International Journal of Food Sciences and Nutrition,

2012; Early Online: 17

LDL cholesterol-lowering effects of grape extract used as a dietary

supplement on healthy volunteers

NOEMI YUBERO1, MARISA SANZ-BUENHOMBRE1, ALBERTO GUADARRAMA2,

N3, EIDER LARRARTE4, & CARLOS MORO2
SONIA VILLANUEVA2, JUAN M. CARRIO
1
Research and Development Department, Esdor Cosmeticos, Valladolid, Spain, 2Research and Development Department, Bodega
Matarromera, Valladolid, Spain, 3Chief of Service of Endocrinology and Nutrition, Hospital Nisa Pardo Aravaca, Madrid,
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Spain, and 4TECNALIA, Health Division, Parque Tecnologico de A lava, Minano, Spain

Abstract
The aim of this study was to evaluate the effects of Eminolw, the polyphenol-rich grape extract supplement (700 mg), on
cardiovascular risk and oxidant stress indicators in a sample of volunteers. A randomized, double-blind, placebo-controlled
clinical trial was performed over 56 days and included 60 volunteers. Thirty volunteers took 700 mg of the grape extract,
Eminolw (E), and 30 took the placebo (P). On comparison of the results, a decrease in total cholesterol (E: 213.77 ^ 4.1 mg/dl
and P: 245.57 ^ 4.1 mg/dl; p 0.01) and LDL cholesterol (E: 142.17 ^ 3.1 mg/dl and P: 165.13 ^ 3.1 mg/dl; p 0.02) levels
as well as an increase in antioxidant capacity (E: 65.63 ^ 5.8 mmol TE/mg and P: 57.80 ^ 7.7 mmol TE/mg; p , 0.01) and
vitamin E (E: 11.46 ^ 0.5 mg/ml and P: 9.06 ^ 0.5 mg/ml; p 0.018) was observed. This result indicates that the grape extract
For personal use only.

Eminolw modulated the lipid profile in terms of cardiovascular risk indicators, lowering total blood cholesterol and LDL
cholesterol levels.

Keywords: grape extract, polyphenols, cholesterol, LDL cholesterol, antioxidant capacity

Introduction
Polyphenols are component parts of the secondary
metabolism of vegetables. Most of them are potent
antioxidant substances, thanks to their ability to
donate electrons, and thus to eliminate free radicals
(Van Acker et al. 1995). In vitro studies have
demonstrated that many polyphenols provide more
antioxidant power than vitamins E and C. Polyphenols
are found in fruits and vegetables, for instance, apples,
grapes and onions, and in beverages such as tea and
wine (Quideau et al. 2011), being therefore a common
constituent of the daily human diet. The polyphenols
ingested in the diet primarily belong to four families:
flavonoids, phenolic acids, stilbenes and lignans
(Perez-Jimenez et al. 2010).
Polyphenols are considered to contribute towards
the prevention of several illnesses such as degenerative
(Thomas et al. 2009), inflammatory (Sakurai et al.
2010) and cardiovascular (Manach et al. 2005; Lecour
and Lamont 2011; Wu and Hsieh 2011) diseases,
among others. So far, many studies have demonstrated
a relationship between the consumption of polyphe-
nols in the diet and various chronic pathologies such as
cancer (Yang et al. 2001; Rajendran et al. 2011),
diabetes (Bashmakov et al. 2011) and osteoporosis
(Shen et al. 2008).
Many studies have focused on cardiovascular
diseases since they are among those with the highest
prevalence in developed countries and their incidence
is gradually increasing in developing countries. Several
epidemiological studies have shown that there is an
inverse relationship between the intake of polyphenols,
or food rich in polyphenols characteristic of the
Mediterranean diet, and the risk of suffering from
cardiovascular diseases (Mukamal et al. 2002;

Correspondence: Noem Yubero, Research and Development Department, Esdor Cosmeticos, S.L., Ctra. San Bernardo, s/n. Valbuena de
Duero 47359, Valladolid, Spain. Tel: 34 902430170. Fax: 34 302430189. E-mail: imasd@esdor.es

ISSN 0963-7486 print/ISSN 1465-3478 online q 2012 Informa UK, Ltd.

