© The Authors, 2026, Published by the Universidad del Zulia*Corresponding author: joseglanza82@gmail.com
Keywords:
Venezuelan cocoa
Proximal analysis
Minerals
Cadmium,
ICP-OES
Cata
Cuyagua
Proximate and mineral composition of venezuelan cocoa beans from Cata and Cuyagua
Composición proximal y mineral de almendras de cacao venezolano de Cata y Cuyagua
Composição proximal e mineral de amêndoas de cacau venezuelano de Cata e Cuyagua
José Gregorio Lanza
1,2*
Rev. Fac. Agron. (LUZ). 2026, 43(1): e254313
ISSN 2477-9407
DOI: https://doi.org/10.47280/RevFacAgron(LUZ).v43.n1XIII
Food technology
Associate editor: Dra. Gretty R. Ettiene Rojas
University of Zulia, Faculty of Agronomy
Bolivarian Republic of Venezuela.
¹National Institute of Nutrition - Physicochemical Analysis
Laboratory, Caracas, Venezuela.
²Normalización Chile LAN Consultores, Santiago, Chile.
Received: 01-10-2025
Accepted: 10-01-2026
Published: 30-01-2026
Abstract
Venezuelan cocoa is renowned for its high sensory quality;
however, the presence of heavy metals may restrict its access to
highly regulated international markets such as the European Union.
This study aimed to evaluate the proximate composition and
mineral content of cocoa beans (Theobroma cacao L.) collected
from Cata and Cuyagua (Aragua, Venezuela) during the main
harvest seasons of 2013 and 2014. Thirty samples (15 per year)
were analyzed. Proximate composition was determined using
ocial AOAC methods, while mineral elements were quantied
following microwave-assisted acid digestion and ICP-OES analysis.
Results showed that moisture content was signicantly higher in
2013 compared to 2014 (p<0.001), whereas fat, protein, ash, and
carbohydrate contents did not dier between years. Signicant
interannual dierences were observed in mineral composition,
with higher concentrations of Ca, Cu, Fe, Mg, and Mn in 2013,
and increased levels of K and Na in 2014. Cadmium exhibited a
non-signicant decrease between years (1.62-1.04 mg.kg
-1
), while
chromium remained low and stable. Arsenic and mercury were
detected only at trace levels, with no signicant dierences between
years. These results provide reference values for cocoa produced in
Cata and Cuyagua, conrm interannual variability in proximate and
mineral composition, and emphasize the importance of continuous
monitoring of cadmium and other metals to ensure compliance with
international regulations.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
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2-6 |
Resumen
El cacao venezolano es reconocido por su alta calidad sensorial;
sin embargo, la presencia de metales pesados puede limitar su acceso
a mercados internacionales exigentes, como la Unión Europea.
El objetivo de este estudio fue evaluar la composición proximal y
mineral de almendras de cacao (Theobroma cacao L.) provenientes
de las localidades de Cata y Cuyagua (Aragua, Venezuela),
correspondientes a las cosechas principales de 2013 y 2014. Se
analizaron 30 muestras (15 por año), determinándose la composición
proximal mediante métodos ociales AOAC, mientras que los
elementos minerales se cuanticaron tras digestión ácida asistida
por microondas y análisis por ICP-OES. Los resultados evidenciaron
una mayor humedad en las muestras de 2013 respecto a 2014
(p<0,001), sin diferencias signicativas en grasa, proteína, cenizas ni
carbohidratos. En la composición mineral se observaron variaciones
interanuales signicativas, destacando mayores concentraciones de
Ca, Cu, Fe, Mg y Mn en 2013, y valores superiores de K y Na en
2014. El cadmio mostró una disminución no signicativa entre años
(1,62-1,04 mg.kg
-1
), mientras que el cromo se mantuvo en niveles
bajos y estables. Arsénico y mercurio se detectaron únicamente en
trazas, sin diferencias relevantes. Estos resultados aportan valores de
referencia para el cacao de Cata y Cuyagua, conrman la variabilidad
interanual de su composición proximal y mineral, y refuerzan la
necesidad de un monitoreo continuo de cadmio y otros metales para
asegurar el cumplimiento de las regulaciones internacionales.
Palabras clave: cacao venezolano, análisis proximal, minerales,
cadmio, ICP-OES, Cata, Cuyagua.
