Food Chemistry 168 (2015) 203210

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Inuence of olive oil on carotenoid absorption from tomato juice

and effects on postprandial lipemia
Sara Arranz a,b, Miriam Martnez-Hulamo b,c, Anna Vallverdu-Queralt b,c, Palmira Valderas-Martinez a,b,
Montse Illn c, Emilio Sacanella a,b, Elvira Escribano d, Ramon Estruch a,b, Rosa Ma. Lamuela-Raventos b,c,
a

Department of Internal Medicine, Institut dInvestigacions Biomdiques August Pi i Sunyer (IDIBAPS), Hospital Clinic, Faculty of Medicine, University of Barcelona, Barcelona, Spain
CIBER CB06/03 Fisiopatologa de la Obesidad y la Nutricin, (CIBERobn), Institute of Health Carlos III, Madrid, Spain
Nutrition and Food Science Department, XaRTA, INSA, School of Pharmacy, University of Barcelona, Barcelona, Spain
d
Department of Pharmacy and Pharmaceutical Technology, Biopharmaceutics and Pharmacokinetics Unit, School of Pharmacy, University of Barcelona, Barcelona, Spain
b
c

a r t i c l e

i n f o

Article history:
Received 16 December 2013
Received in revised form 4 June 2014
Accepted 8 July 2014
Available online 15 July 2014
Keywords:
cis/trans carotenoids
Lipid prole
Lycopene
Tomato juice

a b s t r a c t
The potential benets of tomato-rich diets for the cardiovascular system have been related to plasma
concentrations of carotenoids. In addition, the bioavailability of carotenoids from foods depends on their
chemical structure, processing and the food matrix. Our aim was to evaluate the effect of adding oil to
tomato juice (not treated with heat) on the bioavailability of plasma carotenoids and postprandial lipid
response. In a randomized, controlled, crossover feeding trial, eleven healthy volunteers were assigned to
receive a single ingestion of 750 g of tomato juice (TJ) containing 10% of rened olive oil/70 kg body
weight (BW) and 750 g of TJ without oil/70 kg BW on two different days. All lycopene isomers increased
signicantly in subjects consuming TJ with oil, reaching the maximum concentration at 24 h. LDL
cholesterol and total cholesterol decreased signicantly 6 h after the consumption of TJ with oil, which
signicantly correlated with an increase of trans-lycopene and 5-cis-lycopene, respectively.
2014 Elsevier Ltd. All rights reserved.

1. Introduction
Several studies have observed that a regular intake of tomato
products may have a signicant impact on human health
(Burton-Freeman, Talbot, Park, Krishnankutty, & Edirisinghe,
2012; Ghavipour et al., 2012). The consumption of P7 servings/
week of tomato-based products has been associated with a 30%
relative risk reduction for cardiovascular disease (CVD) (Sesso,
Liu, Gaziano, & Buring, 2003) and a reduction of coronary and
inammatory biomarkers (Riso et al., 2006; Sesso, Wang, Ridker,
& Buring, 2012). These potential benets of tomato-rich diets on
vascular health have been attributed to the high concentrations

Abbreviations: AUC024, area under the concentrationtime curve from time zero
(0) to 24 h; BMI, body mass index; BP, blood pressure; BW, body weight; CVD,
cardiovascular disease; Cmax/AUC024, absorption rate; HDL-c, high density lipoprotein cholesterol; HPLC-MS/MS, high pressure liquid chromatographymass
spectrometry in tandem; LDL-c, low density lipoprotein cholesterol; MTBE, methyl
tert-butyl ether; PTFE, polytetrauoroethylene; SD, standard deviation; TJ, tomato
juice; TC, total cholesterol.
Corresponding author at: Nutrition and Food Science Department, XaRTA, INSA,
School of Pharmacy, University of Barcelona, Avda. Joan XXIII, 08028 Barcelona,
Spain. Tel.: +34 93 4034843; fax: +34 93 4035931.
E-mail address: lamuela@ub.edu (Rosa Ma. Lamuela-Raventos).
http://dx.doi.org/10.1016/j.foodchem.2014.07.053
0308-8146/ 2014 Elsevier Ltd. All rights reserved.