DOI: 10.3109/09637486.2012.753040
 2 N. Yubero et al.
Mennen et al. 2004). Likewise, it has been observed
that moderate wine consumption may help to prevent
or even lower cardiovascular risk (Di Castelnuovo
et al. 2002), probably due to the high polyphenol
content. This relationship between moderate wine
consumption and benefits to cardiovascular health
has grown in significance since the discovery of the
so-called French Paradox in the 1990s (Renaud and
de Lorgeril 1992).
However, most studies just consider the presence
of a limited number of polyphenols representing the
wide range of molecules found in the diet (Neveu
et al. 2010; Perez-Jimenez et al. 2010). The results of
these studies on volunteers have not always indicated
a reduction in cardiovascular risk or a decrease in
target variables such as LDL cholesterol or total
cholesterol (Boots et al. 2008).
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In this study, the lowering effect of grape
polyphenols used as a dietary supplement on LDL
cholesterol levels is assessed in a sample of healthy
volunteers. The grape extract Eminolw comes from
grape skin after the vinification process. During red
wine production, some of the polyphenols contained
in the grape pass into the wine, nevertheless, some are
held in the skin, which can be used afterwards.
To further evaluate the benefits of Eminolw grape
For personal use only.
extract, a sample of healthy individuals was given either
Eminolw or a placebo, both by oral administration,
in order to assess the primary variables involved in
the risk of developing cardiovascular diseases.
Materials and methods
Grape extract
The grape extract used in this trial was obtained from
grape pomace. This extract, registered as Eminolw,
was obtained by means of a unique and new system
developed and patented by Grupo Matarromera
(ES 2 309 032). The extract comes 100% from
the red grape Vitis vinifera variety Tempranillo,
harvested from vineyards located in the central region
of Spain, specifically the Ribera de Duero Designation
of Origin, in Castile and Leon.
The composition of the grape extract Eminolw is
included in Table I, and it is compared with the same
Table I. Grape extract. Polyphenols characterized in Eminolw.
Polyphenolic Eminolw Fresh grape Eminolw/grape
compounds (mg/g) skin (mg/g) skin ratio
Total flavonols 2.61 0.636
Myricetin 1.23 0.153
Quercetin 1.40 0.155
Catechin 2.34 0.0128 183
Epicatechin 0.77 0.0154
Epigallocatechin 1.63 0.0075 217
Catechin gallate 0.11 0.0096
Ellagic acid 2.42
amount of polyphenols in fresh grape skin, also
showing the relationship between Eminolw and fresh
grape skin.
Nutritional information
In order to find out the nutritional composition of
the grape extract, the main components of the extract
were calculated using the following methods:
. Water: internal method based on method number
731 Loss on Drying United States Pharmacopeia
(USP) 34.
. Sodium: internal method based on method number
851 Spectrophotometry and Light-scattering
United States Pharmacopeia (USP) 34.
. Calcium: internal method based on method number
851 Spectrophotometry and Light-scattering
United States Pharmacopeia (USP) 34.
. Vitamin A: internal method based on method
number 621 Chromatography United States
Pharmacopeia (USP) 34.
. Vitamin C: internal method based on method
number 621 Chromatography United States
Pharmacopeia (USP) 34.
Clinical trials with volunteers
Individuals. A sample of 60 healthy volunteers aged
between 34 and 65 was studied. It was a representative
population of healthy men and women with normal
lifestyles and diets.
The main recruitment criteria were as follows:
appropriate cultural level and understanding of the
clinical study; body mass index (BMI) of between 19
and 30 kg/m2; not having taken any hypolipidaemic
drugs or any other medicines to treat hypercholester-
olaemia for 4 weeks prior to the start of the trial; LDL
cholesterol level of between 130 and 190 mg/dl, in the
event two or more cardiovascular risk factors are
detected, LDL cholesterol level of between 100 and
190 mg/dl; triglyceride level no higher than 350 mg/dl;
not having presented any cardiovascular ischaemic
events within the last 6 months; in women, not being
pregnant or lactating; not presenting hypersensitivity
to any of the product components of the trial; not
having taken part in any other clinical trial for the
3 months prior to the start of the study.
During the study period, volunteers were given a
typical Mediterranean diet.
4
Study design. An exploratory, randomized, double-
8 blind, placebo-controlled, parallel-group, clinical-
9 nutritional study was conducted in order to assess
the health properties and tolerability of the
50 polyphenolic extract Eminolw on different
11
cardiovascular risk and oxidative stress indicators by
using a sample of healthy volunteers during an
experimental period of 56 days (8 weeks).

