the roasting degree on coffee brew melanoidin properties and formation mechanisms was studied. Coffee brew fractions differing in molecular weight (Mw) were isolated from green and light-, medium-, and dark-roasted coffee beans. Isolated fractions were characterized for their melanoidin, nitrogen, protein, phenolic groups, chlorogenic acid, quinic acid, caffeic acid, and sugar content. It was found that the melanoidin level in all fractions correlated with both the nitrogen and the protein content. The melanoidin level also correlated with the phenolic groups' level and ester-linked quinic acid level. It was concluded that proteins and chlorogenic acids should be primarily involved in melanoidin formation. Initial roasting, from green to light-roasted beans, especially led to the formation of intermediate Mw (IMw) melanoidins when compared to high Mw (HMw) melanoidins. Indications were found that this IMw melanoidin formation is mainly due to Maillard reactions and chlorogenic acid incorporation reactions between chlorogenic acids, sucrose, and amino acids/protein fragments. Additionally, it was found that prolonged roasting predominantly led to formation melanoidins with a high Mw. Furthermore, arabinogalactans seem to be relatively more involved in melanoidin formation than galactomannans. It was hypothesized that chromophores may be formed or attached through the arabinose moiety of arabinogalactan proteins (AGP). Finally, it could be concluded that galactomannans are continuously incorporated in AGP-melanoidins upon roasting.

The antioxidative properties of coffee brew fractions were studied using electron spin resonance spectroscopy using 2,2,6,6-tetramethyl-1-piperidin-1-oxyl (TEMPO) and Fremy's salt (nitrosodisulfonate) as stabilized radicals. TEMPO was scavenged by antioxidants formed during roasting and not by chlorogenic acid, whereas Fremy's salt was scavenged by all antioxidants tested including chlorogenic acid. The stabilized radical TEMPO allowed the exclusive measurement of roasting-induced antioxidants. The roasting-induced antioxidant activity of coffee brews increased with increasing degree of roast, and most of these antioxidants were formed during the initial roasting stage. The majority of these roasting-induced antioxidants were present in the high molecular weight fractions, indicating that the formation of these antioxidants preferably occurs at specific high molecular weight structures, likely being arabinogalactan and/or protein moieties which might be part of the melanoidin complex. It was found that chlorogenic acids most probably do not lose their antioxidant activity and phenolic characteristics upon incorporation in coffee melanoidins. The parameter fast reacting antioxidants (FRA) was introduced as an alternative for the antioxidative potential. FRA levels showed that coffee fractions rich in roasting-induced antioxidants exposed their antioxidant activity relatively slowly, which must be a consequence of its complex structure. Finally, the melanoidin content and the roasting-induced antioxidant activity showed a positive and linear correlation for the coffee brew fractions, showing that roasting-induced antioxidants are present within melanoidins. This is the first time that the formation of roasting-induced antioxidants could be directly correlated with the extent of Maillard reaction and melanoidin formation in a complex product such as coffee.

Analysis of low molecular weight (LMw) coffee brew melanoidins is challenging due to the presence of many non-melanoidin components that complicate analysis. This study focused on the isolation of LMw coffee brew melanoidins by separation of melanoidins from non-melanoidin components that are present in LMw coffee brew material. LMw coffee fractions differing in polarity were obtained by reversed-phase solid phase extraction and their melanoidin, sugar, nitrogen, caffeine, trigonelline, 5-caffeoylquinic acid, quinic acid, caffeic acid, and phenolic groups contents were determined. The sugar composition, the charge properties, and the absorbance at various wavelengths were investigated as well. The majority of the LMw melanoidins were found to have an apolar character, whereas most non-melanoidins have a polar character. The three isolated melanoidin-rich fractions represented 56% of the LMw coffee melanoidins and were free from non-melanoidin components. Spectroscopic analysis revealed that the melanoidins isolated showed similar features as high molecular weight coffee melanoidins. All three melanoidin fractions contained approximately 3% nitrogen, indicating the presence of incorporated amino acids or proteins. Surprisingly, glucose was the main sugar present in these melanoidins, and it was reasoned that sucrose is the most likely source for this glucose within the melanoidin structure. It was also found that LMw melanoidins exposed a negative charge, and this negative charge was inversely proportional to the apolar character of the melanoidins. Phenolic group levels as high as 47% were found, which could be explained by the incorporation of chlorogenic acids in these melanoidins.

