Inhibitory effects and mechanisms of phenolic compounds in rapeseed oil on advanced glycation end product formation in chemical and cellular models in vitro.

Sinapic acid (SA), canolol (CAO) and canolol dimer (CAO dimer) are the main phenolic compounds in rapeseed oil. However, their possible efficacy against glycation remains unclear. This study aims to explore the impacts of these substances on the formation of advanced glycation end products (AGEs) based on chemical and cellular models in vitro. Based on fluorescence spectroscopy results, three chemical models of BSA-fructose, BSA-methylglyoxal (MGO), and arginine (Arg)-MGO showed that SA/CAO/CAO dimer could effectively reduce AGE formation but with different abilities. After SA/CAO/CAO dimer incubation, effective protection against BSA protein glycation was observed and three different MGO adducts were formed. In MGO-induced HUVEC cell models, only CAO and CAO dimer significantly inhibited oxidative stress and cell apoptosis, accompanied by the regulation of the Nrf2-HO-1 pathway. During the inhibition, 20 and 12 lipid mediators were reversed in the CAO and CAO dimer groups compared to the MGO group.

In Vitro Antiglycation and Methylglyoxal Trapping Effect of Peppermint Leaf (Mentha × piperita L.) and Its Polyphenols.

The most significant reactive α-dicarbonyl RCS involved in the pathomechanism of glycation and related diseases is methylglyoxal (MGO). Hyperglycemia promotes the generation of MGO and leads to the formation of advanced glycation end products (AGEs). Therefore, MGO trapping and glycation inhibition appear to be important therapeutic targets in prediabetes, diabetes, and in the early prevention of hyperglycemic complications. Peppermint leaf is commonly used as herbal tea, rich in polyphenols. Eriocitrin, its predominant component, in a double-blind, randomized controlled study reversed the prediabetic condition in patients. However, the antiglycation activity of this plant material and its polyphenols has not been characterized to date. Therefore, the aim of this study was to evaluate the ability of a peppermint leaf dry extract and its polyphenols to inhibit non-enzymatic protein glycation in a model with bovine serum albumin (BSA) and MGO as a glycation agent. Peppermint polyphenols were also evaluated for their potential to trap MGO in vitro, and the resulting adducts were analyzed by UHPLC-ESI-MS. To relate chemical composition to glycation inhibitory activity, the obtained peppermint extract was subjected to qualitative and quantitative analysis. The capability of peppermint leaf polyphenols to inhibit glycation (27.3-77.2%) and form adducts with MGO was confirmed. In the case of flavone aglycones, mono- and di-adducts with MGO were observed, while eriodictyol and eriocitrin effectively produced only mono-adducts. Rosmarinic acid and luteolin-7-O-glycosides did not reveal this action. IC50 of the peppermint leaf dry extract was calculated at 2 mg/mL, equivalent to a concentration of 1.8 µM/mL of polyphenols, including ~1.4 µM/mL of flavonoids and ~0.4 µM/mL of phenolic acids. The contribution of the four major components to the anti-AGE activity of the extract was estimated at 86%, including eriocitrin 35.4%, rosmarinic acid 25.6%, luteolin-7-O-rutinoside 16.9%, luteolin-7-O-ß-glucuronoside 8.1%, and others 14%. The effect of peppermint dry extract and polyphenols in inhibiting MGO-induced glycation in vitro was comparable to that of metformin used as a positive control.

Comprehensive analysis of the anti-glycation effect of peanut skin extract.

Advanced glycation end-products (AGEs) are produced during protein glycation and associated with diabetic complications. Peanut skin is rich in procyanidins, which may be used as an inhibitor of glycation. This study evaluated the potential anti-glycation effect of peanut skin extract (PSE) and dissected the underlying mechanism. PSE could effectively inhibit the formation of AGEs in BSA-Glc and BSA-MGO/GO models, with 44%, 37% and 82% lower IC50 values than the positive control (AG), respectively. The inhibitory effect of PSE on BSA glycation might be ascribed to its binding interaction with BSA, attenuated formation of early glycation products and trapping of reactive dicarbonyl compounds. Notably, PSE showed a remarkably stronger inhibitory effect on Amadori products than AG. Furthermore, three new types of PSE-MGO adducts were formed as identified by UPLC-Q-TOF-MS. These findings suggest that PSE may serve as an inhibitor of glycation and provide new insights into its application.