All volunteers received verbal and written infor-
mation about the nature and purpose of the study,
and all of them gave written consent for participation.
This study was undertaken in accordance with
the Helsinki II declaration and was approved by the
Ethical Committee at Hospital Universitario Puerta de
Hierro (Majadahonda-Madrid) on 24 March 2010.
The promoter of this study was Grupo Matarromera
and the trial was performed under the scientific
co-ordination of doctors Juan Manuel Carrion Pastor
and Eider Larrarte Lazaro.
Volunteers were randomized into two groups, and
each was given either a 700 mg Eminolw supplement
(two Esdor capsules of 350 mg) or 700 mg of placebo
(two maltodextrin capsules of 350 mg). Both products
were supplied in 350 mg capsule form, and volunters
were instructed to take two capsules at breakfast over
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the 56 days.
Volunteers filled in a questionnaire on their dietary
habits and various parameters related to cholesterol
and antioxidant capacity were recorded before starting
the trial (T0), after 28 days (T28) and after 56 days
(T56). During the clinical trial, neither the volunteer,
nor the researcher knew the type of supplement
administered to the volunteer.
Sample collection and analyses of endogenous parameters
For personal use only.
Sample collection. Blood samples were extracted from
volunteers and collected in the initial visit (T0), after
28 days or the intermediate visit (T28) and after
56 days or the final visit (T56) in order to analyse the
various endogenous parameters.
Analyses
Biomarkers of antioxidant status and oxidative stress.
. Oxygen radical absorbance capacity (ORAC): The
plasma antioxidant capacity was determined by
means of the commercial fluorescence method
called OxiSelecte Oxygen Radical Antioxidant
Capacity (ORAC) Activity Assay (Catalogue
Number STA-345) from Cell Biolabs, following
manufacturers instructions.
. LDL oxidation: Anti-oxLDL autoantibodies were
determined using a commercially available ELISA
kit (Biomedica, Vienna, Austria).
. Vitamin E: Plasma vitamin E levels were deter-
mined using a reagent kit for the HPLC analysis of
vitamins A and E in serum/plasma from Chrom-
Systems following manufacturers instructions.
. Ascorbic acid: Plasma vitamin C levels were deter-
mined using a commercially available Ascorbate
Assay kit item num. 700420 from Cayman Chemical
Company following manufacturers instructions.
. Tumoral necrosis factor (TNF-a): Anti-TNF-a anti-
bodies were determined using a commercially avai-
lable ELISA kit (R&D systems, Inc., Minneapolis,
MN, USA)
LDL cholesterol-effects of grape extract 3
. GGT: Gamma-glutamyl transferase (g-GT) levels
were determined using a commercially available
kit: quantitative determination of gGT In Vitro
Diagnostic IVD, Ref. MI41288, from Spinreact
following manufacturers instructions.
. GOTand GPT: The glutamic transaminase enzymes,
serum glutamic-oxalacetic (GOT) and serum
glutamic-pyruvic (GPT) levels were determined
using a commercially available kit: quantitative deter-
mination of transaminases GOT and GPT IVD,
Ref. 1001165 (GOT) & Ref. 1001175 (GPT) from
Spinreact following manufacturers instructions.
Plasma cholesterol levels: total cholesterol, Low Density
Lipoprotein LDL cholesterol, High Density Lipoprotein
HDL cholesterol and LDL/HDL ratio.
. Cholesterol: Plasma cholesterol levels were calculated
using a commercially available kit: quantitativedeter-
mination of cholesterol IVD from SPINREACT
following manufacturers instructions.
. LDL cholesterol: Plasma LDL cholesterol levels
were calculated using a commercially available kit:
quantitative determination of LDL cholesterol
IVD from SPINREACT following manufacturers
instructions.
. HDL cholesterol: Plasma HDL cholesterol levels
were calculated using a commercially available kit:
quantitative determination of HDL cholesterol
IVD from SPINREACT following manufacturers
instructions.
Data analysis
The sample size was calculated to detect differences
in cholesterol levels after treatment with 700 mg of
Eminolw.
Intra-group time variations in the parameters were
analysed by applying a generalized linear model for
repeated measurements. As for inter-group variations,
a Students t-test was carried out. In every hypothesis
testing process, when the p value was lower than 0.05,
significant differences were considered.
Results
Nutritional information
Nutritional information is given in Table II. Energy
intake from the recommended daily dose of Eminolw
(700 mg/day) is just 2.97 kcal. The composition of
the Eminolw grape extract shows extremely high
amounts of calcium (3.06 mg/700 mg) and dietary
fibre (0.13 g/700 mg).
Clinical-nutritional trial
Of the 60 volunteers who participated in this study,
29 were men and 31 were women aged between 34
and 64 (mean age 51 ^ 7 years old). No significant
 4 N. Yubero et al.

