International Journal of Food Science and Technology 2012 1

Original article
In vitro bioaccessibility of coenzyme Q10 in enriched yoghurts

Pınar Ercan & Sedef Nehir El*

Department of Food Engineering, Engineering Faculty, Ege University, 35100 Bornova, Izmir, Turkey
(Received 9 September 2011; Accepted in revised form 12 March 2012)

Summary In this study, the aim was to determine the bioaccessibilities of coenzyme Q10 (CoQ10) in yoghurt samples
produced using enriched skim milk with different coenzyme Q10 preparations. First, emulsified coenzyme
Q10, c-cyclodextrin ⁄ coenzyme Q10 complex and nanoparticle coenzyme Q10 were prepared with the
reference coenzyme Q10 standard. These coenzyme Q10 preparations were added into the milk to produce
coenzyme Q10 enriched yoghurt samples. Nanoparticle coenzyme Q10 was obtained as in spherical shape
with 176.00 ± 50.62 nm diameter. Coenzyme Q10 bioaccessibility was found as 50.59 ± 1.88% in control
yoghurt. The yoghurt enriched nanoparticle coenzyme Q10 had the highest coenzyme Q10 bioaccessibility
(73.81 ± 1.61%) among the produced yoghurts (P < 0.01). Coenzyme Q10 bioaccessibilities were also
found as 63.75 ± 0.91% and 46.83 ± 1.27% in yoghurts enriched with emulsified coenzyme Q10, and
c-cyclodextrin ⁄ coenzyme Q10 complex, respectively.
Keywords Bioaccessibility, coenzyme Q10, enrichment, nanoparticle coenzyme Q10, c-cyclodextrin ⁄ coenzyme Q10 complex.

ability to produce coenzyme Q10. However, this ability

Introduction
Coenzyme Q10 (CoQ10, ubiquinol 10 and ⁄ or ubiqui-
none 10) is a fat-soluble, vitamin-like substance present
in nearly all tissues and it is a redox molecule, which
exists in both a biochemically reduced form (ubiquinol-10)
and an oxidised form (ubiquinone-10) in biological
tissues (Crane, 2001; Kubo et al., 2008). The name
ubiquinone means ‘ubiquitous quinone’ and relates to
the presence of the substance in all cells. The name
coenzyme Q refers to the chemical structure and in
coenzyme Q10, the human form; the molecule contains
one quinone group and 10 isoprenyl units. Chemically,
CoQ10 is designated 2,3-dimethoxy-5-methyl-6-decapre-
nyl-1,4-benzoquinone (Overvad et al., 1999; Parkhideh,
2008). Coenzyme Q10 plays the essential role of electron
carrier and proton translocator during cellular respira-
tion and ATP production and protects numerous
cellular membranes and plasma lipoproteins against
free radical-induced damage (Thanatuksorn et al.,
2009). Some studies stated that coenzyme Q10 was
effective in cardiomyopathy, hypertension, angina pec-
toris and atherosclerosis, cancer, neurodegenerative
disorders, periodontal diseases and diabetes (Ankola
et al., 2007; Pravst et al., 2010).
Coenzyme Q10 is supplied from two sources; endog-
enous synthesis and exogenous sources (foods and
supplements) (Overvad et al., 1999). Humans have the
*Correspondent: Fax: +90 232 3427592;
e-mail: sedef.el@ege.edu.tr
starts declining at the age of 20 and coenzyme Q10
amounts in humans decrease more rapidly after the age
of 40 (Itagaki et al., 2010). Also, genetic mutation,
cancer and statin-type drugs can cause a decrease in
coenzyme Q10 amount in serum or tissue; therefore,
exogenous sources of coenzyme Q10 are important
(Crane, 2001). Meat, poultry and fish are the main
coenzyme Q10 rich foods in accordance with a high
content of mitochondria in muscle tissue (Kagan &
Quinn, 2001; Ercan & El, 2011). Ercan & El (2011)
determined coenzyme Q10 contents in beef heart, liver
and muscle as 109.97, 33.34, 23.47 lg g)1, respectively.
The average dietary of coenzyme Q10 is about 3–6 mg
(Mattila & Kumpulainen, 2001; Kubo et al., 2008;
Žmitek et al., 2008). The suggested daily intake of
coenzyme Q10 from exogenous sources varies from 30
to 100 mg for healthy people (Pravst et al., 2009). Even
with large amounts of coenzyme Q10 rich food products
like beef heart in the diet; it would be difficult supply
100 mg day)1 (Crane, 2001). Consuming functional
food fortified with coenzyme Q10 can be beneficial for
additional intake of exogenous coenzyme Q10. The
intake can be increased by the fortification of food
products, but until recently, it was not easily achievable
because of its lipophilicity (Pravst et al., 2010).
Coenzyme Q10 from the diet is absorbed from the small
intestinal tract, and it is better absorbed in the presence
of a fatty meal (Mason, 2005; Žmitek et al., 2008).
Emulsification and micelle formation is required for the
absorption of fats and this process is facilitated by

