Characterization of fumonisin B(1)-glucose reaction kinetics and products

The reaction of fumonisin B(1) with the reducing sugar D-glucose can block the primary amine group of fumonisin B(1) and may detoxify this mycotoxin. A method to separate hundred milligram quantities of fumonisin B(1)-glucose reaction products from the excess D-glucose with a reversed-phase C(18) cartridge was developed. Mass spectrometry revealed that there were four primary products in this chain reaction when fumonisin B(1) was heated with D-glucose at 65 degrees C for 48 h: N-methyl-fumonisin B(1), N-carboxymethyl-fumonisin B(1), N-(3-hydroxyacetonyl)-fumonisin B(1), and N-(2-hydroxy, 2-carboxyethyl)-fumonisin B(1). The N-(1-deoxy-D-fructos-1-yl) fumonisin B(1) (fumonisin B(1)-glucose Schiff's base) was detected by mass spectrometry when fumonisin B(1) was heated with D-glucose at 60 degrees C. The nonenzymatic browning reaction of fumonisin B(1) with excess D-glucose followed apparent first-order kinetics. The activation energy, E(a), was 105.7 kJ/mol. Fumonisin B(1) in contaminated corn could precipitate the nonenzymatic browning reaction with 0.1 M D-glucose at 60 and 80 degrees C.     

Effect of pH and temperature on comparative antioxidant activity of nonenzymatically browned proteins produced by reaction with oxidized lipids and carbohydrates

The antioxidative activity of nonenzymatically browned bovine serum albumin (BSA) produced by reaction with ribose (RI), hydroperoxides of methyl linoleate oxidation (HP), and secondary products of methyl linoleate oxidation (SP), at different pHs (4, 7, and 10) and temperatures (25, 37, 50, 80, and 120 degrees C), was studied to compare the antioxidative effects of carbohydrate- and oxidized lipids-modified proteins. The modified proteins (RIBSA, HPBSA, and SPBSA) were tested for antioxidative activity (at 100 ppm) in soybean oil using the thiobarbituric acid-reactive substances (TBARS) assay. All of them decreased significantly (p < 0.05) the TBARS formation in the oil and exhibited different effectiveness as a function of the temperature and the pH of the medium. In addition, there was a good correlation between the antioxidative activity of the protein and the amino acid losses produced during the nonenzymatic browning. These results are in agreement with an analogous and complimentary contribution of both Maillard and oxidized lipid/protein reactions to the antioxidative activity produced in foods during processing and storage.     

Effect of the pyrrole polymerization mechanism on the antioxidative activity of nonenzymatic browning reactions

The present investigation was undertaken to study how the antioxidative activity (AA) of nonenzymatic browning reactions is changing at the same time that the browning (by the pyrrole polymerization mechanism) is being produced. The antioxidative activities of eight model pyrroles (pyrrole, 1-methylpyrrole, 2,5-dimethylpyrrole, 1,2,5-trimethylpyrrole, 2-acetylpyrrole, 2-acetyl-1-methylpyrrole, pyrrole-2-carboxaldehyde, and 1-methyl-2-pyrrolecarboxaldehyde) as well as the browning reaction of 2-(1-hydroxyethyl)-1-methylpyrrole (HMP) and the dimer (DIM) produced during HMP browning were determined. The results obtained suggest that the AAs observed in nonenzymatic browning reactions are the result of the AAs of the different oxidized lipid/amino acid reaction products formed. Thus, the different pyrrole derivatives produced in these reactions had different AAs, and the highest AAs were observed for alkyl-substituted pyrroles without free alpha-positions. Because some of these pyrrole derivatives are implicated in nonenzymatic browning production and this browning production implies the loss of hydroxyl groups and the transformation of some pyrroles with one type of substitution into others, changes in AA during browning production were observed, and the resulting DIM derivative was more antioxidant than HMP or higher polymers. These results explain the AA observed in fatty acid/protein mixtures after slight oxidation and suggest that, when the pyrrole polymerization mechanism is predominant, slightly browned samples may be more antioxidant than samples in which nonenzymatic browning has been highly developed.     

