Food Research International 67 (2015) 349–355

The microbial degradation of onionflavonol glucosides and their roasting products by the
human gut bacteria Eubacterium ramulus and Flavonifractor plautii
Katrin Ulbrich a, Nicole Reichardt b, Annett Braune c, Lothar W. Kroh d, Michael Blaut c, Sascha Rohn
a,⁎
a Institute of Food Chemistry, Hamburg School of Food Science, University of Hamburg, Grindelallee 117, D-20146 Hamburg,
Germany b Microbiology Group, Rowett Institute of Nutrition and Health, University of Aberdeen, Greenburn Road, Bucksburn,
Aberdeen AB21 9SB, UK c Department of Gastrointestinal Microbiology, German Institute of Human Nutrition
Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, D-14558 Nuthetal, Germany d Chair of Food Chemistry and Analysis,
Institute of Food Technology and Food Chemistry, Technische Universität Berlin, Gustav Meyer Allee 25, D-13355 Berlin,
Germany
a r t i c l e i n f o a b s t r a c t Article history: Received 10 October 2014 Accepted 25 November 2014 Available online 3
December 2014
Keywords: Onions Quercetin glycosides Microbial degradation Gastrointestinal microbiota Thermal induced degradation
⁎ Corresponding author. Tel.: +49 40 42838 7979; fax: +49 40 42838 4342.
E-mail address: rohn@chemie-uni-hamburg.de (S. Rohn).
Flavonoids are important constituents of the human diet. One source for flavonols, a major subclass of the flavo- noids, is onion.
It contains high amounts of quercetin glycosides, primarily quercetin-3,4′-di-O-glucoside (QDG) and
quercetin-4′-O-monoglucoside (Q-4′-MG). Due to their high reactivity flavonols are susceptible to thermal degradation as used in
food processing. Especially boiling and roasting influence the flavonoid content of food products. Quercetin and several of its
glycosides may serve as substrates for human gut bacteria. For example, Eubacterium ramulus and Flavonifractor plautii are
capable of cleaving the aglycone quercetin to form 3,4- dihydroxyphenylacetic acid (DHPAA) and phloroglucinol which to some
extent can be degraded further. The aim of this study was to find out whether E. ramulus and F. plautii are also capable of
degrading Q-4′-MG and QDG by and to investigate the influence of a thermal treatment (roasting) of the onion glucosides on the
subse- quent microbial degradation. In this study,E. ramuluswas capable of degrading Q-4′-MG and QDG, whileF. plautii was
not. Roasting of QDG at 180 °C for 5 min led to the formation of quercetin with Q-4′-MG and quercetin-3-O- monoglucoside
(Q-3-MG) as intermediates. Roasting accelerated the microbial degradation of Q-4′-MG and QDG. In the case of F. plautii,
microbial degradation was induced by quercetin which was formed during roasting and is a preferred substrate of this organism.
© 2014 Elsevier Ltd. All rights reserved.
1. Introduction
Flavonoids are polyphenolic compounds found in significant amounts in plants, especially in fruits and vegetables, and are
therefore important constituents of the human diet. In recent years they have gained attention due to their antioxidant,
antibacterial, and even anticarcinogenic proper- ties, which has led to an enormous increase in research for their cancer and CVD
preventive potential (Del Rio et al., 2013; Scalbert, Manach, Morand, Remesy, & Jimenez, 2005). However, bioavailability of
flavonoids is still discussed controversially, as diverse mechanisms may lead to an uptake of flavonoids, either by passive
diffusion or actively by trans- porters (Day, Gee, DuPont, Johnson, & Williamson, 2003; Hollman, de Vries, van Leeuwen,
Mengelers, & Katan, 1995). When taking a closer look at the multitude of absorption studies, recovery rates of flavonoids in
plasma and urine in most cases make up far less than 50% of the actual intake, depending on the chemical structure of the
flavonoids and the food matrix (Arts, Sesink, Faassen-Peters, & Hollman, 2004; Manach, Williamson, Morand, Scalbert, &
Remesy, 2005; Williamson & Manach,
2005). Consequently, the majority of flavonoids reach the colon with its diverse microbiota (Hooper & Gordon, 2001).