Resumo
O cacau venezuelano é reconhecido por sua elevada qualidade
sensorial; entretanto, a presença de metais pesados pode restringir
seu acesso a mercados internacionais altamente regulados, como o da
União Europeia. Este estudo teve como objetivo avaliar a composição
proximal e mineral de amêndoas de cacau (Theobroma cacao L.)
provenientes das localidades de Cata e Cuyagua (Aragua, Venezuela),
correspondentes às principais colheitas de 2013 e 2014. Foram
analisadas 30 amostras (15 por ano). A composição proximal foi
determinada por métodos ociais da AOAC, enquanto os elementos
minerais foram quanticados após digestão ácida assistida por micro-
ondas e análise por ICP-OES. Os resultados indicaram que o teor
de umidade foi signicativamente maior em 2013 em comparação
a 2014 (p<0,001), enquanto os teores de gordura, proteína, cinzas
e carboidratos não diferiram entre os anos. Diferenças interanuais
signicativas foram observadas na composição mineral, com
maiores concentrações de Ca, Cu, Fe, Mg e Mn em 2013, e níveis
mais elevados de K e Na em 2014. O cádmio apresentou redução
não signicativa entre os anos (1,62-1,04 mg.kg
-1
), enquanto o cromo
manteve-se em níveis baixos e estáveis. Arsênio e mercúrio foram
detectados apenas em níveis traço, sem diferenças signicativas. Esses
resultados fornecem valores de referência para o cacau produzido em
Cata e Cuyagua, conrmam a variabilidade interanual da composição
proximal e mineral e reforçam a necessidade de monitoramento
contínuo do cádmio e de outros metais, visando ao atendimento das
regulamentações internacionais.
Palavras-chave: cacau venezuelano, composição proximal, minerais,
cádmio, ICP-OES, Cata, Cuyagua.
Introduction
Venezuelan cacao (Theobroma cacao L.) is internationally
recognized for its genetic and sensory quality, being considered
one of the most appreciated origins for the production of ne and
aromatic chocolates. Coastal regions such as Cata and Cuyagua, in
the state of Aragua, are part of the corridor of Creole and Trinidadian
cocoas, characterized by distinctive aromatic attributes and artisanal
production of high commercial value (Portillo et al., 2012; Katz et
al., 2011).
In recent years, the presence of heavy metals in cocoa has become
very relevant due to the regulatory requirements of international
markets. The European Union established maximum limits for
cadmium (Cd) in cocoa products by Regulation (EU) No. 488/2014,
which were subsequently consolidated in Regulation (EU) 2023/915.
These thresholds, which vary between 0.60 mg.kg
-1
in cocoa powder
and 0.80 mg.kg
-1
in chocolates with ≥ 50 % cocoa, have prompted the
study of the content of Cd and other toxic elements in cocoa beans
produced in Latin America, a region that concentrates more than 70
% of the world’s production (Gramlich et al., 2018).
The mineral composition of cocoa not only has regulatory
implications, but also nutritional and technological ones. Elements
such as K, Mg, Ca, Fe, and Mn are relevant for nutritional quality and
for processes such as fermentation and drying, while the presence of
Cd, Pb, Hg, and As represents a risk to food safety and can aect the
export of grains (Gramlich et al., 2018; García-Porras et al., 2025).
In Venezuela, the rst systematic studies on metals in cocoa included
the analyses of Portillo et al. (2012) in cocoa porcelain from the
South of the Lake. Subsequently, Lanza et al. (2016) reported levels
of Cd, Cu, Ni, Cr and Fe in hybrid cocoas and porcelain from Santa
Bárbara del Zulia, establishing initial reference values for the country.
More recently, Padilla et al. (2025) conrmed the presence of Cd
in Venezuelan cocoa beans, with concentrations that in some cases
exceeded the limits established by the European Union, providing
current evidence of the magnitude of the problem.
At the regional level, research in Ecuador, Peru, and Colombia
has documented signicant variations in Cd levels in soils and
cocoa beans, mainly associated with edaphic factors, agronomic
management, and cultivar genetics (Arévalo-Gardini et al., 2017;
Barraza et al., 2017; Meter et al., 2019). These ndings reinforce the
need to generate local information to understand the interannual and
regional variability of mineral content in Venezuelan cocoa.
In this context, the present study aimed to evaluate the proximal
and mineral composition of roasted cocoa beans collected in Cata and
Cuyagua during the 2013 and 2014 harvests, with emphasis on the
levels of Cd and other minerals of nutritional and regulatory interest,
comparing the results with international reference values.
Materials and methods
Study Location
Samples of cocoa beans were collected in the towns of Cata and
Cuyagua, Aragua state (Venezuela), during the months of September
to December 2013 and 2014, corresponding to the main harvest.
Sample Preparation
The cocoa beans were roasted at 120 °C for 30 minutes, hulled
manually and ground into a homogeneous our. Subsequently, they
were preserved in airtight containers until analysis.