doi:10.1111/j.1365-2621.2012.03061.x
 2012 The Authors. International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
2 Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El
secretions from the pancreas and bile in the small
intestine (Žmitek et al., 2008). Coenzyme Q10, which is
taken up by intestinal mucous cells, is incorporated into
chylomicrons after intestinal absorption, transported via
the lymphatic system to the circulation, and incorpo-
rated into very low-density lipoprotein cholesterol and
low-density lipoprotein cholesterol in the liver and then
concentrated in the tissues (Kagan & Quinn, 2001;
Mason, 2005). The long side chain of the 10-isoprenoid
units in the molecule and high molecular weight of
coenzyme Q10 (863 Da) make it poorly water soluble.
So, coenzyme Q10 is very poorly absorbed in the
gastrointestinal tract (Ankola et al., 2007; Žmitek et al.,
2008; Hu et al., 2011). The FDA has defined bioavail-
ability as the rate and extent to which the active
substances or therapeutic moieties contained in a drug
are absorbed and become available at the site of action.
This definition also applies to active substances present
in foods. Another term that is commonly used is
bioaccessibility, which is defined as the fraction of an
ingested nutrient that is released from food matrix and is
available for absorption in the gut after digestion
(typically based on in vitro procedures) (Prada &
Aguilera, 2007; Žmitek et al., 2008; Ercan & El, 2011).
In the past few years, there has been an extensive
effort to improve the oral bioavailability of coenzyme
Q10 and various formulations for improvement of the
oral bioavailability of coenzyme Q10 have been inves-
tigated, including molecular complexes, emulsions and
liposomal systems (Thanatuksorn et al., 2009). The need
for more information on the bioavailability of coenzyme
Q10 is strongly stressed. McClements (2010) stated that
a bioactive lipophilic component like coenzyme Q10
might be incorporated within a beverage or food that
could easily be consumed by drinking or eating because
this increases their palatability, desirability and bioac-
tivity. Itagaki et al. (2010) investigated the effect of
grapefruit juice on the transport of coenzyme Q10 by
Caco-2 cells. According to this study, the combined
administration of coenzyme Q10 and grapefruit juice
could enhance coenzyme Q10 absorption, because
grapefruit juice is reported to inhibit the function of
P-glycoprotein that efflux transport of coenzyme Q10 is
mediated by P-glycoprotein in Caco-2 cells. Pravst et al.
(2009) studied the stability of coenzyme Q10 in some
fortified products that were enriched by water-soluble
inclusion complex of coenzyme Q10 and b-cyclodextrin
(Q10Vital); milk, kefir, curd and fruit syrup have been
investigated and the coenzyme Q10 level in studied
products was determined to be stable. In addition, no
changes in sensorial, microbiological, physical and
chemical properties were observed. However, to date,
no published information exists in comparison with the
bioaccessibility of coenzyme Q10 between enriched
foods with different coenzyme Q10 preparations.
International Journal of Food Science and Technology 2012
International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
The aim of this study was to prepare emulsified
coenzyme Q10, c-cyclodextrin ⁄ coenzyme Q10 complex
and nanoparticle coenzyme Q10 preparations with the
goal of increasing the coenzyme Q10 bioaccessibility. In
addition, the study aimed to produce enriched coenzyme
Q10 yoghurts with adding these preparations and to
compare the bioaccessibility of coenzyme Q10 in the
enriched yoghurts.
Material and methods
Materials
Skim milk and sunflower oil were purchased at different
times from three hypermarkets in Izmir, Turkey.
Yoghurt culture (Lactobacillus bulgaricus, Streptococcus
thermophilus DSM Delvo-Yog CY-121) was kindly
supplied from Ege University Agriculture Faculty Milk
Technology Department, Turkey.
Chemicals
Coenzyme Q10 standard (analytic-C9538), lipase
(L3126), bile extract (B8631), pepsin (P700), pancreatine
(P8096), c-cyclodextrin (C4892), poly DL-lactide-co-
glycolide (PLGA) (P2191) and didodecyldimethylam-
monium bromide (DMAB) 98% (359025) were pur-
chased from Sigma. Commercial coenzyme Q10
standard (MOLEM78392972) was purchased from
Molecula. 2-propanol (HPLC-8175) was purchased
from LabScan. Tween 20 (S4393684) was purchased
from Merck. Monoglyceride (Rikemol type PUS 90%)
was supplied from Yilmaz Chemistry, Turkey. All
reagents were analytical grade.
Coenzyme Q10 analysis
Extraction
Extraction of samples was performed using a solvent
extraction method according to Mattila & Kumpulainen
(2001). Sample (7 g for control yoghurt and 1 g for
enriched yoghurts) was taken into an extraction tube.
Eight millilitres of ethanol was added into the sample
followed by homogenisation with an IKA Ultra-turrax
T25 homogenizer(IKA-Werke GmbH & Co. KG,
Staufen, Germany) (87.12 g, 5 min). After that, 20 mL
n-hexane [high-performance liquid chromatograph
(HPLC) grade] was added into a tube and mixed
vigorously. The tube was centrifuged to separate the
layers (6600 g, 5 min). Hexane layer was saved and the
lower layer was re-extracted twice with 5 mL ethanol
and 20 mL n-hexane. The combined n-hexane layer was
evaporated at 30–40 C. Extracted enriched yoghurts
were dissolved in 5 mL 2-propanol, and extracted
control yoghurt was dissolved in 2 mL 2-propanol.
 2012 The Authors

