Maillard Reaction Intermediates Research Articles

Influence of Hydroxycinnamic Acids on the Maillard Reaction of Arabinose and Galactose beyond Carbonyl-Trapping.

Hydroxycinnamic acids, known for their health benefits and widespread presence in plant-based food, undergo complex transformations during high-temperature processing. Recent studies revealed a high browning potential of hydroxycinnamic acids and reactive Maillard reaction intermediates, but the role of phenolic compounds in the early stage of these reactions is not unambiguously understood. Therefore, we investigated the influence of caffeic acid and ferulic acid on the nonenzymatic browning of arabinose, galactose, and/or alanine, focusing on the implications on the formation of relevant early-stage Maillard intermediates and phenol-deriving products. Contrary to previous assumptions, hydroxycinnamic acids were found to promote nonenzymatic browning instead of solely trapping reactive intermediates. This was reflected by an intense browning, which was attributed to the formation of heterogeneous phenol-containing Maillard products. Although, caffeic acid is more reactive than ferulic acid, the formation of reactive furan derivatives and of heterogeneous phenol-containing colorants was promoted in the presence of both hydroxycinnamic acids.

Maillard reaction intermediates in Chinese Baijiu and their effects on Maillard reaction related flavor compounds during aging

This study investigated the Maillard reaction in Baijiu and the effects of extended aging in the presence of Maillard reaction intermediates (MRIs) on aromatic compounds, particularly focusing on heterocyclic changes. MRIs with different aroma types in Baijiu aged 1–18 years and force-aged for 6 weeks were determined. Results revealed that MRIs in soy sauce aroma–type Baijiu were significantly more abundant than those in other types of Baijiu. Changes in MRIs were observed and compared in aging and forced-aging Baijiu. Additionally, the distribution and variation of heterocycles in Baijiu were examined, which revealed an increase in N-heterocycle levels but a decrease in S- and O-heterocycle levels to a certain extent. The results of this study demonstrate that the Maillard reaction during the aging of Baijiu influences heterocycle concentrations, thereby improving flavor of aged Baijiu. Research into heterocycles and the Maillard reaction may help elucidate the aromatic evolution of Baijiu with aging and provide guidance for Baijiu storage.

Browning reactions of hydroxycinnamic acids and heterocyclic Maillard reaction intermediates – Formation of phenol-containing colorants

Non-enzymatic conversion of phenolic compounds plays an important role during thermal processing of plant-based food such as coffee, cocoa, and peanuts. However, the more prominent Maillard reaction is mainly studied at a mechanistic level for carbohydrates and amino compounds to clarify reactions that contribute to (‘classic') melanoidin formation, but the role of phenolic compounds in such reactions is rarely discussed yet. To understand their contribution to non-enzymatic browning, reactions between ubiquitous phenolic acids, such as caffeic acid and ferulic acid, and prominent heterocyclic Maillard intermediates, namely furfural, hydroxymethylfurfural, and pyrrole-2-carbaldehyde were investigated. Following incubation under roasting conditions (220 °C, 0–30 min), heterogenous products were characterized by high-resolution mass spectrometry, and, after isolation, by nuclear magnetic resonance spectroscopy. By this, color precursors were identified, and it was shown that in addition to aromatic electrophilic substitution, nucleophilic and condensation reactions are key mechanisms contributing to the formation of phenol-containing melanoidins.

Contribution of Hydroxycinnamic Acids to Color Formation in Nonenzymatic Browning Reactions with Key Maillard Reaction Intermediates.

The Maillard reaction is a vital part of food processing, involving a vast number of complex reaction pathways, resulting in high-molecular-weight colorants. So far, studies have been focused on the conversion of carbohydrates and amino compounds, but the literature elaborating the contribution of phenolic compounds to the formation of the colored end-products is still rare. The aim of this study was to characterize early reactions, underlying the formation of phenol-containing melanoidins. For this purpose, binary model systems of the prominent phenolic compounds caffeic acid and ferulic acid combined with α-dicarbonyl compounds typically formed in the Maillard reaction such as glyoxal, methylglyoxal, and diacetyl were analyzed after heat treatment. High-resolution mass spectrometry revealed that decarboxylation, aromatic electrophilic substitution, and nucleophilic addition are important reaction steps that lead to colored heterogeneous oligomers. Polymerization was favored for phenolic compounds with a high electron density in the aromatic system and for α-dicarbonyl compounds carrying aldehyde functions.

