Chemical basis and self-assembly mechanism of submicroparticles forming in chrysanthemum tea infusion.

Submicroparticles are important components generally existed in chrysanthemum tea infusion, but their functionality, chemical composition, structure and self-assembly mechanism are unclear due to lack of suitable preparation method and research strategy. This study showed that submicroparticles promoted the intestinal absorption of phenolics in chrysanthemum tea infusion by comparison of chrysanthemum tea infusion, submicroparticles-free chrysanthemum tea infusion and submicroparticles. Submicroparticles efficiently prepared by ultrafiltration mainly consisting of polysaccharide and phenolics accounted for 22% of total soluble solids in chrysanthemum tea infusion. The polysaccharide, which was determined as esterified pectin with a spherical conformation, provided spherical skeleton to form submicroparticles. A total of 23 individual phenolic compounds were identified in submicroparticles with the total phenolic content of 7.63 µg/mL. The phenolics not only attached to the external region of spherical pectin by hydrogen bonds, but also got into hydrophobic cavities of spherical pectin and attached to the internal region by hydrophobic interactions.

Combination of Lycium barbarum L. and Laminaria japonica polysaccharides as a highly efficient prebiotic: Optimal screening and complementary regulation of gut probiotics and their metabolites.

The combination of polysaccharides is an effective way to develop prebiotics with stable performance during processing and digestion for human wellness. However, there is little information on optimal screening and complementary regulation of compound polysaccharides. This study aimed to optimally select a combination of Lycium barbarum L. polysaccharide (LBP) and Laminaria japonica polysaccharide (LJP) as a highly efficient prebiotic to regulate the gut probiotics and their metabolites. Two LBPs characterized as rhamnogalacturonan I enriched pectins and two LJPs characterized as fucoidans were obtained by enzyme-assisted acid extraction at moderate and dramatic temperatures and combined in pairs to obtain 4 groups containing 4 proportional combinations. All combinations showed better prebiotic effects than individual LJP. The combination of LBP and LJP extracted at 50 °C at a ratio of 4:1 exhibited the strongest prebiotic effect. The optimal compound polysaccharide achieved superior effect and complementary function via LBP-targeted proliferation of Bifidobacterium, Lactobacillus, and Bacteroides and production of SCFAs and non-SCFA health-associated metabolites, LJP-targeted accumulation of butyrate-producing bacteria and corresponding metabolites, as well as synergistic effect of LJP and LBP at exact proportion. Our study provided theoretical and methodological guidance for optimal screening of compound polysaccharides as new prebiotics.

Rhamnogalacturonan I-Enriched Pectin, Flavonoids, and Alkaloids from Lotus Leaf Infusion in Regulating Glycolipid Absorption and Metabolism: Isolation, In Vitro Bioactivity Verification, and Structural Characterization.

Lotus leaf is effective in regulating glycolipid absorption and metabolism, but the roles of small-molecule compounds and polysaccharides are unknown. In this study, the small-molecule compounds including flavonoids, alkaloids, and polysaccharides were gradually isolated from lotus leaf infusion by multi-column chromatography and applied to in vitro activity verification and structural characterization. Although flavonoids and alkaloids were effective in inhibiting pancrelipase and α-glucosidase, polysaccharides more effectively bounded bile acids, inhibited cholesterol micelle solubility, and stimulated the growth of Bifidobacterium than lotus leaf infusion. Polysaccharides, presented as spherical conformation in water, were identified as rhamnogalacturonan I-enriched (93%) low-ester pectin with multiple branches mainly composed of arabinan, arabinogalactan-type II, and galactan formed by →3)-Galp-(1→, →5)-Araf-(1→ and →4)-Galp-(1→ residues. Polysaccharides, which were a key constituent of lotus leaf infusion in regulating glycolipid absorption and metabolism, should be paid more attention and developed as a functional food ingredient.

Pectin-type polysaccharide from galangal: An efficient emulsifier to construct the emulsion-based delivery system for galangal flavonoids.