Black Tea Theaflavin Detoxifies Metabolic Toxins in the Intestinal Tract of Mice.

SCOPE: This study is to determine the in vivo efficacy of black tea theaflavin (TF) to detoxify two metabolic toxins, ammonia and methylglyoxal (MGO), in mice METHODS AND RESULTS: Under in vitro conditions, TF is able to react with ammonia, MGO, and hydrogen peroxide to produce its aminated, MGO conjugated, and oxidized products, respectively. In TF-treated mice, the aminated TF, the MGO conjugates of TF and aminated TF, and the oxidized TF are searched using LC-MS/MS. The results provide the first in vivo evidence that the unabsorbed TF is able to trap ammonia to form the aminated TF; furthermore, both TF and the aminated TF have the capacity to trap MGO to generate the corresponding mono-MGO conjugates. Moreover, TF is oxidized to dehydrotheaflavin, which underwent further amination in the gut. By exposing TF to germ-free (GF) mice and conventionalized mice (GF mice colonized with specific-pathogen-free microbiota), the gut microbiota is demonstrated to facilitate the amination and MGO conjugation of TF. CONCLUSION: TF has the capacity to remove the endogenous metabolic toxins through oxidation, amination, and MGO conjugation in the intestinal tract, which can potentially explain why TF still generates in vivo efficacy while showing a poor systematic bioavailability.

Analysis of α-dicarbonyl compounds in coffee (Coffea arabica) prepared under various roasting and brewing methods.

This study investigated the correlations of α-dicarbonyl compounds (α-DCs), including glyoxal (GO), methylglyoxal (MGO) and diacetyl (DA), formed in coffee prepared under various roasting and brewing methods. The levels of α-DCs in Brazilian coffee beans (Coffea arabica) ranged from 28.3 to 178 µg/mL. Concentration ranges of GO, MGO and DA were 1.31-6.57, 25.5-159 and 1.50-12.9 µg/mL, respectively. The level of α-DCs increased with high roasting temperature, long roasting time, small coffee bean particles, mineral water and espresso brewing. In correlation analysis, the roasting temperature-time showed strong negative correlations with α-DCs in espresso (-0.886) and cold-brew coffee (-0.957). In espresso coffee, there was a strong negative correlation between the α-DCs and coffee bean particle size (-0.918).

Investigation of the Phytochemical Composition, Antioxidant Activity, and Methylglyoxal Trapping Effect of Galega officinalis L. Herb In Vitro.

Galega officinalis L. has been known for centuries as an herbal medicine used to alleviate the symptoms of diabetes, but its comprehensive chemical composition and pharmacological activity are still insufficiently known. The current study involved the qualitative and quantitative phytochemical analysis and in vitro evaluation of the antioxidative and methylglyoxal (MGO) trapping properties of galega herb. Ultra high-performance liquid chromatography coupled with both the electrospray ionization mass spectrometer and diode-array detector (UHPLC-ESI-MS and UHPLC-DAD) were used to investigate the composition and evaluate the anti-MGO capability of extracts and their components. Hot water and aqueous methanol extracts, as well as individual compounds representing phytochemical groups, were also assessed for antioxidant activity using DPPH (2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) assays. Quercetin and metformin were used as a positive control. We confirmed the presence of tricyclic quinazoline alkaloids, guanidines, flavonoids, and hydroxycinnamic acids (HCAs) in galega extracts. The polyphenolic fraction was dominated by mono-, di-, and triglycosylated flavonols, as well as monocaffeoylhexaric acids. The in vitro tests indicated which G. officinalis components exhibit beneficial antioxidative and MGO trapping effects. For galega extracts, flavonols, and HCAs, a potent antiradical activity was observed. The ability to trap MGO was noted for guanidines and flavonoids, whereas HCA esters and quinazoline alkaloids were ineffective. The formation of mono-MGO adducts of galegine, hydroxygalegine, and rutin in the examined water infusion was observed.

