journal of functional foods 12 (2015) 526529

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Polyphenol antioxidants in commercial

chocolate bars: Is the label accurate?

Joe A. Vinson *, Matthew J. Motisi

Department of Chemistry, Loyola Science Center, University of Scranton, 925 Ridge Row, Scranton, PA
18510, USA

A R T I C L E I N F O A B S T R A C T

Article history: Polyphenols in cocoa have been shown to be responsible for beneficial health effects, es-
Received 20 November 2014 pecially in the cardiovascular area. Many people consume their chocolate in the form of
Received in revised form 8 bars, readily available in retail stores. The only information that may be useful to the con-
December 2014 sumer in choosing a healthier bar, with the exception of the nutrients, is the % cocoa solids
Accepted 10 December 2014 on the label. We have examined the polyphenols in commercial bars by use of two anti-
Available online 25 December 2014 oxidant assays and corrected that value for non-fat cocoa solids, the source of the polyphenols
in the chocolate. We also separated and analyzed by HPLC the two major monomeric poly-
Keywords: phenol antioxidants, epicatechin and catechin. We found a significant and linear relationship
Polyphenols between label % cocoa solids and the antioxidant assays as well as the sum of the mono-
Antioxidants mers. Consumers can thus rationally choose chocolate bars based on % cocoa solids on the
Chocolate label.
% cocoa solids 2014 Elsevier Ltd. All rights reserved.
Epicatechin
Catechin

2006). Consuming polyphenols in a cocoa muffin and dark

1. Introduction
Until the 1990s, chocolate had been regarded as nothing more
than an unhealthy sweet. A 1994 publication showed that the
saturated fat stearic acid in chocolate did not raise serum lipids
(Kris-Etherton & Mustad, 1994). In 1999, a blinded analysis study
showed that dark chocolate bars contained significantly more
of the health-promoting polyphenols than milk chocolate
(Vinson, Proch, & Zubik, 1999). In 2006, an article was pub-
lished that showed that cocoa inhibited atherosclerosis in an
animal model and cocoa was found to be the third leading
source of polyphenol antioxidants in the US diet (Vinson et al.,
chocolate bits produced a post-prandial human in vivo anti-
oxidant effect in spite of the presence of fat and sugar pro-
oxidants in the products (Vinson et al., 2006). Now people are
thinking of chocolate as a functional food.
There has been a recent spate of research regarding the car-
diovascular health benefits of chocolate, as a current PubMed
search found over 4700 citations for chocolate. A meta-
analysis of 114,000 subjects found that the highest chocolate
consumption reduced the risk of heart disease and stroke
deaths by over 30% (Buitrago-Lopez et al., 2001). Further-
more, a clinical study showed that even a very small dose (6 g)
of dark chocolate produced a small but significant decrease in