Then, all the extracts were stored at )40 C until HPLC
analysis.
Determination
Analysis was performed using a method according to
Mattila & Kumpulainen (2001). The analytical HPLC
system consisted of a Hewlett-Packard (HP) 1050 HPLC
equipped with a Waters 486 UV detector and a Vydac
201TP54 C18 column (5 lm, 25 cm · 4.6 mm). The
mobile phase consisted of methanol, 2-propanol and
ethanol (70:15:15, all HPLC grade); the flow rate was
0.8 mL min)1, and 0.2 AUFS. The HPLC analysis was
performed at room temperature. The wavelength used
for identification and quantification of the coenzyme
Q10 was 275 nm. For preparing the mobile phase,
150 mL of 2-propanol and 150 mL of ethanol were
added to 700 mL methanol. The mobile phase was
filtered with blue ribbon filter paper and degassed for
10 min. Coenzyme Q10 standard stock solutions were
prepared by dissolving 10 mg coenzyme Q10 in 100 mL
of ethanol. Working standard solutions (five levels of
concentration: 2.5, 10, 20, 40, 55 lg mL)1) were in the
range 2.5–55 lg mL)1. The working standard solutions
were filtered with 0.2 lm PTFE membrane filters prior
to the HPLC analysis and injected to HPLC. The
analysis was done in triplicate for each standard.
Calibration curves were constructed by plotting peak
areas versus concentrations of coenzyme Q10, and
regression equations were calculated. The sample
extracts were filtered with 0.2 lm PTFE membrane
filters prior to the HPLC analysis and injected to HPLC.
Coenzyme Q10 contents of samples were calculated
using calibration curves. y = 4.5247x + 27.328,
R2 = 0.9985, y = peak area (mAU · s), x=concentra-
tion of coenzyme Q10 (lg).
In vitro digestion
Samples were subjected to simulated gastric and small
intestinal digestion according to Bhagavan et al. (2007).
Sample was diluted with 20 mL 120 mm NaCl before
adjusting pH to 3.0 with 1N HCl. Porcine pepsin was
added to the samples, and final concentration was
adjusted to 2 mg mL)1. The volume was adjusted to
40 mL with NaCl. Nitrogen was sprayed to the samples.
Then, the samples were immediately sealed and incu-
bated in a shaking water bath at 37 C, 90 r.p.m. for
60 min. The gastric phase was terminated by adding 1N
sodium bicarbonate to increase the pH to 6. Porcine
pancreatin, lipase and bile extract were added. Final
concentrations of porcine pancreatin, lipase and bile
extract should be 0.4, 0.2 and 2.4 mg mL)1, respectively.
The pH was adjusted to 6.9 with 1N NaOH and volume
increased to 50 mL. Nitrogen was sprayed to the
samples. Then, the samples were immediately sealed
and incubated in a shaking water bath at 37 C for
 2012 The Authors
International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El 3
120 min. Tubes were placed in an ice bath, and aliquots
of the sample were placed in tubes, sealed and centri-
fuged at 10000 · g at 4 C for 35 min. The supernatants
from centrifugation were filtered and stored at )40 C
under nitrogen until coenzyme Q10 analysis. Coenzyme
Q10 bioaccessibility was calculated as percentage of
coenzyme Q10 content of in vitro digested sample (S)
and coenzyme Q10 content of sample (C) ratio.
Bioaccessibility (% ) ¼ S=C  100
S = Coenzyme Q10 content of in vitro digested
sample (lg CoQ10).
C = Coenzyme Q10 content of sample (before in