Methylglyoxal-hydroimidazolones (MG-Hs) instead of Nɛ-(carboxymethyl)-l-lysine (CML) is the major advanced glycation end-product during drying process in black tea.

This study was to examine the formation of advanced glycation end-products (AGEs) in black tea during drying process at 90, 120, and 150 °C for 1 h. Nine AGEs including Nɛ-(carboxyethyl)-l-lysine (CEL), Nɛ-(carboxymethyl)-l-lysine (CML), three isomers of methylglyoxal-hydroimidazolones (MG-Hs), three isomers of glyoxal-hydroimidazolones (GO-Hs), and argpyrimidine were quantified by using HPLC-MS/MS with isotope-labelled internal standard. Results showed that each AGE during the drying process of 150 °C was significantly higher (p < 0.05) than at 90 and 120 °C, and argpyrimidine was only found in the treatment of 150 °C. MG-H1/3 was first quantified as the major AGE during drying at 120-150 °C, the content respectively reached to (39.66 ± 2.61) µg/g and (58.88 ± 1.76) µg/g after 1 h drying, where CML content only had (19.86 ± 1.02) µg/g and (23.71 ± 1.40) µg/g. This study indicated that arginine derived-AGEs are the key components of black tea AGEs.

5-Hydroxymethylfurfural accumulation plays a critical role on acrylamide formation in coffee during roasting as confirmed by multiresponse kinetic modelling.

This study aims to investigate in depth the mechanism of acrylamide formation in coffee during roasting. For this purpose, a comprehensive kinetic model including the elementary steps for acrylamide formation was proposed. The changes in sucrose, reducing sugars, free amino acids, asparagine, acrylamide, 3-deoxyglucosone, methylglyoxal, glyoxal, and 5-hydroxymethylfurfural were monitored in coffee during roasting at 200, 220 and 240 °C. Dominant pathways of complex reactions leading to acrylamide were unravelled by means of multiresponse kinetic modelling approach. The results of the model indicated that sucrose degrades into glucose and a reactive fructofuranosyl cation. Interestingly, glucose takes part mostly in the formation of intermediates, glyoxal and especially 3-deoxyglucosone rather than acrylamide formation. On the other hand, fructofuranosyl cation contributed mostly to the formation of 5-hydroxymethylfurfural which was found to be the most important intermediate precursor of acrylamide formed in coffee during roasting.

Effects of hydroxycinnamic acids on the reduction of furan and α-dicarbonyl compounds.

The effects of hydroxycinnamic acids such as cinnamic acid (CNA), p-coumaric acid(CMA), caffeic acid (CFA), and chlorogenic acid (CGA) on the reduction of furan in canned-coffee model systems (CCMS) containing α-dicarbonyls [glyoxal (GO) or methylglyoxal (MGO)] were investigated. The concentration of furan in CCMS containing GO, which was 59.76 µg/L, was reduced by the addition of CFA and CGA to 48.31 µg/L and 41.38 µg/L, respectively; similarly, the furan concentration in model system containing MGO was 45.79 µg/L, and this decreased to 35.41 µg/L (by CFA) and 32.65 µg/L (by CGA), respectively. In addition, the effects of hydroxycinnamic acids on the trapping of GO and MGO were determined. CFA and CGA greatly reduced the concentration of GO to 303.51 µg/L and 267.80 µg/L, respectively (compared to 515.79 µg/L in the control), whereas that of MGO was decreased to 207.01 µg/L and 219.14 µg/L (compared to 417.14 µg/L in the control). The trapping of α-dicarbonyls such as GO and MGO by CFA and CGA could be closely related to furan reduction in CCMS.