The incorporation of chlorogenic acids (CGAs) and their subunits quinic and caffeic acids (QA and CA) in coffee brew melanoidins was studied. Fractions with different molecular weights, ionic charges, and ethanol solubilities were isolated from coffee brew. Fractions were saponified, and the released QA and CA were quantified. For all melanoidin fractions, it was found that more QA than CA was released. QA levels correlated with melanoidin levels, indicating that QA is incorporated in melanoidins. The QA level was correlated with increasing ionic charge of the melanoidin populations, suggesting that QA may contribute to the negative charge and consequently is, most likely, not linked via its carboxyl group. The QA level correlated with the phenolic acid group level, as determined by Folin-Ciocalteu, indicating that QA was incorporated to a similar extent as the polyphenolic moiety from CGA. The QA and CA released from brew fractions by enzymes confirmed the incorporation of intact CGAs. Intact CGAs are proposed to be incorporated in melanoidins upon roasting via CA through mainly nonester linkages. This complex can be written as Mel=CA-QA, in which Mel represents the melanoidin backbone, =CA represents CA nonester-linked to the melanoidin backbone, and -QA represents QA ester-linked to CA. Additionally, a total of 12% of QA was identified in coffee brew, whereas only 6% was reported in the literature so far. The relevance of the additional QA on coffee brew stability is discussed.

The reaction of malondialdehyde with casein was studied in aqueous solution to evaluate the impact of this lipid oxidation product on food protein modification. By using multiresponse modeling, a kinetic model was developed for this reaction. The influence of temperature and pH on protein browning and malondialdehyde degradation was evaluated. The hypothesis that one malondialdehyde unit leads to the cross-linking of two casein-bound lysine residues was in accordance with the data. At higher malondialdehyde concentrations, a different reaction mechanism was operative, probably involving a dihydropyridine cross-link. The results obtained were compared with the reaction of casein with 2-oxopropanal, a well-studied alpha-dicarbonyl compound. The reaction of casein with 2-oxopropanal followed a different reaction pathway. Comparison of the degree of browning of casein by reaction with malondialdehyde and 2-oxopropanal showed a considerably higher degree of browning induced by malondialdehyde. This research has shown that kinetic modeling is a useful tool to unravel reaction mechanisms. Clearly, the contribution of lipid oxidation products, such as malondialdehyde, to protein modification (both in food and in vivo) can be substantial and needs to be taken into account in future studies.

The formation of acrylamide in potato crisps was fitted by empirical mathematical models. Potato slices were fried under the same experimental conditions for different times. Besides the content of precursors in the raw potato slices, acrylamide and water content in the potato crisps were quantified after predetermined times (2-6 min). The temperature developments in the surrounding oil and outer cell layer of the potato slices were monitored, giving more insight in the frying process and making future comparisons between studies possible. The pattern found for the formation of acrylamide, which was similar to earlier studies, was fitted to three empirical models. Statistical methods were used to compare the performance of the models, with the "Logistic-Exponential" and "Empirical" model performing equally well. The obtained model parameters were in the range of earlier reported studies, although this comparison is not unequivocal as the experimental conditions differed between studies. The precision of parameter estimates was problematic; this should be improved by better experimental design. Nevertheless, the approach of this study will make it possible to truly compare acrylamide formation patterns and model parameters in the future, with the ability to develop a tool to predict acrylamide formation in potato crisps.

Birch pollen allergy is predominantly caused by the major allergen Bet v 1 and can lead to crossreactions with homologous proteins in food. Two major cross-reactive food allergens are Dau c 1 from carrot and Api g 1 from celery, which have never been purified from their natural source. Here, we describe a non-denaturing purification method for obtaining natural Bet v 1, Dau c 1 and Api g 1, comprising of ammonium sulfate precipitation, hydrophobic interaction chromatography and size exclusion chromatography. This method resulted in 98-99% pure isoform mixtures for each allergen. Characterization of these isoform mixtures with Q-TOF MS/MS clearly showed earlier reported isoforms of Bet v 1, Dau c 1 and Api g 1, but also new isoforms. The presence of secondary structure in the three purified allergens was demonstrated via circular dichroism and showed high similarity. The immune reactivity of the natural allergens was compared with recombinant proteins by Western blot and ELISA and showed similar reactivity.