Maillard Reaction Process and Characteristic Volatile Compounds Formed During Secondary Thermal Degradation Monitored via the Change of Fluorescent Compounds in the Reaction of Xylose-Corn Protein Hydrolysate.

Until now, no effective method has been found to monitor the Maillard reaction process for complex protein hydrolysates. Dynamic changes in the concentration of α-dicarbonyl compounds, fluorescence intensity, and browning degree were investigated during the Maillard reaction of corn protein hydrolysates. When the fluorescence intensity reached the peak, deoxyosones would continue to be increased by ARP's degradation. However, the reaction node with the highest fluorescence intensity coincided with the turning point of the browning reaction, and the subsequent browning rate remarkably increased. Therefore, the change in fluorescence intensity could be used to monitor the degradation of ARP and the formation of browning melanoidin at different stages of the Maillard reaction of complex systems, thus effectively indicating the process of the Maillard reaction. When Maillard reaction intermediates (MRIs) with maximum fluorescent compounds were heated, the most abundant pyrazines were subsequently achieved. However, furan compounds would be progressively increased during the thermal process of MRIs with continuously enhanced browning.

Formation of fluorescent substances and correlation with process flavor changes of dehydrated Maillard reaction intermediates of peanut protein hydrolysates

Maillard reaction intermediates (MRIs) derived from peanut protein hydrolysates (PPH) and xylose (Xyl) were prepared using the water bath combined with spray drying and vacuum drying. The monitoring of free amino content, browning precursors, and taste characteristics was carried out throughout the thermal treatment at low temperature, where the Maillard reaction process was slowed down, and the formation and degradation of MRIs were dynamically balanced at low conversion rates. The reaction between amino compounds and the reducing sugars contributed to the increased taste characteristics during the early phase of Maillard reaction. Fluorescence intensity was reduced after reaching its maximum value and browning intensity was significantly increased during vacuum drying, which confirmed that the Maillard reaction entered into an advanced stage. Vacuum drying also led to a significant increase in the total amount of volatile compounds in the subsequent thermal processing reaction (from 212.93 ng/g to 564.91 ng/g), which demonstrated that the higher fluorescence intensities resulted in the more accumulation of MRIs and better capabilities of flavor formation.

Effects of chlorogenic acid on the formations of furan, acrylamide, α-dicarbonyls and volatile compounds in canned coffee model systems

It was demonstrated that some antioxidants, especially chlorogenic acid (CGA), can trap glyoxal and methylglyoxal, which are Maillard reaction intermediates and precursors of furan, leading to the reduction of furan. This study investigated the effects of CGA on the changes in furan, acrylamide, α-dicarbonyls, and volatile compound levels in 11 types of canned coffee model systems (CCMSs) containing sugars and amino acids after thermal treatments. Furan decreased (3.77–34.7%) in all model systems treated with CGA. However, acrylamide decreased in sugar-only models but increased in sugar/amino acid models. α-Dicarbonyls were generally reduced with CGA treatment but increased in the sucrose/asparagine model. A total of 116 volatile compounds were identified in three types of CCMSs. Overall, volatile compounds were decreased by the addition of CGA. The trapping action of CGA might affect the reduction of furan and some volatiles but exhibits conflicting results for acrylamide.

Colorants and Antioxidants Deriving from Methylglyoxal and Heterocyclic Maillard Reaction Intermediates.

