Formation Of Pyrazines Research Articles

ChemInform Abstract: Vinyl Isonitriles in Radical Cascade Reactions: Formation of Cyclopenta-Fused Pyridines and Pyrazines.

The 4 + 1 radical annulation reaction of vinyl isonitriles with iodoalkynes or iodonitriles affording cyclopenta-fused pyridines and pyrazines respectively and the first example of an intramolecular radical addition to an aryl isonitrile are described.

Formation of Pyrazines in Maillard Model Systems of Lysine-Containing Dipeptides

Whereas most studies concerning the Maillard reaction have focused on free amino acids, little information is available on the impact of peptides and proteins on this important reaction in food chemistry. Therefore, the formation of flavor compounds from the model reactions of glucose, methylglyoxal, or glyoxal with eight dipeptides with lysine at the N-terminus was studied in comparison with the corresponding free amino acids by means of stir bar sorptive extraction (SBSE) followed by GC-MS analysis. The reaction mixtures of the dipeptides containing glucose, methylglyoxal, and glyoxal produced 27, 18, and 2 different pyrazines, respectively. Generally, the pyrazines were produced more in the case of dipeptides as compared to free amino acids. For reactions with glucose and methylglyoxal, this difference was mainly caused by the large amounts of 2,5(6)-dimethylpyrazine and trimethylpyrazine produced from the reactions with dipeptides. For reactions with glyoxal, the difference in pyrazine production was rather small and mostly unsubstituted pyrazine was formed. A reaction mechanism for pyrazine formation from dipeptides was proposed and evaluated. This study clearly illustrates the capability of peptides to produce flavor compounds that can differ from those obtained from the corresponding reactions with free amino acids.

Aroma compounds generated from thermal reaction of l-ascorbic acid with l-cysteine

The reaction of l-ascorbic acid with l-cysteine in heated aqueous solution (141 ± 1 °C) at five different pH values (5.00, 6.00, 7.00, 8.00, or 9.00) for 2 h, resulted in the formation of a complex mixture of aroma volatiles. The volatile compounds generated were analysed by SPME–GC–MS. The results gave 43 aroma compounds. The reaction between l-ascorbic acid and l-cysteine led mainly to the formation of alicyclic sulphur compounds, thiophenes, thienothiophenes, thiophenones, thiazoles and pyrazines, most of which contain sulphur. Many of these volatiles had meaty flavour. The origin of many of the compounds was explained. The studies showed that thienothiophenes and thienones were formed mainly at acidic pH. In contrast, higher pH values could promote the production of thiophenes, thiazoles and pyrazines.

Pyrazines: occurrence, formation and biodegradation

Pyrazines are a class of compounds that occur almost ubiquitously in nature. Pyrazines can be synthesised chemically or biologically, and are used as flavouring additives. The major formation of pyrazines occurs during heating of food. There is very little information available on the degradation of these compounds. In humans and animals, pyrazines are excreted as glucuronates or bound to glutathione via the kidney after hydroxylation, but the pyrazine ring is not cleaved. Bacteria have been isolated, which are able to use various substituted pyrazines as a sole carbon and energy source. In a few cases, the initial metabolites have been characterised; however, the mechanism of ring cleavage and the further degradation pathways are still unknown and await further investigation.

Isotope Labeling Studies on the Origin of 3,4-Hexanedione and 1,2-Butanedione in an Alanine/Glucose Model System

Although the importance of alpha-dicarbonyl compounds as reactive intermediates in the Maillard reaction and as precursors of heterocyclic and odor-active compounds is well-established, however, the detailed origin of many alpha-dicarbonyl compounds such as 3,4-hexanedione and 1,2-butanedione still remains unknown. Using glucose and glyoxal with labeled [(13)C-1]alanine, [(13)C-2]alanine, [(13)C-3]alanine, and [(15)N]alanine, the mechanism of their formation was investigated using the label incorporation pattern of the pyrazines derived through the Strecker reaction. Taking into account the non-oxidative mechanism of pyrazine formation, the data indicated that all of the ethyl-substituted pyrazines identified in the glyoxal/alanine model system incorporated C-2' and C-3' atoms of alanine, and not that of free acetaldehyde, as the ethyl group carbon atoms. This was achieved through spiking experiments using unlabeled acetaldehyde in the presence of labeled alanine. Furthermore, the data also indicated the occurrence of a chain elongation process of sugar-derived alpha-dicarbonyl compounds assisted by alanine. On the basis of the proposed mechanism, the glyoxal interaction with alanine through a decarboxylative aldol addition reaction can lead to the formation of 1,2-butanedione with the terminal ethyl carbon atoms originating from C-2' and C-3' atoms of alanine, and the similar interaction of 1,2-butanedione with a second molecule of alanine can lead to the formation of 3,4-hexanedione with both terminal ethyl carbon atoms originating from C-2' and C-3' atoms of alanine.

