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The quantification of hydrolysable polyphenols such as gallic, ellagic acid and vescalagin by HPLC-DAD is classically run after methanol extraction as a reference solvent. Water extraction is usually discarded because of a lower obtention of total polyphenol content compared to methanol extraction. In our study, methanol was compared to water extraction in both the total polyphenol content method and the HPLC-DAD analysis. Total polyphenol content in water extraction was lower than in methanol extraction, but water extraction gave better results on HPLC-DAD. In conclusion, total polyphenol content cannot be used as reference to choose the solvent of extraction to quantify some polyphenols by HPLC-DAD because of the specific properties of each polyphenol. Indeed, recovery results obtained on hydrolysable polyphenols with water extraction were better and with a lower variability than following methanol extraction., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)

The study aimed to evaluate the effect of using sourdough or yeast as a leavening agent for French bread making on the Maillard reaction and the bifidogenic effect. Sugars, proteins, and free asparagine were quantified in bread dough before and after fermentation. The levels of the Maillard reaction precursors were very different depending on the leavening agent used, which affected the Maillard reaction during baking. Strecker degradation was favored in the crust of sourdough bread (SB), generating about 7 times more aldehydes than in the crust of yeast bread (YB), thus improving the sensory quality of the bread. In the YB crust, the melanoidization pathway was predominant. The bifidogenic effect of crust and crumb from both breads was evaluated through the in vitro growth of Bifidobacterium adolescentis. SB showed a higher bifidogenic effect, probably due to its composition more favorable for bacteria growth., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Scope: Type 2 diabetes (T2D) induces organ damage associated with glycation, among other metabolic pathways. While therapeutic strategies have been tested to reduce the formation and impact of glycation products, results remain equivocal. Anti-diabetic therapies using probiotics have been proposed, but their effect upon glycation has not been reported. Here, the effects of the bacterial strain Lactobacillus fermentum ME-3 on glycation and T2D-related complications in a mouse model of T2D are investigated., Methods & Results: Wild-type LepR db/+ and diabetic LepR db/db littermates receive a daily gavage of either water or the probiotic ME-3 strain (10 10 CFU). Glycation markers, fructoselysine-derived furosine (FL-furosine) and carboxymethyllysine (CML), are quantified in four major organs and plasma using stable-isotope dilution LC-MS/MS. After 12 weeks of ME-3 treatment, diabetic mice gain less weight and exhibit an apparently improved glucose tolerance. The ME-3 treatment reduces median renal levels of FL-furosine in both genotypes by 12-15%, and renal and pulmonary free-CML in diabetic mice by 30% and 18%, respectively. Attenuated hepatic steatosis and an improved plasma lipid profile are also observed with treatment in both genotypes, while the gut microbiota profile is unchanged., Conclusion: L. fermentum ME-3 has therapeutic potential for reducing the formation/accumulation of some glycation products in kidneys and attenuating some common diabetes-related complications., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Tissue aging is a complex phenomenon involving molecular aging of matrix proteins, which mainly results from their progressive alteration by nonenzymatic post-translational modifications (NEPTMs) such as glycation and carbamylation. These two reactions, which correspond to the binding of reactive metabolites (i.e. reducing sugars and urea-derived cyanate, respectively) on amino groups of proteins, occur during aging and are amplified in various chronic diseases such as diabetes mellitus or chronic renal disease (CKD). Since these reactions target the same functional groups, they can reciprocally compete for protein modification. Determining which NEPTM is predominant in tissues is necessary to better understand their role in the development of long-term complications of chronic diseases. For that purpose, two different murine models were used for reproducing such a competitive context: a CKD-diabetic mice model and a cyanate-consuming mice model. The competition has been evaluated by quantifying glycation and carbamylation products by LC-MS/MS in skin and aorta total extracts as well as in skin type I collagen. The results showed that the simultaneous enhancement of glycation and carbamylation reactions resulted in a decrease of the formation of glycation products (especially Amadori products) whereas the concentrations of homocitrulline, a carbamylation product, remained similar. These results, which have been obtained in both tissues and in purified skin type I collagen, suggest that carbamylation takes precedence over glycation for the modification of tissue proteins, but only in pathological conditions favouring these two NEPTMs. While glycation has been considered for a long time the predominant NEPTM of matrix proteins, carbamylation seems to also play an important role in tissue aging. The existence of competition between these NEPTMs must be taken into account to better understand the consequences of molecular aging of matrix proteins in tissue aging.