Melanoidins from Coffee, Cocoa, and Bread Are Able to Scavenge α-Dicarbonyl Compounds under Simulated Physiological Conditions.

Free amino residues react with α-dicarbonyl compounds (DCs) contributing to the formation of advanced glycation end products (AGEs). Phenolic compounds can scavenge DCs, thus controlling the dietary carbonyl load. This study showed that high-molecular weight cocoa melanoidins (HMW-COM), HMW bread melanoidins (HMW-BM), and especially HMW coffee melanoidins (HMW-CM) are effective DC scavengers. HMW-CM (1 mg/mL) scavenged more than 40% DCs within 2 h under simulated physiological conditions, suggesting some physiological relevance. Partial acid hydrolysis of HMW-CM decreased the dicarbonyl trapping capacity, demonstrating that the ability to react with glyoxal, methylglyoxal (MGO), and diacetyl was mainly because of polyphenols bound to macromolecules. Caffeic acid (CA) and 3-caffeoylquinic acid showed a DC-scavenging kinetic profile similar to that of HMW-CM, while mass spectrometry data confirmed that hydroxyalkylation and aromatic substitution reactions led to the formation of a stable adduct between CA and MGO. These findings corroborated the idea that antioxidant-rich indigestible materials could limit carbonyl stress and AGE formation across the gastrointestinal tract.

Inhibiting the formation of advanced glycation end-products by three stilbenes and the identification of their adducts.

The objective of this work is to investigate the inhibition effects on AGEs formation and the ability of scavenging toxic carbonyls of three stilbenes, resveratrol, oxyresveratrol, and piceatannol. The results showed that the three stilbenes had the activity to inhibit the AGEs formation in BSA-acrolein and BSA-methylglyoxal models, especially piceatannol which showed the strongest inhibition effects on the formation of AGEs with the half maximal inhibitory concentrations (IC50) value of 2.44 mM and 0.19 mM in the BSA-acrolein and BSA-methylglyoxal model, respectively. In addition, the three stilbenes showed the ability to scavenge acrolein and methylglyoxal in pH 7.4 at 37 °C. Eight isolated adducts between three stilbenes and toxic carbonyls further confirmed that these three stilbenes could scavenge acrolein and methylglyoxal by forming adducts successfully. Thus, the present study suggested that the consumption of foods containing stilbenes was beneficial for controlling the amount of reactive carbonyl species and the AGEs formation.

Identification of the antiglycative components of Hong Dou Shan (Taxus chinensis) leaf tea.

The formation of advanced glycation end products (AGEs) may affect life quality and cut lifespan. With the increasingly utilization of new foods and booming development of international trade, large amounts of newly emerging teas have come into our sight. In this study, we evaluated the antiglycative capabilities of five popular herbal teas. Results showed that Chinese Hong Dou Shan (Taxus chinensis; HDS) leaf tea had particularly strong antiglycative activity via scavenging methylglyoxal (MGO) in both chemical (glucose-BSA, fructose-BSA, and MGO-BSA) and cell models (HUVECs). Furthermore, the intracellular AGEs level was alleviated to normal state by the two HDS leaf tea fractions, H-47 and H-57 which were almost as effective as EGCG when added at the same level (50 µg/mL). The effective components against glycation were further separated and tentatively identified as catechin, epicatechin, gallocatechin, epigallocatechin, procyanidin B2, a dihydromyricetin dimer, and a quercetin glucoside by HPLC-DAD and LC-Q-TOF/MS analysis.

Characterization of MK8(H2) from Rhodococcus sp. B7740 and Its Potential Antiglycation Capacity Measurements.