of carotenoids, mainly lycopene, reached in plasma (Mordente

et al., 2011).
The bioaccessibility of carotenoids can be affected by many factors, including the food matrix, processing and cooking methods,
and the interactions, during digestion and absorption, with other
dietary compounds, such as bre, lipids, phytosterols and other
carotenoids (Yonekura & Nagao, 2007; Svelander et al., 2010).
Among dietary factors, heat and mechanical treatments of foods
and the presence of a certain amount of fat in the meal appear to
be critical factors for carotenoid bioaccessibility and bioavailability
in vivo (Fielding, Rowley, Cooper, & ODea, 2005). Structure also
plays an important role in carotenoid bioaccessibility. The trans
isomer is the most usual form of carotenoids found in fresh tomatoes, mainly trans-lycopene, since it is the most stable thermodynamically (Erdman, 2005). By contrast, human plasma and
tissues contain more than 50% of cis-isomers, the most common
and bioavailable forms of these compounds (Unlu et al., 2007).
Most studies on the bioavailability of carotenoids from tomato
products are focussed only on lycopene (Frohlich, Kaufmann,
Bitsch, & Bohm, 2006; Lee et al., 2009; Richelle et al., 2012). However, very little information is available, in the literature, regarding
the absorption and bioavailability of different lycopene isomers and
other carotenoids, and their possible effects on human lipid
metabolism.

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S. Arranz et al. / Food Chemistry 168 (2015) 203210

The aim of this study was to evaluate the effect of the addition
of oil to a tomato juice (TJ) preparation, without cooking, on postprandial absorption and bioavailability of carotenoids from TJ and
tentatively evaluate the plausible effect on lipid metabolism. For
this purpose an open, controlled, randomized, crossover feeding
trial was carried out in 11 healthy adults.
2. Materials and methods
2.1. Samples
The TJ was elaborated with raw tomato fruits (Lycopersicum esculentum L., Royalty variety) at commercial maturity, supplied by CASI
Cooperative (Almeria, Spain). TJ preparation was processed at the
Torribera campus, University of Barcelona (UB, Barcelona, Spain)
by a standardised industrial scale-like manufacturing process.
Rened olive oil was kindly furnished by the Juan Ballester Ross
Company (Tortosa, Spain). Briey, tomatoes were cleaned and sanitized with chlorinated water to remove any impurities and chemical
compounds on the surface. Afterwards, the tomatoes were cleaved
and weighed to obtain the desired amount of juice by crushing in
a blender texturiser at high speed. Then the rened olive oil or the
corresponding amount of water (10%) was added and the resulting
TJ was packaged under vacuum to be frozen at 20 C.

subjects were approved by the Ethics Committee of Clinical Investigation of the University of Barcelona (Spain) and the Institutional
Review Board of the Hospital Clinic in Barcelona. Written informed
consent was obtained from all subjects before inclusion in the trial.
This study has been registered at the London Controlled-trial register with ISRCTN99660610 as the number.
2.4. Dietary assessment
Before each intervention, we used a 24 h food recall questionnaire to assess compliance with the recommended diet. Dietary
intake was converted into nutritional data, using the Professional
Diet Balancer software (Cardinal Health Systems).
2.5. Blood pressure monitoring and biochemistry analysis
Blood pressure (BP), glucose and lipid prole were analysed at
baseline and 6 h after each intervention. BP was measured in the
non-dominant arm, using an automatic oscillometer (Omron 705
CP; Omron Matsusaka Co., Ltd., Matsusaka City, Japan) after
10 min resting in a seated position. Plasma concentration of total
cholesterol (TC) and triglycerides was analysed by enzymatic procedures: HDL and LDL cholesterol after precipitation with phosphotungstic acid and magnesium chloride and blood glucose by
the glucose oxidase method.

2.2. Subjects

2.6. Extraction and isolation of carotenoid compounds from TJ

Eleven healthy subjects (6 men and 5 women), with mean age,

28 3 years and mean BMI 23 2 kg/m2, were recruited. BMI was
obtained by doing measurements of height and weight at the
beginning of intervention. None reported any history of heart disease, homeostatic disorders or other medical conditions. All subjects were non-smokers and were not receiving medication or
vitamin supplements. None had followed any special diet for at
least 4 weeks prior to participating in the study.