The Maillard reaction is well known for producing antioxidant compounds alongside colored substances. Low-molecular-weight antioxidant intermediates such as maltol (MAL) or norfuraneol (NF) are well described, but it is still unclear which of these Maillard intermediates are the precursors of antioxidant and colored melanoidins-the so-called late stage Maillard reaction products. This study aimed to provide novel insights into the correlation between browning potential and antioxidant properties of reaction products formed during the heat treatment of prominent Maillard reaction intermediates. It was achieved by the incubation of binary reaction systems composed of methylglyoxal (MGO) or NF in combination with furfural (FF), MAL, and pyrrole-2-carbaldehyde (PA) at pH 5 and 130 °C for up to 120 min. Overall, it could be shown that the formation of colored products in the binary NF reaction systems was more efficient compared to those of MGO. This was reflected in an increased browning intensity of up to 400% and a lower conversion rate of NF compared to MGO. The colorants formed by NF and FF or PA (~0.34 kDa and 10-100 kDa) were also found to exhibit higher molecular weights compared to the analogue products formed in the MGO incubations (<0.34 kDa and 10-100 kDa). The incorporation of NF into these heterogenous products with FF and PA resulted in the preservation of the initial antioxidant properties of NF (p < 0.05), whereas no antioxidant products were formed after the incubation of MGO.

Formation of Fluorescent Maillard Reaction Intermediates of Peptide and Glucose during Thermal Reaction and Its Mechanism

The dynamic changes in fluorescence intensity of the Maillard reactions of l-alanyl-l-glutamine (Ala-Gln)/Diglycine (Gly-Gly)/glycyl-l-glutamine (Gly-Gln) and glucose were investigated. It was found that the fluorescence intensity would increase with the reaction time; however, it would decrease after longer heating at higher temperatures, which was accompanied by rapid browning. The strongest intensity occurred at 45, 35, and 35 min at 130 °C for Ala-Gln, Gly-Gly, and Gly-Gln systems, respectively. The simple model reactions of Ala-Gln/Gly-Gly and dicarbonyl compounds were selected to reveal the formation and mechanism of fluorescent Maillard compounds. It was confirmed that both GO and MGO could react with peptides to form fluorescent compounds, especially GO, and this reaction was sensitive to temperature. The mechanism was also verified in the complex Maillard reaction of pea protein enzymatic hydrolysates.

The potential meat flavoring generated from Maillard reaction products of wheat gluten protein hydrolysates-xylose: Impacts of different thermal treatment temperatures on flavor

Wheat gluten protein hydrolysates were prepared by Flavourzyme, followed by xylose-induced Maillard reaction at different temperatures (80 °C, 100 °C and 120 °C). The MRPs were subjected to analysis of physicochemical characteristics, taste profile and volatile compounds. The results demonstrated that UV absorption and fluorescence intensity of MRPs significantly increased at 120 °C, suggesting formation of a large amount of Maillard reaction intermediates. Thermal degradation and cross-linking simultaneously occurred during Maillard reaction, while thermal degradation of MRPs played a more predominant role at 120 °C. MRPs exhibited high umami and low bitter taste at 120 °C, accompanied by the high content of umami amino acids and low content of bitter amino acids. Furans and furanthiols with pronounced meaty flavor served as the main volatile compounds in MRPs at 120 °C. Overall, high temperature-induced Maillard reaction of wheat gluten protein hydrolysates and xylose is a promising strategy for the generation of potential plant-based meat flavoring.

Variation of moisture state and taste characteristics during vacuum drying of Maillard reaction intermediates of hydrolyzed soybean protein and characterization of browning precursors via fluorescence spectroscopy

Maillard reaction intermediates (MRIs) derived from enzymatic hydrolysates of soybean protein were prepared by combining dehydration method with spray drying and vacuum drying. Taste characteristics, browning precursors, water activity, and moisture state were monitored during the process of vacuum drying. The water activity was significantly reduced and the proportion of bound water was increased, which could promote the large accumulation of MRIs. The saltiness, umami, and bitterness responses of MRIs were all reduced and the reduction of taste intensity during vacuum drying was probably caused by the reaction of small peptides of taste-active amino acids with glucose. The continuous accumulation of browning precursors during vacuum drying was confirmed by fluorescence spectroscopic analysis and by quantitative analysis of α-dicarbonyl compounds. Subsequent browning reactions further demonstrated that the MRI with the highest fluorescence intensity also had the greatest browning potential.