Menaquinone (MK) has an important role in human metabolism as an essential vitamin (VK2), which is mainly produced through the fermentation of microorganisms. MK8(H2) was identified to be the main menaquinone from Rhodococcus sp. B7740, a bacterium isolated from the arctic ocean. In this work, MK8(H2) (purity: 99.75%) was collected through a convenient and economic extraction process followed by high-speed countercurrent chromatography (HSCCC) purification. Additionally, high-resolution mass spectrometry (HRMS) was performed for further identification and the hydrogenation position of MK8(H2) (terminal unit) was determined using nuclear magnetic resonance (NMR) for the first time. MK8(H2) showed a superior antioxidant effect and antiglycation capacity compared with ubiquinone Q10 and MK4. High-performance liquid chromatography⁻mass spectrometer (HPLC-MS/MS) and molecular docking showed the fine interaction between MK8(H2) with methylglyoxal (MGO) and bull serum albumin (BSA), respectively. These properties make MK8(H2) a promising natural active ingredient with future food and medicine applications.

Cretan tea (Origanum dictamnus L.) as a functional beverage: an investigation on antiglycative and carbonyl trapping activities.

Accumulation of advanced glycation end products (AGEs) in vivo is associated with many chronic disorders such as diabetes, renal failure, aging, and Alzheimer's disease. The aim of this study was to expand the knowledge about the functional properties of Origanum dictamnus L. beverage (Cretan tea) by an investigation about the inhibitory effects on the formation of AGEs and the capacity to trap dicarbonyl compounds. Dittany infusion was characterized for its polyphenolic composition by RP-HPLC-DAD-ESI/MSn and twenty compounds were detected. Its antiglycative property was evaluated by in vitro BSA-sugar (glucose, fructose, and ribose) and BSA-methylglyoxal (MGO) assays, tests for the formation of Amadori products and dicarbonyl compounds, and the direct glyoxal (GO) and MGO trapping capacity. The infusion showed the highest inhibitory effect on the formation of dicarbonyl compounds and AGEs (activity values range from 72-100%) and only a weak effect on the formation of Amadori products, indicating that the antiglycative action occurred primarily during the last two phases of the non-enzymatic glycation reaction. These activities are partially correlated with the antioxidant/antiradical activity, as demonstrated by the scavenger capacity against the ABTS cation and DPPH stable radicals, and the reducing power. The registered high anti-AGE capacity could probably be ascribed to the dittany polyphenolic composition particularly rich in flavone derivatives. These findings support further investigations to study the feasibility of dittany as an antiglycative agent in food or cosmetic preparation.

Phytochemicals, Anti-Inflammatory, Antiproliferative, and Methylglyoxal Trapping Properties of Zijuan Tea.

Zijuan tea (Camellia sinensis var. assamica) is a unique anthocyanin-rich tea cultivar in China. Although chemical component analysis of Zijuan tea and extraction technology of anthocyanins was widely documented, its functional properties have not been extensively explored. In this study, the anti-inflammatory, antiproliferative, and methylglyoxal (MGO) trapping activities of water extract (ZWE) and ethyl acetate extract (ZEE) of Zijuan tea were investigated for the 1st time. Results showed that ZWE and ZEE exhibited inhibitory effects on nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-6 production as well as inducible nitric oxide synthase protein (iNOS) expression in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Moreover, Zijuan tea extracts exerted stronger antiproliferative activity against HCT-116 cells compared with HepG2 and MDA-MB-231 cells, and thus could induce apoptosis in HCT-116 cells in a dose-dependent manner. Furthermore, Zijuan tea extracts were effective in trapping MGO under simulated physiological conditions, and the T1/2 (the time for 50% MGO remaining) values of ZWE and ZEE were 3.69 and 6.20 min, respectively. Additionally, the contents of total phenolics and catechins in ZEE were 685.43 ± 16.00 and 454.96 ± 4.21 mg/g extract, respectively, and in ZWE were 422.59 ± 12.09 and 307.29 ± 0.85 mg/g extract, respectively. Therefore, ZEE exhibited better anti-inflammatory, antiproliferative, and MGO trapping properties than ZWE may be mainly attributed to its higher (P < 0.05) content of total phenolics, expecially catechins. These results suggest that Zijuan tea could be a potential natural resource for the development of functional tea beverage. PRACTICAL APPLICATION: This study revealed that Zijuan tea extracts possessed anti-inflammatory, antiproliferative, and methylglyoxal trapping potentials in vitro. With high anthocyanins and polyphenols, Zijuan tea can be developed into a healthy tea beverage or used as a natural component to reduce the level of methylglyoxal in Maillard reaction.