Chemical and antioxidant properties of casein peptide and its glucose Maillard reaction products in fish oil-in-water emulsions

Maillard reaction products (MRPs) were prepared by reacting casein peptides with different concentrations of glucose at 80 °C for up to 12 h. The chemical properties of MRPs and their effects on lipid oxidation in fish oil-in-water emulsions were investigated. Increasing browning development and absorbance in 294 nm in the MRPs caused an increase in DPPH radical scavenging, but a decrease in iron chelation, which could be related to the loss of free amino groups in the peptides. The MRPs produced with longer reaction time or higher glucose concentrations were less effective in inhibiting lipid oxidation in emulsions at pH 7.0 compared to casein peptides alone. However, the antioxidant activity of MRPs in emulsions at pH 3.0 was not decreased by prolonged heating. The bitterness of MRPs was less than that of the original casein peptides, and bitterness decreased with increasing heating time and glucose concentrations. Therefore, the Maillard reaction was a potential method to reduce the bitterness of casein peptides while not strongly decreasing their antioxidant activity.     

Kinetics of interaction of vanillin with amino acids and peptides in model systems

Model systems were used to study the reaction kinetics of vanillin and pentalysine, lysine, glutathione, cysteine, aspartame, or phenylalanine (molar ratio 1:1) in phosphate buffer. The buffer pH was adjusted to the pK(a)(2) of the available alpha-amino group of each amino acid or peptide. Reductions of vanillin followed first-order kinetics at 55, 65, and 75 degrees C in the presence of each of the amino acids or peptides used. The reaction rates were accelerated as the temperature increased. The rate constants were highest for pentalysine followed by lysine, phenylalanine, glutathione/cysteine, and aspartame. The reduction of phenylalanine followed first-order kinetics, whereas the formation of its reaction product followed zero-order kinetics. The activation energy (E(a)) for the reaction ranged from 5.6 to 14.5 kcal/mol.     

Biochemical and microbial changes during the storage of minimally processed cantaloupe

The effect of storage time on pH, titratable acidity, degrees Brix, organic acids, sugars, amino acids, and color of minimally processed cantaloupe melon (Cucumis melo L. var. reticulatus Naud. cv. Mission) was determined at 4 degrees C and 20 degrees C. Changes in most of the biochemical parameters with storage time were relatively slow at the lower temperature. At 20 degrees C, a 17% loss in soluble solids and a 2-fold increase in acidity occurred after 2 days. Organic acid content also increased considerably with time at this temperature as a result of the production of lactic acid. Oxalic, citric, malic, and succinic acids were the organic acids, and glucose, fructose, and sucrose were the sugars present in the freshly cut cantaloupe. Malic acid concentration decreased concurrently with lactic acid production indicating the possible involvement of anaerobic malo-lactic fermentation along with sugar utilization by lactic acid bacteria. The effect of storage on microbial growth was determined at 4, 10, and 20 degrees C. Gram-negative stained rods grew at a slower rate at 4 degrees C and 10 degrees C than the Gram-positive mesophilic bacteria that dominated microorganism growth at 20 degrees C. Eighteen amino acids were identified in fresh cantaloupe: aspartic acid, glutamic acid, asparagine, serine, glutamine, glycine, histidine, arginine, threonine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine, phenyl alanine, and lysine. The dominant amino acids were aspartic acid, glutamic acid, arginine, and alanine. Total amino acid content decreased rapidly at 20 degrees C, but only a slight decrease occurred at 4 degrees C after prolonged storage. Changes in lightness (L), chroma, and hue at both temperatures indicate the absence of browning reactions. The results indicate the potential use of lactic acid and lactic acid bacteria as quality control markers in minimally processed fruits.     

Kinetics of freshly squeezed orange juice quality changes during ozone processing

Freshly squeezed orange juice samples were ozonated with control variables of gas flow rate (0-0.25 L min (-1)), ozone concentration (0.6-10.0%w/w), and treatment time (0-10 min). Effects of ozone processing variables on orange juice quality parameters of pH, degrees Brix, titratable acidity (TA), cloud value, nonenzymatic browning (NEB), color values ( L*, a*, and b*), and ascorbic acid content were determined. No significant changes in pH, degrees Brix, TA, cloud value, and NEB ( p < 0.05) were found. L*, a*, and b* color values were significantly affected by gas flow rate, ozone concentration, and treatment time. The changes in lightness ( L*) values and total color difference (TCD) values were fitted well to zero-order kinetics, whereas a*, b*, and ascorbic acid degradation followed first-order kinetics. The rate constants for a*, b*, and TCD were linearly correlated with ozone concentration ( R (2) = 0.88-0.99), whereas the rate constants for L* and ascorbic acid were exponentially correlated ( R (2) = 0.94-0.98).     