Ultrasound-Assisted Preparation of Maillard Reaction Products Derived from Hydrolyzed Soybean Meal with Meaty Flavor in an Oil-In-Water System.

In the present work, we prepared Maillard reaction products (MRPs) derived from enzyme hydrolyzed soybean meal with ultrasound assistance in an oil-(oxidized lard)-in-water system (UEL-MRPs) or oil-free system (UN-MRPs), and the effect of ultrasound on the properties of the obtained MRPs was evaluated. The analysis of fatty acids in lard with different treatments showed that ultrasound can generate more unsaturated fatty acids in the aqueous phase. The UV-Vis absorbances of UEL-MRPs, UN-MRPs, and MRPs obtained in an oil-in-water system (EL-MRPs) and MRPs obtained in an oil-free system (N-MRPs) at 294 and 420 nm indicated that ultrasound could increase the amount of Maillard reaction intermediates and melanoids in the final products of the Maillard reaction. This was in line with the result obtained from color change determination-that ultrasound can darken the resultant MRPs. Volatile analysis showed ultrasound can not only increase the number of volatile substances, but also greatly increase the composition of volatile substances in UEL-MRPs and UN-MRPs, especially the composition of those contributing to the flavor of the MRPs, such as oxygen-containing heterocycles, sulfur-containing compounds, and nitrogen-containing heterocycles. Descriptive sensory evaluation revealed that UN-MRPs and UEL-MRPs had the highest scores in total acceptance, ranking in the top two, and UEL-MRPs had the strongest meaty flavor among these four kinds of MRPs. Furthermore, the measurements of antioxidant activities, including DPPH radical-scavenging activity, hydroxyl radical scavenging ability, and ferric ion reducing antioxidant power, were conducted, showing that UN-MRPs exhibited the highest antioxidant activity among all the MRPs.

Molecular Basis for the Simultaneous Enhancement of the Aroma-Generating Capacity and Bioactivity of Maillard Reaction Precursors through Mechanochemistry.

Ball milling at ambient temperatures can accelerate the formation and accumulation of early-stage Maillard reaction intermediates considered important precursors of aromas and antioxidants. In this study, using chemical and biological assays, we explored the potential of sequential milling and heating to enhance the antioxidant and aroma-generating capacity of Maillard model systems. Milling (30 Hz/30 min) followed by dry heating (90 °C/30 min) of glycine or lysine with glucose significantly increased not only the intensity of their aroma-active compounds as analyzed by headspace-gas chromatography/mass spectrometry (HS-GC/MS) but also their free radical scavenging capacity as assessed by 2,2'-azino-bis-(3-ethylbenzothiazoneline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) assays. This was attributed to the increased formation of redox-active endiol moieties and precursors of N,N-dialkyl-pyrazinium radical cation in the lysine system assessed by electrospray ionization-quadrupole time-of-flight/tandem mass spectrometry (ESI-QqTOF/MS/MS) analysis. The test samples also inhibited NO generation and cellular oxidative stress in RAW 264.7 murine macrophage cells, indicating size reduction induced by milling promoted paracellular absorption.

Structural characterization of polar melanoidins deriving from Maillard reaction intermediates – A model approach

Chemical conversions of reducing sugars and amino compounds induce the formation of heterogenous, high-molecular-weight colorants (‘melanoidins’) with widely unknown chemical structures. Model experiments of reactive intermediates have proven to be suitable for unravelling the formation mechanisms of colored reaction products. Here, the active methylene norfuraneol was selected and incubated individually as well as in combination with glyoxal, glycolaldehyde, and acetaldehyde at elevated temperatures. Photometric and chromatographic methods as well as mass spectrometry were used to analyze the colored reaction products and reveal the reactivity of different carbonyls regarding the formation of heterogenous oligomers. Aqueous solutions of norfuraneol and glyoxal exceeded the color formation of all other model reaction systems and it could be shown that the initial reactants as well as their degradation products were incorporated into the colorants. The colored oligomers described herein were composed of carbohydrate-based intermediates of the Maillard reaction and defined as melanoidin precursors or pre-melanoidins.