Table II. Nutritional information for Eminolw grape extract and Furthermore, and in accordance with the anti-
grapes. oxidant activity of the polyphenols contained in the
Nutritional Per 100 g Per daily dose Per 100 g grape extract Eminolw, the plasma antioxidant
information of Eminolw (two capsules) of grape(31) capacity (ORAC) and the blood levels of vitamin E
increased during treatment with the grape extract
Energy 320.92 kcal 2.97 kcal 6767.1 kcal
(1341.45 kJ) (12.42 kJ) (280 kJ) Eminolw, comparing T0 with T56, and with the
Carbohydrates 58.32 g 0.54 g 1516 g volunteers who took the placebo (Table III).
Sugars 4.12 g 0.038 g 1516 g The evolution of the LDL cholesterol/HDL
Total polyphenols 13 g 0.12 g 0.130 0.154 g(32) cholesterol ratio decreased (T0: 3.84 ^ 0.16 mg/dl;
Proteins 0.29 g 0.003 g 0.60.7 g
Fats 2g 0.02 g 0.60.7 g
T56: 3.08 ^ 0.14 mg/dl) whereas the HDL choles-
Dietary fibre 13.57 g 0.13 g 0.4 g terol levels increased (T0: 44.23 ^ 1.60 mg/dl; T56:
Water 4.8 g 0.04 g 8183.2 g 47.10 ^ 1.60 mg/dl) among the volunteers who took
Sodium 12.9 mg 0.12 mg 2 mg 700 mg of the grape extract Eminolw, comparing
Calcium 331 mg 3.06 mg 4 mg
Vitamin C ,5 mg ,0.05 mg 4 mg
T0 with T56 where a significant difference was
Vitamin A ,25 mg ,0.23 mg 3 10 mEq noted. Nevertheless, there was no significant differ-
ence when comparison was made with the group who
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took the placebo.

differences were found between treatments and age
( p 0.971).
A descriptive analysis of the basal variables was Table III. Results from the clinical trial on variables related to lipid
carried out, calculating the mean and the standard profile, oxidative stress and inflammation.
deviation for the quantitative variables and performing
Eminolw Placebo
a frequency analysis for the qualitative variables. P between
As demonstrated in Table III, at the beginning of Average Error Average Error groups
the trial no significant differences were found between
Total cholesterol P 0.01
the group of volunteers who took the grape extract
T0 247.43 5.05 252.63 5.05
For personal use only.