Investigations on the Effect of Amino Acids on Acrylamide, Pyrazines, and Michael Addition Products in Model Systems

Acrylamide and pyrazine formation, as influenced by the incorporation of different amino acids, was investigated in sealed low-moisture asparagine-glucose model systems. Added amino acids, with the exception of glycine and cysteine and at an equimolar concentration to asparagine, increased the rate of acrylamide formation. The strong correlation between the unsubstituted pyrazine and acrylamide suggests the promotion of the formation of Maillard reaction intermediates, and in particular glyoxal, as the determining mode of action. At increased amino acid concentrations, diverse effects were observed. The initial rates of acrylamide formation remained high for valine, alanine, phenylalanine, tryptophan, glutamine, and leucine, while a significant mitigating effect, as evident from the acrylamide yields after 60 min of heating at 160 degrees C, was observed for proline, tryptophan, glycine, and cysteine. The secondary amine containing amino acids, proline and tryptophan, had the most profound mitigating effect on acrylamide after 60 min of heating. The relative importance of the competing effect of added amino acids for alpha-dicarbonyls and acrylamide-amino acid alkylation reactions is discussed and accompanied by data on the relative formation rates of selected amino acid-AA adducts.

The effect of pH on the formation of aroma compounds produced by heating a model system containing l-ascorbic acid with l-threonine/l-serine

Abstract The identification of aroma compounds, formed from the reactions of l -ascorbic acid with l -threonine/ l -serine at five different pH values (5.00, 6.00, 7.00, 8.00, or 9.55) and 143 ± 2 °C for 2 h, was performed using a SPME–GC–MS technique, and further use of LRI. The results showed 35 aroma compounds. The reaction between l -ascorbic acid and l -threonine/ l -serine led mainly to the formation of pyrazines. Many of these were alkylpyrazines, such as 2-methylpyrazine, 2,5-dimethylpyrazine, 2-ethylpyrazine, 2-ethyl-6-methylpyrazine, 2-ethyl-5-methylpyrazine, 3-ethyl-2,5-dimethylpyrazine, 2,3-diethyl-5-methylpyrazine, and 3,5-diethyl-2-methylpyrazine; other compounds identified were furans and aldehydes. More volatiles were generated in l -ascorbic acid with l -threonine systems than in l -ascorbic acid with l -serine systems. The studies showed that furans, such as furfural, 2-furanmethanol, benzofuran, 2,5-furandicarboxaldehyde and 2-furfurylfuran were formed mainly at acidic pH. In contrast, higher pH values could promote the production of pyrazines.

Formation of pyrazines from ascorbic acid and amino acids under dry-roasting conditions

Although the participation of ascorbic acid in Maillard-type reactions has been described, the formation of flavour compounds resulting from the interaction of ascorbic acid with different amino acids has not been reported before. Therefore, the formation of flavour compounds from the model reactions of 20 amino acids with ascorbic acid was studied under dry-roasting conditions. Thirty-six different pyrazines were identified, mostly ethyl and methyl substituted pyrazines. The amounts of pyrazines detected were comparable to those formed from pentose sugars. Lysine was the most reactive amino acid and yielded the highest amounts of alkylpyrazines. The reducing activity of ascorbic acid influenced the reaction mechanism of pyrazine formation and thus the type of pyrazines produced. Addition of a base, such as potassium carbonate, significantly enhanced pyrazine formation from ascorbic acid for most amino acids. The formation of pyrazines from serine and threonine without a carbonyl compound was greatly enhanced by the addition of potassium carbonate as well. Furan was detected in all model systems in relatively low amounts and its formation was not enhanced by the addition of potassium carbonate.