The complex composition of the colonic bacterial community varies between subjects, and is actually unique for each
individual (Blaut, Schoefer, & Braune, 2003). Flavonoids serve as substrates for the human gut microbiota and can be
transformed by various bacterial spe- cies (Blaut et al., 2003). The formation of metabolites, in most cases small phenolic acids,
depends on specific abilities of different bacterial strains to cleave certain chemical bonds. While some species utilize the sugar
moiety of flavonoid glycosides and do not degrade the aglycon (Schneider, Schwiertz, Collins, & Blaut, 1999), others are capable
of cleaving aglycons, but not their corresponding glycosides (Winter, Moore, Dowell, & Bokkenheuser, 1989; Winter, Popoff,
Grimont, & Bokkenheuser, 1991). Some are even capable of degrading both struc- tural components (Blaut et al., 2003).
With regard to vegetable consumption, onion (Allium cepa) is a flavonol-rich food with concentrations of flavonol glycosides
of up to approximately 1 g per kg edible part (Hertog, Hollman, & Katan, 1992; Rhodes & Price, 1996). Most of the 20 to 25
detectable flavonols in onion bulbs are present as glycosides of the aglycon quercetin (Slimestad, Fossen, & Vågen, 2007). The
glucosides quercetin-3,4′-di-O-
http://dx.doi.org/10.1016/j.foodres.2014.11.051 0963-9969/© 2014 Elsevier Ltd. All rights reserved.
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 glucoside (QDG) and quercetin-4′-O-monoglucoside (Q-4′-MG) are dom- inant in the onion flavonol profile. They make up to
approximately 80% of all phenolic compounds of the onion bulb and are generally found in higher concentrations in the outer
onion skin layers (Beesk et al., 2010). So far, in all human intervention studies the impact of food process- ing on the
physiological properties of flavonols has not been taken into account comprehensively, even though most of our plant-derived
food is boiled, fried or deep fried prior to consumption. Although some human intervention studies have been performed with
fried onions, be- cause of its higher acceptability to subjects, only the flavonols but not their degradation products have been
analyzed in these studies (e.g. de Vries et al., 1998; Hollman et al., 1995; Hollman et al., 1997).
However, it has been shown that thermal treatment processes lead to considerable degradation of flavonol glycosides (Hirota,
Shimoda, & Takahama, 1998; Ioku et al., 2001; Lombard, Peffley, Geoffriau, Thompson, & Herring, 2005; Makris & Rossiter,
2001; Price, Bacon, & Rhodes, 1997). Depending on the processing parame- ters (e.g. temperature, water content, pH value,
treatment time) and the chemical structure of the glycosides, smaller degradation products are formed that might have different
physiological properties than the origi- nal flavonol (Buchner, Krumbein, Rohn, & Kroh, 2006; Makris & Rossiter, 2000; Rohn,
Buchner, Driemel, Rauser, & Kroh, 2007).
The aim of the present study was to investigate the microbial degradability of the major onion quercetin glucosides by human
gut bacteria after thermal treatment. For this purpose, QDG was roasted and subjected to fermentation by two species
(Eubacterium ramulus and Flavonifractor plautii) isolated from the human gut microbiota whose ability to degrade flavonoids is
well known. The fate of the flavo- nols and formation of metabolites were analyzed using HPLC-DAD.
2. Materials and methods
2.1. Materials
Quercetin and 3,4-dihydroxyphenylacetic acid (DHPAA) were obtained from Sigma-Aldrich Chemie GmbH, Taufkirchen,
Germany. Quercetin-3-O-glucoside (isoquercitrin) was from Extrasynthese SA, Genay, France. The purity of both compounds
was N97% as checked by HPLC analysis. Quercetin-4′-O-monoglucoside and quercetin-3,4′-di-O- glucoside were isolated from
onions as described (Rohn et al., 2007). HPLC solvents were purchased from Roth (Karlsruhe, Germany), and were of HPLC
grade quality.