Eminolw (two capsules of 350 mg) and the group who T28 232.83 4.60 245.83 4.60
took the placebo with regard to antioxidant capacity T56 213.77 4.07 245.58 4.07
and oxidative stress parameters or biomarkers, such as LDL cholesterol P 0.02
total cholesterol, LDL cholesterol, HDL cholesterol, T0 166.67 3.76 168.40 3.76
T28 154.93 3.51 164.93 3.51
oxidized LDL, plasma antioxidant capacity and T56 142.17 3.12 165.13 3.12
vitamins C and E blood levels. These data confirmed LDL oxidation P 0.06
the homogeneous distribution of the individuals T0 54.69 1.92 56.42 1.92
included in the study. T28 51.41 1.85 55.91 1.85
T56 47.23 1.65 55.38 1.65
After 56 days of intake, the volunteers who took ORAC P , 0.01
700 mg of the grape extract Eminolw presented a signifi- T0 51.23 4.80 56.70 7.71
cantly lower total cholesterol level (213.77 ^ T28 61.10 5.05 57.93 7.74
4.07 mg/dl) compared with the volunteers who took T56 65.63 5.79 57.80 7.71
Vitamin E P 0.018
the placebo (245.57 ^ 4.07 mg/dl), with a significance
T0 9.87 0.44 9.00 0.44
level of p 0.01. Moreover, the decrease in total T28 10.41 0.46 9.05 0.46
cholesterol level was also significant for the volunteers T56 11.46 0.46 9.06 0.46
who consumed the grape extract Eminolw, comparing Vitamin C P negligible
the first visit (T0 247.43 ^ 5.05 mg/dl) and the last T0 9.80 0.41 9.65 0.41
T28 9.88 0.40 9.63 0.40
visit (T56 213.77 ^ 4.07 mg/dl). T56 10.13 0.41 9.64 0.41
With regard to the previous result, LDL LDL/HDL ratio P 0.243
cholesterol levels and oxidized LDL levels also T0 3.84 0.16 3.81 0.16
decreased, as observed in Table III. In the case T28 3.56 0.15 3.70 0.15
T56 3.08 0.14 3.70 0.14
of LDL cholesterol, this decrease was statistically HDL cholesterol P 0.599
significant when comparing the volunteers who T0 44.23 1.60 46.27 1.60
took 700 mg of the grape extract Eminolw T28 44.73 1.60 46.70 1.60
(142.17 ^ 3.1 mg/dl) with the volunteers who took T56 47.10 1.60 46.67 1.60
TNF-a P negligible
the placebo (165.13 ^ 3.1 mg/dl), with a significance
T0 4.63 0.24 4.83 0.24
level of p 0.02. Moreover, it was also observed T28 4.54 0.24 4.86 0.24
that the decrease in LDL cholesterol was statistically T56 4.51 0.24 4.87 0.24
significant when comparing the values from the Fasting glucose P negligible
T0 93.13 2.50 90.27 2.32
first day of Eminolw grape extract consumption (T0:
T28 93.60 2.71 90.27 2.30
54.68 ^ 1.9 mg/dl) with the values from the last day, T56 92.63 2.68 89.70 2.28
after 2 months of treatment (T56: 47.23 ^ 1.6 mg/dl).
 LDL cholesterol-effects of grape extract 5

Table IV. Results from the clinical trial on weight, percentage of With regard to the clinical trial carried out to
body fat and BMI. evaluate the health properties of the grape extract
Eminolw Placebo Eminolw on human health, the results of the study
showed that consuming the grape extract Eminolw
Average Error Average Error over a period of 56 days led to a statistically significant
Weight
decrease in total cholesterol, LDL cholesterol and
T0 73.62 1.54 72.93 1.54 oxidized LDL cholesterol concentrations in plasma,
T28 73.49 1.53 72.83 1.53 which are the main metabolic variables indicating
T56 73.40 1.53 72.66 1.53 cardiovascular risk. This reduction in total cholesterol
% Body fat and LDL cholesterol levels was very likely due to the
T0 23.60 0.71 23.57 0.71
T28 23.51 0.71 23.47 0.71
action mechanisms of polyphenols found in the
T56 23.38 0.70 23.30 0.70 polyphenolic extract which include the inhibition of
BMI the specific uptake/transporters for cholesterol, such as
T0 26.92 0.39 26.46 0.39 Niemann-Pick C1 Like 1 cholesterol transporter
T28 26.90 0.39 26.45 0.39 (Leifert and Abeywardena 2008).
T56 26.86 0.39 26.40 0.39
The decrease in cholesterol levels through the
administration of grape polyphenols had already been
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demonstrated both in works with cellular models and

In the case of other variables such as fasting glucose
or TNF-a, no significant differences were found bet-
ween the trial groups (Eminolw vs. placebo) (Table III).
Regarding weight, body fat percentage or BMI,
no significant differences between the groups were
found either; including the volunteers in the protocol
obviously did not modify their total calorie intake
(Table IV).
Finally, from the results obtained it is worth noting
For personal use only.
animals (Leifert and Abeywardena 2008; Ikeda et al.
2010; Peluzio Mdo et al. 2011). Hence, evidence
already existed relating to the reduction in the risk of
suffering from cardiovascular diseases and the intake
of antioxidants, specifically polyphenols, which are
considered to be cardio-protective elements (Zern and
Fernandez 2005). However, effects on cardiovascular
risk factors had not always been observed in clinical

