International Journal of Food Sciences and Nutrition

ISSN: 0963-7486 (Print) 1465-3478 (Online) Journal homepage: http://www.tandfonline.com/loi/iijf20

Mineral essential elements for nutrition in

different chocolate products

Luciano Cinquanta, Cinzia Di Cesare, Remo Manoni, Angela Piano, Piero

Roberti & Giancarlo Salvatori

To cite this article: Luciano Cinquanta, Cinzia Di Cesare, Remo Manoni, Angela Piano,
Piero Roberti & Giancarlo Salvatori (2016): Mineral essential elements for nutrition in
different chocolate products, International Journal of Food Sciences and Nutrition, DOI:
10.1080/09637486.2016.1199664

To link to this article: http://dx.doi.org/10.1080/09637486.2016.1199664

Published online: 26 Jun 2016.

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 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION, 2016
http://dx.doi.org/10.1080/09637486.2016.1199664

RESEARCH ARTICLE

Mineral essential elements for nutrition in different chocolate products

Luciano Cinquantaa, Cinzia Di Cesareb, Remo Manonic, Angela Pianod, Piero Robertid and Giancarlo Salvatorib
a
Department of Agricultural, Environmental and Food Science, Molise University, Campobasso, Italy; bDepartment of Medicine and
Sciences for Health ‘‘V. Tiberio’’, Molise University, Campobasso, Italy; cARPAm Molise, Campobasso, Italy; dCOSIB, Termoli, Italy

ABSTRACT ARTICLE HISTORY

In this work, the essential mineral nutritional elements in cocoa beans, in chocolates at different Received 7 December 2015
cocoa percentage (60,70,80 and 90%) and in milk chocolate are evaluated. Dark chocolates are Revised 4 June 2016
confirmed as an excellent source of magnesium (252.2 mg/100 g) and iron (10.9 mg/100 g): in Accepted 6 June 2016
chocolate containing 90% cocoa, their content corresponds to, respectively, 67.0% and 80.3 of Published online 27 June
2016
Nutrient Reference Values (NRV) in the European Union. The chocolate containing 90% cocoa is
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also a good source of zinc (3.5 mg/100 g), which is important for the immune system, and selen- KEYWORDS
ium (0.1 mg/100 g). Three main components suitable to explain the mineral concentrations are Chocolate; cocoa; iron;
analyzed by factor analysis. The component 1 can be interpreted as the contribution from the magnesium; mineral;
cocoa beans, owing to the mineral characteristics of the soil in which they have grown; the com- nutritional values
ponent 2 is mainly due to the manipulation and transformation of the cocoa in chocolate, while
the component 3 represents the milk powder.