2.2. Roasting
For the roasting experiments 1 mL of a 1 mM methanolic flavonol glycoside solution was dried under nitrogen. The roasting
was carried out in glass ampoules at 180 °C in a behrotest ET1 thermoblock (Behr Labortechnik, Germany). After roasting,
samples were re-dissolved in methanol for HPLC analysis and in DMSO for the fermentation experiments.
2.3. Organisms
E. ramulus strain wK1 (DSM 16296) was previously isolated from a human fecal sample by Schneider et al. (1999) and F.
plautii strain I2 by Schoefer, Mohan, Schwiertz, Braune, and Blaut (2003). Originally F. plautii was named Clostridium
orbiscindens. Because of its similar bio- chemical properties, phylogenetic position, DNA content and DNA-DNA hybridization
with Eubacterium plautii, they were unified to the species F. plautii (Carlier, Bedora-Faure, Kouas, Alauzet, & Mory, 2010). To
culti- vate these strains, the anoxic techniques of Hungate (1969) and Bryant (1972) were used.
350 K. Ulbrich et al. / Food Research International 67 (2015) 349–355
2.4. Degradation experiments with E. ramulus and F. plautii
Degradation experiments were carried out anaerobically in 16 mL tubes fitted with butyl rubber stoppers and screw caps. The
tubes contained 9.8 mL Wilkins-Chalgren Anaerobe (WCA) broth (Oxoid). The medium was gassed with N
2
/CO
2
(80/20; v/v) and autoclaved at 121 °C for 15 min. After autoclaving, 100 μL of a stock
solution (50 mM each in DMSO) of quercetin, Q-4′-MG, QDG or QDG (roasted for 5 min and 8 min) was added. The media
were inoculated with 100μL of an over- night grown culture of either E. ramulus or F. plautii and incubated anaer- obically at 37
°C in a rotator. Immediately after inoculation and then every 3 h for 24 h a sample of 1 mL was taken and centrifuged at 14,000 x
g for 5 min. Five hundred μL of the supernatant and the pellet were lyophilized, re-dissolved in methanol, centrifuged for 5 min
at 14,000 ×g, and subse- quently analyzed by HPLC.
2.5. Degradation experiments using human fecal slurries
Fecal samples were collected from one healthy volunteer who had no previous history of gastrointestinal disorders and had
not undergone antibiotic therapy within 6 months prior to the study. Fresh fecal sam- ples were diluted 10-fold (w/v) in
pre-reduced phosphate-buffered saline (PBS, containing per L: 8.5 g NaCl, 0.3 g KH
2
PO
4
, 0.6 g Na
2
HPO
4
, 0.1 g peptone, 0.25 g cysteine × HCl) and centrifuged for 1
min at 300 ×g and 4 °C to remove large particles. The supernatant was centri- fuged for 5 min at 14,000 ×g and 4 °C and the
resulting bacterial pellet was washed twice with pre-reduced PBS. Fermentations were carried out in triplicate as described above
for the pure cultures.
2.6. Analysis of the flavonol glycosides and their microbial metabolites using HPLC-DAD
The Agilent HPLC-System consisted of a quaternary pump (1050 se- ries), autosampler (1050 series) and a DAD (1100
series). A gradient elution based on Rohn et al. (2007) using acidified water (A; water/ acetic acid; 99.5/0.5, v/v) and acetonitrile
(B) was carried out on a 250 × 4.6 mm i.d., 5 μm, Fluofix 120E column (Wako Pure Chemical In- dustries, Osaka, Japan)
connected to a 10 × 4.6 mm i.d. guard column of the same material. Gradient elution was performed as follows: 0% B (5 min);
0–4% B in 4 min; 4–2% B in 1 min; 2–4% B in 5 min; 4–8% B in 15 min; 8–22% B in 15 min; 22–28% B in 5 min; 28% B (5
min); 28– 45% B in 10 min; 45–0% B in 1 min; at a flow rate of 1.0 mL/min and a column temperature of 30 °C. The detection
was performed simul- taneously at 325 nm, 350 nm and 280 nm. Spectra were recorded from 200 to 500 nm. All compounds
were identified by their retention times and UV spectra in comparison to reference substances. Calibration curves were used for
quantification.