The extraction of carotenoids was carried out very quickly,

avoiding exposure to light, oxygen, high temperatures and pro-oxidant metals such as iron or copper, in order to minimise autoxidation and cis/trans isomerisation. TJ samples (0.5 g) were
homogenised with 5 ml of ethanol/hexane (4:3 v/v), following a
procedure described elsewhere by Vallverdu-Queralt, MartinezHuelamo, Arranz-Martinez, Miralles and Lamuela-Raventos
(2012). The homogenate was sonicated for 5 min and centrifuged
at 2140g for 15 min at 4 C. The supernatant was transferred into
a ask and the extraction was repeated. The two supernatants
were combined and evaporated under nitrogen ow. Finally, the
residue was reconstituted with methyl tert-butyl ether (MTBE)
up to 1 ml and ltered through a 13 mm, 0.45 lm polytetrauoroethylene (PTFE) lter (Waters, Milford, MA, USA) into an insertamber vial for HPLC analysis. Samples were stored at 80 C prior
to analysis. Extractions were performed in triplicate.

2.3. Study design

The design was similar to previous acute studies performed by
our research group with other foods (Roura et al., 2007, 2008).
The study was an open, controlled, randomized and crossover
feeding trial. The eleven participants underwent a three-day washout period during which they were asked to consume their regular
diet avoiding tomato or processed tomato-based products and foods
containing carotenoids. A list showing permitted and forbidden
foods was provided to the participants, as two sample menus. The
subjects fasted for at least 8 h before the intervention intake. Using
a computer-generated random list of numbers, each volunteer was
assigned to receive: 750 g TJ with rened olive oil and 750 g TJ without rened olive oil/70 kg of body weight (BW) on two different
days. The TJ portion was chosen according to data available in the literature (Gustin et al., 2004; Porrini, Riso, & Testolin, 1998) in order to
achieve, after administration of a single dose, a sufcient carotenoid
plasma level to observe a quantiable effect response.
Blood samples were collected in plasma EDTA tubes before the
test meal and at 3, 6 and 24 h after each intervention. During the
6 h interval, participants rested in the clinical ward in a quiet room
and were allowed to drink only water. In the period from 6 to 24 h,
participants were asked to consume their regular diet, avoiding
tomato or processed tomato-based products and foods containing
carotenoids. Blood samples were immediately centrifuged after
collection at 1500g for 15 min at 4 C and plasma was aliquotted
and stored at 80 C prior to analysis.
This study was conducted according to the guidelines laid down
in the Declaration of Helsinki and all procedures involving human

2.7. Extraction and isolation of carotenoid compounds from human

plasma
Briey, 800 ll of ethanol was added to 800 ll of plasma. After
vortex-mixing for 45 s (s), the plasma was extracted twice with
hexane (2 ml, stabilized with 0.1 g/l of butylated hydroxytoluene),
and the extracts were vortex-mixed for 1 min and centrifuged at
2140g for 5 min at 4 C, following the procedure described by
Olmedilla et al. (1997). The organic phases were removed, pooled,
and evaporated under nitrogen ow. Finally, they were reconstituted with 300 ll of a solution of MTBE, ltered through a
13 mm, 0.22 lm PTFE lter (Waters, Milford, MA, USA) into an
insert-amber vial for HPLC analysis and injected into the HPLC
UV system.
2.8. HPLC separation and HPLCMS/MS identication of plasma
carotenoids
Chromatographic analysis was performed, using an HP 1100
HPLC system (HewlettPackard, Waldbronn, Germany) equipped
with a quaternary pump and an autosampler. The analytes were

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S. Arranz et al. / Food Chemistry 168 (2015) 203210

separated and identied according to the procedure previously

described by Vallverdu-Queralt et al. (2012).
Commercially available carotenoid standards (lutein, transbeta-carotene and trans-lycopene from Extrasynthese, Genay
Cedex, France and alpha-carotene from Carotenature, Ostermundigen, Switzerland) were used to identify and quantify analytes.
The HPLCUV chromatograms were acquired at 450 nm wavelength. The tentative identication of carotenoids and their geometrical isomers (cis-isomers) was done on the basis of the
retention times and absorption spectrum characteristics described
previously by our group (Vallverdu-Queralt et al., 2012) and by
comparison with other references in the literature (Frohlich
et al., 2006). When standards were not available, as in the case of
cis isomers of lycopene, the compounds were quantied by the
peak area of the trans-lycopene standard. Results were expressed
as mg l 1.

2.9. Quality parameters

The quality parameters established for the correct validation of
the HPLC method described for the quantication of carotenoids in
plasma were: recovery, limit of detection, limit of quantication,
accuracy, and precision. Validation was based on the criteria of
the AOAC International (Ofcial Methods of Analysis of AOAC
International; Washington, DC, 2005; Appendices D and E).