Artichoke (Cynara cardunculus L. var. scolymus) waste as a natural source of carbonyl trapping and antiglycative agents.

The role of polyphenolic compounds extractable from artichoke solid wastes in the formation of advanced glycation end products (AGEs) was studied. Outer bracts and stems were extracted using different water-ethanol mixtures and HPLC-DAD analyses indicated aqueous and hydro-alcoholic 20:80 stem extracts as the richest in polyphenols. The samples were characterized in their phenolic composition (using mass spectrometry) and antioxidant capacity. Antiglycative capacity was evaluated by in vitro BSA-sugars (glucose, fructose, and ribose) and BSA-methylglyoxal (MGO) tests, formation of Amadori products assay, direct glyoxal (GO) and MGO trapping capacity. Results indicated both extracts as effective inhibitors of fructosamine formation and antiglycative agents. In particular, aqueous extract showed the best activity in the systems containing glucose and fructose, differently from ethanolic extract, that was demonstrated able to better inhibit AGEs formation when ribose or MGO act as precursors. Ethanolic extract was also shown to be able to trap MGO and GO, with efficiency increasing after 24hours of incubation time. These activities are partially correlated with the antioxidant effect of the extract, as demonstrated by the scavenger capacity against ABTS cation and DPPH stable radicals; this relationship is evident when the model system, containing protein incubated with ribose or MGO, is considered. The different activities of the tested extracts could probably be ascribed to the different composition in chlorogenic acids (CQAs), being aqueous extract richer in 1-CQA, 3-CQA, and 1,3-di-CQA, and ethanolic extract in 5-CQA, caffeic acid, 1,5-di-CQA. These findings support further investigations to study the stability of the different CQAs in simil-physiological conditions and the feasibility of artichoke waste as antiglycative agents in food or pharmacological preparations. CHEMICAL COMPOUNDS: 5-caffeoylquinic acid (PubChem CID 5280633); 3-caffeoylquinic acid (PubChem CID 1794427); 1-caffeoylquinic acid (PubChem CID 10155076); 1,3-di-caffeoylquinic acid (PubChem CID 24720973); 1,5 - di-caffeoylquinic acid (PubChem CID 122685); caffeic acid (PubChem CID 689043); apigenin-7-glucuronide (PubChem CID 5319484); methylglyoxal PubChem CID (880); aminoguanidine hydrochloride (PubChem CID 2734687).

Cellular antioxidant, methylglyoxal trapping, and anti-inflammatory activities of cocoa tea (Camellia ptilophylla Chang).

Cocoa tea (Camellia ptilophylla Chang) is a naturally low caffeine-containing but gallocatechin gallate (GCG)-rich tea cultivar, though its biological activities have not been extensively explored. Herein, we evaluated the in vitro cellular antioxidant, methylglyoxal trapping, and anti-inflammatory activities of water extract of green tea from cocoa tea (CWE) and Yunnan Daye tea (Camellia sinensis) (YWE), and their predominant bioactive components GCG and epigallocatechin gallate (EGCG) for comparative purposes. Primarily, CWE exhibited a higher cellular antioxidant potential than YWE via a cellular antioxidant activity assay, while no significant difference was observed between GCG and EGCG. Moreover, CWE was more potent than YWE in the inhibitory effect on inflammatory responses in lipopolysaccharide-induced RAW 264.7 macrophages, including nitric oxide and interleukin-6 productions. Additionally, CWE showed comparable MGO trapping ability to YWE, although EGCG was more reactive with MGO than with GCG. This study suggested that cocoa tea would be a promising and potential functional beverage as a dietary antioxidant, methylglyoxal trapping, and anti-inflammatory agent.

