isoquercitrin and spiraeoside

The flavonoid quercetin is an antioxidant which occurs in foods mainly as glycosides. The sugar moiety in quercetin glycosides affects their bioavailability in humans. Quercetin-3-rutinoside is an important form of quercetin in foods, but its bioavailability in humans is only 20% of that of quercetin-4'-glucoside. Quercetin-3-rutinoside can be transformed into quercetin-3-glucoside by splitting off a rhamnose molecule. We studied whether this 3-glucoside has the same high bioavailability as the quercetin-4'-glucoside. To that end we fed five healthy men and four healthy women (19-57 y) a single dose of 325 micromol of pure quercetin-3-glucoside and a single dose of 331 micromol of pure quercetin-4'-glucoside and followed the plasma quercetin concentrations. The bioavailability was the same for both quercetin glucosides. The mean peak plasma concentration of quercetin was 5.0+/-1.0 micromol/L (+/-SE) after subjects had ingested quercetin-3-glucoside and 4.5+/-0.7 micromol/L after quercetin-4'-glucoside consumption. Peak concentration was reached 37 +/-12 min after ingestion of quercetin-3-glucoside and 27+/-5 min after quercetin-4'-glucoside. Half-life of elimination of quercetin from blood was 18.5+/-0.8 h after ingestion of quercetin-3-glucoside and 17.7+/-0.9 h after quercetin-4'-glucoside. We conclude that quercetin glucosides are rapidly absorbed in humans, irrespective of the position of the glucose moiety. Conversion of quercetin glycosides into glucosides is a promising strategy to enhance bioavailability of quercetin from foods.

Topics: Administration, Oral; Adult; Area Under Curve; Chromatography, High Pressure Liquid; Female; Half-Life; Humans; Male; Middle Aged; Parasympatholytics; Plant Extracts; Quercetin; Structure-Activity Relationship

Since the intake of quercetin glucosides has healthy benefits, the analysis of quercetin glucosides in food is useful. The electrochemical determination of individual quercetin glucosides (quercetin-3-glucoside (Q3G), quercetin-4'-glucoside (Q4'G), and quercetin-3,4'-diglucoside (Q34'G)) in food is carried out. For the detection of quercetin glucosides, a long-length carbon nanotube electrode offers attractive properties such as well-defined current peaks, high sensitivity, and high reproducibility. Cyclic voltammetry (CV) demonstrates distinct and specific peak currents: the oxidation peaks at +0.37, +0.45, and +0.78 V are assigned to the catechol group in the B-ring of Q3G, the 3-hydroxy group in the C-ring of Q4'G, and the resorcinol group in the A-ring of both Q4'G and Q34'G, respectively. Currents, which are determined by CV, of individual quercetin glucosides at the peak potential are proportional to the concentrations of onion, apple peel, and tartary buckwheat, which show good agreement with those obtained by high-performance liquid chromatography.

Topics: Electrochemical Techniques; Electrodes; Fagopyrum; Food Analysis; Glucosides; Malus; Nanotubes, Carbon; Onions; Oxidation-Reduction; Quercetin; Reproducibility of Results; Sensitivity and Specificity

Quercetin is a plant-derived flavonoid and its cytotoxic properties have been widely reported. However, in nature, quercetin predominantly occurs as various glycosides. Thus far the cytotoxic activity of these glycosides has not been investigated to the same extent as quercetin, especially in animal models. In this study, the cytotoxic properties of quercetin (1), hyperoside (quercetin 3-O-galactoside, 2), isoquercitrin (quercetin 3-O-glucoside, 3), quercitrin (quercetin 3-O-rhamnoside, 4), and spiraeoside (quercetin 4'-O-glucoside, 5) were directly compared in vitro using assays of cancer cell viability. To further characterize the influence of glycosylation in vivo, a novel zebrafish-based assay was developed that allows the rapid and experimentally convenient visualization of glycoside cleavage in the digestive tract. This assay was correlated with a novel human tumor xenograft assay in the same animal model. The results showed that 3 is as effective as 1 at inhibiting cancer cell proliferation in vivo. Moreover, it was observed that 3 can be effectively deglycosylated in the digestive tract. Collectively, these results indicate that 3 is a very promising drug candidate for cancer therapy, because glycosylation confers advantageous pharmacological changes compared with the aglycone, 1. Importantly, the development of a novel and convenient fluorescence-based assay for monitoring deglycosylation in living vertebrates provides a valuable platform for determining the metabolic fate of naturally occurring glycosides.