Radical-assisted melanoidin formation during thermal processing of foods as well as under physiological conditions

Color-generating reactions of protein-bound lysine with carbohydrates were studied under thermal as well as under physiological conditions to gain insights into the role of protein/carbohydrate reactions in the formation of food melanoidins as well as nonenzymatic browning products in vivo. EPR spectroscopy of orange-brown melanoidins, which were isolated from heated aqueous solutions of bovine serum albumin and glycolaldehyde, revealed the protein-bound 1,4-bis(5-amino-5-carboxy-1-pentyl)pyrazinium radical cation (CROSSPY) as a previously unknown type of cross-linking amino acid leading to protein dimerization. To verify their formation in foods, wheat bread crust and roasted cocoa as well as coffee beans, showing elevated nonenzymatic browning, were investigated by EPR spectroscopy. An intense radical was detected, which, by comparison with the radical formed upon reaction bovine serum albumin with glycolaldehyde, was identified as the protein-bound CROSSPY. The radical-assisted protein oligomerization as well as the browning of bovine serum albumin in the presence of glycolaldehyde occurred also rapidly under physiological conditions, thereby suggesting CROSSPY formation to be probably involved also in nonenzymatic glycation reactions in vivo.     

Contribution of pyrrole formation and polymerization to the nonenzymatic browning produced by amino-carbonyl reactions

Recent studies have hypothesized that pyrrole formation and polymerization may be contribute to the nonenzymatic browning produced in both oxidized lipid/protein reactions and the Maillard reaction. To develop a methodology that would allow investigation of the contribution of this browning mechanism, the kinetics of formation of color, fluorescence, and pyrrolization in 4, 5(E)-epoxy-2(E)-heptenal/lysine and linolenic acid/lysine model systems were studied. In both cases similar kinetics for the three measurements were observed at the two temperatures assayed (37 and 60 degrees C), and there was a high correlation among color, fluorescence, and pyrrolization measurements obtained as a function of incubation time. Because the color and fluorescence production in the 4,5(E)-epoxy-2(E)-heptenal/lysine system is a consequence of pyrrole formation and polymerization, the high correlations observed with the unsaturated fatty acid also suggest a contribution of the pyrrole formation and polymerization to the development of color and fluorescence observed in the fatty acid/lysine system. Although the contribution of other mechanisms cannot be discarded, all of these results suggest that when the pyrrole formation and polymerization mechanism contributes to the nonenzymatic browning of foods, a high correlation among color, fluorescence, and pyrrolization measurements should be expected.     

Strecker-type degradation of phenylalanine by methyl 9,10-epoxy-13-oxo-11-octadecenoate and methyl 12,13-epoxy-9-oxo-11-octadecenoate

The reaction of methyl 9,10-epoxy-13-oxo-11(E)-octadecenoate, methyl 12,13-epoxy-9-oxo-11(E)-octadecenoate, 4,5(E)-epoxy-2(E)-heptenal, and 4,5(E)-epoxy-2(E)-decenal with phenylalanine in acetonitrile-water (2:1, 1:1, and 1:2) at 80 degrees C and at different pHs and carbonyl compound/amino acid ratios was investigated both to determine if epoxyoxoene fatty esters were able to produce the Strecker-type degradation of the amino acid and to study the relative ability of oxidized long-chain fatty esters and short chain aldehydes with identical functional systems to degrade amino acids. The studied epoxyoxoene fatty esters degraded phenylalanine to phenylacetaldehyde. The mechanism of the reaction was analogous to that described for epoxyalkenals and is suggested to be produced through the corresponding imine, which is then decarboxylated and hydrolyzed. This reaction also produced a conjugated hydroxylamine, which was the origin of the long-chain pyridine-containing fatty ester isolated in the reaction and characterized as methyl 8-(6-pentylpyridin-2-yl)octanoate. Epoxyoxoene fatty esters and epoxyalkenals exhibited a similar reactivity for producing phenylacetaldehyde, therefore suggesting that nonvolatile lipid oxidation products, which are produced to a greater extent than volatile products, should be considered for determining the overall contribution of lipids to Strecker degradation of amino acids produced during nonenzymatic browning. In addition, the obtained data confirm that, analogously to carbohydrates, lipid oxidation products are also able to produce the Strecker degradation of amino acids.     

