Comparison of Quercetin Pharmacokinetics

Following Oral Supplementation in Humans

Diksha Kaushik, Kevin OFallon, Priscilla M. Clarkson, C. Patrick Dunne, Karen R. Conca, and Bozena Michniak-Kohn

Abstract: The objective of the study was to investigate the absorption of quercetin aglycone in 18 healthy human
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subjects administered via the following oral carrier systems: suspension of quercetin (quercetin QU995 powder in Tang
and spring water), nutritional bars (First StrikeTM ), and chews (RealFXTM Q-PlusTM ). Subjects were divided into
3 groups of 6 individuals each receiving 500 mg quercetin in one of the aforementioned formulations. Blood levels were
monitored immediately pre- and for 32 h postadministration. The concentration of total quercetin in blood samples was
determined by solid phase extraction followed by high-performance liquid chromatography analysis. Pharmacokinetic
parameters were determined by noncompartmental modeling using Kinetica software. The Cmax of quercetin was highest
with RealFXTM Q-PlusTM Chews (1051.9 393.1 g/L) achieved within 3.3 h as compared to that for First StrikeTM
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Bars (698.1 189.5 g/L in 2.3 h) and Tang
suspension (354.4 87.6 g/L in 4.7 h). The results showed no
statistically significant difference in quercetin absorption among groups due to high variability within groups receiving
quercetin from same dosage form. This study represents the first comprehensive evaluation of quercetin absorption from
quercetin fortified oral food products at doses commonly used for quercetin supplementation.
Keywords: AUC, buccal, Cmax , oral, quercetin

Practical Application: The current study describes for the first time, comprehensive evaluation of quercetin PK in humans

from quercetin fortified oral food products at doses commonly used for quercetin supplementation. Owing to quercetins
potent antioxidant and anti-inflammatory actions, quercetin is widely being used as a nutritional supplement. In order to
maximize the bioavailability of quercetin for its use in efficacy studies, it is important to determine its ideal oral carrier
system and route for its delivery. The current research unveils vital information about quercetin supplementation to the
international community, especially to soldiers, athletes, and the dietary supplement industry.

caffeinated chewing gum is now incorporated in some U.S. military ration items (Kamimori and others 2005).
Several studies have examined quercetin uptake and pharmacokinetic (PK) in humans when consumed in form of natural sources (Davalos and others 2006; Erlund and others 2006).
Different studies reporting pharmacokinetics of quercetin have
been summarized in Table 1. However, little is known about the
bioavailability and PK of quercetin when delivered in the form
of nutritional food products at commonly used dietary supplementation doses (Davis and others 2008; Cheuvront and others
2009; Utter and others 2009; Nieman and others 2010). Therefore, the purpose of this study was to examine and compare
the bio-absorption of quercetin through 3 different quercetinfortified carrier systems in humans. Real FX Q-PlusR Soft Chews
(Nutravail Inc., Va., U.S.A.) and a nutrition bar [First StrikeTM bar,
Natick Soldier Research Development and Engineering Center
(NSRDEC), Natick, Mass., U.S.A.] were chosen as the carrier
systems for both buccal and GI delivery. A beverage formulation

R
MS 20120682 Submitted 5/12/2012, Accepted 8/5/2012. Authors Kaushik and of quercetin mixed with Tang (Kraft Foods Inc., Ill., U.S.A.)
Michniak-Kohn are with Ernest Mario School of Pharmacy, Rutgers-The State Univ. suspended in spring water was chosen to represent GI delivery.

Quercetin (3,3,4,5,7-pentahydroxyflavone) (Figure 1) represents the most common flavonoid in the human diet
(Erlund and others 2006) that has been widely investigated for
its anti-inflammatory, antioxidant, antiviral, and anticarcinogenic
attributes (McAnlis and others 1999; Mamani-Matsuda and others
2006; Davis and others 2008; Nieman and others 2010). Owing
to the potential therapeutic benefits of quercetin, it is important
to understand the optimal dosage and carrier system that maximize quercetin uptake via buccal and gastrointestinal (GI) routes in
humans. The buccal absorption offered by chews, bars, chewing
gums, or lozenges offers certain advantages over conventional oral
delivery by rapid and efficient uptake in the blood stream by avoiding first pass metabolism by the liver and intestinal cells (McElnay
and Hughes 2006). Chewing gums have been validated as an efficient caffeine delivery system (Kamimori and others 2002), and

of New Jersey, 145 Bevier Road, Life Sciences Bldg, Piscataway, NJ 08854, U.S.A.
Authors OFallon and Clarkson are with Dept. of Kinesiology, Univ. of Massachusetts,
Muscle Biology and Imaging Laboratory, 30 Eastman Lane, Amherst, MA 01002,
U.S.A. Authors Dunne and Conca are with U.S. Army Natick Soldier Research
Development and Engineering Center, Natick, MA 01760, U.S.A. Direct inquiries
to author Michniak-Kohn (E-mail: michniak@biology.rutgers.edu).