2.7. Statistical analysis
The fermentation experiments were repeated three times and the standard deviation was calculated. A maximum of ±10%
standard deviation from the averaged values was generally obtained. The aver- aged values along with standard deviations are
documented in the re- spective figures.
3. Results and discussion
3.1. Degradation of onion flavonol glucosides by E. ramulus
The degradability of several flavonoids and their glycosides by E. ramulus was studied previously (Braune, Gütschow, Engst,
& Blaut, 2001; Schneider et al., 1999). It had already been known that this organ- ism is capable of degrading flavonoid aglycons
such as quercetin or luteolin as well as some selected flavonol glycosides (Schneider & Blaut, 2000). In the present study it was
tested whether E. ramulus is
capable of degrading the flavonol glycosides present in onion, primarily quercetin-4′-O-monoglucoside and
quercetin-3,4′-di-O-glucoside.
In previous experiments, growing cells of E. ramulus converted 0.5 mM quercetin completely to equimolar amounts of
DHPAA within 10 h of incubation (data not shown), being similar to results from Braune et al. (2001) A pathway for the
degradation of flavonol aglycons (Fig. 1) was proposed by Braune et al. (2001). Accordingly, the degrada- tion of quercetin starts
with a reduction to form taxifolin prior to C-ring fission. After passing through the chalcone structure alphitonin is formed
followed by a decarboxylation step which results in the formation of DHPAA and phloroglucinol as cleavage products; the latter
is degraded further, mainly to short chain fatty acids (Braune et al., 2001). However, cleavage of the aglycon by E. ramulus is
strictly dependent on the presence of free glucose in the medium. Furthermore, E. ramulus is able to utilize the glucose moiety in
the 3-O and the 7-O-position, but not in the 5-O- position (Schneider & Blaut, 2000). However, a utilization of glucose in
4′-O-position (as in Q-4′-MG), even in combination with a glycosylation in 3-O-position (as in QDG), has not been shown so far.
In the present study, 0.5 mM Q-4′-MG was fully degraded within 24 h of incubation with DHPPA as the main product (Fig. 2A).
The concentration of QDG de- creased even more rapidly compared to pure Q-4′-MG (Fig. 2B). After 10 h of incubation, only
trace levels of QDG were detectable. How- ever, formation of nearly equimolar amounts of DHPAA (0.4 mM) only occurred
after 22 h. The fermentation kinetics show that the monoconjugated quercetin-4′-O-monoglucoside (Q-4′-MG) is formed during
the degradation of QDG as an intermediate. Q-4′-MG reaches its concentration maximum after 10 h (Fig. 2B). Quercetin was
only detect- able in trace levels whereas quercetin-3-O-monoglucoside (Q-3-MG) was not detected during the fermentation, at
all. This finding suggests that glucose in 3-O-position is more accessible to the glucosidase activity of E. ramulus compared to
glucose in 4′-O-position. Quercetin as a possible intermediate was described in experiments with cell ex- tracts of E. ramulus
(Schneider et al., 1999). There are two possible expla- nations for the fact that quercetin was only detectable in traces: As the
aglycon seems to be the preferred substrate for E. ramulus, quercetin released during the fermentation of either QDG, Q-4′-MG,
or Q-3-MG, is degraded immediately. Additionally, phenolic compounds are highly re- active substances that might undergo
several chemical reactions such as oxidative degradation or covalent addition to other compounds such as proteins and enzymes
(Kroll, Rawel, & Rohn, 2003). It is also conceivable that the deglycosylation occurs within the cell where the resulting agylcon
undergoes further cleavage without being released into the fer- mentation medium. With regard to the importance of E. ramulus,
it can
Fig. 1. Mechanism of the microbial quercetin degradation by E. ramulus according to Braune et al. (2001).