Role of Maillard Reaction Products as Antioxidants in Washed Cod and Washed Turkey Muscle Oxidized by Added Hemoglobin

AbstractThe ability of Maillard reaction products (MRPs) to inhibit hemoglobin (Hb)‐mediated lipid oxidation is examined in washed cod muscle (WCM) and washed turkey breast muscle (WTM). Lipid peroxides and thiobarbituric acid reactive substances (TBARS) are used as markers of primary and secondary lipid oxidation products, respectively. MRPs inhibits Hb‐mediated lipid oxidation in WCM (p &lt; 0.01) but are weakly inhibitory in WTM. α‐tocopherol concentration is 24‐fold greater in WCM compared to WTM. The elevated α‐tocopherol content of WCM may partly explain the enhanced antioxidant effectiveness of MRPs in WCM, noting that MRPs and α‐tocopherol act synergistically in the scavenging of free radicals. MRPs are more effective in inhibiting lipid oxidation in WCM at pH 6.6 compared to lower pH values. Antioxidant effectiveness of MRPs diminishes when stored refrigerated but not with frozen storage. MRPs obtained by 3 h heating contains more Maillard reaction intermediates than 2 h heating, which better inhibits LOOH formation (p &lt; 0.01). These intermediates decrease faster during refrigerated compared to frozen storage. These studies suggest elevated pH and sufficient endogenous tocopherols are required for low concentrations of MRPs to protect highly unsaturated lipid in washed muscle from oxidation by hemoglobin.Practical Applications: This study shows that the maillard reaction products (MRPs) can be used to inhibit the oxidation of tocopherol‐rich meat products, and the MRPs stored in the frozen condition for a certain period also have antioxidant activity.

Optimized preparation and antioxidant activity of glucose-lysine Maillard reaction products

Reductive substances in Maillard reaction products (MRPs) improve the oxidative stability of food. This study explored the antioxidant activity of the main components of MRPs in glucose-lysine (GL) models. The optimal conditions for preparing the Maillard reaction (MR) were studied by a response surface methodology using FRAP as an evaluation index. The antioxidant activities, including ABTS, DPPH, and FRAP assays were determined. The MRPs obtained from gel chromatography purification were subjected to UV absorbance and fluorescence intensity determined, and MRPs with antioxidant activity were detected by HPLC-MS. The results showed that optimal conditions were at a reaction temperature of 110 °C, a reaction time of 120 min, an initial of pH 10, and n (Glc: Lys) = 1:1. MR intermediates, browning pigments, and fluorescent compounds were related to antioxidant activity. Pearson correlation analysis further confirmed that the content of the main components (m/z 309) was significantly positively correlated with the antioxidant activity of MRPs.

Accelerated Dissipation of Free and Immobilized Water Facilitating the Intramolecular Dehydration of N-Xylosamine and Conversion Improvement of the Amadori Rearrangement Product of Aspartic Acid-Xylose Reaction.

Compared to the method of aqueous Maillard reaction at atmospheric pressure tandem vacuum concentration, a coupling dehydration method combining spray drying and vacuum drying was used to increase aspartic acid-xylose conversion to the Amadori rearrangement product (ARP). The water activity and moisture states were found as effective indicators to characterize the degree of dehydration of Maillard reaction intermediates and efficient formation of ARP. During the vacuum drying process, the water activity of the product powder decreased significantly. Because the formation of ARP was accompanied by intramolecular dehydration, combining spray drying and vacuum drying increased the proportion of bound water in the vacuum-dried product. Free water was easily dissipated via dehydration, which then converted the immobilized water continuously to free water, and the decreased immobilized water further converted the bound water to immobilized water. The reduction in bound water contributed to the intramolecular dehydration of N-substituted d-xylosamine, which would further be transformed to be the ARP through an intramolecular rearrangement. The yield of ARP was increased from 1.68 to 21.53% after spray drying. The ARP yield was substantially increased up to 77.9% by subsequent vacuum drying.