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2012 Institute of Food Technologists
doi: 10.1111/j.1750-3841.2012.02934.x


C

Further reproduction without permission is prohibited

Materials and Methods

Materials
Food grade quercetin (99.5% pure and supplied by Quercegen Pharma LLC under license from Merck, Brazil. QU995),
Vol. 77, Nr. 11, 2012 r Journal of Food Science H231

H: Health, Nutrition, &

Food

Introduction

Comparison of quercetin PK . . .
TangR dry mix, First StrikeTM Bars, and Real FXTM Q-PlusTM
Chews/Q-chews were provided by NSRDEC. The compositions
of the quercetin-fortified formulations are listed in Table 2. Note:
The FDA has affirmed the Generally Recognized as Safe status of
quercetin as an ingredient in various foods and beverage products
at levels up to 500 mg/serving in different foods and beverages
(letter to Quercegen Pharma, LLC Nov. 22, 2010 GRAS Notice
GRN000341). Fisetin (99.9% purity), -glucuronidase (Type
HP-2 from Helix pomatia) were obtained from Sigma Aldrich.
The extraction procedure was done using a 20 Position Vacuum Extraction manifold and Oasis HLB cartridge (Waters, Mass.,
U.S.A.).

Methods
In vitro testing of quercetin
HPLC analysis of quercetin. Quercetin concentrations were
determined using a high-performance liquid chromatography
(HPLC) method (Agilent Technologies, Calif., U.S.A.) with ultraviolet detection at 370 nm with 20 L injection volume at 1.0 mL
flow rate using methanol:water (80:20) mobile phase adjusted to
pH 3.72. The Phalanx C18 RP column (5 m and 250 4.6 mm)
(Nest Group Inc., Mass., U.S.A.) was used for chromatographic
separation and maintained at 30 C. The method was tested for
linearity, precision, and accuracy.
Solubility of quercetin in various solvents. The solubility of
quercetin was determined in 2 solvents, namely, deionized water adjusted to acidic pH with acetic acid (4.0) and methanol. The
procedure involved dissolving excess amount of quercetin in 20
mL of the solvents and shaking for 1 wk until no change in solubility measurement (peak area of quercetin at 370 nm) was observed.
The supernatant was diluted with mobile phase and analyzed by
HPLC. No stability experiments were performed as quercetins
stability at room temperature has been well documented at pH
4.0 (Moon and others 2008) and in methanol (Liu and others
2006).
Quercetin content in RealFXTM Q-PlusTM Chews and First StrikeTM
Bars. The procedure for determination of quercetin content in
bars and chews involved weighing triplicate 1 g samples (randomly chosen within the bar or chew) from each bar or chew,
dissolving and swirl mixing in 10 mL of methanol followed by
homogenization. The suspension was then incubated at 20 C
overnight, vortexed, sonicated and 1 mL aliquot was then centrifuged. The resulting supernatant was then diluted and analyzed

H: Health, Nutrition, &

Food
Figure 1Structure of quercetin (3,3,4,5,7-pentahydroxyflavone).

H232 Journal of Food Science r Vol. 77, Nr. 11, 2012

by HPLC. The average weights of quercetin fortified bars and

chews were also determined and tested for content uniformity.
In vivo human pharmacokinetics
Subjects. Eighteen healthy men and women aged 18 to 25 y
were recruited from the Amherst, Massachusetts community. The
study was approved by the University of Massachusetts Amherst
Institutional Review Board (IRB) and Department of Defense
IRB and written informed consent was obtained from each subject prior to participation. At the informed consent visit, all subjects were instructed to engage in their normal physical activity
behaviors up to 72 h prior to day 1 and then refrain from activities other than those required for daily living. In addition, a list
of quercetin-containing foods and dietary supplements was provided, and subjects were asked to refrain from excessive [(based
on the estimated dietary intake of flavonoid-rich food sources by
U.S. adults, reported by Chun and others (2007)] consumption of
quercetin-containing foods for 7 d prior to and during the study.
Subject characteristics (Table 3) were gathered at the morning visit
on day 1. Health status was determined by a brief health history
questionnaire, and a 7-d physical activity recall was recorded on
an International Physical Activity Questionnaire (IPAQ). Subjects
were healthy, sedentary to recreationally active and use of oral
contraceptives was permitted.
Clinical study experimental design. The study duration was 2 consecutive days in which subjects reported to the laboratory in a
fasted (12 h) state each morning. This testing took place at the
University of Massachusetts Amherst campus. On day 1, subjects
were given a single 500 mg dose of quercetin in the carrier vehicle, and blood levels were monitored at specific time points over a
12-h period. On day 2, subjects made 3 visits to the laboratory (once every 4 h) for blood sampling. Strict control over
macronutrient intake and meal timing was maintained each
day by providing subjects with Meals Ready to Eat (MRETM
NSRDEC, Natick, MA). Dietary intake (caloric content and nutrient composition) and meal timing were standardized across
subjects by providing identical MRE menu items at identical feeding times and monitored by the investigator via direct
observation.
Supplementation. Subjects were randomized to ingest one of
3 quercetin carriers: (1) a beverage formulation containing 500
mg quercetin powder mixed in 26.9 g TangR dry mix suspended in 250 mL spring water (6 subjects), (2) quercetin fortified
First StrikeTM bar (6 subjects), or (3) quercetin in 2 RealFXTM
Q-PLUSTM chews (6 subjects). The 3 quercetin carrier systems
were provided to subjects in such a manner that each individual
received 500 mg dose of quercetin.
Blood collection. Subjects reported to the laboratory having fasted
for a minimum of 12 h and were required to remain in the laboratory for the first 12-h period (day 1) of data collection. A venous
catheter was inserted into a forearm vein distal to the antecubital
space of the elbow joint. Blood samples (approximately 12 mL)
were collected into K2 EDTA vacutainers at baseline, 15, 30, and
45 min after ingestion; and 1, 2, 3, 4, 6, 8, 12, 24, 28, and 32 h after ingestion. Samples were immediately centrifuged for 15 min at
1000 g at room temperature. Plasma was separated from collection tubes and 1 mL aliquots were transferred to NuncR cryotube
vials containing 100 L 10% ascorbic acid solution to maintain
quercetin stability (Ishii and others 2003; Moon and others 2008).
Samples were then vortexed for 30 s and stored at 80 C prior to
analysis.
Analysis of quercetin in plasma matrix. Analysis of quercetin in
plasma samples was conducted using an Agilent 1100 HPLC

