Maillard Processes Research Articles

PROPERTIES AND GENERALIZED SCHEME FOR PRODUCTION OF MILK POWDER

Presently, the functional qualities of milk proteins have led to milk powders, being mostly regarded as food components. When milk powders are maintained, numerous physicochemical problems develop, the most of them being generated by the lactose glass transition. They have significant effects on milk powders physical (flow ability) properties. First, powder particle surface microstructure and chemical composition are altered by lactose crystallization. As a result, milk powder's capacity travel is reduced. Milk proteins solubility is harmed because their structural integrity has been destroyed. Additionally, caking and particle collapse occur, and these processes primarily decrease the physical properties of milk powders(Flow capability and density). The unfolding of proteins, which reduces solubility and may also be encouraged by the mechanical stresses that appear. Lastly, enzymatic and chemical reactions (Maillard reaction) occur in order, implementing molecular mobility. Milk powder's solubility is mostly decreased by protein interactions and aggregations, which are enhanced by the Maillard process and oxidation. Emulsifying and foaming characteristics are also decreased by the Maillard process. Additionally demonstrated that milk components, time, and their physical state. The main factors that have been considered are the relative humidity and temperature of the storage

Analysis of Glyoxal- and Methylglyoxal-Derived Advanced Glycation End Products during Grilling of Porcine Meat.

The reaction of the N6-amino group of lysine residues and 1,2-dicarbonyl compounds during Maillard processes leads to advanced glycation end products (AGEs). In the present work, we deliver a comprehensive analysis of changes of carbohydrates, dicarbonyl structures, and 11 AGEs during the grilling of porcine meat patties. While raw meat contained mainly glyoxal-derived N6-carboxymethyl lysine (CML), grilling led to an increase of predominantly methylglyoxal-derived AGEs N6-carboxyethyl lysine (CEL), N6-lactoyl lysine, methylglyoxal lysine dimer (MOLD), and methylglyoxal lysine amide (MOLA). Additionally, we identified and quantitated a novel methylglyoxal-derived amidine compound N1,N2-di-(5-amino-5-carboxypentyl)-2-lactoylamidine (methylglyoxal lysine amide, MGLA) in heated meat. Analysis of carbohydrates suggested that approximately 50% of the methylglyoxal stemmed from the fragmentation of triosephosphates during the heat treatment. Surprisingly, N6-lactoyl lysine was the major AGE, and based on model incubations, we propose that approximately 90% must be explained by the nonenzymatic acylation of lysine through S-lactoylglutathione, which was quantitated for the first time in meat herein.

Comprehensive Analyses of Carbohydrates, 1,2-Dicarbonyl Compounds, and Advanced Glycation End Products in Industrial Bread Making.

The technology of bread making is characterized by three major steps: dough mixing, proofing, and baking. To follow the course of Maillard processes in an authentic food matrix, the complete manufacturing process of wheat bread rolls was assessed along all production steps with the quantitation of sugars, furfurals, 1,2-dicarbonyl compounds, and advanced glycation end products (AGEs). As a result, the AGE profile was significantly enlarged to more than 12 structures, and comprehensive mechanistic insights were provided. The analyses of five major German bread types including wheat, brown, rye bread, pumpernickel, and crispbreads led to AGE contents of 69-149 mg/kg bread or 984-1857 mg/kg protein. Major lysine protein modifications were carboxymethyl, carboxyethyl, and formyl lysine and pyrraline. Arginine was mainly modified by methylglyoxal (MGO) to give imidazolinones. A major part of MGO was confirmed to stem from microbial metabolism.