Galangal is rich in flavonoids and polysaccharides but underutilized. In this study, galangal flavonoids and polysaccharides (GP-HN and GP-UN) were obtained by segmented extraction, used for chemical composition determination/structural characterization, and constructed for the emulsion delivery system. The results showed that galangin accounted for 71.45 % of total flavonoids. GP-HN and GP-UN were prepared by enzymatic-assisted high-temperature and ultrasonic extraction, which were low-molecular-weight pectin-type polysaccharides mainly constructed by galacturonic acid, galactose, and arabinose. GP-UN was the best emulsifier due to interfacial activities, emulsifying properties, interfacial resistance to bile salts displacement abilities, and anti-lipid digestion abilities of GPs. GP-UN emulsion could stably deliver flavonoids. This study presented a method for orderly reorganizing flavonoids and polysaccharides, guiding for utilization of whole bioactive components in galangal.

The hypoglycemic and hypolipemic potentials of Moringa oleifera leaf polysaccharide and polysaccharide-flavonoid complex.

In this study, Moringa oleifera leaf (MOL) flavonoids (MOLF) with strong α-glucosidase inhibitory activity and MOL polysaccharides (MOLP) with strong cholic acid-binding capacity were efficiently prepared by two-stage extraction method and mixed in a certain proportion for development of MOL highly-processed products with hypoglycemic and hypolipemic potentials. Quercetin-3-O-glucoside (6.86%) and kaempferol-3-O-glucoside (4.02%) were identified as the main components of MOLF. MOLP constructed by galactose, arabinose, rhamnose and galacturonic acid possessed the strongest effects on delaying glucose diffusion and dialysis, delaying starch digestion, binding bile acids and inhibiting cholesterol micelle solubility, being the best MOL highly-processed products for regulating carbohydrate and lipid digestion and absorption. MOLF and MOLP had synergistic effect on delaying glucose diffusion and dialysis, delaying starch digestion and binding bile acids, while MOLF impaired the inhibitory effect of MOLP on cholesterol micelle solubility. Compared with MOL primary-processed products including MOL powder and de-phenolic MOL powder, MOL highly-processed products including MOLP and MOLF-MOLP complex possessed stronger hypoglycemic/hypolipemic potentials.

Xanthine oxidase inhibitory activity and antihyperuricemic effect of Moringa oleifera Lam. leaf hydrolysate rich in phenolics and peptides.

ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. leaf (MOL), a rich source of protein and phenolics, was traditionally used to treat various diseases including headaches, fevers, sore throat and dyslipidemia. Recently, MOL was reported to possess antioxidant, anti-dyslipidemia and hepato-renal protective activities, indicating that MOL could become a potential agent to improve metabolic disorders associated with hyperuricemia. The antihyperuricemic effect of MOL hydrolysate (MOLH) with high contents of phenolics and peptides remains unknown. AIM OF THE STUDY: The aim of this study is to investigate xanthine oxidase (XO) inhibitory activity of MOLH, to clarify phenolic and peptide profiles of MOLH, and to evaluate possible mechanism underlying the antihyperuricemic effect of MOLH. MATERIALS AND METHODS: MOLH was prepared by enzymatic hydrolysis using commercial trypsin. XO inhibitory activity was determined by XO reaction-UPLC-MS coupling method. The chemical profiles of the phenolic and peptide fractions of MOLH were determined by UPLC-QTOF-MS/MS. The antihyperuricemic effect of MOLH was evaluated in a potassium oxonate-induced hyperuricemic rat model at doses of 200 and 500 mg/kg. Serum uric acid (UA), urea nitrogen, creatinine (CRE), triglyceride (TG), total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol levels, serum XO activity, liver malondialdehyde (MDA) equivalent level, renal tumor necrosis factor-α and interleukin-1ß levels, and protein expression of renal urate-anion transporter 1, glucose transporter 9 and ATP-binding cassette transporter G2 were determined. RESULTS: The phenolic and peptide fractions played key roles in inhibiting XO activity and blocking uric acid production. Five flavonoids and sixteen polypeptides were identified in the phenolic and peptide fractions of MOLH, respectively. MOLH (200 and 500 mg/kg) could effectively reduce the serum UA level of hyperuricemic rats (p < 0.001) by regulation of serum XO activity (p < 0.05 at 200 mg/kg, p < 0.01 at 500 mg/kg) and renal urate transporters. Besides, MOLH could improve metabolic disorders associated with hyperuricemia by its multiple actions on liver MDA (p < 0.001), serum CRE (p < 0.05 at 500 mg/kg) and serum TG (p < 0.001). CONCLUSION: The results provided scientific evidence that MOLH rich in phenolics and peptides ameliorated hyperuricemia and metabolic disorders. This study validated the potential use of MOLH for regulation of hyperuricemia.