Antiradical efficiency of Maillard reaction mixtures in a hydrophilic media

The Maillard reaction (MR) has a clear impact in food science, nutrition, and medical research. Free radical scavenging capacities of several MR mixtures made from single combinations of glucose or lactose and amino acids (gly, his, lys, trp, met, and cys) were evaluated by using the N,N-dimethyl-p-phenylenediamine radical cation assay. Medium-roasted coffee brew was used as reference of a thermally processed food. A novel approach has been applied in order to get more information about the kinetic behavior of the radical scavenging properties of MR mixtures in a watery environment. Antiradical efficiency (AE) concept has been applied, and it takes into consideration the reaction time, apart from the amount of antioxidant necessary to decrease by 50% the radical initial concentration (EC(50)). Cysteine and histidine reveal as powerful amino acids to exert a high AE in the MR mixtures. No relationship between AE parameter and browning was observed.     

Influence of the maillard reaction on the allergenicity of rAra h 2, a recombinant major allergen from peanut (Arachis hypogaea), its major epitopes, and peanut agglutinin

The influence of thermal processing and nonenzymatic browning reactions on the IgE-binding activity of rAra h 2 was studied and compared to findings recently reported for the allergen's natural counterpart. ELISA experiments as well as inhibition assays revealed that thermal treatment of rAra h 2 in the presence of reactive carbohydrates and carbohydrate breakdown products induces a strong increase of the IgE-binding activity, thus collaborating with the data reported for the natural protein isolated from peanuts. To localize the Ara h 2 sequences responsible for the formation of highly IgE-affine glycation sites, model peptides have been synthesized mimicking sequences which contain possible targets for glycation as well as the immunodominant epitopes. Immunological evaluation of these peptides heated in the absence or presence of reducing sugars and carbonyls, respectively, revealed that neither the two lysine residues of Ara h 2 nor its N-terminus are involved in the formation of IgE-affine structures by Maillard reaction. Also, the cysteine-containing major epitope 3 (aa 27-36) was found to lose its IgE-binding capacity upon heating. By contrast, the overlapping major epitopes 6 and 7, which do not contain any lysine or arginine moieties, showed a distinct higher level of IgE binding when subjected to Maillard reaction, thus giving the first evidence that nonbasic amino acids might be accessible for nonenzymatic glycation reactions and that these posttranslational modifications might induce increased IgE binding of the glycated Ara h 2. Analogous experiments were performed with peanut agglutinin, considered in the literature as a minor allergen. ELISA experiments revealed that the majority of tested sera samples from peanut-sensitive patients showed a high level of IgE binding to the lectin even after heat treatment. In contradiction to published data, nonenzymatic browning reactions seem to deteriorate the IgE affinity of the lectin.     

Total Protein and Amino acid Compositions in the Acorns of Turkish Quercus L. Taxa

Total protein content and level of 14 amino acid in mature acorns of 20 Quercus taxa from Turkey were studied. The range of total protein amounts between 2.75 and 8.44% were detected among taxa. Similar values in related species and variety level were observed. The amino acid profiles for each taxon were characteristically different and high variability of individual amino acid concentration was present at variety, subspecies and species levels. Different amino acid concentrations, relative percentages and critical values of some amino acids are thought to be additional considerable parameters for diagnosis of Quercus. Generally higher total quantity of amino acids in section Quercus and lower values in section Ilex were observed. When the essential and non-essential amino acids are expressed as a relative percent of total protein indicating quality of proteins, the ratios varied significantly among taxa (p < 0.05). But, no significant difference at section level were detected. Major amino acids were aspartic acid and glutamic acid showing the largest variations and the lowest levels were detected for methionine. Amino acid concentrations ranged from 1665 for aspartic acid to 13 mg/100 g dry wt. for methionine. All taxa had relatively higher amounts of leucin, lysin and valine than other essential amino acids. The highest quantity of protein and amino acids was recorded for Q. infectoria ssp. boissieri and the lowest for Q. pontica. The level of all essential amino acids in examined taxa was not sufficient compared to FAO scoring pattern for children (1985). But, threonine and valine in Q. infectoria ssp. boissieri and isoleucine in Q. petraea ssp. iberica show remarkable concentrations to the requirements. All examined essential amino acids among taxa generally provide adequate levels for adults according to FAO standard.     