Effects of the Deacetylation Degree of Chitosan on 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) Formation in Chemical Models and Beef Patties

The effects of the deacetylation degree (DD) of chitosan on heterocyclic aromatic amine formation were investigated in chemical models and beef patties. The results in model systems showed that at lower addition levels (10 mg), chitosan with 85% DD showed the strongest inhibitory effect against 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) formation, while chitosan with a higher DD (95%) or a lower DD (72 and 50%) did not show any significantly inhibitory effect. Further mechanism study showed that chitosan addition reduced the content of Maillard reaction intermediates including phenylacetaldehyde and the aldol condensation product but increased the PhIP precursor creatinine residue in the chemical model, indicating that chitosan at least partially competed with creatinine to react with phenylacetaldehyde to inhibit PhIP formation. In roast beef patties, 0.15% (w/w) chitosan (85% DD) significantly reduced the formation of PhIP, MeIQx, 4,8-DiMeIQx, Harman, and Norharman by 56.21, 33.32, 31.35, 25.14, and 28.12%, respectively. Moreover, chitosan significantly inhibited the formation of aldehydes in roast beef patties, further confirming the above-mentioned inhibition mechanism. However, the addition of chitosan might promote fatty acid oxidation. In addition, chitosan addition below 0.15% (w/w) had no significant effect on the textural properties of the roast samples.

Key Aspects of Amadori Rearrangement Products as Future Food Additives.

Flavor is one of the most important factors in attracting consumers and maximizing food quality, and the Maillard reaction (MR) is highly-involved in flavor formation. However, Maillard reaction products have a big drawback in their relatively low stability in thermal treatment and storage. Amadori rearrangement products (ARPs), MR intermediates, can alternatively act as potential flavor additives for their better stability and fresh flavor formation ability. This review aims to elucidate key aspects of ARPs’ future application as flavorings. The development of current analytical technologies enables the precise characterization of ARPs, while advanced preparation methods such as synthesis, separation and drying processes can increase the yield of ARPs to up to 95%. The stability of ARPs is influenced by their chemical nature, pH value, temperature, water activity and food matrix. ARPs are associated with umami and kokumi taste enhancing effects, and the flavor formation is related to amino acids/peptides of the ARPs. Peptide-ARPs can generate peptide-specific flavors, such as: 1,6-dimethy-2(1H)-pyrazinone, 1,5-dimethy-2(1H)-pyrazinone, and 1,5,6-trimethy-2(1H)-pyrazinone. However, further research on systematic stability and toxicology are needed.

Identification of the Maillard reaction intermediates as divalent iron complexes in alanine/glucose/FeCl2 model system using ESI/qTOF/MS/MS and isotope labelling technique

Due to their high reactivities and short half-lives, the detection of Maillard reaction intermediates is relatively difficult to achieve in a single analytical run. In this study, the formation of Maillard reaction intermediates from heated alanine/glucose mixtures (110 ​°C for 2 ​h) was investigated through their complexation with divalent iron using electrospray ionization/quadrupole time-of-flight mass spectrometry and isotope labeling techniques. Analysis of the mixtures indicated that this approach allows the simultaneous detection of many important labile and reactive Maillard reaction intermediates along with unreacted alanine and glucose in addition to various other Maillard reaction products, such as glyceraldehyde, erythrose, ribose, acetol, glycolaldehyde, fructosamine, glucosone, osones, deoxyosones, and Amadori products. Some osones and deoxyosones also formed their corresponding Schiff bases with alanine. The above mentioned Maillard reactions intermediates were detected either as binary metal complexes with alanine or with other enediol generating species including self-complexation adducts and they formed positively charged ions such as [M + H]+, [M + Na]+, [M + K]+, [M ​+ ​Fe35Cl]+, and [M ​+ ​Fe37Cl]+, that can be detected using the positive ionization mode.