Carboxymethyllysine Content Research Articles

Estimation of Advanced Glycation End Products in Selected Foods and Beverages by Spectrofluorimetry and ELISA.

Advanced glycation end products (AGEs) are formed within the body as a part of normal metabolism and are also the by-products of cooking food. The elevated levels of AGEs in the body are considered pathogenic. The modern diets contain high levels of AGEs which are getting incorporated into the body AGEs pool and contribute to post-diabetic and age-related complications. The objective of the present study is to estimate the cross-linked AGEs (AGE-fluorescence) and the more stable carboxymethyl-lysine (CML) by spectrofluorimetry and ELISA in 58 kinds of foods in India. It was evident from the results that the foods cooked at higher temperatures showed high levels of AGEs. Among the studied foods, the highest fluorescence was observed in Biscuits 2 (362 AU), and the highest level of carboxymethyl lysine (CML) was found in Soya milk (659.3 ng/g). However, there was less correlation between the AGE-fluorescence and the CML content of the food samples. Processed food such as tomato sauce, chilli sauce, and cheese, along with western foods like chicken nuggets, pizza, and biscuits like Biscuits 2, are known to contain high levels of AGEs. In the present study a preliminary database of AGE-fluorescence and CML content of 58 foods was developed, which is the first attempt among Indian foods. Furthermore, elaborated database can be developed including maximum consumed foods in India which will help in suggesting a better diet for the diabetic population.

Liquid Chromatography with Tandem Mass Spectrometry Analysis of Carboxymethyl Lysine in Indonesian Foods.

There is little data on directly measured carboxymethyl lysine (CML) content in Indonesian foods. This study aimed to generate a database of CML values in foods commonly consumed in West Java and West Sumatra. The results were to be used to update our previous estimated CML values. CML values in food samples were measured using high-pressure liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). Food protein content was analyzed by Kjeldahl's method or inferred from the nutrition facts' label. A total of 210 food samples were examined, with the food groups of meat and poultry (1.06 mg CML/100 g edible food), and starchy foods (0.21 mg/100 g edible food) having the highest and lowest mean CML levels, respectively. We found that the foods with the top three highest CML content were fried starch dough (cimol), fried fish crackers, and chicken gulai. The mean of the estimated values (0.80 mg CML/100 g edible food) was higher than the directly measured values (0.66 mg CML/100 g edible food), [p < 0.035]. Conclusion: This database provides information on CML values in Indonesian foods, and can be further used to make a guide policy for the selection of foods to reduce non-communicable diseases. Further measurements are needed on Indonesian dishes to complete the database.

The effects of using sheep tail fat and cooking time on carboxymethyl-lysine formation and some quality characteristics of heat-treated sucuk.

The study's aim was to determine the effect of using sheep tail fat (STF) on carboxymethyl-lysine (CML) content and other properties of heat-treated sucuk (HTS), a type of semi-dry fermented sausage. Three mixtures were prepared: 100% beef fat (BF), 50% BF + 50% STF, and 100% STF. After production (fermentation, heat treatment, and drying), the samples were cooked at 180°C for 0, 1, 3, and 5 min to determine the effect of cooking time on CML, thiobarbituric acid reactive substance (TBARS), total sulfhydryl, and carbonyl contents. The lowest pH value (5.50) was observed in the presence of STF. The most oleic acid (46.02%) was observed in the 100% STF group. The score of taste and general acceptability decreased with increasing STF. Using STF had no significant effect on TBARS, total sulfhydryl, carbonyl, or CML content. These parameters were affected by cooking time. The mean CML content increased from 55.77 to 72.90 μg/g after 5 min of cooking. CML correlated more strongly with TBARS than sulfhydryl or carbonyl.