Comparison of quercetin PK . . .
Table 1Summary of quercetin pharmacokinetics in healthy human subjects reported in literature.
Dose (mg/d)f

Plasma pharmacokinetics

Onion

68

Apple sauce

98

Quercetin-3-rutinoside

100

Quercetin-3-rutinoside

94

Quercetin-4-glucoside

94

Quercetin-3-rutinoside

98

Quercetin-4-glucoside

100

Healthy subjectsd (n = 10)

Random cross over
Single ingestion

Quercetin aglycone

500

Healthy subjects e,g (n = 12)

Random cross over
Single ingestion

Quercetin-3-rutinoside

200

Buck wheat tea

200

Quercetin-4-glucoside

100

Onion

100

Cmax = 224 44 g/L

T1/2 = 28 92 h
Tmax = 0.7 1.1 h
AUC(036h) = 2330 849 gh/L
Cmax = 92 19 g/L
T1/2 = 23 32 h
Tmax = 2.5 0.7 h
AUC(036h) = 1061 375 gh/L
Cmax = 90 93 g/L
T1/2 = not calculated
Tmax = 9.3 1.8 h
AUC(036h) = 983 978 gh/L
Cmax = 54 12 g/L
T1/2 = 28.1 6.4 h
Tmax = 6.0 1.2 h
AUC(0) = 1117 211 gh/L
Cmax = 1057 181 g/L
T1/2 = 21.6 1.9 h
Tmax = less than 0.5 h
AUC(0) = 5678 725 gh/L
Cmax = 1526 315 g/L
T1/2 = 18.5 0.8 h
Tmax = 0.6 0.2 h
AUC(036h) = 5775 876 gh/L
Cmax = 1345 212 g/L
T1/2 = 17.7 0.9 h
Tmax = 0.45 0.08 h
AUC(036h) = 5276 730 gh/L
Cmax = 15.4 ng/mL
T1/2 = 3.47 h
Tmax = 3 h
AUC(0) = 62.5 gh/L
Cmax = 0.32 0.34 g/mL
T1/2 = 11.8 3.1 h
Tmax = 6.98 2.94 h
AUC(036h) = 2.5 2.2 gh/mL
Cmax = 0.64 0.67 g/mL
T1/2 = 10.3 3.5 h
Tmax = 4.32 1.83 h
AUC(036h) = 3.8 3.9 gh/mL
Cmax = 2.12 1.63 g/mL
T1/2 = 11.9 4.0 h
Tmax = 0.70 0.31 h
AUC(024h) = 8.4 9.1 gh/mL
Cma= 2.31 1.46 g/mL
T1/2 = 10.9 4.1 h
Tmax = 0.68 0.22 h
AUC(024h) = 9.7 6.9 gh/mL

Study design

Source

Healthy subjects (n = 9)
Random cross over
Single ingestion
a,g

Healthy subjectsb,g (n = 9)
Random cross over
Single ingestion

Healthy subjectsc,h (n = 9)
Random cross over
Single ingestion

a
Hollman and others (1997).
b
Hollman and others (1999).
c
Olthof and others (2000).
d
Moon and others (2008).
e
Graefe and others (2001).
f
Dose given as quercetin equivalents
g
Results given as mean SD.
h

ingested during 1 d.