Introduction catechins, and possibly procyanidins and metabolites

Chocolate, the most popular among cocoa products, is
reckoned as a luxury good, its composition varies, since
many extra ingredients influence the final composition
of the product (Steinberg et al. 2003). Data on con-
sumption of chocolate vary among countries: in the
United States mean chocolate intake has been calcu-
lated as 12 g per day, in Italy 9 g per day and in Brazil
4 g per day; in India 5.5 g per day for adult and 20 g
per day for children (Sepe et al. 2001; Dahiya et al.
2005; Confectionery news 2014). The eight largest
cocoa-producing countries are C^ote d’Ivoire, Ghana,
Indonesia, Nigeria, Cameroon, Brazil, Ecuador and
Malaysia. These countries represent 90% of the world
cocoa production. After harvest, cocoa seeds (35–50
per baccate) and their surrounding sweet fruit pulp are
placed in heaps or boxes and fermented under the
influence of naturally occurring microbes that multiply
using the sugar from the pulp as an energy source
(Afoakwa et al. 2011). The seeds are then dried and
the thin coats are removed from the embryonic tissue,
which is then roasted, and milled into what is referred
to as chocolate liquor. Chocolate has several beneficial
health effects because of its high content of antioxi-
dants: mainly a subclass of flavonoids, called flavanols,
including especially the monomer epicatechin and
(Fisher & Hollenberg 2005). Thus, it is worth recalling
the potential cardiovascular beneficial effect of the fla-
van-3-ols content in chocolate, related to its percentage
of cocoa. In fact, it is found consistent acute and
chronic benefits of chocolate or cocoa on flow-medi-
ated dilatation and promising effects on insulin and
insulin resistance (Hooper et al. 2012). Cocoa is also
an extremely rich source of many essential minerals
and chocolate has the potential to provide significant
amount of minerals in the human diet (Paoletti et al.
2012). As in any other plant food, the mineral content
of cocoa reflects the soil in which it is grown, another
possible source of variability may be contamination
with copper as fungicides. Some heavy metal are essen-
tial nutrients (Cu, Ni, Zn) needed by our body, but all
of them can be harmful if ingested in heavily metal-
contaminated foods. Reports of heavy metals in cocoa
beans and chocolates are of concern to cocoa pro-
ducers (Dos Santos et al. 2005). Consequently, inter-
national legislative bodies, as well as chocolate
manufacturing countries, have introduced new regula-
tions for the protection of the health of their consum-
ers (Dickson-Spillmann & Siegrist 2011). In the present
study, the levels of Ca, P, Mg, K, Fe, Zn, Cu, Se and
other microelements were evaluated in cocoa beans,
different dark chocolates (60, 70, 80 and 90% cocoa)

CONTACT Luciano Cinquanta cinquant@unimol.it Department of Agricultural, Environmental and Food Science. Molise University, Via F. De Sanctis
86100 Campobasso, Italy
ß 2016 Informa UK Limited, trading as Taylor & Francis Group
 2 L. CINQUANTA ET AL.

and milk chocolate. The samples were also examined
with respect to the reference levels of nutrients.
Material and methods
Samples and extraction procedure
Cocoa beans (Theobroma cocoa, var. hybrid) were
picked from Seed Production Division of Cocobod at
Goaso (Ghana); dark chocolates (at 60, 70, 80 and 90%
cocoa) and milk chocolates were collected from the
Dolceamaro srl company (Monteroduni, Italy). The
samples (three of 100 g of each type), have been
grinded and 1 g has been digested with 10 ml of nitric
acid with mineralizer (SCP Science DIGIprep, Quebec
H9X 4B6, Canada), with the following instrumentals
condition: start at 40  C for 15 min; heating at 60  C in
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Statistical analysis
Statistical analysis was performed using the software
SPSS for Windows 21.0 software (SPSS Inc., Chicago,
IL). Statistical evaluation was carried out by means of
exploratory factor analysis, because our hypotheses
regarding the model are not very specific. In particular,
we do not have specific predictions about the size of
the relation of each observed variable to each latent
variable. The cutoff loading value to determine which
elements will be used at the clustering stage was set at
the level >0.70.
Results
In our samples the amount of calcium is significantly
greater in the milk chocolate (180.4 mg/100 g), com-