The protective effects of bexarotene against advanced glycation end-product (AGE)-induced degradation of articular extracellular matrix (ECM)

Osteoarthritis (OA) is a common debilitating disease primarily characterised by excessive loss of the articular ECM, which is composed of up to 95% type II collagen. Among the factors that contribute to the pathogenesis of OA, the natural process of aging is regarded as the most significant risk factor. AGEs, which are extremely resilient to degradation, are produced in the body naturally as a result of the Maillard process of nonenzymatic glycation and are also introduced through diet and tobacco smoke. AGEs have a high affinity for collagen and therefore accumulate in joint tissues, where they induce increased expression of proinflammatory cytokines, chemokines, and degradative enzymes. Additionally, AGEs induce oxidative stress, which further exacerbates the degradative process. Type II collagen is targeted for degradation by matrix metalloproteinases (MMPs), particularly MMP-3 and MMP-13, and AGEs have been shown to trigger increased expression of these MMPs. The role of retinoid and rexinoid receptors as specific treatment targets has been receiving increasing attention. Bexarotene is a retinoid X receptor (RXR) agonist used for the treatment of T-cell lymphoma and other cancers which has displayed a favourable safety profile. Here, we examined the roles of RXR agonism using bexarotene on AGE-induced markers of OA, including oxidative stress, inflammatory response, and MMP-mediated degradation of type II collagen. Furthermore, we demonstrate that bexarotene inhibited phosphorylation of IκBα, thereby suppressing activation of the proinflammatory NF-κB cellular signalling pathway. These findings present a basis for selective targeting of RXR by bexarotene as a potential treatment of OA induced by AGEs.

Variation of the Chemical Composition of Waste Cooking Oils upon Bentonite Filtration

The chemical composition and the color of samples of waste cooking oils (WCOs) were determined prior to and after filtration on two different pads of bentonite differing in particle size. The volatile fraction was monitored by headspace solid-phase microextraction (HS-SPME) coupled with gas-chromatography, while the variation of the composition of the main components was analyzed by 1H NMR. Both techniques allowed the detection of some decomposition products, such as polymers, terpenes, and derivatives of the Maillard process. The analysis of the chemical composition prior to and after bentonite treatment revealed a tendency for the clays to retain specific chemical groups (such as carboxylic acids or double bonds), independent of their particle size. A pair comparison test was conducted in order to detect the sensory differences of the intensity of aroma between the WCO treated with the two different bentonites. In addition, characterization of the bentonite by means of powder X-ray diffraction (XRD) and thermogravimetric measurements (TG) was performed.

Solution to Maillard and grilled steak challenge.

Today’s solution to the ABC Analytical Challenge is unusual, because there is no answer yet. Let us recall the facts. Maillard processes are famous in science and technology, particularly food and medicine. In fact, Maillard processes are said to be responsible for both good and bad [1]: they contribute to the flavour of some cooked food (particularly meat) but also generate glycation products, which can be bad for our health, as is clearly apparent in the proceedings of the Maillard association (IMARS [2]). It is also true that Maillard processes are “fashionable”: in July 2015, a Google Scholar search for “Maillard reaction” led to more than 45,000 answers (250,000 for “Diels-Alder”), including more than 2 000 results for 2015 only. The same growing trend is also apparent from the Pubmed database (Fig. 1). A variety of aspects and models have been studied but, as said in the Maillard Challenge [3], the chemists are not clear what is and what is not a Maillard reaction. Frequently, articles include such statements as “The Maillard reaction is a very complex series of reactions”, which is a way of admitting the general perspective is far from clear. As said in the Maillard Challenge [3], the term “Maillard reaction” should be used to refer to interactions between an amino group and anα-hydroxy carbonyl moiety of a reducing sugar, which produces Amadori or Heyn’s products, depending on the particular sugar (aldose, ketose) [4–6]. A cascade of reactions follows, including many that have been studied outside the food context of Maillard reactions (thermolysis, oxidation, hydrolysis, reduction, aldol condensation, etc.), and the issue is whether such reactions should be included in the “Maillard reaction”. Of course, we could consider whether Maillard reactions include all these reactions, but it would result in a very complex system. It has been proposed that the Maillard reaction should be classified into its early, advanced, and final stages [7], but this proposal was recognized as simplistic. Later [8], it was proposed that the propagation of the Maillard reaction could be described by the formation and interaction of socalled “chemical pools” generated from specific precursors. Intermediates produced during the initial stage of the Maillard reaction arise from three well defined principal precursors: sugars (S), amino acids (A), and Amadori or Heyn’s products (D). The nature and relative ratio of the principal precursors, which constitute the parent mixture (A + S + D) determines the pathway of the Maillard reaction for particular conditions. The proposal to use the term “fragmentation pools” is not convenient, because what was called “advanced Maillard reactions” involved not only fragmentations but also condensations. However, such a proposal has the disadvantage that “Maillard reactions” would include chemical reactions for which there is no Schiff base, for example caramelization [9, 10], hence the proposal is to restrict the Maillard reaction solely to the first step of Schiff base formation. Indeed, the situation is the same as for the discovery of chlorophylls, when the French chemist Joseph-Bienaime Caventou (1795–1877) proposed the name “chlorophylle” for what cooks had, for centuries, called “spinach green” [11, 12]. Caventou only wanted to conduct a new This article is the solution to the Analytical Challenge to be found at Doi: 10.1007/s00216-015-8714-2