Topics: Animals; Flavonoids; Glucosides; Glycosides; Glycosylation; HCT116 Cells; Humans; Molecular Structure; Quercetin; Structure-Activity Relationship; Vertebrates; Zebrafish

Quercetin and quercetin glycosides are widely consumed flavonoids found in many fruits and vegetables. These compounds have a wide range of potential health benefits, and understanding the bioavailability of flavonoids from foods is becoming increasingly important.. This study combined an in vitro digestion, a lactase treatment and the Caco-2 cell model to examine quercetin and quercetin glucoside uptake from shallot and apple homogenates.. The in vitro digestion alone significantly decreased quercetin aglycone recovery from the shallot digestate (p < 0.05), but had no significant effect on quercetin-3-glucoside recovery (p > 0.05). Digestion increased the Caco-2 cell uptake of shallot quercetin-4'-glucoside by 2-fold when compared to the non-digested shallot. Despite the loss of quercetin from the digested shallot, the bioavailability of quercetin aglycone to the Caco-2 cells was the same in both the digested and non-digested shallot. Treatment with lactase increased quercetin recovery from the shallot digestate nearly 10-fold and decreased quercetin-4'-glucoside recovery by more than 100-fold (p < 0.05), but had no effect on quercetin recovery from apple digestates. Lactase treatment also increased shallot quercetin bioavailability to the Caco-2 cells approximately 14-fold, and decreased shallot quercetin-4'-glucoside bioavailability 23-fold (p < 0.05). These Caco-2 cells had lactase activity similar to that expressed by a lactose intolerant human.. The increase in quercetin uptake following treatment with lactase suggests that dietary supplementation with lactase may increase quercetin bioavailability in lactose intolerant humans. Combining the digestion, the lactase treatment and the Caco-2 cell culture model may provide a reliable in vitro model for examining flavonoid glucoside bioavailability from foods.

Topics: Animals; Caco-2 Cells; Chromatography, High Pressure Liquid; Digestion; Fruit; Glucosides; Humans; Hydrogen-Ion Concentration; Lactase; Malus; Onions; Pepsin A; Quercetin; Shallots; Swine

Quercetin is a typical antioxidative flavonoid found in vegetables, which is more commonly present as its glucosides, quercetin-3-glucoside (Q3G) and quercetin-4'-glucoside (Q4'G). The main aim of this study was to estimate the antioxidant activity of Q3G and Q4'G on iron ion-driven lipid peroxidation of the gastrointestinal mucosa. Q4'G markedly suppressed the lipid peroxidation when rat gastrointestinal mucosa homogenates were incubated with Fe(NO3)3 and ascorbic acid. Its effectiveness was greater as compared to that of Q3G and comparable to that of quercetin aglycone. Furthermore, Q4'G yielded higher amounts of quercetin aglycone than Q3G on incubation with the homogenates. However, Q4'G showed a lower chelating activity in comparison to Q3G. These results indicate that Q4'G, even though it has a low chelating activity, because of its efficient conversion to antioxidative aglycone on exposure to the mucosa, can act as a powerful antioxidant on iron ion driven lipid peroxidation in the intestinal mucosa. Thus, vegetables rich in Q4'G, such as onion, are likely to serve as favorable antioxidant sources for suppressing iron-induced oxidative stress in the intestinal tract.