This study aimed to characterize the number of Salmonella on chicken breast filet at the retail level and to evaluate if this number affects the risk of salmonellosis. From October to December 2005, 220 chilled raw filets (without skin) were collected from five local retail outlets in The Netherlands. Filet rinses that were positive after enrichment were enumerated with a three-tube most-probable-number (MPN) assay. Nineteen filets (8.6%) were contaminated above the detection limit of the MPN method (10 Salmonella per filet). The number of Salmonella on positive filets varied from 1 to 3.81 log MPN per filet. The obtained enumeration data were applied in a risk assessment model. The model considered possible growth during domestic storage, cross-contamination from filet via a cutting board to lettuce, and possible illness due to consumption of the prepared lettuce. A screening analysis with expected-case and worst-case estimates for the input values of the model showed that variability in the inputs was of relevance. Therefore, a Monte Carlo simulation with probability distributions for the inputs was carried out to predict the annual number of illnesses. Remarkably, over two-thirds of annual predicted illnesses were caused by the small fraction of filets containing more than 3 log Salmonella at retail (0.8% of all filets). The enumeration results can be used to confirm this hypothesis in a more elaborate risk assessment. Modeling of the supply chain can provide insight for possible intervention strategies to reduce the incidence of rare, but extreme levels. Reduction seems feasible within current practices, because the retail market study indicated a significant difference between suppliers.

The charge properties of melanoidins in high molecular weight (HMw) coffee brew fractions, isolated by diafiltration and membrane dialysis, were studied. Ion exchange chromatography experiments with the HMw fractions showed that coffee brew melanoidins were negatively charged whereas these molecules did not expose any positive charge at the pH of coffee brew. Fractions with different ionic charges were isolated and subsequently characterized by means of the specific extinction coefficient (K(mix 405nm)), sugar composition, phenolic group content, nitrogen content, and the arabinogalactan protein (AGP) specific Yariv gel-diffusion assay. The isolated fractions were different in composition and AGP was found to be present in one of the HMw fractions. The AGP accounted for 6% of the coffee brew dry matter and had a moderate negative charge, probably caused by the presence of uronic acids. As the fraction that precipitated with Yariv was brown (K(mix 405nm) = 1.2), compared to a white color in the green bean, it was concluded that these AGPs had undergone Maillard reaction resulting in an AGP-melanoidin complex. The presence of mannose (presumably from galactomannan) indicates the incorporation of galactomannans in the AGP-melanoidin complex. As the uronic acid content in the more negatively charged melanoidin-rich, AGP-poor HMw fractions decreased, it was hypothesized that acidic groups are formed or incorporated during melanoidin formation.

The composition of high molecular weight (HMw) coffee melanoidin populations, obtained after ethanol precipitation, was studied. The specific extinction coefficient (K(mix)) at 280, 325, 405 nm, sugar composition, phenolic group content, nitrogen content, amino acid composition, and non-protein nitrogen (NPN) content were investigated. Results show that most HMw coffee melanoidins are soluble at high ethanol concentrations. The amino acid composition of the HMw fractions was similar, while 17% (w/w) of the nitrogen was NPN, probably originating from degraded amino acids/proteins and now part of melanoidins. A strong correlation between the melanoidin content, the NPN, and protein content was found. It was concluded that proteins are incorporated into the melanoidins and that the degree of chemical modification, for example, by phenolic groups, determines the solubility of melanoidins in ethanol. Although the existence of covalent interaction between melanoidins and polysaccharides were not proven in this study, the findings suggest that especially arabinogalactan is likely involved in melanoidin formation. Finally, phenolic groups were present in the HMw fraction of coffee, and a correlation was found with the melanoidin concentration.

In the present study emulsions were made with various potato protein preparations, which varied in protease inhibitor and patatin content. These emulsions were characterized with respect to average droplet size, plateau surface excess, and the occurrence of droplet aggregation. Droplet aggregation occurred only with potato protein preparations that contained a substantial amount of protease inhibitors and could be prevented only at pH 3. The average droplet size of the emulsions made with potato proteins appeared to be related to the patatin content of the preparation used. Average droplet size was found to be dominated by the patatin-catalyzed lipolytic release of surface active fatty acids and monoglycerides from the tricaprylin oil phase during the emulsification process. Addition of monoglycerides and especially fatty acids, at concentrations representative of those during emulsification, was shown to cause a stronger and much faster decrease of the interfacial tension than that with protein alone and to result in a drastic decrease in droplet size. The patatin used was shown to have a lipolytic activity of 820 units/g with emulsified tricaprylin as the substrate. Because of the droplet aggregating properties of the protease inhibitors, the patatin-rich potato preparations seem to be the most promising for food emulsion applications over a broad pH range, provided the lipolytic activity can be diminished or circumvented.