trials (Hansen et al. 2005). This fact should be applied

that there were no significant differences in terms of to both the origin and the composition of the
safety variables when comparing the groups who took polyphenolic extract from the grape. Thus, it was
or did not take the grape extract Eminolw and widely known that moderate variations may be seen in
comparing the beginning (T0) and the end (T56) of the polyphenolic composition and content of grapes
the treatment. The safety variables analysed were depending on the crop, as well as influence from the
creatine and transaminases (GPT, GGT and GOT) geographical location and the climatic conditions
levels (Table V). (Rathel et al. 2007; Preedy et al. 2011). Our study
proves that the extracted and concentrated polyphe-
nols in the grape extract Eminolw, found in the skin of
Discussion grapes cultivated in central Spain, in the Ribera del
This study comprises a clinical trial that has been Duero Designation of Origin, have the capacity of
performed in order to assess the effect of the grape lowering total cholesterol, LDL cholesterol and
extract Eminolw on human health, mainly on lowering oxidized LDL cholesterol levels. On the other hand,
the total cholesterol and LDL cholesterol. the results presented in the INTERHEART study
support the theory stating that the antioxidants
consumed in a normal diet protect against coronary
Table V. Results from the clinical trial on creatine and
diseases (Yusuf et al. 2004) and, therefore, the
transaminases (GPT, GGT and GOT).
consumption of complex blends of antioxidants
Eminolw Placebo found in vegetables may probably be more beneficial
than the ingestion of large doses of only one
Average Error Average Error
antioxidant (Boots et al. 2008; Khan et al. 2011).
GPT Oxidized low-density lipoproteins, atherogenic
T0 14.78 0.75 15.70 1.03 lipoproteins that play an important role in pro-
T28 15.00 0.75 15.80 1.05 inflammatory reactions, are key elements in the
T56 15.15 0.76 16.03 1.07
GGT
atherosclerosis development process (Khan et al.
T0 17.63 0.88 16.73 0.97 2011). This is due to the fact that they are particles
T28 17.47 0.91 16.83 1.01 that favour the response of macrophages in the arteries
T56 17.77 0.95 17.07 1.03 and thus promote atherosclerosis processes (Fraley
GOT and Tsimikas 2006). Our results showed a statistically
T0 15.20 0.68 15.90 0.91
T28 15.28 0.70 15.93 0.94
significant decrease in the plasma oxidized LDL
T56 15.57 0.72 16.13 0.96 cholesterol concentration in the volunteers who took
the grape extract Eminolw compared with those who
 6 N. Yubero et al.
took the placebo. This reduction in plasma-oxidized
LDL cholesterol could be explained by the fact that
some ingested polyphenols may combine with LDL
cholesterol particles in the plasma, thus preventing
their oxidation (Rios et al. 2003).
Moreover, the results of the study proved a trend in
the significance that a bigger n would probably be
significant in the plasma HDL cholesterol concen-
tration on the volunteers who took the grape extract
Eminolw. The intra-group difference was statistically
significant; however, the inter-group difference was
not statistically significant. HDL cholesterol has an
anti-inflammatory function, so an increase in the HDL
cholesterol and a decrease in the plasma LDL
cholesterol would favour the inhibition of atheroma-
tous plaque formation. The increase in HDL
cholesterol due to polyphenol intake has been widely
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described (Ohtsuki et al. 2003). Hence, it is well
known that polyphenols, such as genistein, can
increase the expression of the main protein component
of HDL, apolipoprotein A1, on rats (Lamon-Fava
2000). So, the flavonoids contained in the grape
extract Eminolw were supposed to be absorbed and a
slight upward trend was observed in the HDL
cholesterol levels from the beginning of the adminis-
tration period until the end of the study. However, this
For personal use only.
upward trend is not statistically significant in this
study, so another study would be necessary using more
volunteers to confirm this trend in HDL cholesterol.
The results of the trial showed a rise in plasma
antioxidant capacity in volunteers who took the grape