Stable nanoparticles prepared by heating electrostatic complexes of whey protein isolate-dextran conjugate and chondroitin sulfate.

A simple and green method was developed for preparing the stable biopolymer nanoparticles with pH and salt resistance. The method involved the macromolecular crowding Maillard process and heat-induced gelation process. The conjugates of whey protein isolate (WPI) and dextran were produced by Maillard reaction. The nanoparticles were fabricated by heating electrostatic complexes of WPI-dextran conjugate and chondroitin sulfate (ChS) above the denaturation temperature and near the isoelectric point of WPI. Then, the nanoparticles were characterized by spectrophotometry, dynamic laser scattering, zeta potential, transmission electron microscopy, atomic force microscopy, and scanning electron microscopy. Results showed that the nanoparticles were stable in the pH range from 1.0 to 8.0 and in the presence of high salt concentration of 200 mM NaCl. WPI-dextran conjugate, WPI, and ChS were assembled into the nanoparticles with dextran conjugated to WPI/ChS shell and WPI/ChS core. The repulsive steric interactions, from both dextran covalently conjugated to WPI and ChS electrostatically interacted with WPI, were the major formation mechanism of the stable nanoparticles. As a nutrient model, lutein could be effectively encapsulated into the nanoparticles. Additionally, the nanoparticles exhibited a spherical shape and homogeneous size distribution regardless of lutein loading. The results suggested that the stable nanoparticles from proteins and strong polyelectrolyte polysaccharides would be used as a promising target delivery system for hydrophobic nutrients and drugs at physiological pH and salt conditions.

Conjugation of gluten hydrolysates with glucosamine at mild temperatures enhances antioxidant and antimicrobial properties

Gluten represents one of the principal by-products of the wheat starch industry. Peptides obtained by wheat hydrolysis can be used for specific functional and biological activities, albeit at relatively low yields. Although the Maillard reaction (glycation) is widely used to increase functionality of proteins, its main disadvantage is the production of undesirable compounds due to high processing temperature. In this research, functional and biologically active glycopeptides were obtained from gluten. Alcalase or Flavourzyme proteases were used to hydrolyse gluten protein, and the resulting peptides were conjugated with glucosamine by enzymatic glycosylation, using transglutaminase, or through glycation. Both reactions were performed at mild temperatures (25 or 37 °C). The formation of glycopeptides depended mostly on the glycation process, as demonstrated by MALDI-TOF-MS. The bioactivities of the conjugated hydrolysates were compared to the native hydrolysates. Although a reduction in the anti-ACE activity was detected, improved DPPH scavenging activity and enhanced antimicrobial activity against Escherichia coli were observed in the glycated Alcalase-derived hydrolysates and in the glycated Flavourzyme-derived hydrolysates, respectively. This study showed that mild conditions are an alternate approach to the traditional Maillard process conducted at elevated temperatures in creating conjugated gluten hydrolysates with enhanced bioactivities.