The purpose of this study was to improve the micronutrient quality of indigenous African infant flour using traditional techniques available in the region. Response surface methodology was used to study the effect of duration of soaking, germination, and fermentation on phytate and phenolic compounds (PC), pH, viscosity, and the in vitro solubility (IVS) of iron and zinc in infant sorghum flour. The phytate and the PC concentrations of the flour were significantly modified as a result of the duration of germination and fermentation and their mutual interaction. These modifications were accompanied by a significant increase in % IVS Zn after 24 h of sprouting. Except for the interaction of soaking and fermentation, none of the processing parameters exerted a significant effect on the % IVS Fe. The viscosity of the porridge prepared with the flour decreased significantly with the duration of germination, making it possible to produce a porridge with high energy and nutrient density. The use of germination in combination with fermentation is recommended in the processing of cereals for infant feeding in developing countries.

This paper discusses the importance of the Maillard reaction for food quality and focuses on flavour compound formation. The most important classes of Maillard flavour compounds are indicated and it is shown where they are formed in the Maillard reaction. Some emphasis is given on the kinetics of formation of flavour compounds. It is concluded that the essential elements for predicting the formation of flavour compounds in the Maillard reaction are now established but much more work needs to be done on specific effects such as the amino acid type, the pH, water content and interactions in the food matrix. It is also concluded that most work is done on free amino acids but hardly anything on peptides and proteins, which could generate peptide- or protein-specific flavour compounds.

Seventy-six farmers' varieties of sorghum from Benin were distinguished by amplified fragment length polymorphism (AFLP) and clustered into 45 distinct genotypes. The genotype clusters were evaluated for their Fe, Zn, and phytate concentrations to assess the impact of genetic and environmental effects on the composition of the grains and to identify farmers' varieties with high potential Fe and Zn availability. The Fe concentration of the grains ranged from 30 to 113 mg/kg with an average of 58 mg/kg. The Zn concentration ranged from 11 to 44 mg/kg with an average of 25 mg/kg. The phytate concentration of the grain ranged from 0.4 to 3.5% with a mean of 1.2%. The grain-Fe and grain-Zn did not show consistent linkage to genetic variation, but varied significantly across field locations, suggesting a predominant environmental impact. The phytate concentration of the grains appeared to be environmentally as well as genetically determined. No varieties provide adequate Zn to meet nutritional requirements of sorghum consumers. The most promising varieties for Fe supply were tokogbessenou, mahi swan, biodahu, saï maï, mare dobi, sakarabokuru, and chabicouma, as they showed a [phytate]/[Fe] ratio of <14, which is the critical value above which Fe availability is strongly impaired. These varieties could therefore be recommended for the preparation of food products such as dibou, in which processing methods have only a slight diminishing effect on phytate levels. Further research is needed to test these varieties for the stability of [phytate]/[Fe] molar ratio across various environmental conditions.

A kinetic model for the formation of acrylamide in a glucose-asparagine reaction system is pro-posed. Equimolar solutions (0.2 M) of glucose and asparagine were heated at different temperatures (120-200 degrees C) at pH 6.8. Besides the reactants, acrylamide, fructose, and melanoidins were quantified after predetermined heating times (0-45 min). Multiresponse modeling by use of nonlinear regression with the determinant criterion was used to estimate model parameters. The proposed model resulted in a reasonable estimation for the formation of acrylamide in an aqueous model system, although the behavior of glucose, fructose, and asparagine was slightly underestimated. The formation of acrylamide reached its maximum when the concentration of sugars was reduced to about 0. This supported previous research, showing that a carbonyl source is needed for the formation of acrylamide from asparagine. Furthermore, it is observed that acrylamide is an intermediate of the Maillard reaction rather than an end product, which implies that it is also subject to a degradation reaction.

Kinetic data are reported describing the stability of various classes of polyphenolic antioxidants in an apple juice enriched in these compounds as a function of storage temperature and oxygen concentration. The most thermally sensitive compounds were the various quercetin glycosides and epicatechin, whereas phloridzin and chlorogenic acid were more stable. The quercetin glycosides showed differences in their stability: quercetin galactoside approximately quercetin rhamnoside > quercetin glucoside/rutinoside > quercetin arabinoside. The effect of the presence of oxygen on the degradation rates was clear for only quercetin and to a lesser extent for epicatechin. Accelerated shelf-life testing of enriched apple juice during 4 days at 80 degrees C showed decreases in the antioxidant activity of 20-40%. The parameters obtained were used to predict the stability at different storage conditions. Calculations showed that polyphenolic antioxidants and antioxidant activity of enriched apple juice will be quite stable at ambient or refrigerated storage conditions up to half a year.