Topics: Animals; Antioxidants; Intestinal Mucosa; Iron; Lipid Peroxidation; Male; Quercetin; Rats; Rats, Wistar

Quercetin is an important dietary flavonoid with putative beneficial effects in the prevention of cancer and CVD. The in vivo bioactivity of quercetin depends on its bioavailability, which varies widely between foods. We used an in situ rat intestinal perfusion model to study whether differential small intestinal hydrolysis of the sugar moiety of five naturally occurring quercetin glycosides determines the small intestinal uptake and subsequent biliary excretion of quercetin. After 30 min perfusion, a decrease of intact quercetin glycoside in perfusate was observed for quercetin-3-O-ss-glucoside (20.9 (sem 1.4) micromol/l) and quercetin-4'-O-ss-glucoside (23.5 (sem 1.6) micromol/l), but not of quercetin-3-O-ss-galactoside, quercetin-3-O-ss-rhamnoside and quercetin-3-O-alpha-arabinopyranoside. Appearance of free quercetin in perfusate and conjugated quercetin metabolites (quercetin, isorhamnetin, and tamarixetin) in portal and peripheral plasma and bile were also significantly greater after treatment with quercetin-3-O-ss-glucoside or quercetin-4'-O-ss-glucoside compared with any of the other glycosides. Thus, the type of sugar moiety is a major determinant of the small intestinal absorption of quercetin glycosides, but the position (3 or 4') of the glucose moiety does not further influence absorption. The poor bioavailability of important dietary quercetin glycosides has implications for their in vivo bioactivities.

Topics: Animals; Bile; Biological Availability; Diet; Glycosides; Intestinal Absorption; Intestine, Small; Male; Perfusion; Portal Vein; Quercetin; Rats; Rats, Wistar

Recent experimental data point to an interaction of dietary flavonol monoglucosides with the intestinal Na-dependent glucose transporter 1 (SGLT1). To investigate this interaction in more detail, we performed experiments with SGLT1-containing brush-border-membrane vesicles (BBMV) from pig jejunum. The flavonol quercetin-3-O-glucoside (Q3G) concentration-dependently inhibited Na-dependent uptake of radioactively labelled d-glucose into BBMV. Uptake of l-leucine was not inhibited by Q3G, indicating a specific interaction of the glucoside with SGLT1. Whereas the maximal transport rate of concentration-dependent initial glucose uptake was not altered in the presence of Q3G, the constant for half-maximal glucose uptake was increased, suggesting a competitive type of inhibition of glucose uptake by Q3G. Trans-stimulation experiments suggested the transport of Q3G via SGLT1. In addition, Q3G decreased the Na-independent diffusive uptake of glucose into BBMV. Other flavonoids were also tested for their inhibitory effect on d-glucose uptake. Among the tested quercetin glycosides, only the 4'-O-glucoside (Q4G) also inhibited Na-dependent glucose uptake into BBMV, whereas the 3-O-galactoside, the 3-O-glucorhamnoside and the aglycone quercetin itself were ineffective. Glucosides of some other flavonoid classes such as naringenin-7-O-glucoside, genistein-7-O-glucoside and cyanidin-3,5-O-diglucoside were ineffective as well. Thus, dietary quercetin monoglucosides, for example, Q3G and Q4G, have an impact on intestinal nutrient transporters such as SGLT1 and related systems.

Topics: Animals; Biological Transport; Diet; Glucose; Glucosides; Intestinal Mucosa; Jejunum; Leucine; Membrane Glycoproteins; Microvilli; Monosaccharide Transport Proteins; Quercetin; Sodium-Glucose Transporter 1; Swine