vitro digestion) (lg CoQ10).
Methods for coenzyme Q10 preparations
Three different methods for improvement of the bioac-
cessibility of CoQ10 were investigated, preparing an
emulsified CoQ10, c-cyclodextrin ⁄ coenzyme Q10 com-
plex and nanoparticle coenzyme Q10 products.
Emulsified coenzyme Q10
Emulsified coenzyme Q10 was prepared according to
Thanatuksorn et al. (2009) with some modifications.
Sunflower oil, 8 g ⁄ 100 g w ⁄ w skim milk aqueous solu-
tion and monoglyceride were used for emulsified coen-
zyme Q10 form. Sixty milligram coenzyme Q10 was
dissolved in 0.56 g sunflower oil. Skim milk aqueous
solution (1.46 g) and monoglyceride (170 mg) were
added to the mixture. Then, 7.75 g water was added
and the mixture was homogenised at 64.01 g for 20 min
(IKA Ultra-turrax T25) and then stored at )40 C.
c-Cyclodextrin ⁄ coenzyme Q10 complex
c-Cyclodextrin ⁄ coenzyme Q10 complex was prepared
according to the patent of Moldenhauer & Jan (2005).
Two hundred milligrams of c-cyclodextrin were dis-
solved in 4.5 mL of water at 70 C. Forty milligram of
coenzyme Q10 in 0.5 mL of diethyl ether were added to
the solution, and then homogenisation was carried out
in an IKA Ultra-turrax T25 at 711.2 g for 30 s. The
mixture was cooled on a magnetic stirrer at room
temperature within 30 min and then dried in a drying
oven at 55 C. The material was dissolved in 4 mL water
and stored at )40 C.
Nanoparticle coenzyme Q10
Nanoparticle coenzyme Q10 was performed using an
emulsion–diffusion–evaporation technique according to
Ankola et al. (2007) with some modifications. Four
hundred milligram of PLGA (50:50) was dissolved in
16 mL of ethyl acetate at room temperature and stirred
for 2 h. The organic phase was emulsified with 40 mL of
an aqueous phase containing stabiliser (1% w ⁄ v
International Journal of Food Science and Technology 2012
4 Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El
DMAB). The resulting o ⁄ w emulsion was stirred at
room temperature for 1 h and then homogenised with
IKA Ultra-turrax T25 homogenizer at 1600.2 g for
5 min and with Bandelin Sonopuls ultrasonic homoge-
nizer (Bandelin GmbH & Co. KG, Berlin, Germany) at
50% amplitude for 2 min. To this emulsion, 240 mL
water was added with constant stirring and nanopre-
cipitation resulted. Stirring was continued at
1000 r.p.m., 40 C for evaporation of ethyl acetate.
Nanoparticles were isolated using a centrifuge at
10000 g at 10–15 C and washed three times with
deionised water. Nanoparticles were dissolved in 6 mL,
1% Tween 20 solution and stored at )40 C.
Measurement of nanoparticle size
Scanning electron microscope (SEM – Phillips XL-30S
FEG, Izmir Institute of Technology, Center for Mate-
rials Research, Izmir, Turkey) was used to image and
determine the shape of the nanoparticles. The particle
sizes of the nanoparticles were measured by Olympus
Soft Imaging Solutions measure IT 5.1- Measurement
Software (Olympus Soft Imaging Solutions GmbH,
Münster, Germany).
Efficiency of coenzyme Q10 preparations
Entrapment efficiency is the percentage of ratio of
coenzyme Q10 initially taken for loading to that actually
incorporated into the emulsion structure, nanoparticles
and c-cyclodextrin complex. It was determined for the
free coenzyme using HPLC. The percentage of efficiency