Carboxymethyl lysine content in traditional Indian foods

Carboxymethyl lysine (CML), an advanced glycation end product (AGE), is formed in food systems due to processing conditions employed. Dietary AGEs, when ingested, add to the endogenous AGEs pool and can have health consequences. Hence, in the present work, traditionally consumed foods from India were determined for bound and free CML levels formed by various cooking methods. Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) with multiple reaction monitoring (MRM) was used to estimate the levels of CML for various food categories. In each category among the products tested, milk peda (Indian sweet) had a high amount of CML (230.4 ± 14.65 µg/g). Some of the food products which had significant amounts of CML are Panipuri (24.12 ± 0.35 µg/g), Curd rice (16.70 ± 2.05 µg/g), Pakoda (27.35 ± 4.71 µg/g), and roasted peanut (25.77 ± 2.79 µg/g). Correlating dietary CML consumption with various health parameters would help predict disease outcomes based on eating habits in the long run.

Effect of edible oil type on the formation of protein-bound Nε-(carboxymethyl)lysine in roasted pork patties

Protein-bound Nε-(carboxymethyl)lysine (CML), an advanced glycation end product within meat products, poses a potential health risk to humans. The objective of this study was to explore the impact of various edible oils on the formation of protein-bound CML in roasted pork patties. Eleven commercially edible oils including lard oil, corn oil, palm oil, olive oil, flaxseed oil, blended oil, camellia oil, walnut oil, soybean oil, peanut oil, and colza oil were added to pork tenderloin mince, respectively, at a proportion of 4 % to prepare raw pork patties. The protein-bound CML contents in the pork patties were determined by HPLC-MS/MS before and after roasting at 200 °C for 20 min. The results indicated that walnut oil, flaxseed oil, colza oil, olive oil, lard oil, corn oil, blended oil, and palm oil significantly reduced the accumulation of protein-bound CML in pork patties, of which the inhibition rate was in the 24.43 %–37.96 % range. Moreover, the addition of edible oil contributed to a marginal reduction in the loss of lysine. Meanwhile, glyoxal contents in pork patties were reduced by 16.72 %–43.21 % after roasting. Other than blend oil, all the other edible oils restrained protein oxidation in pork patties to varying degrees (between 20.16 % and 61.26 %). In addition, camellia oil, walnut oil, and flaxseed oil increased TBARS values of pork patties by 2.2–8.6 times when compared to the CON group. After analyzing the fatty acid compositions of eleven edible oils, five main fatty acids (palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid) were selected to establish Myofibrillar protein-Glucose-fatty acids systems to simulate the roasting process. The results showed that palmitic acid, oleic acid, linoleic acid, and linolenic acid obviously mitigated the formation of myofibrillar protein-bound CML, exhibiting suppression rates ranging from 10.38 % to 40.32 %. In conclusion, the addition of specific edible oil may curb protein-bound CML production in roasted pork patty by restraining protein or lipid oxidation, reducing lysine loss, and suppressing glyoxal production, which may be attributed to the fatty acid compositions of edible oils. This finding provides valuable guidance for the selection of healthy roasting oils in the thermal processing of meat products.

The Effects of Sheep Tail Fat, Fat Level, and Cooking Time on the Formation of Nε-(carboxymethyl)lysine and Volatile Compounds in Beef Meatballs.

This study aimed to determine the effects of fat type (sheep tail fat (STF) and beef fat (BF)), fat levels (10, 20, or 30%), and cooking time (0, 2, 4, and 6 min, dry heat cooking at 180 °C) on the carboxymethyl lysine (CML) content in meatballs. pH, thiobarbituric acid reactive substance (TBARS), and volatile compound analyses were also performed on the samples. The use of STF and the fat level had no significant effect on the pH value. The highest TBARS value was observed with the combination of a 30% fat level and STF. CML was not affected by the fat level. The highest CML content was determined in meatballs with STF at a cooking time of 6 min. In the samples cooked for 2 min, no significant difference was observed between STF and BF in terms of the CML content. STF generally increased the abundance of aldehydes. Aldehydes were also affected by the fat level and cooking time. A PCA provided a good distinction between groups containing STF and BF regardless of the fat level or cooking time. Pentanal, octanal, 2,4-decadienal, hexanal, and heptanal were positively correlated with CML.