Physicochemical Characterization of Hizikia fusiforme Polysaccharide and Its Hypoglycemic Activity via Mediating Insulin-Stimulated Blood Glucose Utilization of Skeletal Muscle in Type 2 Diabetic Rats.

In the current study, a functional polysaccharide fraction (HFP) was obtained from Hizikia fusiforme by ultrasound-assisted enzymatic extraction, and its structural characterization and hypoglycemic activity and potential molecular mechanism were investigated. The results indicated that HFP with high uronic acid was a heterogeneous polysaccharide composed of six monosaccharides. Congo red test explained that HFP had no triple helix conformation. AFM analysis revealed that HFP was spherical particle with flame-like aggregates and multiple strands closely arranged. Rheological analysis showed that HFP exhibited shear-thinning flow behavior. HFP significantly ameliorated diabetes-related symptoms and serum profiles and increased muscle glycogen storage in rats. HFP administration at 400 mg/kg body weight/day displayed greater advantages than metformin in controlling the levels of fasting blood glucose, triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bile acid (TBA) of diabetic rats. Intervention of HFP up-regulated markedly the expression of AMPK-α, GLUT4, PI3K and Akt in skeletal muscle of diabetic rats at the mRNA and protein levels, revealing hypoglycemic effects of HFP may be related closely to improving insulin resistance and mitochondrial function of skeletal muscle.

Action mechanisms and interaction of two key xanthine oxidase inhibitors in galangal: Combination of in vitro and in silico molecular docking studies.

Galangal extract (GE)-based hypouricemic functional food is under-developed due to ambiguous quality control standard that is closely associated with action mechanisms and interaction of key xanthine oxidase (XO) inhibitors (kaempferide and galangin) in GE. In terms of kinetics analysis, fluorescence quenching and molecular docking, kaempferide and galangin showed similar docking posture to xanthine in molybdopterin center, and formed flavonol-XO complexes driven by hydrogen bonding, hydrophobic interaction and van der Waals force, competitively inhibiting XO. Kaempferide, had stronger binding affinity for XO and three more hydrogen bonds with XO than galangin, interacting with critical amino acid residues (Arg880 and Glu802) in catalysis reaction of XO and showing stronger XO inhibitory activity than galangin. The combination of kaempferide and galangin enhanced their binding affinities for XO, showing synergistic inhibition on XO at optimal molar ratio 1:4 that could be quality control standard for GE. This study provided new insights into structure-XO inhibitory activity relationship of methoxylated flavonoids and quality control standard for GE-based hypouricemic functional food.

Identifying mechanisms underlying the amelioration effect of Chrysanthemum morifolium Ramat. 'Boju' extract on hyperuricemia using biochemical characterization and UPLC-ESI-QTOF/MS-based metabolomics.

This study was aimed at evaluating the prospect of edible chrysanthemum extract as a potential substance for the prevention and treatment of hyperuricemia. Chrysanthemum morifolium Ramat. 'Boju' extract (CBE), which had the strongest xanthine oxidase inhibitory activity, showed a significant hypouricemic effect on potassium oxonate-induced hyperuricemic rats through inhibiting serum xanthine oxidase activity, regulating renal uric acid transport-related protein (ABCG2, URAT1 and GLUT9) expression and blood lipid levels, and protecting renal function. Serum metabolomics based on UPLC-ESI-QTOF/MS was used to illustrate mechanisms underlying the amelioration effect of CBE on hyperuricemia. A total of 35 potential biomarkers were identified. CBE prevented the pathological process of hyperuricemia by regulating 16/17 biomarkers associated with tryptophan, sphingolipid, glycerophospholipid and arachidonic acid metabolisms. CBE could alleviate hyperuricemia-related diseases including chronic kidney disease, hyperlipidemia and inflammation via reducing indoxyl sulfate, lysophosphatidylcholines and arachidonic acid levels, exhibiting its applicability and superiority in the treatment of hyperuricemia.