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3-6 |
Quality control
In the proximate analyses (moisture, fat, protein, ash and
carbohydrates), each sample was processed in triplicate. Additionally,
random duplicates were included in each batch of analyses, which
allowed us to verify the accuracy (≥ 90 %). Recovery standards
were applied where appropriate, especially in the determinations of
fats (AOAC 920.39, 2012) and proteins (AOAC 960.52, 2012), by
fortifying samples with reference standards.
In the mineral analyses, quality control included the preparation of
multi-elemental calibration curves from certied standards, ensuring
a minimum linearity of R² ≥ 0.99 in the range of interest. Certied
Recovery Standards (MRCs), such as NIST SRM 2384 (Baking
Chocolate, 100 % Cocoa Beans) and ERM-BD512 (Dark Chocolate),
were also used to verify accuracy and ensure comparability in similar
matrices (table 2). With each batch, reagent targets were analyzed
to rule out possible contaminations during sample preparation and
digestion, sample duplicates were included in each run to evaluate
analytical repeatability.
Statistical analysis
Results were expressed as mean ± standard deviation of three
independent determinations (triplicates). Descriptive statistics were
applied for each proximal and mineral component. Dierences
between years (2013 - 2014) were assessed using a one-factor
ANOVA, followed by the Tukey HSD multiple comparison test. In
cases where the averages between years for each mineral or proximal
component were directly compared, a Welch’s t-test was also applied
for independent samples (Welch’s t-test). Statistical signicance was
established at p ≤ 0.05, considering stricter levels of evidence (p ≤ 0.01
and p ≤ 0.001) to highlight highly signicant dierences. All analyses
were performed with SPSS v.22.0 software (IBM Corp., Armonk,
NY, USA), previously verifying the assumptions of normality and
homogeneity of variances.
It is important to note that for this study, roasted beans were used
as the basis for the analyses, in order to guarantee the homogeneity of
the matrix and the elimination of residual moisture, avoiding variations
during milling and acid digestion. This approach remains consistent
with previous research conducted by Lanza et al. (2016), in which the
grains were subjected to a similar heat treatment (100 °C, 30 min).
The inclusion of roasting also allows for improved comparability with
studies of proximal and mineral composition in derived products,
where cocoa is usually processed before consumption.
In total, 30 samples were analyzed (15 samples per year), each
consisting of 1 kg of roasted almonds. Each sample was analyzed
in triplicate, so that the values reported correspond to averages and
standard deviations calculated from three independent determinations.
Proximate analysis
The proximate composition was determined by applying ocial
AOAC methods: moisture (925.10), ash (923.03), protein by
Kjeldahl method (984.13), fat (963.15), and carbohydrate calculated
by dierence. Each determination was made in triplicate, including
random duplicates as an internal precision check.
Determination of minerals
The elements As, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na and Ca were
quantied after microwave-assisted acid digestion, using a Milestone
Start D digester (Milestone, Italy), with a mixture of nitric acid (65
%) and hydrogen peroxide (30 %). The analysis was performed by
optical emission spectrometry with inductively coupled plasma
(ICP-OES, Perkin Elmer Optima 5300 DV), following the ocial
AOAC 999.11 method. To ensure traceability and accuracy, certied
reference materials were used: NIST SRM 2384 (Baking Chocolate,
100% cocoa beans) and ERM-BD512 (Dark Chocolate), as well as
certied multi-elemental standards (Certipur®, Merck). Detection
limits (LOD) ranged from 0.01 to 0.05 mg.kg
-1
, depending on the
element.
Table 1 summarizes the methods used for the determination of the
proximal and mineral composition in cocoa beans below.
Table 1. Summary of the methods used for the determination of the proximate and mineral composition in cocoa beans.
Parameter Method / Standard Technique Instrument / Conditions Observations
Humidity AOAC 925.10 Oven drying 105 °C to constant weight Triplicate; Random duplicates
Ashes AOAC 923.03 Mue incineration 550 °C to constant weight -
Protein AOAC 984.13 Kjeldahl (N × 6.25) Automatic digestion and distillation Applied Recovery Standard
Fat AOAC 963.15 Soxhlet Petroleum ether as a solvent Recovery 80 -120 %
Carbohydrates Calculated By dierence 100 - (moisture + fat + protein ashes) -
As, Cd, Cr, Cu, Fe, Hg,
K, Mg, Mn, Na, Ca
AOAC 999.11
ICP-OES After microwave-
assisted acid digestion
Digestor Milestone Start D; ICP-OES
Perkin Elmer Optima 5300 DV; mezcla
HNO₃ 65 % + H₂O₂ 30 %
MRC NIST SRM 2384, ERM-
BD512; LOD: 0.01 - 0.05 mg.kg
-1
Table 2. Summary of Acceptance Criteria for Quality Controls.