Maillard Reactions in Hyperthermophilic Archaea: Implications for Better Understanding of Non-Enzymatic Glycation in Biology

Maillard reactions are an unavoidable feature of life that appear to be damaging to cell and organisms. Consequently, all living systems must have ways to protect themselves against this process. As of 2012, several such defense mechanisms have been identified. They are all enzymatic and were found in mesophilic organisms. To date, no systematic study of Maillard reactions and the relevant defense mechanisms has been conducted in thermophiles (50°C-80°C) or hyperthermophiles (80°C-120°C). This is surprisingly because Maillard reactions become significantly faster and potent with increasing temperatures. This review examines this neglected issue in two well-defined sets of hyperthermophiles. My analysis suggests that hyperthermophiles cope with glycation stress by several mechanisms: • Absence of glycation-prone head groups (such as ethanoalamine) from hyperthermophilic phospholipids • Protection of reactive carbohydrates and labile metabolic intermediates by substrate channeling. • Conversion of excess reactive sugars such as glucose to non-reactive compounds including trehalose, di-myo-inositol-phosphate and mannosylglycerate. • Detoxification of methylglyoxal and other ketoaldehydes by conversion to inert products through a variety of reductases and dehydrogenases. • Scavenging of the remaining carbonyls by nucleophilic amines, including a variety of novel polyamines. Disruption of the Maillard process at its early stages, rather than repair of damage caused by it at later stages, appears to be the preferred strategy in the organisms examined. The most unique among these mechanisms appears to be a polyamine-based scavenging system. Undertaking research of the Maillard process in hyperthermophiles is important in its own right and is also likely to provide new insights for the control of these reactions in humans, especially in diseases such as diabetes mellitus.

Modeling of the process of moisture loss during the storage of dried apricots

Moisture content is a reference parameter for dried food because the growth of most microorganisms is inhibited below certain water activity levels. In addition, it has a determining influence on the evolution of important parameters, such as color and flavor, and on other properties and deterioration reactions, such as texture, oxidation processes and nutritional value. During the storage of some dried fruits, moisture is produced due to Maillard reactions and exchanged with the surrounding environment through the packaging. The evolution of dried foods during their shelf life depends on the storage conditions. The aim of this study is to analyze the evolution of the moisture content in dried apricots packaged in different types of containers, namely glass and thermosealed polypropylene trays. The samples were stored at constant temperatures: 5, 15, 25 and 35 °C and were analyzed periodically over a period of 12 months. The sorption isotherms of apricots used in this study were also determined. In order to model how the moisture evolved, an empirical kinetic model was tested. This model considers both water transfer from the fruit and also water production as a result of the Maillard processes. The explained variance was higher than 95% in the samples stored in trays, which were thermosealed with film.

Glycoxidative stress and cardiovascular complications in experimentally-induced diabetes: effects of antioxidant treatment.

Diabetes mellitus (DM) is a common metabolic disease, representing a serious risk factor for the development of cardiovascular complications, such as coronary heart disease, peripheral arterial disease and hypertension. Oxidative stress (OS), a feature of DM, is defined as an increase in the steady-state levels of reactive oxygen species (ROS) and may occur as a result of increased free radical generation and/or decreased anti-oxidant defense mechanisms. Increasing evidence indicates that hyperglycemia is the initiating cause of the tissue damage in DM, either through repeated acute changes in cellular glucose metabolism, or through long-term accumulation of glycated biomolecules and advanced glycation end products (AGEs). AGEs are formed by the Maillard process, a non-enzymatic reaction between ketone group of the glucose molecule or aldehydes and the amino groups of proteins that contributes to the aging of proteins and to the pathological complications of DM. In the presence of uncontrolled hyperglycemia, the increased formation of AGEs and lipid peroxidation products exacerbate intracellular OS and results in a loss of molecular integrity, disruption in cellular signaling and homeostasis, followed by inflammation and tissue injury such as endothelium dysfunction, arterial stiffening and microvascular complications. In addition to increased AGE production, there is also evidence of multiple pathways elevating ROS generation in DM, including; enhanced glucose auto-oxidation, increased mitochondrial superoxide production, protein kinase C-dependent activation of NADPH oxidase, uncoupled endothelial nitric oxide synthase (eNOS) activity, increased substrate flux through the polyol pathway and stimulation of eicosanoid metabolism. It is, therefore, not surprising that the correction of these variables can result in amelioration of diabetic cardiovascular abnormalities. A linking element between these phenomena is cellular redox imbalance due to glycoxidative stress (GOS). Thus, recent interest has focused on strategies to prevent, reverse or retard GOS in order to modify the natural history of diabetic cardiovascular abnormalities. This review will discuss the links between GOS and diabetes-induced cardiovascular disorders and the effect of antioxidant therapy on altering the development of cardiovascular complications in diabetic animal models.