Classification of edible chrysanthemums based on phenolic profiles and mechanisms underlying the protective effects of characteristic phenolics on oxidatively damaged erythrocyte.

In this study, phenolic profiles of chrysanthemums derived from five main species were determined for characterization of rationality of their application in tea, beverages and functional foods. A total of 41 phenolics including 3 phenolic acids, 17 flavones, 9 flavanones, 1 dihydroflavonol, 4 flavonols, 4 chalcones and 3 aurones were identified. The contents of 22 characteristic phenolics were simultaneously determined. Chrysanthemum morifolium cv. 'Qiju' (with abundant phenolics) and Coreopsis tinctoria Nutt. (with unique and abundant flavonoid aglycones and glycosides), exhibited excellent cellular antioxidant activities and strong market potentials. Chlorogenic acid, 3,5-dicaffeoylquinic acid and luteolin-7-O-glucoside largely contributing to cellular antioxidant activity of 'Qiju' by forming protective membrane around erythrocyte should be markers for quality control of 'Qiju'. Okanin, the gut microbial-produced metabolite of marein, possessed strong protective effect on oxidatively damaged erythrocyte via incorporating in membrane and entering cytoplasm. Okanin and marein should be markers for quality control of C. tinctoria.

In Vitro Digestion and Fermentation of Three Polysaccharide Fractions from Laminaria japonica and Their Impact on Lipid Metabolism-Associated Human Gut Microbiota.

Our previous study has proved that the three polysaccharide fractions from L. japonica (LP-A4, LP-A6, and LP-A8) had significantly different structure characterization. Herein, we conducted in vitro simulated digestion and fermentation to study the digestive mechanism of LP-As. The results of gastrointestinal digestion indicated that LP-A6 and LP-A8 would be easier to trap the enzyme molecules for their denser interconnected macromolecule network compared with LP-A4. Fermentation of LP-As by human gut microbiota, especially for LP-A8, generated a large amount of short-chain fatty acids (SCFAs), which could upregulate the abundance of Firmicutes ( Lachnoclostridium and Eubacterium). The high content of sulfate and highly branched sugar residue of LP-A8 might help it be easily used by Firmicutes in gut microbiota of hyperlipidemic patients. Functional analysis revealed that the increased metabolic activities of glycerophospholipid metabolism, ether lipid metabolism, and fatty acid metabolism induced by LP-A8 treatment were closely associated with metabolic syndromes and hyperlipidemia.

Interaction of ß-conglycinin with catechin-impact on physical and oxidative stability of safflower oil-in-water emulsion.

The study aimed at improving the antioxidant activity of ß-conglycinin to enhance the oxidative and physical stabilities of safflower oil-in-water emulsion stabilized by ß-conglycinin. Heating promoted binding affinity and antioxidant activity of ß-conglycinin. Catechin and chlorogenic acid showed higher binding affinities towards unheated (or heated) ß-conglycinin than caffeic acid and quercetin. The enhancement efficiencies of the phenolics on the antioxidant activity of unheated (or heated) ß-conglycinin decreased in the order of catechin > quercetin > chlorogenic acid > caffeic acid. Hydrophobic force and hydrogen bonding were the important binding forces for the selected phenolics to ß-conglycinin. The complexation with catechin has no side effect on interfacial behavior and emulsifying property of ß-conglycinin. The use of heated ß-conglycinin-catechin complex as an emulsifier for preparing safflower oil emulsion effectively improved the oxidative and physical stabilities of the emulsion treated with lipoxygenase through inhibition of lipid oxidation, protein carbonyl formation and sulfhydryl loss.

Identification of the free phenolic profile of Adlay bran by UPLC-QTOF-MS/MS and inhibitory mechanisms of phenolic acids against xanthine oxidase.