2.10. Pharmacokinetic analysis

Pharmacokinetic parameters were determined by means of a
non-compartmental analysis, using the WinNonlin software version 6.3 (2012 Certara, L.P.). The peak plasma concentration [Cmax
(mg/l)] was determined directly from the individual observed lycopene concentrationtime data. Area under the concentrationtime
curve from time zero (0) to 24 h [AUC024 (mg h/l)] was determined by the linear trapezoidal method. The absorption rate
parameter [Cmax/AUC024 (h 1)] was also calculated.
2.11. Statistical analysis
Statistical analysis was performed using the SPSS Statistical
Analysis System (version 17.0; SPSS Inc, Chicago, IL). Data are
presented as means and standard deviation (SD). Statistical differences between the two interventions, as well as between different
time periods, were analysed by the nonparametric statistical Friedman test for between-group comparisons and the Wilcoxon test for
paired comparisons. Statistical signicance was set at p < 0.05.
Associations between changes in carotenoid concentrations after
6 h and changes in lipid prole were calculated by the Spearmans
rank correlation coefcient (signicance level (alpha): 0.05).

3. Results
3.1. Carotenoid content in tomato juice
Table 1 shows the content of the main carotenoids identied in
the TJ administered during interventions. Fig. 1 shows the HPLC
MS/MS chromatogram of the carotenoid prole of TJ. The major
compound was beta-carotene (65.8 mg), followed by trans- and
13-cis-lycopene, (46.6 and 15.9 mg, respectively). The sums of
lycopene isomers and beta-carotene contributed to 52% and 46%
of the total intake of carotenoids, respectively. Moreover, translycopene of TJ represented 63% of the total lycopene content while
total cis-lycopene isomers represented 37%.

Table 1
Carotenoid content in dose of tomato juice administered during the intervention.
Compound

(mg/dose administered)a

cis-Lutein
trans-Lutein
alpha-Carotene
beta-Carotene
13-cis-Lycopene
9-cis-Lycopene
trans-Lycopene
5-cis-Lycopene
Total carotenes
Total lycopene isomers
Total carotenoids

0.53 0.08
0.52 0.08
0.06 0.01
65.8 9.55
16.0 2.32
5.96 0.86
46.7 6.77
5.50 0.80
66.7 23.1
74.1 10.8
141 20.4

Values are expressed as means SD.

a
Dose administered: 750 g/70 kg body weight.

In addition, carotenoid concentrations in the rened olive oil

were also analysed (data not shown) to verify that there was no
presence of these compounds.
3.2. Validation results
The HPLC method for quantication of plasma carotenoids has
been validated by the criteria of the AOAC International.
The recovery is the detector response obtained from an amount
of the analyte added to and extracted from the biological matrix,
compared to the detector response obtained for the same concentration of the pure authentic standard. The results showed comparable values of recovery in plasma samples, with values between
85% and 115% (109% for lutein, 88% for alpha-carotene, 108% for
beta-carotene and trans-lycopene and 99% for 5-cis-lycopene).
Limit of detection (LOD) was estimated for a signal-to-noise
ratio of 3 from the chromatograms of spiked blank plasma samples
at the lowest analyte concentration tested. Similarly, the limit of
quantication (LOQ) was determined for a signal-to-noise ratio of
10. Spiked plasma samples at ve different concentrations were
prepared in triplicate to establish the LOD and LOQ in the HPLC
system. The LODs for lutein, alpha-carotene, beta-carotene, translycopene and 5-cis-lycopene were 0.095, 0.013, 0.024, 0.142 and
0.238, respectively. Similarly, LOQs for lutein, alpha-carotene,
beta-carotene, trans-lycopene and 5-cis-lycopene were 0.13, 0.11,
0.07, 0.17 and 0.31, respectively.
Accuracy is the closeness of agreement between the measured
value and the accepted true or reference value. Experiments
were evaluated by repetitively spiking the biological matrix with
known levels of analyte standards. The accuracy of the method
was expressed as percentage of relative error. The accuracy of
the method was acceptable at each concentration in plasma: 99
114% for the low, 95115% for the medium, and 94109% for the
high concentrations.
Precision expresses the closeness of agreement among a series
of measurements obtained from multiple testing of a homogeneous test sample under the methods established conditions. Precision was determined by independently processing spiked plasma
samples at three different levels. Results showed acceptable precision of the proposed HPLC method with relative standard deviation
(RSD) values lower than 15% for all carotenoids.
3.3. Carotenoid content in plasma
All participants followed a three-day washout period before
each intervention, during which the main foods containing carotenoids were restricted. Therefore, baseline samples were assumed
to be representative of the minimum concentration of carotenoids
in plasma. No differences were observed for any carotenoid at
baseline between the two interventions.