Combination of 1H NMR and chemometrics to discriminate manuka honey from other floral honey types from Oceania.

Manuka honey is a product produced essentially in New Zealand, and has been widely recognised for its antibacterial properties and specific taste. In this study, 264 honeys from New Zealand and Australia were analysed using proton NMR spectroscopy coupled with chemometrics. Known manuka markers, methylglyoxal and dihydroxyacetone, have been characterised and quantified, together with a new NMR marker, identified as being leptosperin. Manuka honey profiling using 1H NMR is shown to be a possible alternative to chromatography with the added advantage that it can measure methylglyoxal (MGO), dihydroxyacetone (DHA) and leptosperin simultaneously. By combining the information from these three markers, we established a model to estimate the proportion of manuka in a given honey. Markers of other botanical origins were also identified, which makes 1H NMR a convenient and efficient tool, complementary to pollen analysis, to control the botanical origin of Oceania honeys.

Phytochemicals from Camellia nitidissima Chi inhibited the formation of advanced glycation end-products by scavenging methylglyoxal.

The objective of this study was to investigate the inhibitory effects of Camellia nitidissima Chi (CNC) on the advanced glycation end-product (AGE) formation. CNC was extracted with ethanol and further separated into dichloromethane, ethyl acetate, n-butanol, and water soluble fractions. Ethyl acetate fraction had the highest total phenolic and quercetin content compared with other fractions. Sixteen phenolic compounds were identified using HPLC Triple TOF MS/MS. Bovine serum albumin (BSA)-glucose assay showed that dichloromethane and ethyl acetate fraction inhibited AGE formation by 88.1% and 87.5% at 2.5mg/mL. BSA-methylglyoxal assay showed that ethyl acetate fraction inhibited 54.1% AGE formation while dichloromethane fraction inhibited 28.1%. Over 96.0% of methylglyoxal was scavenged by different fractions within 12h. Both mono- and di-methylglyoxal quercetin adducts were identified after incubating quercetin with methylglyoxal using HPLC-ESI-MS(n). The results in this study suggest that CNC extracts inhibited AGEs formation in part through scavenging methylglyoxal by phenolic compounds.

[Inhibitory effects of extracts from Moutan Cortex on formation of advanced glycation end-products (AGEs) in vitro and specificity screening of its potential active components].

In this study, bovine serum albumin (BSA)/methylglyoxal (MGO) non-enzymatic glycosylation reaction system was used for the evaluation of the inhibitory effects of Moutan Cortex extracts on the formation of AGEs. The HPLC-LC-ESI-MS/MS technology was adopted to test and indentify active components in Moutan Cortex against AGEs formation. The different concentrations of extracts (crude herb concentration 50, 100, 150, 200, 250 g•L⁻¹) from Moutan Cortexwas determined by fluorospectrophotometry, indicating an activity against AGEs formation in different concentrations of extracts, the inhibition ratio were (36.2±5.3)%, (43.5±6.2)%, (55.4±7.8)%, (68.6±6.7)%, (70.4±8.2)%, respectively after 6-day reaction in a dose dependent manner. Besides, the forming speed of AGEs tended to be steady after 24 h reaction. The HPLC technology was used to analyze chromatograms before and after the incubation of Moutan Cortex and methylglyoxal, identify changes in five chromatographic peaks and show decrease or increase in chromatographic peaks. These substances were trigalloyl glucose, tetragalloyl glucose, galloylpaeoniflorin, hexagalloyl glucose and benzoylpaeoniflorin after LC-ESI-MS/MS identification. Extracts from Moutan Cortex showed the remarkable inhibitory effects against formation of AGEs in BSA/glucose system. Furthermore, these potential active components might be associated with the efficacy of Moutan Cortex on treatment of diabetic nephropathy, which enriches basic studies for Moutan Cortex and provides ideas and reference basis for subsequent studies.