Acrylamide and Pyrazine Formation in Model Systems Containing Asparagine

The effect of different sugars and glyoxal on the formation of acrylamide in low-moisture starch-based model systems was studied, and kinetic data were obtained. Glucose was more effective than fructose, tagatose, or maltose in acrylamide formation, whereas the importance of glyoxal as a key sugar fragmentation intermediate was confirmed. Glyoxal formation was greater in model systems containing asparagine and glucose rather than fructose. A solid phase microextraction GC-MS method was employed to determine quantitatively the formation of pyrazines in model reaction systems. Substituted pyrazine formation was more evident in model systems containing fructose; however, the unsubstituted homologue, which was the only pyrazine identified in the headspace of glyoxal-asparagine systems, was formed at higher yields when aldoses were used as the reducing sugar. Highly significant correlations were obtained for the relationship between pyrazine and acrylamide formation. The importance of the tautomerization of the asparagine-carbonyl decarboxylated Schiff base in the relative yields of pyrazines and acrylamide is discussed.

The Aroma Side of the Maillard Reaction

The Maillard reaction in food produces, among others, a diversity of sensory-active compounds (aroma, taste, color). The resulting key aroma compounds are often present only in trace concentrations of 1 microg/kg to 1 mg/kg. Nevertheless, they contribute to the respective flavor because of their low odor-perception thresholds. While Maillard intermediates, such as Amadori compounds and deoxyosones, are formed at percentage levels during model reactions, the yield of aroma compounds, in particular nitrogen and sulfur-containing ones, is often as low as 0.001-0.01 mol%, thus indicating their formation through chemical side reactions. The elucidation of the relevant precursors in food and the identification of previously unknown intermediates can throw light on these minor pathways. Also, model reactions with isotopically labeled precursors are of great value in gaining insight into the relevant formation mechanisms. Several examples of these studies are illustrated including work to elucidate the role of the solvent glycerol in the formation of pyrazines, trials to reveal the relative significance of 4-hydroxy-5-methyl-3(2H)-furanone as intermediate in the reaction between ribose and cysteine, and experiments to assess the proportional contribution of the precursors cysteine, xylose, and thiamine to the formation of the resulting aroma compounds in the thermal reaction.

Formation of Pyrazines and a Novel Pyrrole in Maillard Model Systems of 1,3-Dihydroxyacetone and 2-Oxopropanal

Alkylpyrazines are a very important class of Maillard flavor compounds, but their mechanism of formation is complex and consists of different pathways. The model reaction of 20 different amino acids with 1,3-dihydroxyacetone, as a precursor of 2-oxopropanal, was studied by means of SPME-GC-MS to investigate the involvement of the amino acid side chain in the substitution pattern of the resulting pyrazines. 2,5-Dimethylpyrazine was quantitatively the most important pyrazine formed from all of the amino acids. The amino acid side chain is not involved in its formation. The substituents of other less abundant pyrazines resulted mainly from the incorporation of the Strecker aldehyde or aldol condensation products in the intermediate dihydropyrazine. The importance of different reaction mechanisms was evaluated, taking into account the pattern of pyrazines identified. In the solvent extracts of aqueous model reactions of 2-oxopropanal with amino acids, the main reaction product was not a pyrazine but a novel pyrrole. This pyrrole was identified as 2,5-diacetyl-3-methyl-1 H-pyrrole by means of spectral analysis, secured by chemical synthesis. A reaction mechanism for its formation was proposed and evaluated. The influence of various reaction conditions on the formation of 2,5-diacetyl-3-methyl-1 H-pyrrole and 2,5-dimethylpyrazine in the model reaction of alanine with 2-oxopropanal was studied. These results underscore the importance of the ratio of the different reagents and the presence of water in the resulting flavor formation in the Maillard reaction.

Mechanisms of Alkylpyrazine Formation in a Potato Model System Containing Added Glycine

The use of glycine to limit acrylamide formation during the heating of a potato model system was also found to alter the relative proportions of alkylpyrazines. The addition of glycine increased the quantities of several alkylpyrazines, and labeling studies using [2-13C]glycine showed that those alkylpyrazines which increased in the presence of glycine had at least one 13C-labeled methyl substituent derived from glycine. The distribution of 13C within the pyrazines suggested two pathways by which glycine, and other amino acids, participate in alkylpyrazine formation, and showed the relative contribution of each pathway. Alkylpyrazines that involve glycine in both formation pathways displayed the largest relative increases with glycine addition. The study provided an insight into the sensitivity of alkylpyrazine formation to the amino acid composition in a heated food and demonstrated the importance of those amino acids that are able to contribute an alkyl substituent. This may aid in estimating the impact of amino acid addition on pyrazine formation, when amino acids are added to foods for acrylamide mitigation.