15 min; stay at 60  C for 15 min; heating to 90  C in
20 min. The digested samples have been cooled and
brought to a volume of 50 ml with bidistilled water
and analyzed at the optical ICP (Inductively Coupled
Plasma).
Chemicals and reagents
Ultrapure nitric acid for trace analysis, sulfuric acid
(96%) and standard mono element in nitric acid 2% were
purchased from Sigma Aldrich (20151-Milan, Italy).
Analysis of minerals
The determination of metals (Al, Ba, K, Ca, Cr, Co,
Cu, Fe, Mg, Mn, Mo, Na, Ni, P, Se, Zn) in cocoa and
chocolate samples was carried out with the technique
of nitric mineralization and the analysis by spectropho-
tometry plasma emission (Varian ICP 710, OES,
Inductively Coupled Plasma Optical Emission
Spectrometers, Palo Alto, CA 94304-1038).
pared to all the other types of chocolate investigated
(Table 1). Hundred grams of this product fulfill the
22.5% of the daily intake for adults according to the
Nutrient Reference Values (Table 2). The content of
calcium increases from 64.4 mg/100 g to 90.8 mg/100 g,
as the cocoa content increases from 60 to 90%. The
Magnesium content in samples vary from a minimum
of 52.3 mg/100 g in milk chocolate to a maximum of
252.2 mg/100 g in chocolate containing 90% cocoa
(Table 1), corresponding to 67.0% NRV (Table 2).
Similarly, the highest content in Potassium is observed
in chocolate containing 90% cocoa (720.1 mg/100 g),
corresponding to about 36.0% NRV (Table 2). The
amount of some micronutrients (aluminum, barium,
boron, cobalt, molybdenum) in different samples is
reported in Table 1 (Sager 2012). The highest sodium
values are observed in milk chocolate: 72.9 mg/100 g,
and the lowest in chocolate containing 80% cocoa:
3.3 mg/100 g (Table 1). In the samples analyzed signifi-
cantly higher Copper content is observed in chocolate
containing 90% cocoa: 2.0 mg/100 g (Table 1),

Table 1. Mean (±SD) of some elements (mg/100 g) in different chocolate and cocoa beans.
Nutrient Milk chocolate Dark chocolate 60% Dark chocolate 70% Dark chocolate 80% Dark chocolate 90% Cocoa Beans
Ca 180.43a ± 3.00 64.33b ± 1.01 79.51c ± 8.14 72.41bc ± 3.10 90.83d ± 4.92 70.14bc ± 2.97
Mg 52.28a ± 2.03 158.78b ± 7.33 192.23b ± 16.87 198.76b ± 10.88 252.21c ± 21.58 290.64c ± 55.15
P 198.91a ± 2.72 221.80a ± 7.48 279.90b ± 14.14 297.63b ± 9.37 396.51c ± 24.76 515.84d ± 37.66
K 379.05a ± 10.94 465.55b ± 11.22 540.43c ± 30.11 565.88c ± 20.17 720.11d ± 49.91 720.22d ± 37.35
Na 72.91a ± 1.48 5.20 ± 0.11 4.20bc ± 0.45 3.30c ± 1.27 3.50bc ± 0.53 3.40bc ± 0.36
Cu 0.31a ± 0.00 1.43b ± 0.04 1.83c ± 0.08 1.78c ± 0.10 2.02c ± 0.12 3.12d ± 0.26
Fe 1.19a ± 0.03 9.73b ± 0.02 9.84b ± 0.29 11.24c ± 0.25 10.89c ± 0.13 3.20d ± 0.09
Se 0.06a ± 0.01 0.08a ± 0.00 0.09b ± 0.02 0.10b ± 0.01 0.10b ± 0.07 0.07a ± 0.05
Zn 0.94a ± 0.04 2.24b ± 0.08 3.19c ± 1.01 2.85bc ± 0.14 3.52c ± 0.16 4.88d ± 0.33
Cr 0.01a ± 0.00 0.04ac ± 0.00 0.07bc ± 0.02 0.07bc ± 0.02 0.08b ± 0.03 0.01a ± 0.00
Mn 0.31a ± 0.00 1.65b ± 0.06 1.98b ± 0.11 1.85b ± 0.07 2.05b ± 0.15 3.01c ± 0.55
Al 0.05a ± 0.01 0.56b ± 0.02 0.77b ± 0.29 1.38c ± 0.04 1.32c ± 0.02 0.69b ± 0.36
Ba 0.10a ± 0.00 0.54b ± 0.00 0.64c ± 0.03 0.68c ± 0.05 0.80d ± 0.04 0.33e ± 0.04
Ni 0.07a ± 0.00 0.36b ± 0.01 0.49bc ± 0.05 0.49bd ± 0.02 0.61cd ± 0.03 0.60cd ± 0.16
Mo (mg/kg) 0.06a ± 0.02 0.09ab ± 0.02 0.12bce ± 0.06 0.19c ± 0.02 0.07d ± 0.03 0.07ae ± 0.05
Co (mg/kg) 0.10a ± 0.02 0.33b ± 0.03 0.43c ± 0.06 0.46c ± 0.04 0.58d ± 0.04 0.26b ± 0.06
a,b,c,d,e
Scheffè test, different letters within same row indicate significant difference (p < 0.01).
 INTERNATIONAL JOURNAL OF FOOD SCIENCES AND NUTRITION 3