Contents and Correlations of Nε-(carboxymethyl)lysine, Nε-(carboxyethyl)lysine, Acrylamide and Nutrients in Plant-Based Meat Analogs.

High temperatures applied in the production of plant-based meat analogs (PBMA) lead to the occurrence of Maillard reactions, in which harmful compounds Nε-(carboxymethyl)lysine (CML), Nε-(carboxyethyl)lysine (CEL) and acrylamide are formed. However, little research has focused on these compounds in PBMA. In this study, the contents of CML, CEL and acrylamide in 15 commercial-sold PBMA were determined by an ultra-high performance liquid chromatograph coupled with a triple quadrupole tandem mass spectrometer (UHPLC-QqQ-MS/MS). Nutrients (protein, amino acids, fatty acids and sugars) which are related to the formation of these compounds were also studied. The results showed that CML, CEL and acrylamide contents were in the range of 16.46-47.61 mg/kg, 25.21-86.23 mg/kg and 31.81-186.70 μg/kg, respectively. Proteins account for 24.03-53.18% of PBMA. Except for Met + Cys, which is the limiting amino acid of most PBMA, all other indispensable amino acids met the requirements for adults. Besides, PBMA had more n-6 fatty acids than n-3 fatty acids. A correlation analysis showed that proteins and the profiles of amino acid and fatty acid had little influence on CML but significant influence on CEL and acrylamide. The results of the present study can be used as a reference to produce PBMA with higher amounts of nutrients and lower amounts of CML, CEL and acrylamide.

Nϵ -(carboxymethyl)lysine in bakery products: A review.

The purpose of this review is to draw attention to the Nϵ -(carboxymethyl)lysine (CML) content of bakery products with respect to their formation during baking and their health effects. Phenolic components added to the formulation in bakery products significantly reduce the formation of CML. Among the phenolic components, ferulic acid showed the most significant lowering effect on CML. Among the flavanones tested in the model cookie system, dihydromyricetin exhibited the strongest CML-reducing effect. The addition of fat-, sugar-, and protein-rich ingredients to the formulations of bakery products generally increases the CML content in these products. In addition, the addition of components that have a water activity-reducing effect, such as dietary fiber, and the high temperature in baking also increase the formation of CML. Therefore, the food industry should also focus on optimizing food production to minimize CML formation while maintaining the safety and organoleptic properties of bakery products. PRACTICAL APPLICATION: The CML level in foods is likely to increase 200 times with an increase in cooking temperature. The addition of protein and fat to bakery product formulations can increase CML formation. The addition of glucose in cakes can produce higher levels of CML than fructose, refined sucrose, or unrefined sucrose. Phenolic compounds have a reducing effect on CML formation in bakery products.

Aerobic Exercise Training Reduces Atherogenesis Induced by Low-Sodium Diet in LDL Receptor Knockout Mice.

This study investigated the efficacy of aerobic exercise training (AET) in the prevention of dyslipidemia, insulin resistance (IR), and atherogenesis induced by severe low-sodium (LS) diet. LDL receptor knockout (LDLR KO) mice were fed a low-sodium (LS) (0.15% NaCl) or normal-sodium (NS; 1.27% NaCl) diet, submitted to AET in a treadmill, 5 times/week, 60 min/day, 15 m/min, for 90 days, or kept sedentary. Blood pressure (BP), plasma total cholesterol (TC) and triglyceride (TG) concentrations, lipoprotein profile, and insulin sensitivity were evaluated at the end of the AET protocol. Lipid infiltration, angiotensin II type 1 receptor (AT1), receptor for advanced glycation end products (RAGE), carboxymethyllysine (CML), and 4-hydroxynonenal (4-HNE) contents as well as gene expression were determined in the brachiocephalic trunk. BP and TC and gene expression were similar among groups. Compared to the NS diet, the LS diet increased vascular lipid infiltration, CML, RAGE, 4-HNE, plasma TG, LDL-cholesterol, and VLDL-TG. Conversely, the LS diet reduced vascular AT1 receptor, insulin sensitivity, HDL-cholesterol, and HDL-TG. AET prevented arterial lipid infiltration; increases in CML, RAGE, and 4-HNE contents; and reduced AT1 levels and improved LS-induced peripheral IR. The current study showed that AET counteracted the deleterious effects of chronic LS diet in an atherogenesis-prone model by ameliorating peripheral IR, lipid infiltration, CML, RAGE, 4-HNE, and AT1 receptor in the intima-media of the brachiocephalic trunk. These events occurred independently of the amelioration of plasma-lipid profile, which was negatively affected by the severe dietary-sodium restriction.