Two hypotheses on absorption mechanisms of flavonoid glucosides across the small intestine have been proposed: active uptake of the quercetin glucoside by the sodium-dependent glucose transporter (SGLT1) with subsequent deglycosylation within the enterocyte by cytosolic beta-glucosidase, or luminal hydrolysis of the glucoside by lactase phlorizin hydrolase (LPH) and absorption by passive diffusion of the released aglycone. To test the above hypotheses we employed phlorizin (as an inhibitor of SGLT1) and N-(n-butyl)-deoxygalactonojirimycin (as an inhibitor of the lactase domain of LPH) in a rat everted-jejunal sac model. Quercetin-4'-glucoside mucosal hydrolysis was 10 times greater than quercetin-3-glucoside hydrolysis in the absence of inhibitors (449 and 47 nmol g(-1) tissue, respectively), despite the similar amounts (13+/-4 and 9+/-1 nmol g(-1), respectively) being transferred to the serosal compartment during the 15 min incubation. Apical hydrolysis of both quercetin glucosides was significantly reduced in the presence of NB-DGJ (80%), and transfer of quercetin (measured as quercetin metabolites) to the serosal solution was also significantly reduced (40-50%). In the presence of phlorizin, transfer of metabolites to the serosal solution was only reduced in the case of quercetin-4'-glucoside. Evidently the mechanism of absorption of quercetin-4'-glucoside involves both an interaction with SGLT1 and luminal hydrolysis by LPH, whereas quercetin-3-glucoside appears to be absorbed only following hydrolysis by LPH.

Topics: Analysis of Variance; Animals; Intestinal Absorption; Intestine, Small; Lactase-Phlorizin Hydrolase; Male; Membrane Glycoproteins; Monosaccharide Transport Proteins; Quercetin; Rats; Rats, Wistar; Sodium-Glucose Transporter 1

Recently it has been postulated that flavonoid glucosides may be absorbed by the small intestine via the SGLT-1. Therefore, we applied an in vitro mucosal uptake method to investigate mucosal uptake of the non-metabolisable glucose analogue methyl-alpha-D-glucopyranoside (MDG) as influenced by quercetin-3-glucoside (isoquercitrin) and quercetin-4'-glucoside (spiraeosid). We found that both glucosides significantly inhibited SGLT-1-mediated MDG uptake whereas the aglycon quercetin or the quercetin-3-rhamnoglucoside (rutin) were ineffective. Calculated apparent kinetic parameters (Km, Vmax) of initial Na+-dependent MDG uptake by the jejunal mucosa indicate a competitive type of inhibition.

Topics: Alanine; Animals; In Vitro Techniques; Intestinal Absorption; Intestinal Mucosa; Jejunum; Kinetics; Membrane Glycoproteins; Methylglucosides; Monosaccharide Transport Proteins; Quercetin; Rats; Rats, Wistar; Rutin; Sodium; Sodium-Glucose Transporter 1

The nature of quercetin conjugates present in blood after consumption of quercetin glucosides is still unclear. In this study, we analyzed plasma of volunteers that had consumed 325 micromol of either quercetin-3-glucoside or quercetin-4'-glucoside as an oral solution. Quercetin metabolites were extracted with acetonitrile/phosphoric acid and these extracts were analyzed using a high performance liquid chromatography with Coularray detection that distinguishes between the glucuronidated and the glucosylated forms of quercetin. No intact quercetin glucosides and only trace amounts of aglycone were found in human plasma, irrespective of the glucoside ingested. This was confirmed by spiking the plasma with glucoside standards. The major components in plasma had the same retention time as quercetin glucuronide standards. These plasma components disappeared after treatment of the plasma with bovine liver beta-glucuronidase, under reformation of quercetin, and showed the same oxidation pattern as the glucuronides. These results suggest that after consumption of quercetin glucosides, quercetin glucuronides are major metabolites in plasma.