system with Phalanx C18 (5 m and 250 4.6 mm) RP column. The injection volume was maintained at 20 L with detection wavelength of 370 nm. Methanol:water (80:20, pH 3.72)
was used as the mobile phase that was pumped at 0.5 mL/min.
The analytical method in plasma matrix was tested for linearity,
precision, intraday, interday variability, recovery, lower limit of
quantification, and detection as indicated in Table 4.
Quercetin extraction from plasma matrix. The quercetin was extracted by solid phase extraction (SPE) using procedures reported
in the literature (Erlund and others 1999; Chen and others
2005). Briefly, to plasma samples containing 10% W/V ascorbic acid, 0.78 M sodium acetate buffer (pH 4.8), 40 L of -

glucuronidase/sulfatase, and 100 g/mL of fisetin (HPLC internal

standard) were added. The extraction was performed on an Oasis
HLB cartridge using methanol as the eluent. The eluate containing quercetin was then evaporated to dryness under nitrogen and
reconstituted in 300 L of methanol followed by HPLC analysis.
Noncompartmental PK modeling. The PK modeling was performed using Kinetica software version 4.4.1. In absence of any
intravenous data, a noncompartmental, extravascular model was
used to calculate the PK parameters. The model inputs involved
500 mg of quercetin (dose of quercetin in oral carrier systems) and
the time and concentration values for each subject. The software
generated PK outputs as shown in Table 6.

Vol. 77, Nr. 11, 2012 r Journal of Food Science H233

H: Health, Nutrition, &

Food

Results given as mean SEM (standard error of mean).

Comparison of quercetin PK . . .
Table 2Composition of various quercetin fortified supplements.

R

Quercetin fortified Tang

dry mix

Quercetin
-Tang dry mix
Sugar, Fructose, Citric Acid, Calcium Phosphate, Contains Less Than 2% of Orange Juice
Solids, Natural Flavor; Ascorbic Acid (Vitamin C), Vitamin E Acetate, Niacinamide, Vitamin
B6, Vitamin A Palmitate, Riboflavin (Vitamin B2), Beta Carotene; Maltodextrin, Sucralose,
Acesulfame Potassium and Neotame (Sweeteners), Guar and Xanthan Gums, Artificial Color,
Yellow 5, Yellow 6, Butyl hydroxyl anisole
-Water
Quercetin
-First StrikeTM Bar
Raspberry filling (fructose, maltodextrin, water, raspberry concentrate, food starch, modified
carrageenan, natural flavors, malic acid), maltodextrin, corn syrup, dried cranberries, crisp
corn (degermed yellow corn meals, sugar, malt extract, salt, calcium carbonate, mono and
diglycerides), apple nuggets (dried apple pieces, flavor, red 40, blue 1), partially hydrogenated
cottonseed/soyabean oil, whey protein concentrate, apple powder, rice bran concentrate,
glycerin, fructose, natural and artificial flavors, lecithin, vitamin premix (ascorbic acid,
DL-alpha-tocopherol acetate, niacinamide, pyridoxine hydrochloride, riboflavin, thiamine
mononitrate, folic acid, cholecalciferol, cyanocobalamin, zinc oxide), ascorbyl palmitate,
natural mixed tocopherols, red 40, blue 1
Quercetin
- Sugar, corn syrup, ascorbic acid, soy lecithin, natural and artificial flavors, palm oil, corn
starch, glycerin, xylitol, carrageenan, niacinamide, sucralose, sunflower oil, FD&C yellow 6,
FD&C yellow 5, folic acid.

Quercetin fortified First StrikeTM Bar

RealFXTM Q-PlusTM Chews

Table 3Subject characteristics.

Variablea
Height (m)
Weight (kg)
Age (y)
a

RealFXTM
Q-PlusTM Chews
(N = 6)

Quercetin
R
fortified Tang
suspension
(N = 6)

First
StrikeTM Bar
(N = 6)

1.7 0.07
60.8 10.9
20.3 1.4

1.7 0.1
65.7 4.9
19.5 1.1

1.8 0.1
71.7 10.7
20.5 1.1

Data are shown as means standard deviation.

All PK results were statistically analyzed using SPSS software

(SPSS 16, SPSS, Inc., Chicago, Ill., U.S.A.) through one-way
ANOVA and multiple comparisons were performed using Tukey
HSD (honestly significantly different) post hoc method at 95% confidence interval (CI).

Results and Discussion

H: Health, Nutrition, &

Food

In vitro studies. The main objective of this study was to compare

the absorption of quercetin through 3 quercetin-fortified carriers. The selection of supplementation/dosage forms (Cheuvront
and others 2009; Davis and others 2010; Ganio and others 2010)
and quercetin dose amounts (Moon and others 2008; Davis and
others 2010) for the study were based on dosage forms and doses
previously used in human clinical trials.
The quantification of quercetin in each carrier system and
quercetin solubility measurements were performed using a validated HPLC assay method (Table 4). Since the solubility
determination of quercetin in various solvents showed highest solubility of quercetin in methanol (8.5640 0.0754
mg/mL), methanol was selected as solvent for extraction of
quercetin carrier systems. The evaluation of quercetin fortified supplements, namely, First StrikeTM Bars and Real FXTM
Q-PLUSTM chews (Table 5) showed negligible overages beyond
their labeling of 500 mg and 250 mg of quercetin, respectively.
A known quantity of (500 0.04 mg) quercetin was added to
each TangR beverage and overages were negligible. On determination of content uniformity, the variability in quercetin content
within chews and bars showed RSD (relative standard deviation)