Background: Early (furosine) and advanced (carboxymethyllysine, CML) products of glycation (AGEs) have been reported as increased in plasma, tissues, and organs of diabetic people, indicating a direct link between glycation and type 2 diabetes (T2D). While murine models present some of the characteristics observed in diabetic humans, their pertinence as models of glycation, particularly for T2D, remains poorly described. The aim of this study was to characterize and compare glycation in several organs of two commonly studied murine models of T2D using stable isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS)., Methods: Defining parameters of type 2 diabetes including body weight, fasting glycaemia, and glucose intolerance were measured in three different C57BL6 mouse models of T2D-the genetic LepR db/db (db/db) model and two diet-induced obesity (DIO) models-and their respective controls. Furosine, free, and protein-bound CML were quantified in kidneys, lungs, heart, and liver by LC-MS/MS., Results: The obesity, hyperglycaemia, and glucose intolerance in db/db mice was accompanied by an increase of furosine and protein-bound CML levels in all organs relative to controls. The DIO models took several months to become obese, exhibited less severe hyperglycaemia and glucose intolerance, while glycation products were not significantly different between these groups (with the exception of furosine in liver and CML in lungs)., Conclusions: The db/db model better reflected the characteristics of human T2D compared with the DIO models and exhibited greater formation and accumulation of both furosine and protein-bound CML in all of the organs tested here., (© 2018 John Wiley & Sons, Ltd.)

The goals of this study were to evaluate the effect of baking time on the Maillard reaction products (MRPs) generated in the crust of the traditional French baguette and to estimate their impact on its sensory characteristics, its acrylamide content, and its bifidogenic effect. Melanoidins, volatile compounds, and acrylamide were evaluated in the crust of traditional French baguettes baked between 12 and 22 min at 225 °C. The increase in melanoidins was positively correlated to the baking time, while volatile compounds only increased until 18 min. The acrylamide content was estimated to be below 18 μg/kg, which confirms the findings of EFSA that bread is not a main contributor to dietary acrylamide. A descriptive sensory analysis showed that the baking time positively affected the sensory quality of the crust. The consumer test revealed the same trend and the panelists favorably judged the well-baked baguettes based on a better crust flavor and crispness. The bifidogenic effect of the crust and the crumb from the baguettes baked 22 min was evaluated on the in vitro growth of Bifidobacterium adolescentis. The results demonstrated that the crumb and the crust had exactly the same bifidogenic impact, therefore not caused by melanoidins., Practical Application: The consumption of bread in France has decreased since 2007, although bread is considered by French people as "healthy" and essential to maintain a balanced diet. The current study evaluated the global qualities of the French baguette in order to highlight its high sensory quality and its beneficial effect by inducing a possible growth of bifidobacteria, even in well-baked baguettes. These findings allow the French bakery industry to develop an argument to promote its technical know-how and to help consumers choose healthier and tastier bread. Moreover, this study provided some recommendations of baking processes to maintain a high sensory quality of the French baguette and limit the formation of undesirable compounds, such as acrylamide, in the crust., (© 2018 Institute of Food Technologists®.)

