Chemical conversion of alpha-amino acids into alpha-keto acids by 4,5-epoxy-2-decenal

The comparative formation of phenylalanine and phenylpyruvic acid in the reaction of 4,5-epoxy-2-decenal with phenylalanine was studied to determine whether epoyalkenals may also degrade amino acids without producing their decarboxylation. Both compounds were produced in the reaction to an extent that depended on the reaction pH, the amount of lipid oxidation product, and the reaction time and temperature. The optimum pH was 3 for producing both carbonyl derivatives, and the amount of both compounds increased linearly with the amount of epoxyalkenal present in the reaction mixture. In addition, phenylpyruvic acid was produced to a higher extent than phenylacetaldehyde at 37 degrees C. However, at 60 degrees C the degradation of phenylpyruvic acid was observed and phenylacetaldehyde was usually found to a higher extent than the alpha-keto acid in the overnight-incubated reaction mixtures. The degradation of phenylpyruvic acid produced benzaldehyde and phenylacetaldehyde. All these results suggest that epoxyalkenals can not only degrade amino acids by a Strecker-type mechanism but convert them into their corresponding alpha-keto acids. This new reaction may be an alternative chemical route for the formation in foods of alpha-keto acids, which can later participate in the generation of important amino acid-derived flavor compounds.     

Strecker-type degradation produced by the lipid oxidation products 4,5-epoxy-2-alkenals

Strecker degradation is one of the most important reactions leading to final aroma compounds in the Maillard reaction. In an attempt to clarify whether lipid oxidation products may be contributing to the Strecker degradation of amino acids, this study analyzes the reaction of 4,5-epoxy-2-alkenals with phenylalanine. In addition to N-substituted 2-(1-hydroxyalkyl)pyrroles and N-substituted pyrroles, which are major products of the reaction, the formation of both the Strecker aldehyde phenylacetaldehyde and 2-alkylpyridines was also observed. The aldehyde, which was produced at 37 degrees C-as could be determined by forming its corresponding thiazolidine with cysteamine-and pH 6-7, was not produced when the amino acid was esterified. This aldehyde is suggested to be produced through imine formation, which is then decarboxylated and hydrolyzed. This reaction also produces a hydroxyl amino derivative, which is the origin of the 2-alkylpyridines identified. All these data indicate that Strecker-type degradation of amino acids is produced at 37 degrees C by some lipid oxidation products. This is a new proof of the interrelations between lipid oxidation and Maillard reaction, which are able to produce common products by analogue mechanisms.     

Strecker type degradation of phenylalanine by 4-hydroxy-2-nonenal in model systems

The reaction of 4-hydroxy-2-nonenal, an oxidative stress product, with phenylalanine in acetonitrile-water (2:1, 1:1, and 1:2) at 37, 60, and 80 degrees C was investigated to determine whether 4-hydroxy-2-alkenals degrade amino acids, analogously to 4,5-epoxy-2-alkenals, and to compare the reactivities of both hydroxyalkenals and epoxyalkenals for production of Strecker aldehydes. In addition to the formation of N-substituted 2-pentylpyrrole and 2-pentylfuran, the studied hydroxyalkenal also degraded phenylalanine to phenylacetaldehyde with a reaction yield of 17%. The reaction mechanism is suggested to be produced through the corresponding imine, which is then decarboxylated and hydrolyzed. This reaction also produced a conjugated amine, which both may be one of the origins of the produced 2-pentyl-1H-pyrrole and may contribute to the development of browning in these reactions. 4-Hydroxy-2-nonenal and 4,5-epoxy-2-decenal degraded phenylalanine in an analogous extent, which is likely a consequence of the similarity of the degradation mechanisms involved. These results suggest that different lipid oxidation products are able to degrade amino acids; therefore, the Strecker type degradation of amino acids produced by oxidized lipids may be quantitatively significant in foods.     

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.     