Dietary advanced glycation end products and cancer risk in Japan: From the Takayama study.

Few large epidemiological studies have evaluated the association between dietary advanced glycation end products (AGEs) and cancer risk. We evaluated the relationship between dietary AGE intake and the incidence of total cancer and site‐specific cancers in a population‐based prospective study in Japan. Participants were 14,173 men and 16,549 women who were 35 years of age or older in 1992. Dietary intake was assessed via a validated food frequency questionnaire. Intake of the AGE N ε‐carboxymethyl‐lysine (CML) was estimated using databases of CML content in foods determined using ultraperformance liquid chromatography–tandem mass spectrometry. Cancer incidence was confirmed through regional population‐based cancer registries. During a mean follow‐up period of 13.3 years, 1954 men and 1477 women developed cancer. We did not observe a significant association between CML intake and the risk of total cancer in men or women. In men, compared with the lowest quartile of CML intake, the hazard ratios of liver cancer for the second, third, and highest quartiles were 1.69 (95% CI: 0.92–3.10), 1.48 (95% CI: 0.77–2.84), and 2.10 (95% CI: 1.10–3.98; trend p = 0.04). Conversely, a decreased relative risk of male stomach cancer was observed for the second and highest quartiles of CML intake versus the lowest quartile, with hazard ratios of 0.73 and 0.67, respectively (trend p = 0.08). Our finding on the potential harmfulness of consuming AGEs on liver cancer risk is intriguing and warrants further study.

A novel formation pathway of Nε-(carboxyethyl)lysine from lactic acid during high temperature exposure in wheat sourdough bread and chemical model

Nε-(carboxymethyl)lysine (CML) and Nε-(carboxyethyl)lysine (CEL) have been the most extensively studied advanced glycation end-products (AGEs) in foods. Their formation mechanism, especially the latter, has not been clearly delineated in fermented food. In this work, the relative contents of CEL and CML were evaluated in a sourdough-bread and a silica solid chemical model. Lactic acid (LA) content in the sourdough increased with fermentation time that was accompanied by an increase in CEL, but not CML content in the bread. The role of LA as a precursor for CEL was supported by a positive significant correlation between LA and CEL contents, and further analysis using isotope-labeled LA (LA-13C3) revealed that the three carbon atoms of LA were incorporated into CEL. These findings for the first time indicate LA may function as a precursor to promote CEL formation in sourdough bread that merits further investigation.

Nɛ-(carboxymethyl)lysine, Available Lysine, and Volatile Compound Profile of Biscuits Enriched with Grape by-Product During Storage.

We investigated the changes in Nɛ-(carboxymethyl)lysine (CML) and available lysine content, antioxidant properties, volatiles, and oxidation products of biscuits enriched with grape by-product (GP), stored for six months under a modified atmosphere of 0%/30%/70% O2/CO2/N2 and in air. Fresh GP-formulated biscuits showed lower concentrations of CML (89%), available lysine (40%), and pyrazines (75%), but higher antioxidant capacities (~ sixfold), furans (12-fold), and lipid-derived compounds (three-fold) than the control. Although ~ 15% higher losses of Maillard-type volatiles were identified in the air atmosphere during storage, lipid oxidation was ~ 30% less pronounced in the modified atmosphere. A significant correlation of 0.994 between the reduction in CML and the available lysine suggest further CML reactions with the ɛ-NH2 group of amino acids. Significant correlations (of -0.550 to -0.980) between oxidation products, antioxidant capacities, and changes in CML content during storage suggest that these parameters might be involved in the CML elimination mechanism.