Storage stability of ascorbic acid incorporated in edible whey protein films

The stability of ascorbic acid (AA) incorporated in whey protein isolate (WPI) film and the related color changes during storage were studied. No significant loss of AA content was found in any films prepared from pH 2.0 casting solution stored at 30% relative humidity (RH) and 22 °C over 84 days. Total visible color difference (ΔE*(ab)) of all films slowly increased over storage time. The ΔE*(ab) values of pH 3.5 films were significantly higher than those of pH 2.0 films. The stability of AA-WPI films was found to be mainly affected by the pH of the film-forming solution and storage temperature. Oxidative degradation of AA-WPI films followed Arrhenius behavior. Reduction of the casting solution pH to below the pK(a1) (4.04 at 25 °C) of AA effectively maintained AA-WPI storage stability by greatly reducing oxidative degradation, whereas anaerobic and nonenzymatic browning were insignificant. The half-life of pH 2.0 AA-WPI film at 30% RH and 22 °C was 520 days.     

Nonenzymatic browning in food models in the vicinity of the glass transition: effects of fructose,glucose, and xylose as reducing sugar

Effects of a reducing sugar, fructose, glucose, or xylose, and glass transition on the nonenzymatic browning (NEB) rate in maltodextrin (MD), poly(vinylpyrrolidone) (PVP), and water systems were studied. Glass transition temperatures (T(g)) were determined using DSC. Water contents were determined gravimetrically, and NEB rates were followed at several temperatures spectrophotometrically at 280 and 420 nm. Reducing sugar did not affect water contents, but xylose reduced the T(g) of the solid models. Sugars showed decreasing NEB reactivity in the order xylose > fructose > glucose in every matrix material. The NEB reactivity and temperature dependence of the single sugars varied in different matrices. The NEB rates of the solid models increased at temperatures 10-20 degrees C above the T(g), and nonlinearity was observed in Arrhenius plots in the vicinity of T(g). The temperature dependence of nonenzymatic browning could also be modeled using the WLF equation.     

Nonenzymatic browning of lactose and caseinate during dry heating at different relative humidities

Dry mixtures of lactose and caseinate were heated at 60 degrees C for up to 96 h at different relative humidities (RHs) ranging from 29 to 95%. The resulting nonenzymatic browning was studied by determining lactulosyl lysine formation in the caseinate (as measured by the conversion to furosine), amount of reacted lactose, loss of lysine, color formation, and fluorescent intensity. For each measurement, the maximum reaction occurred at intermediate RHs. While there is general agreement between the results obtained by different methods, discrepancies are understandable given the complex nature of nonenzymatic browning. It was shown that the degradation of the Amadori product, lactulosyl lysine, increased with RH. Moreover, the Maillard reaction, as opposed to caramelization of lactose, was the major pathway at all RHs. Visible browning occurred when the destruction of Amadori product became dominant, and interactions between sugar fragments and caseinate were not the rate-limiting steps in the nonenzymatic browning.     