Strecker degradation of phenylalanine initiated by 2,4-decadienal or methyl 13-oxooctadeca-9,11-dienoate in model systems

The reaction of 2,4-decadienal and methyl 13-oxooctadeca-9,11-dienoate with phenylalanine was studied to determine if alkadienals and ketodienes are able to produce the Strecker-type degradation of amino acids to the corresponding Strecker aldehydes. When reactions were carried out at 180 degrees C, both carbonyl compounds degraded phenylalanine to phenylacetaldehyde, among other compounds. The yield of the phenylacetaldehyde produced depended on the reaction pH and increased linearly with both the amount of the lipid and the reaction time. The yield of this conversion was approximately 8% when starting from decadienal and approximately 6% when starting from methyl 13-oxooctadeca-9,11-dienoate, and the reaction rate was lower for the ketone than for the aldehyde. Simultaneous to these reactions, the lipid was converted into pyrrole, pyridine, or aldehyde derivatives as a result of several competitive reactions. In particular, 9-14% of the decadienal was converted into hexanal under the assayed conditions. All these reactions are suggested to be produced as a consequence of the oxidation of the alkadienal or the ketodiene to the corresponding epoxyalkenal or unsaturated epoxyketone, which were identified in the reaction mixtures by GC-MS. All these results suggest that alkadienals and ketodienes, which are quantitatively important secondary lipid oxidation products, can degrade amino acids to their corresponding Strecker aldehydes. Therefore, under appropriate conditions, these products are not final products of the lipid oxidation and can participate in carbonyl-amine reactions analogously to other lipid oxidation products with two oxygenated functions.     

Alkylpyridiniums. 1. Formation in model systems via thermal degradation of trigonelline

Trigonelline is a well-known precursor of flavor/aroma compounds in coffee and undergoes significant degradation during roasting. This study investigates the major nonvolatile products that are procured after trigonelline has been subjected to mild pyrolysis conditions (220-250 degrees C) under atmospheric pressure. Various salt forms of trigonelline were also prepared and the thermally produced nonvolatiles analyzed by thin layer chromatography, liquid chromatography-electrospray ionization tandem mass spectrometry, and (1)H and (13)C nuclear magnetic resonance. Results revealed the decarboxylated derivative 1-methylpyridinium as a major product of certain salts, the formation of which is positively correlated to temperature from 220 to 245 degrees C. Moreover, trigonelline hydrochloride afforded far greater amounts of 1-methylpyridinium compared to the monohydrate over the temperature range studied. Investigations into other potential quaternary amine products of trigonelline also indicate nucleophilic substitution reactions that lead to dialkylpyridiniums, albeit at concentration levels approximately 100-fold lower than those recorded for 1-methylpyridinium.     

Studies on the aroma of five fresh tomato cultivars and the precursors of cis- and trans-4,5-epoxy-(E)-2-decenals and methional

Three tasty (BR-139, FA-624, and FA-612) and two less tasty (R-144 and R-175) fresh greenhouse tomato cultivars, which significantly differ in their flavor profiles, were screened for potent odorants using aroma extract dilution analysis (AEDA). On the basis of AEDA results, 19 volatiles were selected for quantification in those 5 cultivars using gas chromatography-mass spectrometry (GC-MS). Compounds such as 1-penten-3-one, ( E, E)- and ( E, Z)-2,4-decadienal, and 4-hydroxy-2,5-dimethyl-3(2 H)-furanone (Furaneol) had higher odor units in the more preferred cultivars, whereas methional, phenylacetaldehyde, 2-phenylethanol, or 2-isobutylthiazole had higher odor units in the less preferred cultivars. Simulation of the odor of the selected tomato cultivars by preparation of aroma models and comparison with the corresponding real samples confirmed that all important fresh tomato odorants were identified, that their concentrations were determined correctly in all five cultivars, and that differences in concentration, especially of the compounds mentioned above, make it possible to distinguish between them and are responsible for the differential preference. To help elucidate formation pathways of key odorants, labeled precursors were added to tomatoes. Biogenesis of cis- and trans-4,5-epoxy-( E)-2-decenals from linoleic acid and methional from methionine was confirmed.     

Characterization of model melanoidins by the thermal degradation profile

Different types of model melanoidins were thermally degraded, with subsequent identification of the volatiles produced, to obtain and compare the thermal degradation profile of various melanoidins. At first, the volatiles produced from heated glucose/glycine standard melanoidins were compared with glucose/glutamic acid and L-(+)-ascorbic acid/glycine standard melanoidins. In the headspace of heated glucose/glycine melanoidins, mainly furans, were detected, accompanied by carbonyl compounds, pyrroles, pyrazines, pyridines, and some oxazoles. Heating of L-(+)-ascorbic acid/glycine melanoidins resulted in relatively more N-heterocycles, while from glucose/glutamic acid melanoidins no N-heterocycles were formed. In a second part, a chemical treatment was applied to glucose/glycine melanoidins prior to the thermal degradation. Acid hydrolysis was performed to cleave glycosidically linked sugar moieties from the melanoidin skeleton. Nonsoluble glucose/glycine melanoidins were also subjected to an oxidation. The results indicate that the thermal degradation profile is a useful tool in the characterization of different types of melanoidins.     