Table 2. Percentage of elements dietary intake from 100 g of chocolate compared with NRV.
Milk chocolate Dark chocolate 60% Dark chocolate 70% Dark chocolate 80% Dark chocolate 90% Recommen-dations
Nutrient % NRV % NRV % NRV % NRV % NRV NRV Adults (mg/day)
Ca 22.5 8.0 9.9 9.0 11.3 800
Mg 13.9 42.3 51.2 53.1 67.2 375
P 28.4 31.7 39.9 42.5 56.6 700
K 18.9 23.3 27.0 28.3 36.0 2000
Cu 31.8 143.8 183.3 178.5 202.1 1
Fe 8.5 69.5 70.3 80.3 77.8 14
Se 103.2 159.6 163.4 182.0 192.4 0.055
Zn 9.4 22.5 31.9 28.6 35.2 10
Cr 30.7 106.7 174.7 198.0 206.2 0.04
Mn 15.8 82.8 99.2 92.6 102.7 102

corresponding to about 202.1% NRV (Table 2).
Content of Fe in chocolate containing 80% cocoa is
about 11.2 mg/100 g, corresponding to about 80.3%
NRV (Table 2). In our research, chocolates containing
80% and 90% cocoa are also a good source of selenium,
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Calcium, the most abundant mineral in the body, is
required for vascular contraction and vasodilatation,
muscle function, nerve transmission, intracellular sig-
naling and hormonal secretion. Milk, yogurt and
cheese are rich natural sources of calcium, broccoli and

equal to about 0.10 mg/100 g, value far away by UL
(upper limit ¼ 0.3 mg/day) (EFSA 2016). In our sam-
ples, the highest values of Zinc are observed in choc-
olate containing 90% cocoa and the lowest
concentration in milk chocolate, with values respect-
ively of 3.52 mg/100 and 0.94 mg/100 g. The Zn content
observed in chocolate 90% cocoa corresponds to about
35.2% NRV (Table 2). In the samples studied, the
amount of Chromium3þ varies from 0.01 mg/100 mg in
milk chocolate to 0.08 mg/100 g in chocolate containing
90% cocoa (Table 1). The content observed in chocolate
containing 90% cocoa, corresponds to about 206.2%
NRV (Table 2). Finally, the lowest concentration in
Manganese is observed in milk chocolate and the high-
est in 90% cocoa-containing chocolates (Table 1), with
values, respectively, equal to 0.31 mg/100 g and 2.05 mg/
100, corresponding to about 102.7% NRV (Table 2).
Discussion
The chocolate manufacturers often receive very hetero-
geneous batches of cocoa liquors, for these reasons,
they have only rough expectations of the qualitative
parameters by country of origin. In recent years, vari-
ous handicraft companies are trying to buy the cocoa
beans directly from the producers. In this context, our
goal is to contribute to characterize the different raw
materials (here, the minerals present followed by phe-
nols present) and study the evolution of such com-
pounds in chocolate-based products.
The human body utilizes minerals for the proper
composition of the bone and blood and maintenance
of the normal cell function. The Regulation (EU) No
1169/2011 of the European Parliament and of the
Council of 25/X/2011 on the provision of food infor-
mation to consumers gives the daily reference intakes
for vitamins and minerals (adults) and their NRVs.
spinach provides calcium but its bioavailability is poor
(Borchers et al. 2000). The susceptibility of calcium to
form complexes with oxalates and phytates interferes
with its absorption. Our results show that milk choc-
olate could be an excellent source of calcium (Chekri
et al. 2012). Magnesium is involved in catalyzing multi-
tude biological reactions, including protein synthesis,
transmission of nerve impulse, muscle relaxation,
energy production, and bone and teeth adsorption.
Chocolate is a strong source for Mg (up to 252.2 mg/
100 g), with values similar to whole-meal bread and
potatoes. Cocoa contains 4-5 times more Mg than
peas, white wheat, corn and rice (Spiegel & Sager
2008). This component can be interpreted as the con-
tribution from the cocoa beans indeed. Typically, foods
rich in protein are a good source of phosphorus with
about 15 mg P/g protein (Anderson et al. 2006). The
chocolate is a good source of phosphorus, providing
on an average about 200 mg/100 g of protein
(Carnovale & Marletta 2001), and its concentration
increases with cocoa percentage, as well as Potassium
content. Potassium is essential for maintaining cellular
osmolality and membrane potentials, thus playing a
role in vascular tone and other biochemical pathways
related to cardiovascular health. A diet high in proc-
essed foods and low in fresh fruits and vegetables is
often high in sodium that is a huge problem through-
out the world, as it contributes to blood pressure eleva-
tion, along with increased risk of cardiovascular
diseases (Strazzullo et al. 2009). The minimum intake
level necessary for proper bodily function is estimated
to be as little as 200–500 mg/day (Shin et al. 2011;
Taylor et al. 2011). Data from around the world sug-
gest that the population average sodium consumption
is well above the value recommended by the 2002 Joint
(WHO/FAO) Expert Consultation (WHO Library
2012): 2 g sodium/day (equivalent to 5 g salt/day).
 4 L. CINQUANTA ET AL.