Comparison of the ion exclusion chromatographic method with the Monier-Williams method for determination of total sulfite in foods

Experimental data comparing the alkali extraction/ion exclusion chromatographic method with the Monier-Williams method for determination of total sulfite are presented in (a) enzymatic and nonenzymatic browning systems, (b) vegetables containing naturally occurring sulfite, and (c) a carbohydrate-type food additive, erythorbic acid. Excellent agreement, with a linear correlation coefficient of 0.99, was observed in fresh potato samples homogenized with sulfite and allowed to react for different time intervals (enzymatic browning system). A good overall correlation was observed in dehydrated, sulfited apple samples heated for different times (nonenzymatic browning system); however, as heating time increased, higher results were obtained by the Monier-Williams method than by the alkali extraction/ion exclusion chromatographic method. The results of determining sulfite in the alkali trapping solution following acid distillation or acid treatment without heat suggested that this deviation was due to a fraction of sulfite bound to the browning reaction products in such a way that it was released by acid distillation but not by alkali extraction or acid treatment without heat. Similar behavior was demonstrated in cabbage with naturally occurring sulfite, which was released by acid distillation but not by alkali extraction or acid treatment without heat. The ion exclusion chromatographic method could overcome interference by the volatile caramelization reaction products in the Monier-Williams determination of erythorbic acid.     

Characterization of polyphenol oxidase from the Manzanilla cultivar (Olea europaea pomiformis) and prevention of browning reactions in bruised olive fruits

The crude extract of the polyphenol oxidase (PPO) enzyme from the Manzanilla cultivar (Olea europaea pomiformis) was obtained, and its properties were characterized. The browning reaction followed a zero-order kinetic model. Its maximum activity was at pH 6.0. This activity was completely inhibited at a pH below 3.0 regardless of temperature; however, in alkaline conditions, pH inhibition depended on temperature and was observed at values above 9.0 and 11.0 at 8 and 25 degrees C, respectively. The thermodynamic parameters of substrate oxidation depended on pH within the range in which activity was observed. The reaction occurred according to an isokinetic system because pH affected the enzymatic reaction rate but not the energy required to carry out the reaction. In the alkaline pH region, browning was due to a combination of enzymatic and nonenzymatic reactions that occurred in parallel. These results correlated well with the browning behavior observed in intentionally bruised fruits at different temperatures and in different storage solutions. The use of a low temperature ( approximately 8 degrees C) was very effective for preventing browning regardless of the cover solution used.     

Quantitation of furan and methylfuran formed in different precursor systems by proton transfer reaction mass spectrometry

Furan has recently received attention as a possibly hazardous compound occurring in certain thermally processed foods. Previous model studies have revealed three main precursor systems producing furan upon thermal treatment, i.e., ascorbic acid, Maillard precursors, and polyunsaturated lipids. We employed proton transfer reaction mass spectrometry (PTR-MS) as an on-line monitoring technique to study furan formation. Unambiguous identification and quantitation in the headspace was achieved by PTR-MS/gas chromatography-mass spectrometry coupling. Ascorbic acid showed the highest potential to generate furan, followed by glyceryl trilinolenate. Some of the reaction samples generated methylfuran as well, such as Maillard systems containing alanine and threonine as well as lipids based on linolenic acid. The furan yields from ascorbic acid were lowered in an oxygen-free atmosphere (30%) or in the presence of reducing agents (e.g., sulfite, 60%), indicating the important role of oxidation steps in the furan formation pathway. Furthermore, already simple binary mixtures of ascorbic acid and amino acids, sugars, or lipids reduced furan by 50-95%. These data suggest that more complex reaction systems result in much lower furan amounts as compared to the individual precursors, most likely due to competing reaction pathways.     

Free amino acids of tronchuda cabbage (Brassica oleracea L. Var. costata DC): influence of leaf position (internal or external) and collection time

The free amino acid profile of 18 samples of tronchuda cabbage ( Brassica oleracea L. var. costata DC) leaves, harvested at three different months, was determined by HPLC/UV-vis. The tronchuda cabbage leaves total free amino acid content varied from 3.3 to 14.4 g/kg fresh weight. Generally, arginine was the major compound, followed by proline, threonine, glutamine, cysteine, and glutamic acid. This study indicates that free amino acids are not similarly distributed: in external leaves, proline and arginine were the major free amino acids, while in internal ones, arginine was the main free amino acid, followed by threonine, glutamine, and cysteine. Significant differences were observed for valine, proline, arginine, leucine, cysteine, lysine, histidine, and tyrosine contents. The levels of some free amino acids were significantly affected by the collection period. In external leaves, this occurred with glutamic acid, serine, valine, leucine, cysteine, and ornithine contents, while in internal leaves, it occurred with aspartic acid, arginine, and total contents.