Origin of carbohydrate degradation products in L-Alanine/D-[(13)C]glucose model systems

Maillard model systems consisting of labeled D-[(13)C]glucoses and L-[(13)C]alanines have been utilized to identify the origin of carbon atoms in glycolaldehyde, pyruvaldehyde, 1-hydroxy-2-propanone (acetol), 2,3-butanedione, 3-hydroxy-2-butanone, 2,3-pentanedione, and compounds containing C(5) and C(6) intact glucose carbon chains. The origin of carbon atoms in glycolaldehyde and pyruvaldehyde was inferred from the analysis of label incorporation pattern of methyl and dimethylpyrazines. The origin of carbon atoms in the remaining compounds was determined by direct analysis. The data indicated that glycolaldehyde incorporated intact C5-C6 and C1-C2 carbon chains of glucose. Acetol and pyruvaldehyde incorporated intact C1-C2-C3 and C4-C5-C6 carbon chains of glucose. On the other hand, 2, 3-butanedione and 3-hydroxy-2-butanone incorporated intact C3-C4-C5-C6 carbon chain of glucose. In addition, analysis of compounds containing intact glucose C(5) carbon chains have indicated that glucose in the presence of L-alanine can lose either C-1 atom to produce a pentitol moiety responsible for the formation of furanmethanol or it can lose the C-6 atom to produce a pentose moiety responsible for the formation of furfural. Plausible mechanisms, consistent with the observed label incorporation, were proposed for the formation of sugar degradation products.     

Carbohydrate and amino acid degradation pathways in L-methionine/D-[13C] glucose model systems

Maillard model systems consisting of labeled D-[(13)C]glucoses, L-[(15)N]methionine, and L-[methyl-(13)C]methionine, have been utilized to identify the amino acid and carbohydrate fragmentation pathways occurring in the model system through Py-GC/MS analysis. The label incorporation analyses have indicated that the carbohydrate moiety produces 1-deoxy- and 3-deoxyglucosones and undergoes C(2)/C(4) and C(3)/C(3) cleavages to produce glycolaldehyde, tetrose, and C(3)-reactive sugar derivatives such as acetol, glyceraldehyde, and pyruvaldehyde. Glycolaldehyde was found to incorporate C-1, C-2 (70%) and C-5, C-6 (30%) glucose carbon fragments, whereas the tetrose moiety incorporates only C-3, C-4, C-5, C-6 glucose carbon atoms. In addition, the major source of reactive C(3) fragments was found to contain C-4, C-5, C-6 sugar moiety. On the other hand, methionine alone also generated Strecker aldehyde as detected by its condensation product with 3-(methylthio)propylamine. Plausible mechanisms were proposed for the formation of the interaction products between sugar and amino acid degradation products on the basis of the label incorporation patterns.     

Binding of 2-nonanone and milk proteins in aqueous model systems

Interactions of the model flavor compound 2-nonanone with individual milk proteins, whey protein isolate (WPI), and sodium caseinate in aqueous solutions were investigated. A method to quantify the free 2-nonanone was developed using headspace solid-phase microextraction followed by gas chromatography with flame ionization detection. Binding constants (K) and numbers of binding sites (n) for 2-nonanone on the individual proteins were calculated. The 2-nonanone binding capacities decreased in the order bovine serum albumin > beta-lactoglobulin > alpha-lactalbumin > alpha s1-casein > beta-casein, and the binding to WPI was stronger than the binding to sodium caseinate. All proteins appeared to have one binding site for 2-nonanone per molecule of protein at the flavor concentrations investigated, except for bovine serum albumin, which possessed two classes of binding sites. The binding mechanism is believed to involve predominantly hydrophobic interactions.     

Heat-induced, metal-catalyzed oxidative degradation of quercetin and rutin (Quercetin 3-O-rhamnosylglucoside) in aqueous model systems

The oxidative degradation of quercetin and rutin in phosphate buffer solutions, pH 8.0, at 97 degrees C, was studied by means of UV-vis spectroscopy and reversed-phase high-performance liquid chromatography (HPLC). The effect of the transition metal ions Fe(2+) and Cu(2+) on degradation rate and browning development was also assessed. It was shown that both flavonols are very labile to thermally induced degradation under oxidative conditions. Fe(2+) and Cu(2+) caused an increase in the degradation rate, as well as an increase in browning (A(420)). Significant differences were observed in the degradation mechanisms, as implied by HPLC analyses. It is postulated that metal ions promote flavonol oxidation through reactive oxygen species formation, whereas increases in browning could be ascribed to oxidation and metal-polyphenol interactions.     

