Prevention of quercetin precipitation in red wines: a promising enzymatic solution

Flavonols are known for causing undesirable deposits in both red and white wines. Among flavonols, quercetin is widely considered the principal factor determining this phenomenon. One of the most accredited hypotheses claims that glycosylated derivatives of quercetin undergo hydrolysis of the glycosylic bond during the fermentation and the wine ageing, releasing quercetin aglycone, which is much less soluble in water solution and causes the precipitation. Our work describes the dynamics of quercetin-derived deposition in Chianti wines and purposes a new method, based on the enzymatic quercetin glycoside hydrolysis of the glycosidic bond, to prevent the unpleasant deposit formation during the wine ageing. In our study, forty-four monovarietal wines obtained from 7 different Italian grape varieties were compared in the content of total quercetin-3-glycosides (rutin, quercetin-3-glucuronide, quercetin-3-glucoside) and quercetin aglycone. The data confirmed the literature revealing Sangiovese as the richest in quercetin. We tested then, in a Sangiovese wine, four fining agents (PVPP, PVPP/PVI, bentonite and a vegetal protein) for quercetin removal, showing that only the PVPP had a modest aglycone removal activity. Then, the kinetics of deposit formation was studied in three Chianti wines which differed in the initial content of quercetin aglycone. This investigation highlighted that the chemical equilibrium of quercetin changes over time as the turbidity slowly increases, as previously documented. The comparison of the three dynamics also permitted us to conclude that different wines show a different ability to keep in solution quercetin. Finally, a new approach for deposit prevention was studied by a precocious Chianti wine treatment with a pectolytic enzyme having secondary glycosidase activity. This enzyme significantly accelerated the hydrolysis of glycosylated quercetins into their aglycone, which could enhance the deposition before bottling, without serious wine colour depletion. Our study represents the first evidence of the promising potential of using the pectolytic enzyme with secondary glycosidase activity to prevent quercetin deposit during Chianti ageing, in a way that is compatible with organic wine production.

In Vitro Expanded Bioaccessibility of Quercetin-3-Rutinoside and Quercetin Aglycone from Buckwheat Biscuits Formulated from Flours Fermented by Lactic Acid Bacteria

The expanded bioaccessibility of rutin (Ru) and quercetin (Q) from buckwheat biscuits (BBs) formulated from liquid-state fermented flours by selected lactic acid bacteria (LAB) were determined after gastrointestinal digestion. Fermentation of buckwheat flours caused a LAB-dependent variation in Ru and Q content. BBs baked at 220 °C for 30 min showed lower content of Ru and Q, and no correlation was found between the content of these compounds in fermented flours and BBs. The expanded bioaccessibility of Ru from BBs was low when its content in the soluble and insoluble fractions remaining after digestion in vitro was taken into account. Contrary results were found for Q bioaccessibility which had an index greater than 1, indicating the high Q bioaccessibility from BBs. Since very low Q content was noted in the insoluble fraction remaining after BBs digestion, the high Q bioaccessibility was determined to be due to its concentration in the soluble fraction.

Composition of rabbit caecal microbiota and the effects of dietary quercetin supplementation and sex thereupon

The purpose of this study was to add to the current understanding of rabbit caecal microbiota. This involved describing its microbial composition and linking this to live performance parameters, as well as determining the effects of dietary quercetin (Qrc) supplementation (2 g/kg feed) and sex on the microbial population. The weight gain and feed conversion ratio of twelve New Zealand White rabbits was measured from 5 to 12 wk old, blood was sampled at 11 wk old for the determination of serum hormone levels, and the rabbits were slaughtered and caecal samples collected at 13 wk old. Ion 16S^TM metagenome sequencing was used to determine the microbiota profile. The dominance of <em>Firmicutes</em> (72.01±1.14% of mapped reads), <em>Lachnospiraceae</em> (23.94±1.01%) and <em>Ruminococcaceae</em> (19.71±1.07%) concurred with previous reports, but variation both between studies and individual rabbits was apparent beyond this. Significant correlations between microbial families and live performance parameters were found, suggesting that further research into the mechanisms of these associations could be useful. Negative correlations with the caecal flavonoid content were found, but the latter was not affected by diet, and the effects of quercetin supplementation on the microbiota were very limited, possibly due to the absorption of the quercetin-aglycone from the gastrointestinal tract prior to the caecum. Nonetheless, <em>Clostridiales Family XIII. Incertae Sedis</em> was more abundant in the quercetin-supplemented rabbits (Control: 0.003±0.003%; Qrc: 0.020±0.000; <em>P</em>=0.005), as was the genus <em>Anaerofustis</em> (Control: 0.000±0.002; Qrc: 0.010±0.002; <em>P</em>=0.003). Serum cortisol levels were higher in females, and several microbial families differed between the sexes. Most were more abundant in female rabbits, including the most abundant, the family <em>Eubacteriaceae</em> (Male: 2.93±0.40; Female: 4.73±0.40; <em>P</em>=0.01).