Topics: Acetonitriles; Biological Availability; Chromatography, High Pressure Liquid; Glucosides; Glucuronides; Intestinal Absorption; Kinetics; Phosphoric Acids; Quercetin

Lactase phlorizin hydrolase (LPH; EC 3.2.1.62) is a membrane-bound, family 1 beta-glycosidase found on the brush border of the mammalian small intestine. LPH, purified from sheep small intestine, was capable of hydrolysing a range of flavonol and isoflavone glycosides. The catalytic efficiency (k(cat)/K(m)) for the hydrolysis of quercetin-4'-glucoside, quercetin-3-glucoside, genistein-7-glucoside and daidzein-7-glucoside was 170, 137, 77 and 14 (mM(-1) s(-1)) respectively. The majority of the activity occurred at the lactase and not phlorizin hydrolase site. The ability of LPH to deglycosylate dietary (iso)flavonoid glycosides suggests a possible role for this enzyme in the metabolism of these biologically active compounds.

Topics: Animals; beta-Galactosidase; Flavonoids; Flavonols; Glycosides; Intestinal Absorption; Intestinal Mucosa; Intestine, Small; Isoflavones; Kinetics; Lactase; Lactase-Phlorizin Hydrolase; Lactose; Mammals; Microvilli; Phlorhizin; Quercetin; Sheep; Substrate Specificity

Flavonoids are polyphenolic plant secondary metabolites with antioxidant and other biological activities potentially beneficial to health. Food-borne flavonoids occur mainly as glycosides, some of which can be absorbed in the human small intestine; however, the mechanism of uptake is uncertain. We used isolated preparations of rat small intestine to compare the uptake of the quercetin aglycone with that of some quercetin glucosides commonly found in foods, and investigated interactions between quercetin-3-glucoside and the intestinal hexose transport pathway. The nature of any metabolism of quercetin and its glucosides during small intestinal transport in vitro was determined by HPLC. The presence of quercetin-3-glucoside in the mucosal medium suppressed the uptake of labeled galactose by competitive inhibition and stimulated the efflux of preloaded galactose. Quercetin-3-glucoside and quercetin-4'-glucoside, but not quercetin-3,4'-diglucoside, were transported into everted sacs significantly more quickly than quercetin aglycone. Intact quercetin glucosides were not detected in mucosal tissue or within the serosal compartment, but both free quercetin and its metabolites were present, mainly as quercetin-3-glucuronide and quercetin-7-glucuronide. Evidently, quercetin derived from quercetin-3-glucoside passes across the small intestinal epithelium more rapidly than free quercetin aglycone. Monoglucosides of quercetin interact with the sodium-dependent glucose transporter. During passage across the epithelium, quercetin-3-glucoside is rapidly deglycosylated and then glucuronidated.

Topics: Analysis of Variance; Animals; Biological Transport; Chromatography, High Pressure Liquid; Galactose; Intestine, Small; Male; Monosaccharide Transport Proteins; Quercetin; Rats; Rats, Wistar

The efficiency of intestinal absorption and metabolic conversion of quercetin aglycone and its glucosides, quercetin-4'-O-beta-D-glucoside (Q4'G), quercetin-3-O-beta-D-glucoside (Q3G), and quercetin-3,4'-di-O-beta-D-glucoside (Q3,4'G), was estimated by using Caco-2 cell monolayers as an intestinal epithelial cell model. Aglycone was significantly lost from the apical side, resulting in the appearance of free and conjugated forms of quercetin and those of isorhamnetin in the cellular extracts. In the basolateral solution, the conjugated form of quercetin was predominant and increased with the elapse of incubation. As compared with quercetin aglycone, none of the quercetin glucosides were absorbed efficiently from apical side. However, Q4'G yielded conjugated quercetin and isorhamnetin in basolateral solution at higher amounts than Q3G or Q3,4'G. Lipophilicity of Q4'G was found to be higher than that of Q3G or Q3,4'G. This suggests that lipophilicity contributes to the relatively efficient absorption of Q4'G. It is likely that the occurrence of hydrolysis enhances the efficiency of intestinal absorption and metabolic conversion of dietary quercetin glucosides.

Topics: Caco-2 Cells; Cell Extracts; Cell Polarity; Flavonols; Humans; Intestinal Absorption; Intestinal Mucosa; Quercetin