Formation Pathways and Opioid Activity Data for 3‐Hydroxypyridinium Compounds Derived from Glucuronic Acid and Opioid Peptides by Maillard Processes

The kinetics of formation and identity of the reaction products of the glucuronic acid with three representative opioid peptides were investigated in vitro. Peptides were conjugated with glucuronic acid either in solution or under dry-heating conditions. From the incubations performed in solution N-(1-deoxy-D-fructofuranos-1-yluronic acid)-peptide derivatives (Amadori compounds) were isolated, whereas from the dry-heated reactions products containing the 3-hydroxypyridinium moiety at the N-terminal of the peptide chain were obtained. Experiments performed under mild dry-heating conditions (40 degrees C) in model systems based on Leu-enkephalin and glucuronic acid, and in environment of either 40% or 75% relative humidity, revealed that the higher level of humidity promoted a process that enhanced 3-hydroxypyridinium compound generation. The mechanism of 3-hydroxypyridinium formation is discussed. In comparison with their respective parent peptides, the N-(1-deoxy-D-fructofuranosyl-uronic acid) derivatives of the opioid peptides showed three- to 11-fold lower mu- and delta-receptor-binding affinities and agonist potencies in the functional assays, likely as a consequence of the steric bulk introduced at the N-terminal amino group. The further decrease in opioid activity observed with the 3-hydroxypyridinium-containing peptides may be due to the lower pK(a) of the 3-hydroxypyridinium moiety and to delocalization of the positive charge in the pyridinium ring system.

Free Radicals in the Maillard Reaction

This article reviews the study of free radical intermediates in the Maillard reaction from a historical perspective. Early observation of free radicals in biological systems led other workers to discover similar radicals in non‐enzymic Maillard processes such as those observed in foods. Emphasis is placed on a discussion of the chemistry of radical species detected in Maillard systems and their possible relevance to food flavor and color.

Formation pathways for lysine–arginine cross-links derived from hexoses and pentoses by Maillard processes

α-Dicarbonyl compounds are crucial intermediates in the cross-linking of proteins by reducing sugars in the course of the Maillard reaction. The novel dideoxyosones N6-(2,3-dihydroxy-5,6-dioxohexyl)lysine (9a,b) and N6-(2-hydroxy-4,5-dioxopentyl)lysine (12) were unequivocally identified via their triazine derivatives and these α-diketo compounds established as precursors of the major in vivo cross-links glucosepane 11 and pentosidine 16, respectively. This new dideoxyosone class is formed in high yield in the model incubations and the underlying reaction sequence will improve our understanding of protein cross-linking and of Maillard chemistry in general.

Formation Pathways for Lysine-Arginine Cross-links Derived from Hexoses and Pentoses by Maillard Processes: UNRAVELING THE STRUCTURE OF A PENTOSIDINE PRECURSOR