Quantitative determination of Nε-(carboxymethyl)lysine in sterilized milk by isotope dilution UPLC-MS/MS method without derivatization and ion pair reagents

This study developed a sensitive and easy method to directly analyze Nε-(carboxymethyl)lysine (CML) by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Acetonitrile and water were used as the binary mobile phase without ion pair reagents, while BEH Amide column was applied to detect CML. The target substance without derivatization could be well retained and separated after the pretreatment conditions were optimized. Oasis MCX solid phase extraction (SPE) cartridge showed the best recovery rate of CML 96.7 % and purification ability among the tested SPE cartridges. There was no obvious difference in pretreatment abilities with or without protein precipitation on the CML quantitative analysis. The recovery rates 97–98 % for CML were achieved by UPLC-MS/MS, as well as the detection limit and quantification limit were 0.05 mg/kg and 0.15 mg/kg sample, respectively. This method was suitable for quantifying CML content with high recovery rate and low detection and quantification limit in sterilized milk.

Characterizing changes in Maillard reaction indicators in whole milk powder and reconstituted low-temperature pasteurized milk under different preheating conditions.

The chemical properties and quality of milk powder and reconstituted milk can be changed by preheating treatment during milk powder processing. The effect of preheating conditions (low-temperature pasteurization [LT], high-temperature pasteurization [HT], extended shelf-life [ESL], ultra-high-temperature [UHT], and in-container sterilization [CS]), on furosine, furfural compounds, and advanced glycation end products were investigated in milk powders and reconstituted milk. The results revealed that Maillard reaction indicators were affected by preheating intensity; contents of furosine, 5-hydroxymethylfurfural (HMF), 2-furfural, Nε-(carboxymethyl)lysine (CML), and Nε-(carboxyethyl)lysine (CEL) increased, whereas lysine and 5-methyl-furfural (MF) decreased in both milk powder and reconstituted milk with an increase in the intensity of preheating. CML and CEL contents in reconstituted milk increased by 1.7- to 2.4-fold and 1.3- to 1.6-fold, respectibely. The content of CML and CEL in high-preheating powders was 11%-34% and 18%-123% higher than that in LT samples, respectively. Compared with milk powder, furosine was reduced by 22%-55% and MF by 57%-69% in reconstituted milk; HMF increased by 12%-52%, CML and CEL increased 1.7-2.4 times and 1.3-1.6 times, respectively. A superposition effect was observed between preheating intensity and heat process times, which increased the content of Maillard reaction products. PRACTICAL APPLICATION: Our findings revealed the promotion effect of higher preheating conditions on Maillard reaction indicators in milk powder and reconstituted low-temperature pasteurized milk. The results will help reduce some of the harmful Maillard reaction indicators (e.g., advanced glycation end products) formed during heat processing and assist customers in selecting dairy products with low amounts of Maillard reaction products.

Formation of advanced glycation end products in raw and subsequently boiled broiler muscle: biological variation and effects of postmortem ageing and storage

The study aimed to investigate and compare the contents of carboxymethyllysine (CML) in two kinds of broilers during postmortem ageing and storage. The contents of CML in raw and boiled (100°C, 30min) broiler briskets and legs which were from white feather broilers (n=8) and yellow feather broilers (n=8) with ageing and storage at 4°C for 0–168h were determined. Postmortem ageing and storage had a significant (P<0.05) effect on the color and A420nm in both boiled broilers meat. In addition, with the ageing and storage time increasing, CML content in raw white feather broiler brisket meat had no significant (P>0.05) change, while that in boiled brisket meat significantly (P<0.05) increased during 0–6h, then decreased during 6–24h, finally increased again. CML content in leg meat increased significantly (P<0.05) with the ageing and storage time prolonging. But postmortem ageing and storage had no significant (P>0.05) effect on the CML contents in raw/boiled yellow feather broilers. Meanwhile, CML contents in white feather broilers were much higher than that in yellow feather broilers. Thus, white feather broilers can be selected as the research object to study the mechanism of ageing and storage on CML content in the postmortem broiler in the future.