in the coenzyme Q10 preparations was calculated.
Efficiency (% ) ¼ A=B  100
A = Coenzyme Q10 content in the preparation.
B = Coenzyme Q10 added amount into the prepara-
tion.
Production of enriched yoghurt products with coenzyme
Q10 preparations
Milk analyses
Skim milk was used in the yoghurt production, and dry
matter, fat and protein contents were determined
according to AOAC (1995).
Yoghurt products
Crane (2001) reported that to increase the concentration
significantly requires in a day at least 100 mg coenzyme
Q10 that can increase the level in blood to around
2 lg mL)1 or more. Taking into consideration this data,
it was planned that portion yoghurt (250 g) was
enriched with 100 mg coenzyme Q10. The skim milk
was boiled at 80 C for 5 min, and then, the milk was
cooled to room temperature. After heat treatment,
3.33 g of emulsified coenzyme Q10, 2 mL of c-cyclo-
dextrin ⁄ coenzyme Q10 complex and 2 mL of nanopar-
International Journal of Food Science and Technology 2012
International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
ticle coenzyme Q10 were added into the milk in three
different cups for each preparation. The mixtures of
milk and preparations were stirred using magnetic
stirrer for 10 min. Then, 3% cultures were poured into
each mixture and stirred. These mixtures were hold at
42 C for about 2.5–3 h until the pH of yoghurts
reached to 4.7. After incubation, yoghurt samples were
hold at room temperature for half an hour and then
cooled at refrigeration to finish the fermentation pro-
cess. Yoghurt samples were stored at )40 C until
analyses. Coenzyme Q10 loss during yoghurt production
was calculated by this formula:
Loss (% ) ¼ ðP  YÞ=P  100
P = Coenzyme Q10 content of the preparation added
into the milk.
Y = Coenzyme Q10 content after yoghurt produc-
tion.
Statistical analysis
All experiments were performed in triplicate and paral-
lel. Six values for each sample were averaged (n = 6).
SPSS (version 13; SPSS Institute Inc., Chicago, IL,
USA) was used for all statistical analysis. The paired
samples t-test procedure was used to compare the means
of two variables for a single group. When more than one
different group was compared, analysis of variance
(anova) and Tukey’s test were performed for investi-
gating statistically significant differences. A P value of
<0.01 was considered to be significant.
Results and discussion
Coenzyme Q10 preparations
The highest efficiency percentage was found in the c-
cyclodextrin ⁄ coenzyme Q10 complex (85.08 ± 0.32%)
(P < 0.01) (Table 1). Moldenhauer & Jan (2005)
reported that the formulation with c-cyclodextrin was
stable to air and light. Therefore, coenzyme Q10 was not
affected from the light during the preparation of c-
cyclodextrin ⁄ coenzyme Q10 complex.
Because the preparation time of emulsified coenzyme
Q10 was shorter than the preparation time of nanopar-
Table 1 Efficiency of coenzyme Q10 (CoQ10) preparations
CoQ10 preparations % Efficiency
Emulsified CoQ10 73.01 ± 0.16b
Cyclodextrin CoQ10 85.08 ± 0.32a
Nanoparticle CoQ10 67.40 ± 0.24c
Data represent mean ± SD (n = 6).
Different superscript letters are indications of significant differences
(P < 0.01).
 2012 The Authors
 Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El 5