Covalently cross-linked proteins are among the major modifications caused by the advanced Maillard reaction. So far, the chemical nature of these aggregates and their formation pathways are largely unknown. Synthesis and unequivocal structural characterization are reported for the lysine-arginine cross-links N(6)-(2-([(4S)-4-ammonio-5-oxido-5-oxopentyl]amino)-5-[(2S,3R)-2,3,4- trihydroxybutyl]-3,5-dihydro-4H-imidazol-4-ylidene)-l-lysinate (DOGDIC 12), N(6)-(2-([(4S)-4-ammonio-5-oxido-5-oxopentyl]amino)-5-[(2S)-2,3-dihydroxypropyl]-3,5-dihydro-4H-imidazol-4-ylidene)-l-lysinate (DOPDIC 13), and 6-((6S)-2-([(4S)-4-ammonio-5-oxido-5-oxopentyl] amino)-6-hydroxy-5,6,7,7a-tetrahydro-4H-imidazo[4,5-b] pyridin-4-yl)-l-norleucinate (pentosinane 10). For these compounds, as well as for glucosepane 9 and pentosidine 11, the formation pathways could be established by starting from native carbohydrates, Amadori products, and 3-deoxyosones, respectively. Pentosinane 10 was unequivocally proven to be an important precursor of pentosidine 11, which is a well established fluorescent indicator for advanced glycation processes in vivo. The Amadori products are shown to be the pivots in the formation of the various cross-links 9-13. The bicyclic structures 9-11 are directly derived from aminoketoses, whereas 12 and 13 stem from reaction with the 3-deoxyosones. All products 9-13 were identified and quantified from incubations of bovine serum albumin with the respective 3-deoxyosone or carbohydrate. From these results it seems fully justified to expect both glucosepane 9 and DOGDIC 12 to constitute important in vivo cross-links.

Iterated reaction graphs: simulating complex Maillard reaction pathways.

This study investigates a new method of simulating a complex chemical system including feedback loops and parallel reactions. The practical purpose of this approach is to model the actual reactions that take place in the Maillard process, a set of food browning reactions, in sufficient detail to be able to predict the volatile composition of the Maillard products. The developed framework, called iterated reaction graphs, consists of two main elements: a soup of molecules and a reaction base of Maillard reactions. An iterative process loops through the reaction base, taking reactants from and feeding products back to the soup. This produces a reaction graph, with molecules as nodes and reactions as arcs. The iterated reaction graph is updated and validated by comparing output with the main products found by classical gas-chromatographic/mass spectrometric analysis. To ensure a realistic output and convergence to desired volatiles only, the approach contains a number of novel elements: rate kinetics are treated as reaction probabilities; only a subset of the true chemistry is modeled; and the reactions are blocked into groups.

Cross-linking of proteins by maillard processes: characterization and detection of lysine–arginine cross-links derived from glyoxal and methylglyoxal

α-Dicarbonyl compounds, such as glyoxal and methylglyoxal, are crucial intermediates in the browning and cross-linking of proteins by reducing sugars in the course of the Maillard reaction. The cross-linking units 2-ammonio-6-({2-[(4-ammonio-5-oxido-5-oxopentyl)amino]-4,5-dihydro-1H-imidazol-5-ylidene}amino)hexanoate (9) and 2-ammonio-6-({2-[(4-ammonio-5-oxido-5-oxopentyl)amino]-4-methyl-4,5-dihydro-1H-imidazol-5-ylidene}amino)hexanoate (10), designated as GODIC and MODIC, are identified and quantified from glyoxal/methylglyoxal-bovine serum albumin (BSA) incubations. Independent syntheses and unequivocal structural characterization are given for 9 and 10. A protocol was established for their determination by liquid chromatography–mass spectrometry (LC–MS) with electrospray ionization (ESI). BSA and the respective α-dicarbonyl compound were incubated at 37°C, pH 7.4 for 1 week, and the time-dependent formation of 9 and 10 was observed. The maximum value obtained from a solution containing 50g/L BSA and 2mM glyoxal or methylglyoxal after a 7-day incubation period corresponds to an arginine derivatization quota of 13.0±0.32mmol 9/mol Arg or 3.0±0.12 mmol 10/mol Arg. The cross-links 9 and 10 were also detected in a d-glucose–BSA incubation. From these results, it seems justified to assign an important role to 9 and 10 in the cross-linking of proteins in vivo as well as in foodstuffs. In an additional model study, formation of 9 and 10 was compared to that of the imidazolium cross-links GOLD 3 and MOLD 4.