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S. Arranz et al. / Food Chemistry 168 (2015) 203210

Fig. 1. Tomato juice carotenoids identied by HPLCMS/MS (HPLC coupled with triple quadrupole mass spectrometry).

Table 2
Plasma carotenoid concentrations at baseline and 3, 6 and 24 h after the TJ interventions (n = 11).
(mg l

trans-Lutein
cis-Lutein
alpha-Carotene
beta-Carotene
13-cis-Lycopene
9-cis-Lycopene
trans-Lycopene
5-cis-Lycopene
Total lycopene
Total carotenoids

TJ with oil

TJ without oil

0h

3h

6h

24 h

0h

3h

6h

24 h

0.27 0.11a
0.06 0.04a
0.09 0.03a
1.52 0.91a
0.61 0.22a
0.08 0.04a
0.81 0.40a
1.84 0.69a
3.33 1.20a
5.22 1.94a

0.24 0.10a
0.05 0.03a
0.10 0.04a
1.65 1.07a
0.68 0.27a,
0.11 0.08b,
1.24 0.49b,
2.14 0.92a,b,
4.17 1.48a,b,
6.40 1.85b,

0.23 0.10a,b
0.06 0.06a,b
0.10 0.04a
1.57 0.99a
0.71 0.30a,b
0.09 0.04a,b
1.38 0.73b,c
2.28 0.79a,b
4.02 1.84a
5.96 2.35a,b

0.20 0.11b
0.05 0.03b
0.09 0.02a
1.45 0.90a
0.97 0.70b,
0.13 0.09b,
1.62 0.80c,
2.28 1.13b,
4.98 2.46a,b,
7.25 2.34b

0.21 0.09a
0.04 0.02a
0.10 0.02a
1.73 0.88a
0.59 0.30a,b
0.08 0.04a
0.82 0.21a
1.73 0.70a,c
3.34 1.47a
5.48 1.85a,b

0.21 0.02b
0.05 0.02a
0.09 0.02a
1.69 0.94b
0.58 0.20a
0.07 0.03a
0.72 0.27b
1.78 0.64b,c
3.12 1.00a
4.97 1.65a

0.23 0.12c
0.04 0.02a
0.09 0.02a
1.86 1.10a
0.66 0.21b
0.08 0.03a
0.90 0.32a
1.91 0.56a
3.23 1.41a
5.77 1.53b

0.20 0.10a,c
0.04 0.01a
0.09 0.03a
1.71 1.42a,b
0.60 0.34a,b
0.07 0.05a
0.69 0.37b
1.50 0.70c
3.00 1.51a
5.27 2.68a,b

TJ, tomato juice.

Values are expressed as means SD. The non-parametric statistical Wilcoxon test was used for paired comparisons.
Values in a row with different letters are signicantly different (Wilcoxon test, p < 0.05).
Values with asterisks are statistically different between interventions at the same time (Wilcoxon test, p < 0.05).

Table 2 shows the mean plasma concentrations of the main

carotenoids at baseline and 3, 6 and 24 h after the consumption
of TJ, with and without rened olive oil; 5-cis-lycopene and betacarotene were the main compounds in plasma, irrespective of the
TJ consumed.
After TJ with oil consumption, the plasma concentrations of all
lycopene isomers showed a signicant increase over time, observing the maximum concentrations at 24 h. Accordingly, the sum of
all carotenoids showed the highest concentrations at 24 h, mainly
due to the contribution of the sum of lycopene isomers. By contrast, the concentrations of cis- and trans-lutein, alpha-carotene

and beta-carotene remained almost unchanged after both

interventions.
Fig. 2 shows the concentrations of plasma lycopene isomers and
the sum of all carotenoids over the intervention period according
to the TJ consumed. Fig. 2 also shows the changes from baseline
of lycopene and total carotenoid concentrations. The most signicant differences were observed in trans- and 5-cis-lycopene at 3
and 24 h, presenting concentrations up to 40% higher after the
consumption of TJ with oil than after TJ without oil. Both, total
lycopene after 3 and 24 h and total carotenoids, 3 h after the
consumption of TJ with oil, also showed signicantly higher

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S. Arranz et al. / Food Chemistry 168 (2015) 203210

Fig. 2. Plasma concentration and change from baseline of different lycopene isomers and total carotenoids through both interventions. Statistically different (p < 0.05)
between interventions.