Polyphenol Characterization in Red Beverages of Carapa procera (D.C.) Leaf Extracts

The red aqueous beverages of Carapa procera (D.C.) leaf extracts were investigated for their polyphenol contents using HPLC-DAD, HPLC-ESI-MS, and semipreparative HPLC. Polyphenols were extracted, clarified, and concentrated using a multistep process including ultrasound-assisted extraction (UAE), cross-flow microfiltration (CFM), and reverse osmosis (RO). On the basis of analytical and semipreparative chromatographic techniques, 12 phenolic compounds were identified and quantified for the first time: 2 anthocyanins (cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside), 5 phenolic acids (protocatechuic, the three caffeoylquinic isomers, and coumaroylquinic acid), and 5 flavonols (quercetin 3-O-rutinoside, quercetin 3-O-galactoside, quercetin 3-O-glucoside, quercetin aglycone, and kaempferol 3-O-rutinoside). From the concentrated extract, it was possible to recover for anthocyanins (28.4 ± 0.3 µmol L−1 cyanidin equivalents) the two glycosides of cyanidin, for flavonols (1587 ± 3 µmol L−1 quercetin equivalents) the two glycosides of quercetin and kaempferol, and for phenolic acids (3650 ± 10 µmol L−1 gallic acid equivalents) chlorogenic and protocatechuic acids.

Bioconversion of Kaempferol and Quercetin Glucosides from Plant Sources Using Rhizopus spp.

Fermentation with filamentous fungi is known for the ability to convert bioactive compounds. The aim of this research was to investigate the metabolism of glycosidic derivatives of kaempferol and quercetin during fungal fermentation of extracts from cauliflower outer leaves and onion by Rhizopus oryzae and R. azygosporus. The highest release of kaempferol and quercetin was observed after 2 days and 1 day of fermentation with R. oryzae, respectively. It was proposed that glycosidic compounds were initially deglycosylated to form kaempferol-3-glucoside and quercetin-3-glucoside and then further metabolized into their aglycones. Clear differences in conversion efficiency towards the aglycones were observed between the two Rhizopus strains. Although both flavonoids only differ in one hydroxyl group, the metabolism of the glycosides towards their respective aglycones, kaempferol or quercetin, was different. It is concluded that the fermentation with R. oryzae and R. azygosporus could be considered as a way to produce kaempferol and quercetin aglycone from their glycosidic derivatives.

Enrichment and Assessment of the Contributions of the Major Polyphenols to the Total Antioxidant Activity of Onion Extracts: A Fractionation by Flash Chromatography Approach

The present study extensively fractionated crude red onion extract in order to identify the polyphenols which contributed most in the total antioxidant capacity of the onion extract using a flash chromatography system. The flash separations produced 70 fractions which were tested for their total phenol content, total flavonoid content, and antioxidant capacities as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. Out of these 70 fractions, four fractions which were representatives of the four major peaks of the flash chromatograms, were further analysed for their constituent polyphenols using liquid chromatography tandem mass spectrometry (LC-MS/MS). The main contributor of onion antioxidant capacity is quercetin glycoside followed by quercetin aglycone although quercetin aglycone had higher antioxidant capacity than its glycosidic counterparts. High abundance of quercetin glycosides such as quercetin-3,4′-diglucoside and quercetin-4′-glucoside had compensated for their relatively low antioxidant capacities. A Higher degree of glycosylation resulted in lower antioxidant capacity. The fractionation approach also contributed in enrichment of the onion antioxidant polyphenols. A >9 folds enrichment was possible by discarding the early fractions (fractions 1–15) which contained the main bulk of the extracts, predominantly sugars.