Cross-linking of proteins by maillard processes—characterization and detection of a lysine-arginine cross-link derived from d-glucose

Covalently cross-linked proteins are among the major modifications caused by the advanced Maillard reaction. So far, the chemical nature of these aggregates is largely unknown. Investigations are reported on how the cross-linking unit 2-ammonio-6-{2-[(4-ammonio-5-oxido-5-oxopentyl)amino]-6,7-dihydroxy-4,5,6,7,8,8a-hexahydroimidazo[4,5- b]azepin-4-yl} hexanoate ( 7), designated as glucosepan, can be identified and quantified from d-glucose-bovine serum albumin (BSA) incubations. Independent synthesis and unequivocal structural characterization are given for glucosepan 7. A protocol was established for its determination by LC–MS with electrospray ionization (ESI). BSA and d-glucose were incubated at 37 °C, pH 7.4 for eight weeks and the time-dependent formation of 7 was observed. Since glucosepan 7 is unstable under acid proteolytic conditions, BSA was cleaved enzymatically. The maximum value obtained from a solution containing 50 g/L BSA and 100 mM d-glucose after eight weeks incubation time corresponds to an arginine derivatization rate of 1.38±0.07 mmol 7/mol Arg (equivalent to 31.7±1.6 mmol 7/mol BSA). From these results, it seems justified to expect 7 to play an important role in the cross-linking of proteins in vivo as well as in foodstuffs. The structural similarity of glucosepan 7 and pentosidine 1 made it obvious to also look for an eventual parallelism in the respective formation pathways.

Cross-linking of proteins by maillard processes—Model reactions of d-glucose or methylglyoxal with butylamine and guanidine derivatives

Advanced Maillard reaction in proteins leads to formation of covalently cross-linked aggregates the chemical nature of which is largely unknown. From model reactions of methylglyoxal and butylamine with creatine or α-N-acetyl-l-arginine, one main product each was isolated. These two compounds were identified, on the basis of unequivocal spectroscopic evidence, as 2-[(5-butylimino-4-methyl-4,5-dihydro-1H-2-imidazolyl)(methyl)amino]acetic acid and 2-acetylamino-5-[(5-butylimino-4-methyl-4,5-dihydro-1H-2-imidazolyl)amino]pentanoic acid, respectively. Using d-glucose instead of methylglyoxal, two main products each were obtained from reaction with the respective guanidine derivative. The spectroscopic data definitively establish the formation of the diastereoisomeric 2-[(4-butyl-6,7-dihydroxy-4,5,6,7,8,8a-hexahydroimidazo[4,5-b]azepin-2-yl)(methyl)amino]acetic acid from the reaction with creatine, and of the diastereoisomeric 2-acetylamino-5-[(4-butyl-6,7-dihydroxy-4,5,6,7,8,8a-hexahydroimidazo[4,5-b]azepin-2-yl)amino]pentanoic acid from the reaction with α-N-acetyl-l-arginine. All products were isolated in fairly good yield and represent 1:1:1 adducts of the respective reaction partners. Formation of these compounds thus constitutes an efficient reaction pathway for linking primary amines to guanidine derivatives. It seems justified, therefore, to expect cross-linking of proteins by action of reducing carbohydrates to proceed analogously.

Reaction of 3-deoxypentosulose with N-methyl- and N,N-dimethylguanidine as model reagents for protein-bound arginine and for creatine.

Deoxyosones are established key-intermediates in Maillard processes. Due to their dicarbonyl structure, they undergo condensation to form heterocyclic compounds with guanidine derivatives. In biological systems, guanidino functions are present in protein-bound arginine moieties as well as in creatine. The reactivity of such structures towards 3-deoxypentosulose is investigated with N-methyl- and N,N-dimethylguanidine as model substrates. Two diastereoisomers each are isolated from both reactions; they have been characterized unequivocally, respectively, as 4-(2,3-dihydroxypropyl)-2-N-methylamino-2-imidazoline-5-one and 4-hydroxy-5-(2,3-dihydroxypropyl)-2-(N,N-dimethylamino)-5H-imidazole. In aqueous medium as well as in the crystalline state, both diastereoisomer pairs exist in different tautomeric forms.