Background: Coffee substitutes made of roasted chicory are affected by the formation of acrylamide whose main precursor is asparagine. One strategy for limiting the formation of acrylamide is to reduce free asparagine in the chicory roots by lessening the supply of nitrogen in the field. However, decreasing nitrogen fertilizer could affect the formation of the volatile compounds and, consequently, the sensory characteristics of the roasted chicory. The present study aimed to investigate the impact of the nitrogen supply in five commercial varieties on their aroma profile., Results: The addition of 120 kg ha -1 of nitrogen fertilizer in the field resulted in a greater amount of pyrazines in the roasted chicory. Triangle tests were performed to determine the effect of the nitrogen level on the sensory quality of the five varieties. The results revealed that the chicory aroma was modified in two out of five varieties., Conclusion: The results of the present study suggest that a strategy aiming to limit the amount of acrylamide could affect the sensory quality of some varieties of chicory. Further acceptance tests need to be conducted to assess the effect (whether favourable or otherwise) on the sensory quality of the coffee substitutes. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Scope: N ɛ -Carboxymethyl-lysine (CML) is a prominent advanced glycation end-product which is not only found in vivo but also in food. It is known that a percentage of the dietary CML (dCML) is absorbed into the circulation and only partly excreted in the urine. Several studies have tried to measure how much dCML remains in tissues. However obstacles to interpreting the data have been found., Methods and Results: A new protocol which discriminates dCML from native CML (nCML) has been developed. Three CML isotopes with different mass-to-charge ratios were used: nCML N ε -carboxymethyl-L-lysine, dCML N ε -[ 13 C]carboxy[ 13 C]methyl-L-lysine and internal standard N ε -carboxymethyl-L-[4,4,5,5- 2 H 4 ]lysine. Wild-type (n = 7) and RAGE -/- (n = 8) mice were fed for 30 days with either a control, or a BSA-bound dCML-enriched diet. Organs were analyzed for nCML and dCML using liquid chromatography-tandem mass spectrometry. Mice exposed to dCML showed an accumulation in all tissues tested except fat. The rate of deposition was high (81-320 μg dCML /g dry matter) in kidneys, intestine, and lungs and low (<5 μg/g) in heart, muscle, and liver. This accumulation was not RAGE dependent., Conclusion: The kidney is not the only organ affected by the accumulation of dCML. Its high accumulation in other tissues and organs may also, however, have important physiological consequences., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The aim of this study was to develop a white bread with improved nutrient contents and reduced levels of potentially harmful Maillard reaction products such as N(ε)-carboxymethyllysine (CML) and 5-hydroxymethylfurfural (HMF). Assays were carried out through a full factorial experimental design allowing the simultaneous analysis of four factors at two levels: (1) wheat flour extraction rates (ash content: 0.60%-0.72%), (2) leavening agents (bakers' yeast - bakers' yeast and sourdough), (3) prebaking and (4) baking conditions (different sets of time and temperature). The baking conditions affected HMF and CML as well as certain mineral contents. A reduced baking temperature along with a prolonged heat treatment was found to be favourable for reducing both the CML (up to 20%) and HMF concentrations (up to 96%). The presence of sourdough decreased the formation of CML (up to 28%), and increased the apparent amounts of calcium (up to 8%) and manganese (up to 17.5%) probably through acidification of the dough. The extraction rate of flours as well as interactions between multiple factors also affected certain mineral content. However, compounds like folate, thiamine, copper, zinc, iron and phytic acid were not affected by any of the factors studied.

The aim of this study was to test the methods currently in use and to develop a new protocol for the evaluation of melanoidins in bread. Markers of the early and advanced stages of the Maillard reaction were also followed in the crumb and the crust of bread throughout baking, and in a crust model system. The crumb of the bread contained N(ε)-fructoselysine and N(ε)-carboxymethyllysine but at levels 7 and 5 times lower than the crust, respectively. 5-Hydroxymethylfurfural was detected only in the crust and its model system. The available methods for the semi-quantification of melanoidins were found to be unsuitable for their analysis in bread. Our new method based on size exclusion chromatography and fluorescence measures soluble fluorescent melanoidins in bread. These melanoidin macromolecules (1.7-5.6 kDa) were detected intact in both crust and model system. They appear to contribute to the dietary fibre in bread., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