351 K. Ulbrich et al. / Food Research International 67 (2015) 349–355
Fig. 2. A and B. Fermentation of (A) quercetin-4′-O-glucoside (Q-4′-MG) and (B) quercetin-3,4′-di-O-glucoside (QDG) with E.
ramulus. Q = quercetin; DHPAA = 3,4-dihydroxyphenylacetic acid.
be stated that this organism is a strict anaerobe, ubiquitous resident in the human intestinal tract (Simmering, Kleessen, & Blaut,
1999). Simmering et al. (1999) developed an innovative 16S rRNA-targeted oli- gonucleotide probe for fluorescence in situ
hybridization to investigate the occurrence of the flavonoid-degrading bacterium E. ramulus in the human intestinal tract. In
2002, Simmering, Pforte, Jacobasch, & Blaut performed a human intervention study, where they investigated the influence of
different dietary flavonoids on the fecal population of the flavonoid-degrading bacterium E. ramulus. At hand of twenty-eight
healthy subjects they showed that after a significant decrease of the total fecal microbiota during the first 3 days of the
intervention period,
the oral intake of the flavonoids resulted in a significant increase in the fecal E. ramulus population. The relative proportion of E.
ramulus rose from minimally 0.2% to 6.9% of the total microbiota on day 8.
3.2. Degradation of onion flavonol glucosides by F. plautii
C. orbiscindens (referred to as F. plautii), which is capable of cleaving the C-ring of flavonoids, had been known as an
asaccharolytic organism (Winter et al., 1989; Winter et al., 1991). Schoefer et al. (2003) tested the degradability of flavonols
conjugated with diverse sugar moieties at varying positions of the aglycon skeleton and showed that this species is not capable of
degrading flavonols with sugar moieties at the 3-O, 5-O or 7-O-position. Glycosidation at the 4′-O-position was not investigated
in that study (Schoefer et al., 2003).
In the present study it was demonstrated that F. plautii is capable of degrading the aglycon structure quercetin confirming
previous results. However, neither its mono conjugated derivatives Q-4′-MG and Q-3- MG nor QDG were degraded. Their
concentrations remained unchanged throughout the whole fermentation period of 24 h (data not shown). F. plautii is dependent
on other enzyme activities, primarily deglycosyl- ating activities of human tissues such as those found in the small intestine and
liver (Nemeth et al., 2003) and other flavonoid degrading bacteria such as E. ramulus, Enterococcus casseliflavus (Schneider et
al., 1999), Clostridium perfringens (Zhang et al., 2014), or Bacteroides spp. (Bokkenheuser, Shackleton, & Winter, 1987). In
contrast to E. ramulus, E. casseliflavus and Bacteroides spp. deglycosylate the flavonoid only to take advantage of the sugar
moieties. The aglycon is not used any further by these species and becomes available for organisms such as F. plautii.
Schoefer et al. (2003) developed the species-specific 16S rRNA- targeted oligonucleotide probe for fluorescence in situ
hybridization probe “C.orb0179” and tested human fecal samples in order to estimate the prevalence of F. plautii in the human
intestinal tract. It was detected in the feces of 8 from 10 subjects investigated. The cell numbers ranged from 1.87 × 108 to 2.50 ×
109 cells per gram dry feces, corresponding to a mean count of 4.40 × 108 cells per g. The numbers determined are equivalent to
0.12% of the total number of fecal bacteria.
3.3. Thermal degradation of onion flavonol glucosides
According to the literature flavonol glycosides are highly susceptible to a thermal degradation depending on roasting time and
temperature (Rohn et al., 2007). Under the roasting conditions applied here, QDG is strongly degraded. In the present experiment
only 20% of the initial QDG was still detectable after 5 min (Fig. 3). The aglycon quercetin was the major end product of this
degradation process. Prior to the formation of quercetin, deglycosylation of the diglycoside occurs and leads to the intermediary
monoglucosides Q-4′-MG and Q-3-MG (Fig. 3). Whereas Q-4′-MG accumulates during roasting, Q-3-MG is not
352 K. Ulbrich et al. / Food Research International 67 (2015) 349–355
Fig. 3. Roasting of QDG (5 min) followed by the fermentation with E. ramulus.
detectable at all or only at trace levels, because the sugar moiety attached to the C-ring of the flavan skeleton is more susceptible
to the deglycosyl- ation than the sugar conjugated at the B-ring. In Fig. 4 the different path- ways of the thermally induced QDG
degradation are illustrated. To summarize the above mentioned results it can be stated: Step 1 is slower than step 2; step 3 is
slower than steps 1 and 4, with step 4 being the fastest, as no Q-3-MG was detectable in the roasting mixture. The aglycon
quercetin did not undergo further degradation under such conditions and its concentration remained unchanged (Figs. 3 and 4).