Adlay bran free phenolic extract has been previously demonstrated to possess potent xanthine oxidase (XOD) inhibitory activity. The aims of this study were to characterize the free phenolic profile of adlay bran and investigate the structure-activity relationship, underlying mechanism and interaction of phenolic acids as XOD inhibitors. A total of twenty phenolics including ten phenolic acids, two coumarins, two phenolic aldedhyes and six flavonoids were identified in a phenolic compound-guided separation by UPLC-QTOF-MS/MS. Adlay bran free phenolic extract possessed strong XOD inhibitory activity related to hydroxycinnamic acids with methoxyl groups. The hydrogen bonding and hydrophobic interactions were the main forces in the binding of adlay phenolics to XOD. Sinapic acid, identified in adlay bran for the first time, possessed strong XOD inhibitory activity in a mixed non-competitive manner, and synergistic effects with other adlay phenolic acids at low concentrations, and would be a promising agent for preventing and treating hyperuricemia.

Comparison Study on Polysaccharide Fractions from Laminaria japonica: Structural Characterization and Bile Acid Binding Capacity.

Our previous study has suggested that the crude polysaccharide obtained from Laminaria japonica by acid assisted extraction (LP-A) have significant bile acid-binding capacity, which probably ascribed to its specific structure characterization. The relationship between structure characterization and bile acid-binding capacity of the purified LP-A fractions are still unknown. This paper conducted a comparison study on the structure characterization and bile acid-binding capacity of three LP-A fractions (LP-A4, LP-A6, and LP-A8). The results indicated that LP-A4, LP-A6, and LP-A8, characterized as mannoglucan, fucomannoglucan, and fucogalactan, had significantly different structure characterization. Furthermore, the bile acid-binding capacity of LP-A8 was obviously higher than the other fractions, which may be attributed to its highly branched structure, abundant sulfate, fucose, and galactose in chemical composition and denser interconnected macromolecule network in molecular morphology. This study provides scientific evidence for the potential utilization of LP-A8 as an attractive functional food supplement candidate for the hyperlipidemia population.

A comparison study on polysaccharides extracted from Laminaria japonica using different methods: structural characterization and bile acid-binding capacity.

In this study, the structural characterization and bile acid-binding capacity of Laminaria japonica polysaccharides (LP), obtained by seven different extraction methods, were investigated. The results indicated that extraction methods exhibited significant effects on extraction yield, molecular weight, monosaccharide composition and the content of neutral sugar, fucose, uronic acid and sulfate of LP. AFM analysis indicated that LP extracted by different methods exhibited certain different, flexible and worm-like chains with many branches. Rheological measurements showed that the LP, obtained by pressurized hot water extraction and acid assisted extraction, exhibited lower viscosity due to their lower molecular weight, compared to other extracted polysaccharides. The bile acid-binding capacity of acid assisted extracted LP was significantly higher than other LP samples tested, which was probably ascribed to its highly branched structure, low molecular weight, low viscosity and abundant uronic acid and fucose in total monosaccharides. The present study provides scientific evidence and advances in the preparation technology and a method for evaluating hypolipidemic activities of L. japonica polysaccharides.

Enrichment of antioxidants in black garlic juice using macroporous resins and their protective effects on oxidation-damaged human erythrocytes.

The black garlic juice is popular for its nutritive value. Enrichment of antioxidants is needed to make black garlic extract an effective functional ingredient. Five macroporous resins were evaluated for their capacity in adsorbing antioxidants in black garlic juice. XAD-16 resin was chosen for further study due to its high adsorption and desorption ratios. Pseudo-second-order kinetics (qe=625µmol Trolox equiv/g dry resin, k2=0.0001463) and Freundlich isotherm models (ΔH=-10.1547kJ/mol) were suitable for describing the whole exothermic and physical adsorption processes of the antioxidants from black garlic juice on XAD-16 resin. The antioxidants and phenolics were mostly enriched in 40% ethanol fraction by XAD-16 resin column chromatography. The black garlic extract and its fractions could protect erythrocytes against AAPH-induced hemolysis in dose-dependent manners. The pretreatment of AAPH-damaged erythrocytes with 40% ethanol fractions (2.5mg/mL) significantly decreased the hemolysis ratios from 53.58% to 3.79%. The 40% ethanol fraction possessing strong intracellular antioxidant activity could be used as a functional food ingredient.

Effect of Soy Sauce on Serum Uric Acid Levels in Hyperuricemic Rats and Identification of Flazin as a Potent Xanthine Oxidase Inhibitor.