Formation of pyrazines in hydroxyacetaldehyde and glycine nonenzymatic browning Maillard reaction: A computational study

By considering the formation of pyrazines (C 4N 2H 6 and C 4N 2H 4) as one of the possible final Maillard flavour compounds, Density Functional Theory computations at the standard state have been performed on the proposed mechanisms of glyoxal and glycine in the advanced stage of hydroxyacetaldehyde and glycine nonenzymatic browning reaction under different pH conditions. The results reveal that the basic condition is the most favourable for the production of pyrazines ( Pzs), and the aqueous solution is more favourable than that of the gaseous state. The reactions at the isoelectric point of glycine and under neutral conditions are the second and third most favourable for the production of Pzs, respectively. The reaction under acidic conditions is the least feasible for the production of Pzs. Amino acetaldehyde is the most likely precursor of the pyrazine ring in the reaction. Presence of air or oxygen is necessary for the production of 2,3,5,6-tetrahydropyrazine (C 4N 2H 4) from 3,6-dihydropyrazine (C 4N 2H 6). Water is necessary with glyoxal and glycine species for the formation of Pzs and water is produced as a by-product during the formation of Pzs.

Effect of Enzyme Treatments and Drying Temperatures on Methylpyrazine Content in Cocoa (Theobroma Cacao L.) Powder Extract

ABSTRACT: The effects of combining enzyme treatments and heating protocols on pyrazine formation in cocoa powder extracts have been studied. Five commercial enzyme preparations containing protease and carbohydrase activities were initially assessed for their ability to release amino acids and reducing sugars, both of which are substrates for Maillard reactions. The enzyme preparation Flavourzyme was subsequently selected for further study given its ability to liberate both types of substrate. Cocoa powder solutions were treated with 2 doses of Flavourzyme and processed at 2 drying temperatures. The combination of the higher dose of the enzyme (12% w/w enzyme/substrate) and the higher drying temperature (150 °C) resulted in a synergistic increase in the concentration of methyl‐ and dimethylpyrazines. A statistically significant increase in the concentration of tetramethylpyrazine occurred in samples treated at 150 °C that was independent of the enzyme dose assayed. No significant changes in trimethylpyrazine content were detected in treated compared to untreated samples.

Modification of maillard browning in a microwaved glucose/glycine model system by water‐soluble natural antioxidants and foods containing them

AbstractInhibition of pyrazine formation by natural antioxidants and the foods containing them was measured in a microwaved glucose/glycine model system. Inhibition of lipid oxidation by the same materials was assayed in both bulk and emulsion systems. Pyrazines were determined by solid‐phase micro extraction followed by GC. Lipid oxidation volatiles were assayed by polyamide fluorescence produced by either a bulk oil display or a hematin‐ or 2,2′‐azobis‐(2‐amidino=propane) dihydrochloride‐accelerated lecithin or fish oil emulsion. It was shown that (i) the inhibition of pyrazine formation depends on high concentrations of water‐soluble antioxidants; (ii) such antioxidants occur naturally in some foods and are usually polyphenols; (iii) during pyrazine inhibition, oxidized polyphenols show enhanced nonfluorescing browning similar to enzymic browning products; (iv) monophenols, which structurally cannot form quinone polymers on oxidation, inhibit pyrazines with less browning; (v) during the final pyrazine‐forming phase of the Maillard reaction, polyphenolics and reducing agents such as glutathione and ascorbic acid are partially consumed with some nutritional loss; (vi) fruit powders of grape seed, grape skin, and red wine are highly pyrazine‐inhibitory, steeped blueberry strongly so, but plum purees are moderately pro‐pyrazine, and freeze‐dried vegetables strongly pro‐pyrazine; and (vii) black and green tea infusions are highly inhibitory, whereas spices have mixed effects.

Effect of Citric Acid and Glycine Addition on Acrylamide and Flavor in a Potato Model System

Acrylamide levels in cooked/processed food can be reduced by treatment with citric acid or glycine. In a potato model system cooked at 180 degrees C for 10-60 min, these treatments affected the volatile profiles. Strecker aldehydes and alkylpyrazines, key flavor compounds of cooked potato, were monitored. Citric acid limited the generation of volatiles, particularly the alkylpyrazines. Glycine increased the total volatile yield by promoting the formation of certain alkylpyrazines, namely, 2,3-dimethylpyrazine, trimethylpyrazine, 2-ethyl-3,5-dimethylpyrazine, tetramethylpyrazine, and 2,5-diethyl-3-methylpyrazine. However, the formation of other pyrazines and Strecker aldehydes was suppressed. It was proposed that the opposing effects of these treatments on total volatile yield may be used to best advantage by employing a combined treatment at lower concentrations, especially as both treatments were found to have an additive effect in reducing acrylamide. This would minimize the impact on flavor but still achieve the desired reduction in acrylamide levels.