Conversion of phenylalanine into styrene by 2,4-decadienal in model systems

2,4-Decadienal was heated under an inert atmosphere and in the presence of phenylalanine to investigate whether this secondary lipid oxidation product is a final product of lipid oxidation or it reacts with the amino acid. The results obtained showed that, in the presence of the alkadienal, the amino acid was degraded to styrene. This reaction was favored in dry systems at pH approximately 6 and in the absence of oxygen. If oxygen was present, the alkadienal was oxidized and the Strecker degradation of the amino acid was produced. The activation energy for the formation of styrene from phenylalanine was 150.4 kJ/mol. The reaction mechanism is suggested to be produced either by an electronic rearrangement of the imine produced between the aldehyde and the amino acid with the formation of styrene, 2-pentylpyridine, carbon dioxide, and hydrogen, or by Michael addition of the amino compound to the alkadienal followed by beta-elimination to produce the same compounds. Both reaction schemes were supported on the results obtained by studying both the degradation of phenylethylamine and phenylalanine methyl ester produced by 2,4-decadienal, and the formation of ethylbenzene in decadienal/phenylalanine reaction mixtures heated in the presence of platinum oxide. All these results suggest that, analogously to carbohydrates, certain lipid oxidation products may degrade appropriate amino acids to their corresponding vinylogous derivatives.     

Thermally induced degradation of sulfur-containing aliphatic glucosinolates in broccoli sprouts (Brassica oleracea var. italica) and model systems

Processing reduces the glucosinolate (GSL) content of plant food, among other aspects due to thermally induced degradation. Since there is little information about the thermal stability of GSL and formation of corresponding breakdown products, the thermally induced degradation of sulfur-containing aliphatic GSL was studied in broccoli sprouts and with isolated GSL in dry medium at different temperatures as well as in aqueous medium at different pH values. Desulfo-GSL have been analyzed with HPLC-DAD, while breakdown products were estimated using GC-FID. Whereas in the broccoli sprouts structural differences of the GSL with regard to thermal stability exist, the various isolated sulfur-containing aliphatic GSL degraded nearly equally and were in general more stable. In broccoli sprouts, methylsulfanylalkyl GSL were more susceptible to degradation at high temperatures, whereas methylsulfinylalkyl GSL were revealed to be more affected in aqueous medium under alkaline conditions. Besides small amounts of isothiocyanates, the main thermally induced breakdown products of sulfur-containing aliphatic GSL were nitriles. Although they were most rapidly formed at comparatively high temperatures under dry heat conditions, their highest concentrations were found after cooking in acidic medium, conditions being typical for domestic processing.     

Thermal generation of 3-amino-4,5-dimethylfuran-2(5H)-one, the postulated precursor of sotolone, from amino acid model systems containing glyoxylic and pyruvic acids

4,5-Dimethyl-3-hydroxy-2(5H)-furanone (sotolone), a naturally occurring flavor impact compound, can be isolated from various sources, especially fenugreek seeds. It can also be thermally produced from intermediates generated from the Maillard reaction such as pyruvic and ketoglutaric acids, glyoxal, and 2,3-butanedione. A naturally occurring precursor of sotolone, 3-amino-4,5-dimethyl-2(5H)-furanone, was thermally generated for the first time from pyruvic acid and glycine or from glyoxylic acid and alanine model systems. Isotope labeling studies have implicated 4,5-dimethylfuran-2,3-dione as an intermediate that can be converted into 3-amino-4,5-dimethyl-2(5H)-furanone through Strecker-like interaction with any amino acid. Furthermore, these studies have also indicated the presence of two pathways for the formation of 4,5-dimethylfuran-2,3-dione, one requiring pyruvic acid and a formaldehyde source and the other requiring glyoxylic acid and acetaldehyde. Self-aldol condensation of pyruvic acid followed by lactonization and further aldol reaction with formaldehyde can generate the same intermediate as the self-aldol addition product of acetaldehyde with glyoxylic acid followed by lactonization. The pyruvic acid pathway was found to be a more efficient route than the glyoxylic acid pathway. Furthermore, the pyruvic acid/glycine model system was able to generate sotolone in the presence of moisture, and in the presence of ammonia, commercial sotolone was converted back into 3-amino-4,5-dimethyl-2(5H)-furanone.     