This is the first report on the ability of soy sauce to effectively reduce the serum uric acid levels and xanthine oxidase (XOD) activities of hyperuricemic rats. Soy sauce was partitioned sequentially into ethyl acetate and water fractions. The ethyl acetate fraction with strong XOD inhibition effect was purified further. On the basis of xanthine oxidase inhibitory (XOI) activity-guided purification, nine compounds including 3,4-dihydroxy ethyl cinnamate, diisobutyl terephthalate, harman, daidzein, flazin, catechol, thymine, genistein, and uracil were obtained. It was the first time that 3,4-dihydroxy ethyl cinnamate and diisobutyl terephthalate had been identified from soy sauce. Flazin with hydroxymethyl furan ketone group at C-1 and carboxyl at C-3 exhibited the strongest XOI activity (IC50 = 0.51 ± 0.05 mM). According to fluorescence quenching and molecular docking experiments, flazin could enter into the catalytic center of XOD to interact with Lys1045, Gln1194, and Arg912 mainly by hydrophobic forces and hydrogen bonds. Flazin, catechol, and genistein not only were potent XOD inhibitors but also held certain antioxidant activities. According to ADME (absorption, distribution, metabolism, and excretion) simulation in silico, flazin had good oral bioavailability in vivo.

Adsorption and desorption characteristics of adlay bran free phenolics on macroporous resins.

In this study, the adsorption and desorption characteristics of six macroporous resins including XAD-7HP, XAD-16, HP-20, HP-2MGL, SP-207 and SP-825 for enrichment of adlay bran free phenolics were studied. XAD-16, SP-207 and SP-825 were chosen for further study due to their strong adsorption and desorption capacities. XAD-16, SP-207 and SP-825 had similar phenolics adsorption/desorption behaviors. Pseudo-second-order kinetics model and Freundlich isotherm model were suitable for describing the whole exothermic and physical adsorption processes of adlay bran free phenolics on XAD-16, SP-207 and SP-825. After treatment with gradient elution on XAD-16 resin column, the free phenolics were mostly enriched (from 89.61 to 1015.26mg/100g) in 50% ethanol fraction. The oxygen radical absorbance capacity of 50% ethanol fraction was eight times higher than that of the crude extract. Therefore, the production of highly concentrated phenolics might expand the application of adlay bran used as a bioactive ingredient in functional food.

Absorption and desorption behaviour of the flavonoids from Glycyrrhiza glabra L. leaf on macroporous adsorption resins.

The kinetics of adsorption and desorption behaviours of five macroporous resins for enriching flavonoids from Glycyrrhizaglabra L. leaf were investigated. All five resins showed similar and effective adsorption and desorption properties. A pseudo-second-order kinetics model was suitable for evaluating the whole adsorption process. Additionally, two representative resins (XAD-16 and SP825) were chosen for adsorption thermodynamics study. The adsorption of the representative resins was an exothermic and physical adsorption process. Further column chromatography of XAD-16 and SP825 showed that the total flavonoids (from 16.8% to 55.6% by XAD-16 and to 53.9% by SP825) and pinocembrin (from 5.49% to 15.2% by XAD-16 and to 19.8% by SP825) were enriched in 90% ethanol fractions. Meanwhile, the antioxidant capacities and nitrite-scavenging capacities were 2-3times higher than those of the crude extract. The fractions with high flavonoid and pinocembrin contents could be used as biologically active ingredients in functional food.

Bioactive profiles, antioxidant activities, nitrite scavenging capacities and protective effects on H2O2-injured PC12 cells of Glycyrrhiza glabra L. leaf and root extracts.

This study compared the total flavonoid content of Glycyrrhiza glabra L. leaf and root extracts. Results suggested that the total flavonoid content in the leaf extract was obviously higher than that in the root extract. Pinocembrin, the main compound in the leaf extract after purification by column chromatography, showed good antioxidant activity and nitrite scavenging capacity, but moderate inhibitory effect on mushroom tyrosinase. Liquiritin was the main compound in root extract and possessed strong inhibitory effect on mushroom tyrosinase. Both compounds exhibited significant protection effect on H2O2-injured PC12 cells at a low concentration. These results indicate that Glycyrrhiza glabra L. leaf is potential as an important raw material for functional food.