Table 3. Rotated component matrix after factor have grown. Thus, the data partially confirming
analysis of mineral concentrations found in cocoa (for phosphorus, iron and zinc) the significant differen-
beans and different chocolate samples.
ces found due to the effect of cocoa origin (Torres-
Component
Moreno et al. 2015). Iron is a mineral essential for life
1 2 3 and deficiency of Fe in the body could result in
Ca 0.485 0.343 0.776
K 0.925 0.354 0.034
anemia, of course, the form in which iron is present in
Mg 0.909 0.310 0.182 food is a major factor of bioavailability. Approximately
Na 0.560 0.494 0.647
P 0.988 0.017 0.092
20–30% of heme Fe (found only in meats) is absorbed,
Al 0.437 0.798 0.081 whereas absorption of nonheme Fe is in the range of
Ba 0.253 0.889 0.326 5–10%. Among nutrients important to feed babies and
Co 0.341 0.901 0.158
Cr 0.039 0.839 0.198 small children, chocolate contained significantly more
Cu 0.929 0.090 0.345 Fe (25 mg/100 g) than milk, honey and eggs (Falandysz
Fe 0.016 0.865 0.478
Mn 0.842 0.129 0.486 & Kotecka 1994). The iron content of cocoa is higher
Mo 0.236 0.900 0.202 than the level found in other foods as beef or chicken
Ni 0.759 0.436 0.385
Se 0.064 0.550 0.376 liver (Paoletti et al. 2012). The bioavailability of iron is
Zn 0.916 0.094 0.256 quite higher in chocolate with respect to other vegeta-
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bles, because of low levels of phytic acids therein. The