Table 3
Pharmacokinetic parameters of carotene and lycopene isomers after TJ, with and without oil interventions (n = 11).
Cmax (mg/l)
TJ with oil
trans-Lutein
cis-Lutein
alpha-Carotene
beta-Carotene
13-Cis-Lycopene
9-cis-Lycopene
trans-Lycopene
5-cis-Lycopene

0.30 0.11
0.07 0.03
0.10 0.02
1.88 1.19
1.06 0.65
0.13 0.08
1.78 0.64
2.55 0.82

AUC024 (mg h/l)

TJ without oil
0.26 0.11
0.06 0.01
0.10 0.02
2.10 1.32
0.75 0.24
0.10 0.03
0.98 0.29
2.29 0.84

TJ with oil
4.88 2.52
0.96 0.65
1.98 0.62
31.5 18.6
15.9 6.96
1.87 1.08
28.0 13.2
44.7 19.4

Cmax/AUC024 (h
TJ without oil
5.14 2.05
1.08 0.28
2.19 0.56
36.4 20.2
14.9 5.57
1.81 0.65
18.7 6.11
43.5 12.4

TJ with oil

)
TJ without oil

0.07 0.04
0.10 0.07
0.06 0.04
0.06 0.04
0.06 0.04
0.07 0.06
0.07 0.04
0.06 0.04

0.05 0.01
0.05 0.01
0.04 0.01
0.05 0.01
0.05 0.01
0.05 0.01
0.05 0.01
0.05 0.01

Values are means SD. The non-parametric statistical Wilcoxon test was used for paired comparisons.
Cmax, peak plasma concentration; AUC, area under the plasma concentrationtime curve; Cmax/AUC, absorption rate.
Values with asterisks are statistically different between interventions (Wilcoxon test, p < 0.05).

concentrations than after taking the intervention without oil. This

was mainly due to the contribution of 5-cis-lycopene and
trans-lycopene to the total amount of carotenoids.
3.4. Effect of food matrix on the pharmacokinetic parameters
Table 3 summarises the pharmacokinetic parameters of the
eight carotenoids identied in plasma samples after the intake of
either TJ, with or without oil. The compounds with highest values
were 5-cis-, trans- and 13-cis-lycopene isomers and beta-carotene,
although beta-carotene levels remained stable from time 0 to 24 h.
While in juice the predominant isomer forms were the trans, in

plasma the major increase was observed for the cis isomers. Possibly, an isomerisation of trans-lycopene to 5-cis-lycopene occurs
during the absorption process. Moreover, it was also observed that
oil signicantly increased the Cmax values of 13-cis-, trans- and 5cis-lycopene. However, only the trans-lycopene isomer showed a
signicantly higher AUC024 after TJ with oil intervention than after
TJ without oil. With respect to the absorption rate, no statistical
differences were observed in Cmax/AUC024 of lycopene isomers
between interventions; the trend of this kinetic parameter indicates that oil tends to increase the absorption rate. This effect
was more pronounced in the case of cis- and trans-lutein, for which
Cmax/AUC024 was signicantly higher after TJ with oil intervention.

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Table 4
Blood lipids and BP at baseline and 6 h after both interventions (n = 11).
Parameter

Time

TJ with oil

TJ without oil

Triglycerides (mg/dl)

Baseline
6h
Change (%)
Baseline
6h
Change (%)
Baseline
6h
Change (%)
Baseline
6h
Change (%)
Baseline
6h
Change (%)
Baseline
6h
Change (%)
Baseline
6h
Change (%)

81.3 17.9
83.7 37.2
3.7 43.9
160 27.6
150 22.4
6.2 5.1
102 17.9
96.4 16.3
5.5 7.2
41.6 10.9
38.9 10.9,a
3.7 2.8
2.5 0.4
2.6 0.5
0.1 0.2
69.3 7.7
67.0 7.1
3.8 12.4
120.9 17.5
116.0 9.5
1.9 6.3

87.5 28.7
69.7 18.6
21.3 18.4
160 24.8
153 23.2
5.8 4.4
100 16.0
99.3 16.3
1.0 7.4
42.5 11.1
40.5 11.1,b
2.0 2.9
2.4 0.4
2.5 0.3
0.1 0.1
70.2 10.5
69.5 11.1
0.4 7.8
117.5 15.9
117.2 15.3
1.0 10.3

Total cholesterol (mg/dl)

LDL cholesterol (mg/dl)

HDL cholesterol (mg/dl)

LDL/HDL cholesterol ratio

Diastolic BP (mm Hg)

Systolic BP (mm Hg)

BP, blood pressure; TJ, tomato juice.