During the heat treatment of coffee and its substitutes some compounds potentially deleterious to health are synthesized by the Maillard reaction. Among these, N(ε)-carboxymethyl-lysine (CML) was detected at high levels in coffee substitutes. The objective of this study was to evaluate the impact of changes in agricultural practice on the lysine content present in chicory roots and try to limit CML formation during roasting. Of the 24 varieties analyzed, small variations in lysine content were observed, 213 ± 8 mg/100 g dry matter (DM). The formation of lysine tested in five commercial varieties was affected by the nitrogen treatment with mean levels of 176 ± 2 mg/100 g DM when no fertilizer was added and 217 ± 7 mg/100 g DM with a nitrogen supply of 120 kg/ha. The lysine content of fresh roots was significantly correlated to the concentration of CML formed in roasted roots (r = 0.51; p < 0.0001; n = 76).

The fat food group, especially butter, has so far been thought to have a high N(ε)-carboxymethyl-lysine (CML) content. However recent data have challenged this opinion. The objective of this article was to determine not only CML content but also that of 5-hydroxymethylfurfural (HMF) in raw and cooked butters. The first aim of this study was to verify the accuracy of the LC-MS/MS and LC-UV methods used for the quantification of CML and HMF. The tests on fortified butter samples showed recovery values of 72% for CML and 78% for HMF. The amounts of CML in raw and cooked butters were 0.25 ± 0.03 and 2.22 ± 0.56 μg/g, respectively. The level of HMF in cooked butters was 61 ± 40 μg/g. No CML was detected in clarified butter, and no HMF was detected in raw and clarified butters. The results indicate that the contribution of butter alone to the exposure to CML and HMF is very low., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The purpose of this study was to investigate the intake, excretion, and tissue accumulation of carboxymethyl-lysine (CML), after feeding rats a diet containing advanced glycation end products (AGEs) from a glucose-lysine (GL) model system. Rats were distributed into two groups and assigned to a control diet or a diet including 3% heated GL (GL diet) for three months. Feces and urine were collected over the last week. After sacrifice, serum was obtained and some organs were removed for CML analysis. The percentage of fecal CML was 2.5-fold higher in the animals fed the GL diet (33.2 vs 76.5% for control and GL group), whereby total recovery was 91.8% compared with a level of 54.6% in the animals fed the control chow, evidencing the importance of the chemical form and the net quantity of dietary CML on its elimination. We suggest that dietary dicarbonyl compounds from GL diet or dietary CML itself are responsible for CML accumulation in hearts and tendons. The most significant result of the present study is that the regular consumption of dietary AGEs in healthy individuals promotes CML accumulation in some organs.

Sensitive analytical methods were developed and validated for the quantification of acrylamide, N(ε)-carboxymethyl-lysine (CML) and 5-hydroxymethylfurfural (HMF) in 24 commercial coffee substitutes (CSs) and 12 instant coffees (ICs). Acrylamide levels varied widely from 200 to 4940 µg kg(-1) with higher levels in CSs. Only two out of 24 CSs had a level of acrylamide above the indicative value set for this food category by the European Commission (4000 µg kg(-1)). None of the ICs tested in this study exceeded the indicative value set for this foodstuff (900 µg kg(-1)). CML ranged from 0.17 to 47 mg kg(-1) and it increased in proportion to the protein content of the samples. The highest concentrations were found in IC partly due to the relatively high protein content of this food group. HMF was the most abundant neoformed compound (NFC) found in the tested commercial samples. It was found between 0.59 and 13 g kg(-1). Among other food categories IC and CS could appear to be major contributors to the exposure to NFCs if consumed on a daily basis. Further investigations are needed to elucidate the acrylamide formation during processing and to determine the daily intake level of frequent consumers of these products.