Formation of pyrazines and 2-acetyl-1-pyrroline by Bacillus cereus

The production of alkylpyrazines and 2-acetyl-1-pyrroline by different Bacillus cereus strains, which has been previously reported, was studied in detail. B. cereus ATCC 27522 produced the highest amounts of flavour compounds when grown as surface cultures on plate count agar. Pyrazine, methylpyrazine, 2,5-dimethylpyrazine, trimethylpyrazine and 3-ethyl-2,5-dimethylpyrazine were produced in low amounts. Since they were also detected in control flasks, an enzymatic formation was concluded to be unlikely. Only the production of 3-ethyl-2,5-dimethylpyrazine was in all cases significantly different from the control. Detailed precursor studies revealed that the production of 2-acetyl-1-pyrroline by B. cereus ATCC 27522 proceeds via acetylation of 1-pyrroline, a metabolic degradation product of proline and ornithine. Comparison of results obtained from dynamic headspace and simultaneous steam distillation – solvent extraction showed that the use of a non-thermal extraction method is essential to obtain reliable results on the biological formation of these Maillard flavour compounds.

Formation of pyrazines in dihydroxyacetone and glycine Maillard reaction: A computational study

Based on the electronic and Gibb’s free energy changes, the possibility of the formation of 2,5-dimethyl pyrazine ( 25Pz) as one of the probable final products in dihydroxyacetone ( DHA) and glycine Maillard reaction under different pH conditions is described. Mechanisms for the reaction have been proposed following the Hodge-scheme. Density functional computational calculations at the standard state have been performed on the proposed mechanisms of the reaction. Electronic and Gibb’s free energy changes for the formation of different compounds have been estimated by following the total mass balance for different steps of the reaction. The possible order of feasibility for the production of 25Pz has been found as DHA + deprotonated glycine ( DGly) gaseous > DHA + DGly aqueous > DHA + unprotonated glycine ( UGly) gaseous > DHA + glycine zwitterion ( GlyZ) gaseous > DHA + UGly aqueous > DHA + GlyZ aqueous > DHA + protonated glycine ( PGly) aqueous > DHA + PGly gaseous phase reactions. Amino-acetone has been found to be the most likely precursor of the pyrazine ring. Oxidation plays an important role during the production of 25Pz. Water has been found as a by-product during the formation of 25Pz.

Correlation of Acrylamide Generation in Thermally Processed Model Systems of Asparagine and Glucose with Color Formation, Amounts of Pyrazines Formed, and Antioxidative Properties of Extracts

The relations between the formation of acrylamide and color, pyrazines, or antioxidants in an asparagine/d-glucose browning model system under various conditions were investigated. The highest level of acrylamide was produced in the asparagine/glucose (1:3) system heated at 170 degrees C for 30 min (2629 microg/g asparagine). Color intensity increased with temperature and heating time. The formation of pyrazines increased steadily with an increase of temperature (140-170 degrees C) and heating time (15-60 min). Antioxidant formation varied among the samples heated under different conditions. A clear correlation between formation of acrylamide and browning color was obtained. The formation of acrylamide was linearly correlated with the formation of total pyrazines during the initial stages of the Maillard reaction. No obvious correlation between formation of acrylamide and antioxidants was observed. However, excess amounts of asparagine increased the formation of antioxidants, whereas excess amounts of glucose reduced its formation.

Formation of Pyrazines in Aqueous Maltose/Glucose/Fructose-Glutamine Model Systems upon Heating at below 100.DEG.C.

Pyrazines generated from aqueous sugar-glutamine model systems heated at 90°C were investigated quantitatively. To determine trace levels of pyrazines in aqueous matrices, an efficient method using solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GCMS) was developed. A series of alkylpyrazines as well as trace levels of acetylpyrazines and bis-(2-furyl)pyrazines were detected in the model systems by this method. Remarkable difference in the formation of pyrazines between monosaccharide and disaccharide was observed. The yield of acetylpyrazines and bis-(2-furyl)pyrazines from maltose was larger than those from fructose and glucose, while that of alkylpyrazines was less. The pH dependency on the generation of pyrazines in maltose-glutamine model systems was also examined.