Values are expressed as mean SD. The non parametric statistical Wilcoxon test
was used for paired comparisons.
Values with asterisks are statistically different from baseline (Wilcoxon test,
p < 0.05).
Values in a row with different letters are signicantly different (Wilcoxon test,
p < 0.05).

3.5. Blood biochemical parameters

Table 4 shows lipid prole and blood pressure (BP) of the 11
volunteers at baseline and 6 h after both interventions with TJ.
All parameters were within the reference physiological ranges.
Systolic and diastolic BP and the plasma concentrations of HDLcholesterol, LDL-cholesterol, TC, LDL/HDL ratio, and triglycerides at
baseline and 6 h after each intervention were also analysed. A
signicant decrease from baseline was observed in TC (6%), LDLcholesterol (5%) and HDL-cholesterol (3.7%) after the TJ with oil
intervention. Additionally, a signicant decrease was also observed
for triglycerides (20%), after TJ without oil, probably due to the
postprandial metabolism of oil. There were no signicant changes
in BP, or glucose after interventions compared with the baseline.
Changes in plasma concentrations of different carotenoids correlated with changes in cholesterol after 6 h only in the case of
TJ with oil intervention. TC correlated inversely with 5-cis-lycopene (r2 = 0.67; p = 0.023), and LDL-cholesterol correlated inversely with trans-lycopene (r2 = 0.56; p = 0.049). Moreover, a
signicant inverse correlation was observed between HDL-cholesterol and 13-cis-lycopene (r2 = 0.59; p = 0.025) and the sum of all
carotenoids (r2 = 0.48; p = 0.027).

4. Discussion
Several studies have focussed on the bioavailability of lycopene
due to its potent health benets on the cardiovascular system and
certain types of cancer (Bohm, 2012; Ilic, Forbes, & Hassed, 2011).
However, scarce information is available regarding the absorption
of other carotenoids and the different lycopene isomers.
In contrast to other reports related to carotenoids, our study
analysed different carotenoids (lutein, alpha and beta-carotene)
and all the main cis and trans lycopene isomers present in plasma
after the consumption of TJ, with or without oil, to evaluate both,
the absorption of each of these compounds and the postprandial
lipid response.

Interestingly, the addition of rened olive oil (10%) to TJ and its

consumption, without cooking, to avoid any loss of vitamins and
polyphenols, signicantly increased the total amount of carotenoids in plasma, mainly due to the increased concentrations of
all lycopene isomers. Our results are in agreement with previous
studies reporting an improvement of the bioavailability of some
carotenoids in vivo due to the presence of a fatty matrix embedding
tomato components (Goltz, Campbell, Chitchumroonchokchai,
Failla, & Ferruzzi, 2012; Hu, Jandacek, & White, 2000; Unlu et al.,
2007).
The maximum concentrations of carotenoids in plasma, mainly
lycopene isomers, were observed 24 h after the consumption of TJ
with oil while this time seems to decrease when TJ does not
contain oil. The rate of absorption tends to increase when TJ is
consumed with oil, suggesting that oil increases the absorption
rate of lycopene isomers. Moreover, when considering the sum of
lycopene isomers or all carotenoids, two maximum increases in
concentrations were observed, at 3 and 24 h after the consumption
of TJ with oil. This could be related to the postprandial absorption
of triacylglycerol from oil that usually reaches maximum plasma
levels 3 h after meal consumption (Lambert and Parks, 2012).
The absorption process of some carotenoids, such as lycopene,
needs time to reach Cmax and this ranged from 4.2 to 8.4 h, whereas
tmax for total lycopene occurred roughly after 24 h post-administration (15.632.6 h) (Gustin et al., 2004). Notably, the concentrationtime curve from 0 to 24 h does not cover the total kinetic
prole of some carotenoids such as lycopene.
Beta-carotene is the predominant carotenoid in the TJ used in
our study. However, its plasma level does not differ between interventions. Some authors have attributed beta-carotene with the
ability to enhance the absorption of other carotenoids, such as
lycopene (Johnson, Qin, Krinsky, & Russell, 1997). Thus, carotenoid
bioavailability from TJ depends, not only on carotenoid geometrical
isomerization (trans or cis), but also on the presence of other
carotenoids. Similar to our results, where data do not show statistical differences for the two compounds, Roodenburg et al. (2000)
reported that the bioavailabilities of alpha- and beta-carotene were
not inuenced by the presence of fat in the meal.
Regarding cardiovascular outcomes, the modest reduction of TC
observed in our study ( 4.2 2.9%) might be attributed to the
effect of lycopene isomers consumed with TJ (74.09 12.48 mg/
intervention). It has been reported that a 2.8% reduction of TC
may reduce cardiovascular risk by 4% (Prospective Studies Collaboration, Lewington et al., 2007). Moreover, the consumption of TJ
with oil may provide an extra effect due to the reduction of 5.5%
in LDL-cholesterol that is related to the increase of trans-lycopene
in plasma. The reduction of HDL-cholesterol observed in our study,
after the consumption of TJ with oil, was not expected, according to
the results obtained by other authors reporting an increase of HDLcholesterol associated with tomato product consumption and concretely with diets rich in monounsaturated fatty acids (Ahuja,
Pittaway & Ball, 2006). Other authors have associated this increase
of HDL-cholesterol with the presence of virgin olive oil but not
with rened olive oil (Marrugat et al., 2004), attributing this effect
to the phenolic compounds naturally present in virgin olive oil.
However, while those tests were over a long period of time, in
our study we evaluated the immediate response of HDL-cholesterol 6 h after the intake of each intervention. Other possible mechanisms could be involved.
The remarkable reduction of triglycerides associated with TJ
without oil consumption has also been observed by other authors,
who attributed this effect to a tomato-rich diet (Ahuja et al., 2006),
and mainly accredited it to lycopene (Fuhrman, Elis, & Aviram,
1997). In our study, no association was observed with any carotenoid, excluding the correlation observed between lycopene isomers