Changes in harmful Maillard reaction products in low-temperature long-time pasteurization-treated milks reconstituted from whole-milk powders after different storage times

Reconstituted milk treated with low-temperature long-time pasteurization was produced from whole-milk powders stored for 0–18 months. The profiles of initial, intermediate, and advanced stage Maillard reaction products, furosine, furfurals, and advanced glycation end-products in reconstituted milk were investigated. The total lysine content decreased in the reconstituted milk, whereas furosine content increased by 1.8- to 7.4-fold with storage of the corresponding milk powder. 5-(Hydroxymethyl)furfural (HMF) increased by 25–271 % with storage. 2-Furaldehyde (F) and 5-methyl-2-furfural also showed an increasing trend with storage, but 2-furyl methyl ketone (FMC) was detected only in fresh and 4-month-old reconstituted milk powder. The Nε-(carboxymethyl)lysine (CML) content increased by 1.8- to 5.2-fold with storage (from 9.9 ± 1.6 to 28.3 ± 3.3 mg/L). The Nε-(carboxyethyl)lysine content increased by 34–175 %. The increase in the furosine, HMF, F, and CML content in reconstituted milk accelerated between 12 and 18 months of storage. These compounds were more affected by the storage time of the milk powder than other harmful Maillard reaction products.

Can fingernail quality predict bone damage in Type 2 diabetes mellitus? a pilot study.

Type 2 diabetes mellitus (T2DM) adversely affects the normal functioning, intrinsic material properties, and structural integrity of many tissues, including bone. It is well known that the clinical utility of areal bone mineral density (aBMD) is limited to assess bone strength in individuals with T2DM. Therefore, there is a need to explore new diagnostic techniques that can better assist and improve the accuracy of assessment of bone tissue quality. The present study investigated the link between bone and fingernail material/compositional properties in type 2 diabetes mellitus (T2DM). For that, femoral head and fingernail samples were obtained from twenty-five adult female patients (with/without T2DM) with fragility femoral neck fractures undergoing hemi/total hip arthroplasty. Cylindrical cores of trabecular bone were subjected to micro-CT, and lower bone volume fraction was observed in the diabetic group than the non-diabetic group due to fewer and thinner trabeculae in individuals with T2DM. The material and compositional properties of bone/fingernail were estimated using nanoindentation and Fourier Transform Infrared Spectroscopy, respectively. Both bone/fingernails in T2DM had lower reduced modulus (Er), hardness (H), lower Amide I and Amide II area ratio (protein content), higher sugar-to-matrix ratio, and relatively high carboxymethyl-lysine (CML) content compared with non-diabetic patients. Sugar-to-matrix ratio and relative CML content were strongly and positively correlated with HbA1c for both bone/fingernail. There was a positive correlation between bone and fingernail glycation content. Our findings provide evidence that the degradation pattern of bone and fingernail properties go hand-in-hand in individuals with T2DM. Hence, the fingernail compositional/material properties might serve as a non-invasive surrogate marker of bone quality in T2DM; however, further large-scale studies need to be undertaken.