ticle coenzyme Q10, the efficiency of emulsified coen-
zyme Q10 (73.01 ± 0.16%) was found higher than the
efficiency of nanoparticle coenzyme Q10
(67.40 ± 0.24%) (P < 0.01) (Table 1). Hariharan et al.
(2006) reported that 15% estradiol loaded PLGA
nanoparticles prepared by emulsion–diffusion–evapora-
tion method had 68.20% efficiency. Ankola et al. (2007)
studied the effect of initial coenzyme Q10 loading on
entrapment efficiency. They reported that the efficiency
of nanoparticles was 61–83% because of the 5–75%
initial coenzyme Q10 load.
Particle size of nanoparticle coenzyme Q10
Fifteen per cent of coenzyme Q10 loaded PLGA
nanoparticles prepared by emulsion–diffusion–evapora-
tion method was obtained as in spherical shape with
176.00 ± 50.62 nm diameter. SEM image of coenzyme
Q10 nanoparticles is given in Fig. S1. The size distribu-
tion of coenzyme Q10 nanoparticles is presented in
Fig. S2. ‘Nano-sized’ coenzyme Q10 means coenzyme
Q10 that has an average effective particle size of less
than about 1000 nanometers (nm) (Parkhideh, 2008).
Ankola et al. (2007) obtained nanoparticles that had
107.3–131.6 nm particle size because of the amount of
coenzyme Q10 by emulsion–diffusion–evaporation tech-
nique. Nehilla et al. (2008) reported that coenzyme Q10
nanoparticles obtained using 150 mg PLGA had spher-
ical morphology and average 174 nm diameters. Bala
et al. (2005) found that the particle size of 15% ellagic
acid loading PLGA nanoparticles prepared by the
method of emulsion–diffusion–evaporation was
423.3 ± 56.6 nm. Esmaeili et al. (2008a) found that
10% estradiol loaded PLGA nanoparticle prepared by a
emulsion–diffusion method were 150–200 nm in size
with a maximum standard deviation of about 20 nm.
Hariharan et al. (2006) reported that the particle size of
15% estradiol loaded PLGA nanoparticles prepared by
emulsion–diffusion–evaporation method was 150.8 nm
in spherical shape. Esmaeili et al. (2007) found that the
particle size of PLGA nanoparticles prepared by emul-
sification–diffusion method was 200–260 ± 10–24 nm.
Esmaeili et al. (2008b) determined that PLGA nano-
Table 2 Coenzyme Q10 (CoQ10) losses in the production of yoghurts, CoQ10 contents and bioaccessibilities of enriched yoghurts
particles prepared by emulsification–solvent diffusion
method had 189–225 nm in spherical shape.
Milk analyses
The skim milk used in the production of enriched
yoghurts had 8.85 ± 0.01% dry matter content,
0.1 ± 0.00% fat, 8.75 ± 0.01% nonfat solids and
3.43 ± 0.08% protein. Protein content of milk should
be at least 3% and fat content of milk should be <0.5
according to Turkish Food Codex Communiqué on
Fermented Milk Products (2009).
Bioaccesibilities of coenzyme Q10 in the enriched yoghurts
Coenzyme Q10 contents of control and enriched
yoghurts with emulsified coenzyme Q10, c-cyclodex-
trin ⁄ coenzyme Q10 complex and nanoparticle coenzyme
Q10 are given in Table 2. Coenzyme Q10 content of
control yoghurt was found as 1.85 ± 0.13 lg g)1. This
result was in agreement with previous studies. In the
literature, coenzyme Q10 content was found as
2.4 lg g)1 by Mattila & Kumpulainen (2001),
0.26 lg g)1 by Kubo et al. (2008), 1.2 lg g)1 by Weber
et al. (1997). Coenzyme Q10 contents were found as
248.75 ± 3.50, 300.99 ± 3.81 and 228.43 ± 3.78 lg g)1
in yoghurt enriched with emulsified coenzyme Q10,
cyclodextrin ⁄ coenzyme Q10 complex and nanoparticle
coenzyme Q10, respectively. In the production of
yoghurt, coenzyme Q10 preparations were added into
the milk at the stage of inoculation of yoghurt culture.
Fir et al. (2006) reported that when coenzyme Q10
standard was added into the milk prior to fermentation,
micro-organisms consumed more than one-third of
coenzyme Q10 during the fermentation process. There-
fore, in our study, coenzyme Q10 contents were
decreased during yoghurt fermentation. Coenzyme
Q10 losses in the production of yoghurts are given in
Table 2. Coenzyme Q10 losses were found as
14.83 ± 1.35%, 11.56 ± 1.21% and 15.27 ± 1.50%
in yoghurt samples enriched with emulsified coenzyme
Q10, c-cyclodextrin ⁄ coenzyme Q10 complex and nano-
particle coenzyme Q10, respectively.