Research on the impact of Maillard reaction products (MRPs) on microorganisms has been reported in the literature for the last 60 years. In the current study, the impact of an MRP-rich medium on the growth of three strains of Escherichia coli was measured by comparing two classic methods for studying the growth of bacteria (plate counting and optical density at 600 nm) and by tracing MRP utilisation. Early stage and advanced MRPs in the culture media were assessed by quantifying furosine and N (ε) -carboxymethyllysine (CML) levels, respectively, using chromatographic methods. These measures were performed prior to and during bacterial growth to estimate the potential use of these MRPs by Escherichia coli CIP 54.8. Glucose and lysine, the two MRP precursors used in the MRP-rich medium, were also quantified by chromatographic means. Compared to control media, increased lag phases and decreased growth rates were observed in the MRP-rich medium for two out of the three Escherichia coli strains tested. In contrast, one strain isolated from the faeces of a piglet fed on a MRP-rich diet was not influenced by the presence of MRPs in the medium. Overall, CML as well as the products obtained by the thermal degradation of glucose and lysine, regardless of the Maillard reaction, did not affect the growth of the three strains tested. In addition, no degradation of fructoselysine or CML was found in the presence of Escherichia coli CIP 54.8.

Our aim was to investigate carboxymethyl-lysine (CML) intake and excretion after feeding rats with diets containing advanced glycation end-products (AGEs) from bread crust (BC) or its soluble or insoluble fractions, and to identify the factors responsible for the effects observed. CML in serum and different tissues was measured to detect possible accumulations. For 88 days, weanling rats were fed with either a control diet or one containing BC, or its soluble low molecular weight (LMW), soluble high molecular weight (HMW) or insoluble fractions. In the last week of the assay, faeces and urine were collected daily and stored as a 1 week pool. After sacrifice, blood was drawn to obtain serum and some organs were removed. CML analysis was performed by HPLC/MS/MS in diets, faeces, urines, serum and tissues. Faecal excretion of CML was strongly influenced by dietary CML levels and represents the major route of excretion (i.e. 33.2%). However, the urinary elimination of CML was probably limited or saturated, especially when more complex compounds were present in the diet. BC consumption increased CML in the cardiac tissue (170 ± 18 vs. 97 ± 3 μmol per mol lysine for BC and control groups), which correlated with the CML intake. The levels of this AGE in bone were unaffected by the dietary treatment, but in tail tendons CML was greatly increased in the animals that consumed the BC diet (102 ± 13 vs. 51 ± 8 μmol per mol lysine for BC and control groups, P = 0.006), which was associated with the intake of soluble LMW compounds present in BC. Despite the CML accumulation detected in different tissues, serum levels of protein-bound CML were unchanged, indicating the importance of measuring the free CML in this fluid as a real index of dietary CML.

Maillard reaction products (MRPs) are a mixture of compounds generated after the heat treatment of food. High circulating levels of MRPs have been associated with degenerative pathologies such as diabetes, but little is known about their effect on the gut, the main organ in contact with food-derived MRPs. This study was aimed at determining whether repeated low-level exposure to MRPs, generated via two different heat treatments, can contribute to the modulation of experimental colitis in mice. In the first series of experiments, we tested whether pellets rich in MRPs would increase plasmatic and faecal concentration of MRPs. In the second series, we assessed whether two levels of pellet-derived MRPs would be able to modulate chemically-induced inflammation and affect tissue healing. The ingestion of MRPs correlates with the increase of its plasmatic and faecal concentration. Highly treated pellets were proved to significantly protect against inflammation whereas standard or moderately heated pellets had no effect on the inflammatory course. The chemical analysis of the different pellets indicated that high heating generates more melanoidins. There is a correlation between the exposure to highly heated foods and the reduction of murine inflammation, of which the mechanisms remain to be elucidated.