Effect of Quercetin Monoglycosides on Oxidative Stress and Gut Microbiota Diversity in Mice with Dextran Sodium Sulphate-Induced Colitis

The pathogenesis of inflammatory bowel disease (IBD) is linked to an intricate association of environmental, microbial, and host-related factors. This study examined the potential effects of dietary addition of two preparations from onion, one comprising quercetin aglycone alone (Q: 0.15% polyphenols, quercetin aglycone:quercetin monoglycosides, 98:2) and another comprising quercetin aglycone with monoglycosides (Q+MQ: 0.15% total polyphenols, quercetin aglycone:quercetin monoglycosides, 69:31), on dextran sodium sulphate- (DSS-) induced colitis in mice. The results revealed a significant decrease in the body weight gain of the mice with DSS-induced colitis, which was counteracted by the dietary Q or Q+MQ supplementation. Meanwhile, the oxidative stress indicated by myeloperoxidase (MPO), reduced glutathione (GSH), malondialdehyde (MDA), and serum nitrate (NO) concentrations was higher in mice with DSS-induced colitis than in the control group mice, but dietary Q or Q+MQ supplementation counteracted this trend. The colitis mice demonstrated reduced Chao1, angiotensin-converting enzyme (ACE), and Shannon indices and an increased Simpson index, but the colitis mice receiving dietary Q or Q+MQ exhibited higher Chao1, ACE, and Shannon indices and a reduced Simpson index. In conclusion, this research showed that even at a low dose, dietary Q or Q+MQ supplementation counteracts DSS-induced colitis in mice, indicating that Q or Q+MQ may be used as an adjuvant therapy for IBD patients.

Determination of quercetin aglycone in Flos Sophorae Immaturus extract of (Sophora japonica L.) by High Performance Liquid Chromatography

The research has established a method to directly extract and determine free quercetin (aglycone form) from Flos Sophorae Immaturus methanol extract by using a simple HPLC method. Conducting experiment with system HPLC Agilent 1260 Infinity, reverse column ZORBAX SB-C18 (temperature 25oC), flow rate 0.5 ml/min, average pressure 30 and 35 bar, and diode array detector (DAD), we found that these parameters is suitbale: λmax = 370 nm, injection volume is 20 µl, analysis time 16 minutes, mobile phase (% volume) consists of methanol (15%), acetonitril (20%) and solvent C (65%, contains 1% acetic acid, methanol, acetonitril and H2O, 40%, 15% and 45% respectively. After using a combination of irocratic elution and standard addition, retention time of free quercetin in Flos Sophorae Immaturus methanol extract has found to be 8.84 ± 0,05 (min). Relative standard deviation (RSD) of retetion time, peak area and peak height have been less than 1%, this results have indicated that the proposed method has fullfilled the validation parameters such as selectivity/specitifity, precision/repeatability. This study provided useful information for screening activity of quercetin by using different methods.

Bioavailability of Quercetin

Quercetin is generally present as quercetin glycoside in nature and involves quercetin aglycone conjugated to sugar moieties such as glucose or rutinose. Quercetin has been reported to exhibit antioxidative, anti-carcinogenic, anti-inflammatory, anti-aggregatory and vasodilating effects. Unfortunately, quercetin bioavailability is generally poor and several factors affect its bioavailability. Quercetin bioavailability varies widely between individuals. Gender may affect quercetin bioavailability, but there is no clear evidence. There has been little research looking for the effects of age and vitamin C status on bioavailability of quercetin supplements, but there is no research seeking out the effects of age and vitamin C status on bioavailability of food-derived quercetin. Presence of sugar moieties increases bioavailability and differences in quercetin-conjugated glycosides affect bioavailability. For instance, onion-derived quercetin, which is mainly quercetin glucoside, is more bioavailable than apple-derived quercetin, which contains quercetin rhamnoside and quercetin galactoside. Quercetin is lipophilic compound, thus dietary fat enhances its bioavailability. Nondigestible fiber may also improve quercetin bioavailability. Quercetin bioavailability is greater when it is consumed as an integral food component. This study reviews and discusses factors affecting quercetin bioavailability.