From the nutritional point of view, chocolate is not the
preferred source of sodium. Copper introduced in the
diet is essential for cellular respiration, to the produc-
tion of ATP, thermoregulation and it is also involved
in antioxidant defense, as well as in the modulation of
the immune system (Maggini et al. 2007). The total
dietary copper intake by males and females was posi-
tively associated with the consumption of chocolate
foods (Pennington & Young 1990). Analysis of copper
contents in chocolate is also important since copper
compounds are widely used as fungicides against black
pod disease while farming cocoa. The upper limit to
copper is starting from 5 mg/day (EFSA 2016); excess
copper intake can cause stomach pain, nausea and
diarrhea and can cause tissue damage.
For understanding the results of the mineral ana-
lysis, we created a rotated factor matrix table (Table 3).
Factors are rotated so that they are easier to interpret
thus different items are explained or predicted by dif-
ferent underlying factors, and each factor explains
more than one item. By factor analysis, we can infer
three main components (Table 3): component 1 can be
interpreted as the contribution from the cocoa beans,
since the mineral content of cocoa reflects the mineral
characteristics of the soil in which they have grown
(Shittu & Badmus 2009; No€el et al., 2012). Component
2 seems derived mainly from secondary contamination,
owing to the manipulation and transformation of the
cocoa in chocolate (Beckett 2008); while component 3
represents the milk powder in the milk chocolate, in
fact, calcium is part of the component 3 (Table 3).
Magnesium, together with copper, zinc and phos-
phorus is part of component 1, which reflects the min-
eral characteristics of the soil in which the cocoa beans
samples of chocolate at 80% and 90% cocoa are an
excellent source of Iron confirming the upward trend
of the content of iron by increasing the percentage of
cocoa. Iron, Chromium, Aluminum, Barium, Cobalt
and Molybdenum are part of Component 2; these ele-
ments probably may reflect the possible contamination
of the cocoa beans or because of human activities such
as industrial processes to transform cocoa in chocolate.
Selenium is another essential micronutrient due to its
presence in the active site of some enzymes involved in
thyroid hormone metabolism, modulation of the
immune system and defense activity against oxidative
stress (Arthur et al. 2003). Based on the significant role
of selenium in antioxidant enzymes, it is important to
consider the contribution of chocolates containing 80%
and 90% cocoa in its diet intake. Zinc has a wide var-
iety of roles in mammalian system, exerts multiple
functions in the cell and is the key component of the
catalytic site of hundreds of metalloenzymes, playing a
central role in the growth and cell differentiation
(EFSA NDA Panel 2014). Zinc has a considerable
influence on the immune system and the lack of this
mineral causes atrophy of lymphoid organs (Tuerk &
Fazel 2009). Zinc nutritional deficiency is a global
health problem (Osendarp et al. 2003). The content of
zinc in the chocolate with 90% cocoa contributes
approximately one third of the nutritional requirement
by consuming 100 g of such product, while for
Manganese this value rises significantly. Chromium(3þ)
enhances the action of insulin, a hormone critical for
the metabolism and storage of carbohydrate, fat and
protein in the body. Due to lack of data, the European
Food Safety Authority recommended a maximum tol-
erable intake level (upper limit) for the Cr3þ equal to
0.25 mg/day. The content of copper, zinc and

chromium is always far below the safety limits for
human health.
The originality of this research is to try whether the
different mineral content in cocoa beans and chocolate
are explained or predicted by different ‘‘underlying fac-
tors’’. By exploratory factor analysis, using the rotated
factor matrix table, we inferred three main compo-
nents: the geographical origin of cocoa beans, the sec-
ondary contamination until their transformation into
chocolate and the added ingredients. Further tests are
ongoing to confirm such preliminary results.
Conclusions
Dark chocolates are confirmed as an excellent source
of magnesium: in chocolate containing 90% cocoa, it
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corresponds to 67.0% NRV. Similarly, the high per-
centage of cocoa is related to iron content: in chocolate
containing 80% cocoa, the intake corresponds to about
80.3% NRV. The latter is of particular importance for
vegetarians and vegans, because in cocoa derivatives,
the availability of iron is quite high, because of the low
levels of phytic acids therein. The dark chocolate is
also a good source of zinc, an important for the
immune system, and selenium, which is in the active
site of antioxidant enzymes.
Disclosure statement
There is no conflict of interest in this study. Funding was
not needed to carry out the research, and the authors have
no affiliation with the firm involved in this study
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