* Corresponding author. Department of Chemistry, Loyola Science Center, University of Scranton, 925 Ridge Row, Scranton, PA 18510, USA.
Tel.: +1 570 941 7551; fax: +1 570 941 7572.
Email address: joe.vinson@scranton.edu (J.A. Vinson).
http://dx.doi.org/10.1016/j.jff.2014.12.022
1756-4646/ 2014 Elsevier Ltd. All rights reserved.
 journal of functional foods 12 (2015) 526529
blood pressure after 18 weeks in subjects with pre- or stage 1
hypertension (Taubert, Roesen, Lehmann, Jung, & Schmig,
2007). Another investigation found that 10 grams of a 70% cocoa
solid chocolate for 1 month induced multiple cardiovascular
benefits for young healthy subjects, including improvements
to blood pressure, flow mediated dilation, pulse wave veloc-
ity, and arterial stiffness index (Pereira et al., 2014). A different
group investigating the same cardiovascular benefits proved
that polyphenols in cocoa, not nutrients, were responsible for
the beneficial changes (Grassi et al., 2014). They showed in a
double-blind crossover study with healthy subjects that a low
calorie 10 g dose of cocoa containing increasing levels of poly-
phenols caused a doseresponse improvement in the
parameters. Chocolate has also been linked to improvement
of chronic diseases. For instance, dark chocolate (but not milk
chocolate) increased nitric oxide and improved walking au-
tonomy in patients with peripheral artery disease (Loffredo
et al., 2014). Only dark chocolate increased serum epicatechin
and its metabolites, which are the putative bioactives. Choco-
late consumption was also found to decrease mortality
following an initial myocardial infarction (Janszky et al., 2009).
The flavanols contained in chocolate were reviewed as mood
enhancers (Smith, 2013). Another review highlighted the mecha-
nisms of the flavanol metabolites in the brain which slow the
cognitive decline in aging and improve cognitive function in
healthy subjects (Sokolov, Pavlova, Klosterhalfen, & Enck, 2013).
The increased publicity of these studies and the possibil-
ity that chocolate consumption can affect surrogate markers
of heart disease may ultimately lead manufacturers to a health
claim for chocolate. A chocolate bar, rather than cocoa, is the
preferred product consumed in many cultures. What choco-
late bar should the informed consumer purchase, given the large
number of domestic and foreign chocolates available in the mar-
ketplace? At present, the labeling on many chocolate bars lists
the % cocoa solids. Since cocoa solids represent the source of
bioactive polyphenols (flavanols) in the bar, the number may
indicate the level of these compounds in the product. We hereby
present evidence that this is the case.
2. Materials and methods
Commercial chocolate bars (n = 46), both labeled and unla-
beled with % cocoa solids, were obtained from local markets
and the internet. Five manufacturing countries (four from
Europe and the USA) and seven single countries of origin were
represented. Fat varied from 28 to 64%, while cocoa solids listed
on the label ranged from 30 to 100%. There are two common
methods available to analyze the polyphenols in chocolate. We
measured the total amount of polyphenol antioxidants in
chocolate in duplicate as catechin equivalents using two dif-
ferent antioxidant assays, the single reagent Folin method
(Vinson, Proch, & Bose, 2001) and a free radical antioxidant
power (FRAP) method using catechin as the standard. The an-
tioxidant concentration was corrected for label fat content since
nonfat cocoa solids are the origin of the polyphenol antioxi-
dants in chocolate bars. Another more complicated and
expensive method is the HPLC assay of the individual com-
pounds, especially the putative bioactive flavanols catechin and
527
epicatechin, which we used on 17 of the bars. For the HPLC
assay, we used our sample preparation method (Vinson et al.,
1999), followed by gradient elution at 1 ml/min from 100% water/
1% acetic acid to 100% methanol/1% acetic acid over 30 min
with a Phenomenex 25 cm Luna C18 column (Torrance, CA) at
280 nm.
3. Results
The labeled chocolate bars (n = 33) ranged from 30 to 100% cocoa
solids containing 21.690.4 mol/g of Folin catechin equiva-
lents and 24.6189 mol/g FRAP catechin equivalents. Dark
chocolate contained significantly more polyphenol antioxi-
dants than milk chocolate, p < 0.001, as previously shown
(Vinson et al., 1999). Products with % cocoa solids labeling had
significantly more antioxidants than those without labeling
(n = 13), p < 0.001. For single manufacturers with products of
differing % cocoa solids, the antioxidant content always in-
creased with the % cocoa solids. Independent of the type of
chocolate, manufacturer, or country of origin, there was a posi-
tive linear correlation between polyphenol concentration and
% cocoa solids for 31 products for both the Folin and FRAP data
(p < 0.001). One chocolate manufactured by a patented pro-
cessing technique with 72% cocoa solids contained the most
total polyphenols of all chocolate bars. Other published re-
search found linear relations with Folin but used a complicated
analyzed value for non-fat cocoa solids (Cooper et al., 2008;
Miller et al., 2006). The label % cocoa solids were also highly
correlated with the sum of the major flavanols catechin and
epicatechin as measured by HPLC (0.680, p < 0.001) (Cooper et al.,
2008; Miller et al., 2006).
A choice the consumer can make would be whether to buy
a foreign or domestic produced chocolate, with the latter prob-
ably being less expensive. A comparison of 13 labeled domestic
versus 20 foreign chocolate bars showed that there was a sta-
tistically significantly greater amount of Folin and FRAP
antioxidants in the foreign products (p < 0.01). A paired com-
parison of domestic and foreign bars with identical % cocoa
solids, 25, 60 and 72% also showed a significantly greater
amount of antioxidants in the foreign products (p < 0.05).
For consumer information, the data in Fig. 1 with a slope
of almost 1 show doubling the % cocoa solids will double the
amount of antioxidants found in the chocolate bar. While con-
sumers need only look for a dark chocolate with % cocoa solids
to get a high polyphenol antioxidant chocolate, they should
also pay attention to the caloric content. A recent long-term
supplementation study showed a slight but significant weight
gain at the high dose (25 g) of chocolate, but a beneficial blood
pressure lowering effect of the low dose (6 g) with no effect
on weight gain (Desch et al., 2010). In addition, a 6 year epi-
demiological study found that habitual chocolate consumption
may increase body weight in a doseresponse manner
(Greenburg & Buijsse, 2013). It is important for the consumer
to know that dark chocolate consumption promotes more
satiety, lowers the desire to eat something sweet, and sup-
presses energy consumption compared with milk chocolate
(Srensen & Astrup, 2011).
Although detrimental effects, such as weight gain, may arise
from frequent chocolate consumption, there are also many
528 journal of functional foods 12 (2015) 526529
Fig. 1 Correlation of % cocoa solids on label and antioxidant activity (Folin) for commercial chocolate bars.
health benefits that can be attributed to the polyphenols in
the chocolate. The polyphenol content in chocolate bars is
accurately reflected by the % cocoa solids listed on the
labels.
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