Gel network formation of pea legumin (8.4% on a protein basis, pH 7.6) was monitored via dynamic rheological measurements. Gelation was performed in the absence and presence of the thiol-blocking reagent N-ethylmaleimide, at different rates of heating and cooling. Overall, it was shown that pea legumin gel formation was not effected by changes in the heating rate, and the two differently heated samples were unaffected by the addition of 20 mM NEM, which indicated that disulfide bonds were not essential within the network strands of these legumin gels. However, slowly cooling the legumin samples caused disulfide bonds to become involved within the network; this was observed by a large increase in gel strength that was then substantially reduced when repeating the sample in the presence of NEM. These experiments were repeated with soybean glycinin in order to determine whether a common model for gel formation of legumin-like proteins could be built, based upon molecular reasoning. The two proteins were affected in the same way by changes in the conditions used, but when applying a procedure of reheating and recooling the gel networks responded differently. Pea legumin gel networks were susceptible to rearrangements that caused the gels to become stronger after reheating/recooling, yet glycinin gel networks were not. It was concluded that the same physical and chemical forces drove the processes of denaturation, aggregation, and network formation. Each process can therefore be readily targeted for modification based upon molecular reasoning. Pea legumin and soybean glycinin gel networks had structurally different building blocks, however. A model of gelation aimed at texture control therefore requires additional information.

The gelling characteristics of two vicilin fractions from pea (Pisum sativum L.) were compared over a range of pH and salt conditions after preliminary results showed that despite having equal opportunity to unfold, and expose hydrophobic residues, they had different minimum gelling concentrations (at pH 7.6). Furthermore, at this pH one fraction formed turbid gels and the other formed transparent gels. The fraction that formed transparent gels contained a substantial amount of the 70 kDa alpha-subunits of vicilin, and thus it was hypothesized that the highly charged N-terminal extension region on these 70 kDa subunits hinders gelation of this vicilin fraction at pH 7.6 and I = 0.2 due to repulsion of the net negative charge. The experiments designed to test this hypothesis are presented and discussed in this paper and prove that the hypothesis was true, which offers the possibility to control or modify the gelation behavior of vicilin on the basis of information of its subunit composition.

Vicilin, a major globulin protein of pea that has been described as "extremely heterogeneous in terms of its polypeptide composition", was extracted from pea flour under alkaline conditions and subsequently fractionated by salt under acid conditions. This procedure induced the separation of vicilin into two fractions, which, after purification, were called vicilin 1 degrees and vicilin 2 degrees. Vicilin 2 degrees was seen on SDS-PAGE to contain the third globulin protein of pea, convicilin (a band at approximately 70 kDa). Vicilin fractions were thus characterized using gel electrophoresis, differential scanning calorimetry, circular dichroism, and pH-dependent solubility in order to determine whether the convicilin should in fact be considered as a third separate globulin protein of pea. On the basis of the results obtained it was concluded that this distinct polypeptide of the Pisum vicilin gene family should be further denoted as a subunit of the salt extractable protein vicilin. The definition of vicilin heterogeneity should therefore be extended to acknowledge the possible oligomeric inclusion of the 70 kDa polypeptide that is here denoted as the alpha-subunit.

A kinetic model for N-(1-deoxy-D-fructos-1-yl)-glycine (DFG) thermal decomposition was proposed. Two temperatures (100 and 120 degrees C) and two pHs (5.5 and 6.8) were studied. The measured responses were DFG, 3-deoxyosone, 1-deoxyosone, methylglyoxal, acetic acid, formic acid, glucose, fructose, mannose and melanoidins. For each system the model parameters, the rate constants, were estimated by non-linear regression, via multiresponse modelling. The determinant criterion was used as the statistical fit criterion. Model discrimination was performed by both chemical insight and statistical tests (Posterior Probability and Akaike criterion). Kinetic analysis showed that at lower pH DFG 1,2-enolization is favoured whereas with increasing pH 2,3-enolization becomes a more relevant degradation pathway. The lower amount observed of 1-DG is related with its high reactivity. It was shown that acetic acid, a main degradation product from DFG, was mainly formed through 1-DG degradation. Also from the estimated parameters 3-DG was found to be the main precursor in carbohydrate fragments formation, responsible for colour formation. Some indication was given that as the reaction proceeded other compounds besides DFG become reactants themselves with the formation among others of methylglyoxal. The multiresponse kinetic analysis was shown to be both helpful in deriving relevant kinetic parameters as well as in obtaining insight into the reaction mechanism.