Assessment the influence of salt and polyphosphate on protein oxidation and Nε-(carboxymethyl)lysine and Nε-(carboxyethyl)lysine formation in roasted beef patties

The impact of NaCl and tripolyphosphate (TPP)/pyrophosphate (PP) on protein oxidation and Nε-(carboxymethyl)lysine (CML) and Nε-(carboxyethyl)lysine (CEL) formation in roasted beef patties was investigated. The content of CML and CEL in patties treated with salts was approximately 1.1–1.7 and 1.2–3.2 times higher than that of the control samples, respectively. An increase in salt content caused higher oxidation of tryptophan and protein carbonylation with a decrease in Schiff bases (P < 0.05) and a slight decrease in lipid oxidation (P < 0.05). Significant correlations (P < 0.05) between CML, CEL, and protein oxidation measurements was found. The higher salts content, causing less cooking loss and higher moisture content, significantly correlated (P < 0.05) with CML, CEL content, and protein oxidation of the patties. The increase in CML and CEL content and protein oxidation in roasted patties with salts might be related to the pro-oxidation of salts, and also partly due to the temperature changes caused by the water-holding capacity of salts.

Profiles of initial, intermediate, and advanced stages of harmful Maillard reaction products in whole-milk powders pre-treated with different heat loads during 18 months of storage

This study evaluated the chemical changes in five types of whole-milk powders (WMP) with different heating loads during storage. The WMP was preheated using low-heat [low-temperature long-time (LTLT), high-temperature short-time pasteurization (HTST)] and high-heat process [ultra-pasteurization (ESL), ultra-high-temperature (UHT) treatments, and in-bottle sterilization (BS)]. Furosine increased by 2.5–3.0 times in high-heat WMP and 5.7–8.4 times in low-heat WMP during storage. 5-(hydroxymethyl)furfural (HMF) content in high-heat WMP was on average 1.4- to 2.4-fold higher than in low-heat WMP during storage. The increases in the amount of Nε-(carboxymethyl)lysine (CML) and Nε-(carboxyethyl)lysine (CEL) in high-heat WMP were more than that in low-heat WMP (CML, 3.4–4.9 vs 3.1–3.4 times; CEL, 3.4–4.2 vs 2.7–3.0 times). Pyrraline in high-heat WMP increased by 1.8- to 2.1-fold. 2-Furaldehyde, CML, and CEL increased slowly with 12 months of storage and then accelerated. Storage time significantly contributed to more furosine, HMF, CML, and CEL contents in high-heat WMP.

Development of an Isotope Dilution UHPLC-QqQ-MS/MS-Based Method for Simultaneous Determination of Typical Advanced Glycation End Products and Acrylamide in Baked and Fried Foods.

In this work, a stable isotope dilution ultrahigh-performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS) method was developed and validated for simultaneous determination of Nε-(carboxymethyl)lysine (CML), Nε-(carboxyethyl)lysine (CEL), and acrylamide (AA) in baked and fried foods. Ground food samples were extracted with acetone followed by two parallel assays. In assay A, a cleanup procedure based on dispersive solid-phase extraction was conducted for AA, free CML, and CEL analysis using the supernatant. In assay B, a multistep process including reduction, protein precipitation, acid hydrolysis, and solid-phase extraction was conducted for bound CML and CEL analysis using precipitation. The developed method was validated in terms of linearity, sensitivity (limit of detection, LOD; limit of quantitation, LOQ), accuracy, and precision. The results showed that the method had a wide linear range (0.25-500 ng/mL for CML and CEL, 0.5-500 ng/mL for AA), low LOD and LOQ (0.47-0.94 and 1.52-1.91 μg/kg, respectively), and good linearity (R2 > 0.999). The recovery test on baby biscuit and French fries samples showed the recovery rates of 90.2-108.3% for CML, 89.0-106.1% for CEL, and 94.5-112.3% for AA with satisfactory precision (relative standard deviation (RSD) < 10%). Finally, the developed method was successfully applied to 11 baked and fried food samples, and total CML, CEL, and AA contents varied in the ranges of 4.07-35.88 mg/kg, 1.99-14.49 mg/kg, and 5.56-506.64 μg/kg, respectively. Therefore, the isotope dilution UHPLC-QqQ-MS/MS method developed herein is promising for routine analysis of CML, CEL, and AA in baked and fried foods.