Only a few minutes of thermal treatment, which corresponds to a homelike heat treatment of onions, affected the concentrations
of QDG, Q-4′-MG, Q-3- MG, and quercetin, significantly, and thereby possibly influences microbi- al degradation.
3.4. Microbial fermentation of onion flavonol glucosides roasted for 5 min
Compared to the fermentation of unroasted QDG (Fig. 2B), the fermentation of the QDG roasting mixture started with a
different pro- file of flavonol compounds: As mentioned above, after 5 min of roasting, only 20% of the initial QDG amount was
left and intermediary Q-4′-MG as well as quercetin was present (Fig. 3). Quercetin, apparently the most preferred substrate for E.
ramulus, was nearly completely degraded after 3 h of incubation and not detectable anymore after 9 h (Fig. 3). Similar to the
unroasted sample, Q-4′-MG accumulated during the degradation of the QDG roasting mixture. Even after 10 h it was still present
at 50% of its maximum concentration and it was still detectable after 24 h (Fig. 3).
As already mentioned, F. plautii is not capable of degrading glycosyl- ated flavonoids. Indeed, among the compounds formed
during roasting of QDG, only quercetin was degradable. The concentrations of Q-4′-MG, although also formed during roasting,
and of QDG remained unchanged during the incubation (Fig. 5). However, the main fermentation product, DHPAA, was formed
at an almost twofold higher molar concentration compared to that of quercetin. A possible explanation for this might be that the
degradation of phenolic compounds depends not only on the source but also on the region of the colon where they are fermented
(van Dorsten et al., 2012). Additionally, it has to be considered that flavonoids might be also degraded non-microbiologically
during the fermentation. Such reactions are depending on pH value and chemical structure of the phenolic compound: neutral and
alkaline pH-values lead to an oxidative cleavage of the flavonoid skeleton resulting in the formation of several smaller
breakdown products such as protocatechuic acid. Further, also reaction products of the oxidized flavonoid (e.g.
2,2,5,7-tetrahydroxybenzofuran-3-on) can occur (Buchner et al., 2006). These compounds might also act as substrates for the
microorganisms.
3.5. Fermentation of roasted onion flavonol glucosides by human fecal samples
To better mimic the in vivo situation, roasted as well as unroasted QDG were subjected to fermentation by human fecal
slurries (Figs. 6 and 7). At the initial time point of the fermentation of unroasted QDG, trace levels of quercetin, Q-4′-MG, as
well as Q-3-MG were detected. The bacteria in this human fecal sample degraded the QDG completely within 2 h (Fig. 6). In
contrast to the degradation experiments with pure cultures of E. ramulus, Q-4′-MG did not accumulate when in- cubated with
fecal microbiota, but decreased continuously and was completely gone after 2 h. However, Q-3-MG and quercetin were the main
degradation intermediates, whose concentration increased with maxima between 1 and 2 h. After that time point, Q-3-MG was
degrad- ed at a high rate and not detectable anymore after 2 h, whereas querce- tin was gone within 6 h (Fig. 6). From these
results it can be concluded that a complex human gut microbiota consists of several organisms capable of deglucosylating
flavonol glucosides with prevalence for hy- drolysis of the sugar moiety at the 4′-O-position of the aglycon.
When investigating the microbial degradation of a roasted QDG mix with a complex human fecal community, it was observed
that prior to the addition of roasted QDG, traces of Q-4′-MG, Q-3-MG were initially present suggesting that the traditional
Western diet consumed by the donor contains various flavonols. Immediately after adding the roasted QDG mix to the fecal
slurry and subsequent mixing, QDG was not de- tectable anymore, although small amounts had been detectable after roasting
(Fig. 7). In contrast to the degradation of unroasted QDG by the fecal sample, in the QDG roasting mixture all quercetin
glucosides were degraded within 2 h. Quercetin was in this case degraded much faster and totally degraded after 2 h, as well.