Assessment of urea degradation rate in model wine solutions by acid urease from Lactobacillus fermentum

The specific activity (piA) of a whole cell acid urease preparation was assessed in model wine solutions at different levels of malic (M) and lactic acids, metabisulfite, ethanol, and pH by performing a central composite design. M and then pH were found to be the most controlling variables, their effects being practically coincident but of opposite sign. For urea concentrations up to approximately 1 mol m(-3) the ammonium formation rate was assumed of the pseudo-first-order with respect to urea, this being confirmed by two independent validation tests performed at 20 degrees C for as long as 24 h. In the case of real wines the effective pseudo-first-order kinetic rate constants were found to be smaller than those pertaining to the model solutions having the same wine composition and pH by a factor varying from 10 to 1000, this affecting significantly the specific urease treatment costs per liter of wine treated.     

Chemical conversion of phenylethylamine into phenylacetaldehyde by carbonyl-amine reactions in model systems

The chemical conversion of phenylethylamine into phenylacetaldehyde in the presence of lipid oxidation products (LOPs) was studied to investigate the possibility that biogenic amines can be converted into Strecker aldehydes upon processing. Model systems of phenylethylamine and methyl 13-hydroperoxyoctadeca-9,11-dienoate (HP), 2,4-decadienal (DD), 4,5-epoxy-2-heptenal (EH), 4,5-epoxy-2-decenal (ED), 4-oxo-2-hexenal (OH), 4-oxo-2-nonenal (ON), or 4-hydroxy-2-nonenal (HN) were heated for 1 h at 180 °C and pH 3. Although HN and EH did not produce more phenylacetaldehyde than when phenylethylamine was heated alone, all other lipid oxidation products assayed increased the amount of phenylacetaldehyde produced by 300-900%, with ON being the most reactive compound for this reaction. The reaction was mainly produced at acidic pH values (<6) and was dependent upon the concentration of the LOPs involved, and the phenylacetaldehyde produced increased linearly as a function of the time and temperature. The E(a) values for the reactions between phenylethylamine and DD and ON were 54.8 and 53.8 kJ/mol, respectively. The reaction is proposed to take place by the formation of an imine between the phenylethylamine and the LOPs, which is later converted into another imine by an electronic rearrangement. This new imine is the origin of phenylacetaldehyde by hydrolysis. These results show a new pathway for Strecker aldehyde formation. This route provides a potential way to reduce biogenic amine content in foods when they can be thermally processed before consumption.     

Haze formation in model beer systems

The interaction of a haze-active protein (gliadin) and a haze-active polyphenol (tannic acid) was studied in a model beer system in order to investigate the principle mechanisms of haze formation at low temperatures. Low concentrations (g/L) of tannic acid, high concentrations of gliadin, and comparatively high temperatures lead to maximum haze values. When considered on a molar basis, the greatest haze levels are displayed at an approximate 1:1 equivalence of polyphenol and protein. The greater part of haze formation was completed within 0.5 h, irrespective of the concentration of gliadin, the concentration of tannic acid, and the temperature of the model solution.     

Photochemistry of imidacloprid in model systems

The photochemical behavior of the neonicotinoid insecticide imidacloprid was studied with regard to different chemical environments. Different model solvents simulated the structure moieties mainly occurring in waxes and cutin of the plant cuticle. Cyclohexane and cyclohexene substituted saturated and unsaturated hydrocarbon chains, whereas ethanol and 2-propanol were models for primary and secondary alcohol groups of cuticular components. After 5 h of irradiation, imidacloprid was completely degraded in all solvents. With 88-96 mol% 1-[(6-chloropyridin-3-yl)methyl]imidazolidin-2-imine was formed as the main product, whereas 1-[(6-chloropyridin-3-yl)methyl]imidazolidin-2-one was identified as minor product in the range 4-6 mol%. By contrast, besides the photoproducts formed in organic solvents, irradiation of the solid imidacloprid on a glass surface delivered a complex variety of unidentified photoproducts. The nucleophilic addition reaction of the main photoproduct, 1-[(6-chloropyridin-3-yl)methyl]imidazolidin-2-imine, with both cyclohexene oxide and methyl 9,10-epoxystearate as model compounds indicates that epoxidized cutin acids are possible reaction partners for the formation of plant cuticle bound residues of imidacloprid, which could explain the reported findings of nonextractable residues of imidacloprid in plants.