H234 Journal of Food Science r Vol. 77, Nr. 11, 2012

of not more than 4.2% and 6.0%, respectively, indicating uniform

fortification of quercetin within bars and chews (Table 5).
In vivo human pharmacokinetics. The clinical study involved 18
healthy subjects that were divided into 3 groups each group receiving a 500 mg dose of quercetin in one of the carriers. In order
to control for interindividual variability due to meal timing, potential drug-interactions, and exogenous sources of quercetin, subjects
were provided and directly observed ingesting standardized meals,
and agreed to refrain from the use of drugs, over-the-counter
medications, quercetin containing dietary supplements, and foods
rich in quercetin (apples and onions) 7 d prior to and during the
study. The analysis of quercetin in plasma samples was performed
using validated bioanalytical method summarized in Table 4. The
quercetin extraction was performed using SPE and methanol was
used as eluent owing to high solubility of quercetin in methanol
(approximately 8.5640 mg/mL). Preliminary experiments in our
laboratory showed that addition of 10% W/V of ascorbic acid
maintained chemical stability of quercetin at 37 C for at least
36 h and is also reported by Ishii and others (2003). Since the
aim of the study was to compare concentration of total quercetin
absorbed following administration of 3 nutritional supplements of
interest (and not individual isolation and determination of concentrations of quercetin aglycone and quercetin conjugates), an
enzymatic hydrolytic method was used to convert quercetin sulfate and the glucuronide conjugates to quercetin aglycone using
-glucuronidase. The enzymatic hydrolytic method involved addition of 40 L of -glucuronidase/sulfatase to acidified plasma
containing quercetin and fisetin (internal standard) and incubation of mixture for 17 h at 37 C for the complete hydrolysis of
quercetin glucuronides and sulfate conjugates to quercetin. Subsequently the assay for total quercetin was developed and validated
empirically to determine quercetin aglycone.
Noncompartmental PK modeling. The concentration-time profiles
of mean total quercetin in healthy subjects following administration of quercetin carrier systems is presented in Figure 2a.
The PK parameters generated by noncompartmental modeling
using Kinetica software are listed in Table 6. In descending order,
the data indicated that the greatest Cmax (1051.9 393.1 g/L)
was attained within 3 h in subjects that administered RealFXTM

Comparison of quercetin PK . . .
R
Table 5Quercetin content in Q-chews
and First StrikeTM Bar.

Method validation summary in pure solvent matrix

Parameter

Specifications

Calibration rangea

Calibration curve parameters

Lower limit of detection (LLOD)
Lower limit of quantification (LLOQ)
Run time
Retention time
Intraday precision
Interday precision
Recovery (pure solution)

3100000 g/L
(range evaluated)
390100000 g/L
(range with linearity)
R2 0.9873, Slope: 0.0002
20 g/L
390 g/L
8 min
4.1 min
<0.1%
1.0%
9899%

Method validation summary in plasma matrix

Parameter (n = 3)
a,b

Calibration range
Calibration curve parameters
Lower limit of detection (LLOD)
Lower limit of quantification (LLOQ)
Run time
Retention time (quercetin)
Retention time (fisetin)
Intraday accuracy & precisionc
Interday accuracy & precisionc
(3 separate days)
Recovery (quercetin)d & recovery (fisetin)d

Specifications
54000 g/L
R2 0.9870, Slope: 0.0002
3 g/L
5 g/L
15 min
8.5 min
7.4 min
<2.0%
<12.0%
8285% & 9899%

Quercetin stock was prepared in 100% methanol at concentration of 20 g/mL and

stored at 20 C. The experiments in our laboratory showed the stock solution showed
stability at 20 C and at 4 C for period of 6 mo and at least for 1 mo in methanol at
room temperature.
b
The calibration curve standards were freshly prepared in blank human plasma by adding
known amounts of quercetin from quercetin spiking solution made in 100% methanol
to 1 mL plasma containing 10% ascorbic acid; all calibration curves consisted of one
blank sample and 7 calibration points in the concentration range of 5 4000 g/L. Low,
medium, and high concentrations of quercetin selected as QCs were at 50, 500, and
2000 g/L. Calibration curve standards underwent solid phase extraction after addition
of internal standard, fisetin and eluted with methanol and evaporated to dryness and
reconstituted in 300 L methanol. The solid phase extraction of each plasma standard
and reconstitution of sample in less amount of solvent (methanol) after dryness hence
concentrating the sample and improving sensitivity of quercetin in plasma (LLOQ = 5
g/L) as compared to pure solvent matrix (390 g/L). In pure solvent matrix all levels
of calibration curve were prepared and analyzed by dilute and inject method.
c
Inter-assay and intra-assay precision and accuracy were evaluated at all plasma standard
levels and at all plasma QC levels that were prepared by spiking known amounts of
quercetin and fisetin in plasma (n = 3).
d
The recovery was performed by comparing the response (area) of processed QC samples
at concentrations 50 and 2000 g/L with same concentrations of solutions prepared in
pure matrix.
a