extract Eminolw compared to those who took the
placebo. The data suggest that this increase is related
to the absorption levels of the polyphenols present in
Eminolw. Therefore, ingesting the grape extract
Eminolw for several days could cause a cumulative
effect in plasma antioxidant capacity.
Finally, the results of the clinical trial showed that
neither of the two volunteer groups, one having taken
the grape extract Eminolw and the other having taken
the placebo, recorded changes in their weight, BMI or
body fat percentage. This suggests that the calorie
intake from the polyphenolic extract did not signify
additional intake in the calories ingested by every
volunteer. The nutritional profile of the capsules
shown in Table II highlights that two capsules
containing 700 mg of Eminolw in total (the rec-
ommended daily intake) provide 2.97 kcal. This value
is 32 times lower than the 93.8 kcal that would need to
be ingested to reach an equivalent quantity of
antioxidant capacity units from fresh grapes.
Conclusions
In conclusion, a 56-day intervention with 700 mg of
polyphenol-rich grape extract Eminolw modulated the
lipid profile with regard to cardiovascular risk
indicators, lowering total blood cholesterol and LDL
cholesterol levels. Furthermore, it had a positive
influence on antioxidant stress indicators, increasing
the antioxidant capacity in plasma and vitamin E
levels.
Acknowledgements
This research has been carried out within the
framework of the project (SENIFOOD-Industrial
research on diets and foods with specific character-
istics for older people) funded by the CDTI, Ministry
of Economy and Competitiveness (Spain). We thank
the Ministry of Agriculture and Livestock of the
Autonomous Government of Castile and Leon for
the funding awarded to develop the previous studies.
We also thank Quantum Experimental for coordi-
nating the study, and Hospital Txagorritxu (Vitoria-
Spain) and Hospital Puerta de Hierro (Madrid-Spain)
for participating in the study.
Declaration of interest: The authors report no
conflicts of interest. The authors alone are responsible
for the content and writing of the paper.
References
Bashmakov YK, Assaad-Khalil S, Petyaev IM. 2011. Resveratrol
may be beneficial in treatment of diabetic foot syndrome. Med
Hypotheses 77(3):364367.
Boots AW, Haenen RG, Bast A. 2008. Health effects of quercetin:
from antioxidant to nutraceutical. Eur J Pharmacol 585:
325 337.
Di Castelnuovo A, Rotondo S, Iacoviello L, Donati MB, de
Gaetano G. 2002. Meta-analysis of wine and beer consumption
in relation to vascular risk. Circulation 105:28362844.
Fraley AE, Tsimikas S. 2006. Clinical applications of circulating
oxidized low-density lipoprotein biomarkers in cardiovascular
disease. Curr Opin Lipidol 17:502509.
Hansen AS, Marckmann P, Dragsted LO, Finne Nielsen IL, Nielsen
SE, Gronbaek M. 2005. Effect of red wine and red grape extract
on blood lipids, haemostatic factors, and other risk factors for
cardiovascular disease. Eur J Clin Nutr 59:449 455.
Ikeda I, Yamahira T, Kato M, Ishikawa A. 2010. Black-tea
polyphenols decrease micellar solubility of cholesterol in vitro
and intestinal absorption of cholesterol in rats. J Agric Food
Chem 58(15):85918595.
Khan N, Monagas M, Andres-Lacueva C, Casas R, Urp-Sarda` M,
Lamuela-Raventos RM, Estruch R. 2011. Regular consumption
of cocoa powder with milk increases HDL cholesterol and
reduces oxidized LDL levels in subjects at high-risk of
cardiovascular disease. Nutr Metab Cardiovasc Dis 22(12):
1 8.
Lamon-Fava S. 2000. Genistein activates apolipoprotein AI gene
expression in the human hepatoma cell line Hep G2. J Nutr 130:
24892492.
Lecour S, Lamont KT. 2011. Natural polyphenols and cardiopro-
tection. Mini Rev Med Chem 11(14):11911199.
Leifert WR, Abeywardena MY. 2008. Grape seed and red wine
polyphenol extracts inhibit cellular cholesterol uptake, cell
proliferation, and 5-lipoxygenase activity. Nutr Res 28(12):
842 850.
Manach C, Mazur A, Scalbert A. 2005. Polyphenols and prevention
of cardiovascular diseases. Curr Opin Lipidol 16(1):7784.
Mennen LI, Sapinho D, De Bree A, et al. 2004. Consumption of
foods rich in flavonoids is related to a decreased cardiovascular
risk in apparently healthy French women. J Nutr 134:923926.