This can be explained by the so-called cross-feeding. Certain bacteri- al species in the colon only survive by consuming
breakdown products produced by other microbial population groups such as lactic acid or succinate (Scott, Gratz, Sheridan, Flint,
& Duncan, 2013). One strain par- tially degrades the primary energy resource, in this case the sugar moi- ety of the glucosides,
and excretes an intermediate that is used as an energy source by other microorganisms whose growth is stimulated even though
they cannot use the original phenolic compounds as sub- strates. It has been shown that fermentation of polyphenols stimulated
proliferation of Bifidobacteria and decreased the ratio of Firmicutes to Bacteroidetes in an in vitro mixed culture model of human
intestinal microbiota (Parkar, Trower, & Stevenson, 2013). But also vice versa, it seems that the addition of probiotic strains such
as Lactobacillus
Fig. 5. Fermentation of roasted QDG (5 min) with F. plautii.
Fig. 4. Mechanism of the thermal degradation of QDG and respective formation of quercetin.
353 K. Ulbrich et al. / Food Research International 67 (2015) 349–355
plantarum might influence the metabolism of phenolic compounds (Barroso et al., 2013).
4. Conclusion
Roasting conditions led to a thermal degradation of quercetin glyco- sides. The mechanism involves a deglycosylation
(thermohydrolysis) to the corresponding aglycon, the kinetics of which depends on roasting time and temperature. In the case of
QDG, the aglycon is formed via for- mation of Q-4′-MG as monoglycosidic intermediate, which underlines the importance of the
kind and position of the sugar moiety. During the first minutes of the roasting at 180 °C Q-4′-MG accumulates. With regard to
the physiological properties of quercetin glycosides, it is not important whether the 3-O- or the 4′-O-glucoside is ingested
(Olthof, Hollman, Vree, & Katan, 2000), as both have the same absorption rate, but Day et al. (2003) showed that the
4′-O-glucoside is more susceptible to luminal hydrolysis by brush border enzymes such as the lactase phlorizin hydrolase. The
aglycon is subsequently released and absorbed by passive diffusion and/or may exert positive effects on the mucosa as
hypothesized by Murota et al. (2004): They found the 4′-O-glucoside in the intestinal mucosa had a higher antioxidant activity on
iron ion driven lipid peroxidation, even though it has a lower chelating ability compared to Q-3-MG. Thermal processes are often
assessed as critical in food production (e.g. formation of acrylamide, loss of vitamins).
Fig. 6. Fermentation of QDG with a human fecal sample.
Here, it seems that the roasting of the flavonol glycosides has beneficial effects. Compared to the compound predominantly
present in onions, QDG, the more bioavailable Q-4′-MG and quercetin were formed. Even more importantly, both are preferred
substrates for selected microorgan- isms in the colon, leading to the hypothesis that cross-feeding mecha- nisms may support gut
microbiota diversification. Some researchers even proposed flavonoids to have prebiotic effects, as the gastrointestinal
bioconversion of flavonoids might stimulate growth of bacterial species such as Bifidobacteria (Kawabata, Sugiyama, Sakano, &
Ohigashi, 2013; Parkar et al., 2013). Brown et al. (2012) evaluated the effect of a combina- tion of an in vitro-simulated upper
intestinal tract digestion and subse- quent fecal fermentation. They showed that over a physiologically- relevant dose range, the
digested and fermented berry extracts used, still provided significant antigenotoxic, antimutagenic and antiinvasive activity on
colonocytes. With regard to antioxidant activity, it is highly de- pending on the substrate fermented. In the present study,
comparatively higher antioxidant active compounds such as Q-4′-MG and quercetin have been formed as intermediates. When
they arefinally degraded, a de- cline of antioxidant can be proposed, as shown by Peng et al. (2014). However, DHPAA has a
comparatively lower antioxidant capacity, but as it is present in quite amounts, it might also contribute to the overall anti- oxidant
capacity of the colonic lumen (Jaganath, Mullen, Lean, Edwards, & Crozier, 2009).
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