CoQ10 loss in the CoQ10 content CoQ10 content after Bioaccessibility (%)
production (lg CoQ10 per g) in vitro digestion
Samples of yoghurt (%) (lg CoQ10 per g)

Control yoghurt – 1.85 ± 0.13a,D 0.93 ± 0.04b,D 50.59 ± 1.88C

Yoghurt containing emulsified CoQ10 14.83 ± 1.35A 248.75 ± 3.50a,B 158.59 ± 4.20b,B 63.75 ± 0.91B
Yoghurt containing cyclodextrin CoQ10 11.56 ± 1.21B 300.99 ± 3.81a,A 140.93 ± 3.59b,C 46.83 ± 1.27D
Yoghurt containing nanoparticle CoQ10 15.27 ± 1.50A 228.43 ± 3.78a,C 168.60 ± 4.28b,A 73.81 ± 1.61A

Data represent mean ± SD (n = 6).

Different superscript letters within same row are indications of significant differences (P < 0.01).
Different superscript capital letters within same column are indications of significant differences (P < 0.01).

 2012 The Authors International Journal of Food Science and Technology 2012
International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
6 Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El
The simulated gastric ⁄ small intestinal digestion mod-
els are being extensively used at present because they are
rapid, safe and do not have the ethical restrictions of in
vivo methods. In vitro methods either simulate the
digestion and absorption processes (for bioavailability)
or only the digestion process (for bioaccessibility), and
the response measured is the concentration of a nutrient
and other dietary bioactive compound in some kind of
final extract. The digestion process is simulated under
controlled conditions using commercial digestive
enzymes while the final absorption process is commonly
assessed using the coupled simulated gastric ⁄ small
intestinal digestion with Caco-2 cells cultures. The
amount of a nutrient present in the extract after Caco-
2 cell step is assumed as the final amount bioavailable
(Prada & Aguilera, 2007). In our study, bioaccesibilities
of coenzyme Q10 in enriched yoghurts were calculated
with the coenzyme Q10 contents of yoghurts after in
vitro digestion. The bioaccessibility percentages are
given in Table 2. Coenzyme Q10 bioaccessibilities were
found as 50.59 ± 1.88% and 63.75 ± 0.91% in control
yoghurt and enriched with emulsified coenzyme Q10
preparation, respectively. Most of the previous studies
have focused exclusively on the improvement of oral
bioavailability. Thanatuksorn et al. (2009) reported that
practical interest in development of coenzyme Q10
products was generated not only by improvement of
oral bioavailability but also by reduction in production
costs. They concluded that emulsions with fats and
emulsifiers used commonly in the food industry were
one of most suitable approaches to improving bioavail-
ability. Žmitek et al. (2008) determined bioaccessibility
of coenzyme Q10 preparation in soybean oil, where
increased coenzyme Q10 of about 60 per cent. Ozaki
et al. (2010) stated that when the volunteers took the
coenzyme Q10 as bulk powder, no peak was observed in
the plasma concentration, and they also reported that
emulsification of coenzyme Q10 that they prepared
using gum arabic compared with the coenzyme Q10
bulk powder significantly increased bioavailability in
both rats and humans and the emulsified coenzyme Q10
was highly stable under various conditions like heat and
high humidity. Another successful approach was to use
the cyclodextrin complex inclusion to facilitate absorp-
tion from the gastrointestinal tract and to improve
bioaccessibility. Moldenhauer & Jan (2005) have a US
Patent 6861447 on a method for producing a coenzyme
Q10 ⁄ c-cyclodextrin complex. They reported that a