Mukamal KJ, Maclure M, Muller JE, et al. 2002. Tea consumption
and mortality after acute myocardial infarction. Circulation 105:
24762481.
Neveu V, Perez-Jimenez J, Vos F, et al. 2010. Phenol-Explorer: an
online comprehensive database on polyphenol contents in foods,
Database (Oxford) 2010:bap024.
Ohtsuki K, Abe A, Mitsuzumi H, Kondo M, Uemura K, Iwasaki Y,
et al. 2003. Glucosyl hesperidinim proves serum cholesterol
composition and inhibits hypertrophy in vasculature. J Nutr Sci
Vitaminol (Tokyo) 49:447450.
Peluzio Mdo C, Teixeira TF, Oliveira VP, Sabarense CM, Dias CM,
Abranches MV, Maldonado IR. 2011. Grape extract and
a-tocopherol effect in cardiovascular disease model of Apo
E2/2 mice. Acta Cir Bras 26(4):253260.
Perez-Jimenez J, Neveu V, Vos F, Scalbert A. 2010. Systematic
analysis of the content of 502 polyphenols in 452 foods and
beverages: an application of the Phenol-Explorer database.
J Agric Food Chem 58:49594969.
Preedy VR, Watson RR, Patel VB. 2011. Nuts and seeds in health
Int J Food Sci Nutr Downloaded from informahealthcare.com by 77.226.250.121 on 12/19/12
and disease prevention. Part 2: effects to specific Nuts and Seeds.
Academic Press. Elsevier Inc. ISBN: 9780123756886.
Quideau S, Deffieux D, Douat-Casassus C, Pouysegu L. 2011. Plant
polyphenols: chemical properties, biological activities, and
synthesis. Angew Chem Int Ed Engl 50(3):586621.
Rajendran P, Ho E, Williams DE, Dashwood RH. 2011. Dietary
phytochemicals, HDAC inhibition, and DNA damage/repair
defects in cancer cells. Clin Epigenetics 3(1):4.
Rathel TR, Samtleben R, Vollmar AM, Dirsch VM. 2007.
Activation of endothelial nitric oxide synthase by red wine
polyphenols: impact of grape cultivars, growing area and the
vinification process. J Hypertens 25(3):541549.
For personal use only.
LDL cholesterol-effects of grape extract 7
Renaud S, de Lorgeril M. 1992. Wine, alcohol, platelets, and the
French paradox for coronary heart disease. Lancet 339(8808):
15231526.
Rios LY, Gonthier MP, Remesy C, Mila I, Lapierre C, Lazarus SA,
et al. 2003. Chocolate intake increases urinary excretion of
polyphenol-derived phenolic acids in healthy human subjects.
Am J Clin Nutr 77:912918.
Sakurai T, Kitadate K, Nishioka H, Fujii H, Kizaki T, Kondoh Y,
et al. 2010. Oligomerized grape seed polyphenols attenuate
inflammatory changes due to antioxidative properties in
coculture of adipocytes and macrophages. J Nutr Biochem
21(1):47 54.
Shen CL, Wang P, Guerrieri J, Yeh JK, Wang JS. 2008. Protective
effect of green tea polyphenols on bone loss in middle-aged
female rats. Osteoporos Int 19(7):979990.
Thomas P, Wang YJ, Zhong JH, Kosaraju S, OCallaghan NJ,
Zhou XF, Fenech M. 2009. Grape seed polyphenols and
curcumin reduce genomic instability events in a transgenic
mouse model for Alzheimers disease. Mutat Res 661:12.
Van Acker SA, Tromp MN, Haenen GRMM, van der Vijgh WJ, Bast
A. 1995. Flavonoids as scavengers of nitric oxide radical.
Biochem Biophys Res Commun 214:755759.
Wu JM, Hsieh TC. 2011. Resveratrol: a cardioprotective substance.
Ann N Y Acad Sci 1215:1621.
Yang CS, Prabhu S, Landau J. 2001. Prevention of carcinogenesis by
tea polyphenols. Drug Metab Rev 33(3 4):237253.
Yusuf S, Hawken S, Ounpuu S, Dans T, Avezum A, Lanas F, et al.
2004. Effect of potentially modifiable risk factors associated
with myocardial infarction in 52 countries (the INTERHEART
study): case-control study. Lancet 364:937 952.
Zern TL, Fernandez ML. 2005. Cardioprotective effects of dietary
polyphenols. J Nutr 135:22912294.