S. Arranz et al. / Food Chemistry 168 (2015) 203210

and the decrease of LDL-cholesterol and total cholesterol after the

consumption of TJ with oil intervention.
The authors are aware that acute studies are adequate for evaluating absorption and bioavailability of compounds. It is for that
reason that limitations arise when analysing the association
between TJ consumption and cardiovascular outcomes, as well as
when elucidating the involvement of possible mechanisms. In
addition, the contribution and synergistic effect of other bioactive
compounds, such as polyphenols, sterols, vitamin C, folates, and
potassium, in association with carotenoids, could also have an
important role in cardiovascular protection.
5. Conclusions
The consumption of TJ with oil provides a signicant increase to
lycopene exposure over time, mainly due to trans-lycopene and 5cis-lycopene isomers, which clearly make a major contribution to
the total carotenoid concentrations in plasma. Kinetic parameters
provided evidence of how the fatty matrix tends to increase the
absorption rate of some carotenoids, mainly lycopene isomers.
Although beta-carotene is the predominant carotenoid in the
studied TJs, the oil added does not affect its absorption. Thus, carotenoid bioavailability from TJ depends, not only on geometrical
isomerization (trans or cis), but also on chemical structure.
LDL cholesterol and total cholesterol decreased signicantly
after the consumption of TJ with oil, which is associated with an
increase of trans-lycopene and 5-cis-lycopene, respectively.
Further research is required to explore the long-term effects of
different doses of tomato products on cholesterol, BP and other
cardiovascular biomarkers, in both healthy and high cardiovascular
risk populations.
Conicts of interest
None of the authors has any conicts of interest.
Acknowledgements
The authors thank Juan Ballester Ross Company (Tortosa,
Spain) for kindly providing the rened olive oil used in the study.
We express our gratitude for nancial support from CICYT
(AGL2010-22319-C03), from the Spanish Ministry of Science and
Innovation (MICINN). MAPFRE Foundation. The CIBERobn CB06/
03 is an initiative of the Instituto de Salud Carlos III, Spain. S.A.
thanks the postdoctoral Sara Borrell program (CD10/00151) from
the Instituto de Salud Carlos III, Spain. A.V.-Q. and M.M.-H. received
support from MICINN. P.V.-M. is grateful for the APIF predoctoral
fellowship from the University of Barcelona.
The authors responsibilities were as follows: M.M.-H., A.V.-Q.
and P.V.-M. participated in the study design, data collection and
laboratory analyses and assisted in every aspect of the study; S.A.
participated in the study design, data collection, laboratory analyses and wrote the rst draft of the manuscript; M.I. participated in
the elaboration of tomato juices; E.S., R.E. and R.M. L.-R. participated in the organization of the study, data interpretation and
preparation of the manuscript. All authors read and approved the
nal manuscript.
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