formulation with c-cyclodextrin is stable to air and light
and increases the solubility in water as well as the
bioaccessibility. In this study, bioaccessibility of yoghurt
enriched with c-cyclodextrin ⁄ coenzyme Q10 complex
was determined as 46.83 ± 1.27%.
The yoghurt sample enriched with nanoparticle coen-
zyme Q10 had the highest coenzyme Q10 bioaccessibility
(73.81 ± 1.61%) among the yoghurt products
International Journal of Food Science and Technology 2012
International Journal of Food Science and Technology  2012 Institute of Food Science and Technology
(P < 0.01). Bioefficiency of the added bioactive sub-
stances in food matrices depends strongly on the
structural and chemical composition of the respective
matrix and chosen encapsulation system. The release of
the active molecules from the matrix during digestion
and their absorption into blood stream are a function of
micro or nano structure of the food and the encapsu-
lation systems developed by the food and ingredient
industry for delivery of bioactives and micronutrients
(Ubbink & Krüger, 2006; Prada & Aguilera, 2007;
Palzer, 2009). The intestine has special mechanism to
absorb particles, and there may be a size-exclusion
phenomenon in the gastrointestinal absorption of par-
ticles, with 100 nm particles showing a higher uptake
(10- to 250-fold higher) than larger particles (500 -
nm–10 lm) (Hu et al., 2011). Swarnakar et al. (2011)
investigated the development and characterisation of
coenzyme Q10 loaded PLGA nanoparticles (size <
100 nm) by a scalable emulsion–diffusion evaporation
method and coenzyme Q10 nanoparticles showed
improved oral bioavailability (4.28 times) as compared
to free coenzyme Q10. Ankola et al. (2007) reported that
the intestinal uptake of coenzyme Q10 as the developed
nanoparticle formulation was found to be 79%, sug-
gesting that solubility and permeability related problems
of coenzyme Q10 were overcome by nanoparticle
formulation.
Conclusion
Coenzyme Q10 has become an increasingly popular
dietary supplement in recent years. Numerous CoQ10
products are available on the market in the form of
chewable, soft gelatin or capsule. However, the bio-
availability of CoQ10 in these products is very low
because pure CoQ10 is insoluble in water. On the other
hand, CoQ10 added to foods with emulsion, complex or
nanoparticle was more soluble during simulated gastric
and small intestine phases of digestion. Results from the
present study indicated that coenzyme Q10 enriched
foods could be used as important sources of coenzyme
Q10. As a new product approach that had a high
nutritional value was formed, the yoghurts were
enriched with different coenzyme Q10 preparations.
This study demonstrates that coenzyme Q10 could be
released from the matrix of enriched yoghurts. The
highest coenzyme Q10 bioaccessibility was found in
yoghurt enriched with nanoparticle coenzyme Q10.
These enriched yoghurts can increase the daily intake
of coenzyme Q10. A possible extension of this research
would involve further collection of fundamental data to
produce bioavailable CoQ10 products (e.g. nanoemul-
sion). Also, the coupled simulated gastric ⁄ small intesti-
nal digestion with Caco-2 cell model can be studied for
determining the bioavailability ⁄ bioaccessibility of
CoQ10.
 2012 The Authors
 Bioaccessibility of coenzyme Q10 P. Ercan and S. N. El 7

McClements, D.J. (2010). Design of nano-laminated coatings to control

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International Journal of Food Science and Technology  2012 Institute of Food Science and Technology