Q-PlusTM Chews, followed by a Cmax of 698.1 189.5 g/L

within 2 h in subjects administered quercetin-fortified First
StrikeTM Bars, and lowest Cmax (354.4 87.6 g/L) was attained
with quercetin fortified TangR suspension within 5 h. A T1/2 of
8 h was observed in First StrikeTM Bars and quercetin fortified
TangR suspension but a relatively low T1/2 (5.5 h) was observed
in RealFXTM Q-PlusTM Chews. On comparison of AUCtotal , in
descending order, maximum AUC was obtained in subjects that
administered First StrikeTM Bars (5314.8 1432.4 g/L.h2 ) followed by group receiving RealFXTM Q-PlusTM Chews (4147.1
671.8 g/L.h2 ) followed by subjects receiving quercetin fortified
TangR suspension (3845.9 689.8 g/L.h2 ).
The concentration-time profile of quercetin delivered through
various carrier systems, showed a low uptake of quercetin via
TangR as compared to First StrikeTM and Q-ChewTM delivery
carriers, however, no statistical differences (P > 0.05) were observed in absorption profile of quercetin among the 3 carrier
systems. Owing to the fact that quercetin has low solubility

Type of formulation
(n = 3)
RealFXTM Q-PlusTM Chews
Quercetin fortified
First StrikeTM Bar
R
Quercetin containing Tang
Suspensionc

Mean
quercetin
content
(mg) SDa

Relative
standard
deviation,
RSDb (%)

Claim
(mg)

265.1 11.1
531.2 32.1

4.2
6.0

250.0
500.0

N/A

N/A

500 0.04

a
SD is
b
RSD
c

standard deviation.
= 100XSD/Mean.
R
A known quantity of (500 0.04 mg) quercetin was added to each Tang
beverage
R
dry mix suspended in 250 mL spring water).
(26.9 g Tang
N/A: not applicable.

when suspended in TangR suspension as compared to other carrier systems, it was initially expected that TangR carrier system
would depict poor absorption of quercetin as compared to First
StrikeTM and Q-ChewTM carriers. The determination of solubility of quercetin was performed at pH 4 and data indicated that
quercetin is sparingly soluble (0.0019 0.0001 mg/mL) at acidic
pH and only a small amount of quercetin was dissolved in TangR
suspension (pH approximately 4.0). The lack of statistical difference in absorption profile of quercetin in spite of differences in
quercetin solubility among the carrier systems is supported by
finding by Piskula and others (1998) that reported that quercetins
solubility in carrier systems enhances its absorption but at the
same time is not an absorption limiting factor. Thus, the results
suggest that absorption of quercetin was not negatively affected by
solubility in the carrier system.
High variability in the PK profile of subjects within the same
group (receiving the same quercetin fortified carrier) was observed
in the study. For instance, within the treatment group administering RealFXTM Q-PlusTM Chews, certain subjects showed
high plasma quercetin levels (Cmax approximately 2849 g/L),
while others showed relatively low (Cmax approximately 270 g/L)
amounts of quercetin in the blood (Figure 2c). Similarly, in group
receiving First StrikeTM Bars (Figure 2b), UM3 (Cmax approximately 1578 g/L) showed substantially greater absorption of
quercetin than UM12 (Cmax approximately 374 g/L). Likewise, within group receiving quercetin fortified Tang suspension,
UM16 depicted relatively greater (Cmax approximately 682 g/L)
quercetin levels than UM5 (Cmax approximately 89 g/L) (Figure
2a). Interindividual variability is not a rare phenomenon and has
been reported in many studies (Morand and others 1998; Moon
and others 2000; Graefe and others 2001). The interindividual
variability is often linked with polymorphism existing in transporters of subjects within a group (Manach and Donovan 2004).
Quercetin is known to be absorbed by passive diffusion and a pHdependent mechanism mediated by the organic anion transporting
protein B (OATP-B) (Chabane and others 2009). There are reports of polymorphism in OATP-B transporters among individuals
in studies performed in Japanese population (Nozawa and others
2002). The polymorphism in OATP-B transporters may have led
to greater quercetin absorption in certain subjects whereas lower
quercetin absorption in others that received quercetin through
same kind of carrier system.
It was observed that RealFXTM Q-PlusTM Chews and First
StrikeTM Bars receiving subject groups showed higher intragroup
intersubject variability in quercetin absorption as compared to
Tang receiving group. The absorption of quercetin when administered through RealFXTM Q-PlusTM Chews and First StrikeTM

Vol. 77, Nr. 11, 2012 r Journal of Food Science H235

H: Health, Nutrition, &

Food

Table 4 Method validation summary in pure solvent matrix

and plasma matrix.

Comparison of quercetin PK . . .
Table 6PK parameters of quercetin administered via various oral carrier systems.
Parameters
a

Cmax (g/L)
Tmax (h)
c
AUCtotal (g/L.h)
d
Lz (1/h)X102
e
AUMCtotal (g/L.h2 )
f
T1/2 (h)
g
MRT (h)
h
Cl (L/h)
i
Vz (L)
j
Vss (L)
b

R
Tang
(N = 6)

354.4
4.7
3845.9
9.9
48141.3
8.3
12.9
153.8
2011.0
2131.7

87.6
0.3
689.8
0.02
12506.9
1.4
2.0
28.4
664.7
686.1

First StrikeTM (N = 6)
698.1
2.3
5314.8
9.9
66231.8
8.0
11.4
125.4
1255.2
1279.0

189.5
0.5
1432.4
0.01
22169.0
1.3
1.7
27.1
190.6
194.8

RealFX Q-Plus Chews/

Q-ChewTM (N = 6)
1051.9
3.3
4147.1
15.4
32103.6
5.5
7.9
143.4
987.1
1140.0

393.1
0.8
671.8
0.04
5607.5
0.9
1.3
29.7
138.4
286.7

a
Cmax : maximum concentration.
b
Tmax : time to each maximum concentration.
c
AUCtot : area under curve in concentration compared

with time curve, AUCtot = AUC 0t + C t /Lz .

The AUC was computed using the log linear method, trapezoidal when Cn > Cn1 .
d
Lz : smallest (slowest disposition rate constant), slope of the elimination phase of the PK profile.
e
AUMCtot : area under moment curve, AUMCtotal = AUMC0t + (Ct Tt )/Lz + Ct /(Lz 2 ).
f
T1/2 : half life of elimination, ln2/lz.
g
MRT: mean residence time, AUMCtotal /AUCtotal .
h
Cl: clearance, (Dose/AUCtotal ).
i
Vss : apparent volume of distribution in the plasma compartment at steady state, (Dose.MRT)/AUC total .
j
Vz : apparent volume of distribution during terminal phase, Dose/(AUCtotal ..Lz ).
Data are shown as means SEM (standard error of mean).

H: Health, Nutrition, &

Food

Bars occurred by means of both buccal and GI routes. When a

bioactive is absorbed by means of buccal route, saliva greatly affects
the bioavailability of the bioactive. Although saliva facilitates dissolution of the bioactive at the buccal absorption site, high secretion
of saliva while chewing lead to premature swallowing of the
bioactive fortified carrier system and also causes involuntary swallowing of the saliva. This phenomenon limits the absorption of
bioactive through the buccal route (McElnay and Hughes 2006)
and leads to relatively more absorption through the GI route. In
individuals where less saliva is produced while chewing, absorption
of the bioactive is not greatly impacted by involuntary swallowing
of saliva via the buccal route. At the same time, there are individuals
that do not chew enough and tend to swallow the bioactive fortified carrier system leading to more absorption through the GIT as
compared to buccal route. Therefore, it is suggested that the differential mode of administration of RealFXTM Q-PlusTM Chews
and First StrikeTM Bars in terms of chewing/biting/swallowing
of the carrier system by each individual likely contributed to the
variability of quercetin absorption through buccal compared with
the GI route. Future studies are currently being planned to test the
aforementioned hypothesis. It should also be noted that PK profile
of quercetin via TangR carrier showed less intersubject variability
(Figure 2a and 2d). Since quercetin absorption was mainly through
GI via the TangR carrier with minimal involvement of the buccal
component, different chewing and swallowing habits of individuals did not affect subject to subject quercetin absorption profile. In
aqueous carrier systems such as TangR , appreciable buccal absorption of quercetin is not expected because TangR suspension had
very low soluble amounts of quercetin available for buccal absorption and therefore presence of saliva in the oral mucosa could not
facilitate dissolution of quercetin at the buccal absorption site. This
explanation is further supported by observations of fine particles
of quercetin powder accumulated on the lingual tissue of subjects
(yellow color observed on the tongue) immediately following oral
consumption of quercetin via the TangR carrier. Moreover TangR
carrier was a liquid formulation and no chewing or swallowing
was involved while administering the carrier system. This implicates GI tract as the predominant route of quercetin absorption
via the TangR carrier.

H236 Journal of Food Science r Vol. 77, Nr. 11, 2012

Another interesting finding was the quercetin fortified TangR

suspension in spite of being in suspension form showed a trend
(P > 0.05) toward low quercetin bioavailability (AUC 3846
g/L.h) as compared to First StrikeTM Bars (AUC 5314.8 g/L.h)
and RealFXTM Q-PlusTM Chews (AUC 4147 g/L.h). In principle, oral suspensions improve the rate and extent of the absorption of the bioactive because the viscosity of the suspending agent present in the suspension increases with time. This in
turn leads to the improved wetting, increased particle deaggregation, increased particle surface area, improved GI transit time
and greater dispersability of the bioactive, hence, higher absorption of the active (Swarbrick and others 2005). However, when a
phytonutrient/bioactive such as quercetin that is sparingly soluble
in water (0.0019 mg/mL) is suspended in an aqueous carrier in
absence of any suspending agent, rate and extent of GI transit time
is significantly decreased (Piskula and others 1998). Reports by
Hackett and Griffiths (1982) showed that quercetin and catechin
absorption in rat plasma increased with co-administration with an
emulsifying agent (3-palmitoyl-(+)-catechin). Reports by Li and
others (2009) have shown that poor bioavailability of quercetin via
aqueous suspensions than corresponding solid lipid nanoparticle
carrier systems.
The terminal T1/2 in the study ranged from 5 to 8 h with
T1/2 approximately 5 h being for Real FXTM Q-PLUSTM and approximately 8 h for the First StrikeTM Bars and quercetin-TangR
suspension. There were no significant differences (P > 0.05) between the terminal half-life (slope of elimination phase) across
different groups which was expected as these parameters are PK
characteristics of the molecule (quercetin). Similarly, AUMCTotal ,
LZ , Cl, MRT, VSS and VZ did not show significant differences
across the group (P > 0.05) as they represented characteristics of
quercetin PK. The parameters from PK modeling studies are of
physiological importance as they aid in selection of appropriate
dosage regimens and carrier systems while designing quercetin efficacy studies. PK parameters also aid in estimating the extent to
which bioactives are either retained or cleared by the body with
time. The determination of PK parameters for quercetin is important in clinical studies that involve investigation of the effects of
quercetin following quercetin dosing at 12 h intervals over several

Comparison of quercetin PK . . .
A

2500.00

Quercetin via
Tang, N=6

Concentration (ug/l)

2000.00

Quercetin via
First Strike
bar, N=6

1500.00

Quercetin via
RealFX Q-Plus
Chews, N=6

1000.00
500.00

porate measurement of post supplementation plasma quercetin

levels in the study design that will assist in assessing significance of the results. In this study, each quercetin carrier system significantly increased total plasma quercetin levels, relative
to presupplementation values. Similar absorption profiles were
observed among RealFXTM Q-PlusTM chews, First StrikeTM
Bars, and the quercetin fortified TangR suspension, suggesting that these delivery vehicles are equally effective at elevating
plasma quercetin and suitable for use in future human clinical
trials.

0.00

0.25 0.5 0.75

1
2
3
Time (hrs)

12

24

28

32

UM3

2000.00
1500.00

UM11
1000.00

UM12

500.00

UM13
UM15

0.00
0 0.25 0.5 0.75 1

2 3 4 6
Time (hrs)

8 12 24 28 32
UM2

3000.00

Concentration (ug/l)

2500.00

UM4

2000.00

Concentration (ug/l)

Disclaimer
The opinions contained herein are the private views of the
author(s) and are not to be construed as official or as reflecting the
views of the U.S. Army or the Dept. of Defense. Any citations of
commercial organizations and trade names in this report do not
constitute an official Dept. of the Army endorsement of approval
of the products or services of these organizations.

UM8

1500.00
1000.00

UM14

500.00
UM17

0.00
0 0.25 0.5 0.75 1

Funding was provided by U.S. Army Contract #W911QY-07C-0027. The authors express gratitude to Paul Magurie and other
members of Performance Optimization Research Team, Combat
Feeding Directorate, US Army NSRDEC, Natick, MA for their
technical support in providing special First StrikeTM bars.

2 3 4 6
Time (hrs)

12 24 28 32
UM18

800.00

UM5
UM6

600.00

UM7

400.00

UM10
200.00

UM16
0.00

UM1
0 0.25 0.5 0.75 1

2 3 4
Time (hrs)

12 24 28 32

Figure 2(a) Mean total quercetin in plasma following administration of

quercetin via various oral carrier systems. (b) Plasma profile of quercetin in
subjects following administration of First StrikeTM Bar. UM 3, 9, 11, 12, 13,
and 15 represent subjects that received First StrikeTM Bar. (c) Plasma profile of quercetin in subjects following administration of RealFXTM Q-PlusTM
Chews. UM2, 4, 8, 14, 17, and 18 represent subjects that received Q-PlusTM
Chews. (d) Plasma profile of quercetin in subjects following administration
R
of Quercetin fortified Tang
suspension. UM1, 5, 6, 7, 10, and 16 represent
subjects that received Q-PlusTM Chews.

days to maintain appreciable levels of quercetin in blood (Ganio

and others 2010; Nieman and others 2010).

Conclusion
This study is the first to report comparisons of PK parameters
and bio-absorption of total quercetin among different quercetinfortified foods and beverage base following oral consumption in
human volunteers. The high intragroup intersubject variability
in absorption observed in this study indicates that future studies
investigating the effects of quercetin in humans should incor-

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