Quality Control Application Standard/Reference Acceptance criteria
Triplicate Proximal and minerals All Samples CV <10 %
Random duplicates Proximal Analysis Batches Accuracy ≥90 %
Recovery Standard Proximal (fats, proteins)
AOAC 920.39 (2012), AOAC 960.52
(2012)
80 - 120 %
Calibration curve Minerals Certied multi-elementary standards R² ≥ 0,99
Certied Standard (MRC) Minerals NIST SRM 2384, ERM-BD512 80 - 120 %
Reagent Target Minerals Non-sample reagents ≤ Detection limit
Repeatability (duplicates) Minerals Sample Lots CV <10 %
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Rev. Fac. Agron. (LUZ). 2026, 43(1): e264313 January-March ISSN 2477-9409.
4-6 |
Results and discussion
Proximate composition
Proximal analyses (table 3) conrmed values consistent with
those reported for Venezuelan cocoa.
Table 3. Proximate composition of cocoa beans from Cata and
Cuyagua (2013 - 2014).
Parameter 2013 2014
Humidity
**
3.79 ± 0.42
a
2.65 ± 0.68
b
Fat
**
49.69 ± 1.91
a
45.27 ± 3.90
b
Protein
*
13.80 ± 0.79
a
13.56 ± 0.98
a
Ashes
*
3.28 ± 0.33
a
3.27 ± 0.26
a
Carbohydrates
*
29.44 ± 1.85
a
29.25 ± 3.92
a
Values expressed as mean ± SD, (*p < 0.05; **p < 0.01); equal letters indicate that there are
no statistically signicant dierences
Humidity was signicantly higher in 2013 (3.79 %) compared to
2014 (2.65 %) (p < 0.0001). Likewise, fat had a signicantly higher
value in 2013 (49.69 %) compared to 2014 (45.27 %) (p < 0.001). In
contrast, protein, ashes and carbohydrates did not show statistically
signicant dierences between years (p > 0.05). These results
suggest that, although the proximate composition of cocoa remained
relatively stable between both harvest years, moisture and fat showed
relevant variations, possibly associated with climatic or post-harvest
conditions, indicating that they inuenced both water retention and
lipid content. In contrast, protein, ash, and carbohydrates remained
stable between 2013 and 2014, which coincides with what was
previously reported, indicating that the main determinants of these
fractions are genotype and postharvest practices (fermentation and
drying), rather than interannual climatic variation (Arévalo-Gardini
et al., 2017; Gramlich et al., 2018). Recent studies conrm that
fermentation and drying dynamics inuence not only the proximate
composition, but also the bioavailability of nutrients and metals
(García-Porras et al., 2025).
Mineral prole
The mineral prole showed notable dierences between the
harvest years (table 4).
Table 4. Mineral content of cocoa beans harvested in Cata and
Cuyagua (2013–2014).
Mineral
2013
(media ± DE)
2014
(media ± DE)
As* 0.00 ± 0.00
a
0.00 ± 0.00
a
Cd* 1.62 ± 1.08
a
1.04 ± 0.59
a
Cr* 0.22 ± 0.16
a
0.23 ± 0.08
a
Cu* 24.14 ± 6.83
a
19.19 ± 2.40
b
Fe** 83.18 ± 61.54
a
34.57 ± 7.88
b
Hg** 0.19 ± 0.15
a
0.04 ± 0.08
b
K** 4.959.60 ± 632.60
b
7.447.73 ± 227.04
a
Mg** 4.473.60 ± 887.85
a
3.566.20 ± 418.38
b
Mn** 29.4 ± 12.00
a
19.40 ± 8.39
b
Na* 60.84 ± 36.60
b
96.32 ± 20.30
a
Values expressed as mean SD ± (*p < 0.05; **p < 0.01); equal letters indicate that there are no
statistically signicant dierences.
Calcium (Ca) and potassium (K) showed highly signicant
dierences (p<0.0001), with higher concentrations of Ca in 2013
and K in 2014. Similarly, iron (Fe) and magnesium (Mg) levels
were signicantly higher in 2013. while in 2014 they were reduced,
Manganese (Mn) was also higher in 2013, while sodium (Na) reached
higher values in 2014.
Copper (Cu) showed a signicant decrease in 2014 compared to
2013, while mercury (Hg), although present in low concentrations,
was higher in 2013 and practically undetectable in 2014 (p<0.001).
On the other hand, arsenic (As) was not detected at quantiable
levels (values below the detection limit in both campaigns), and both
cadmium (Cd) and chromium (Cr) remained at low levels with no
signicant dierences between years (p>0.05).
Although several of the elements analysed correspond to
essential minerals, only cadmium has maximum levels specically
established for cocoa products in the European Union. For other
potentially toxic elements, such as As, Hg and Cr, there are currently
no specic regulatory limits for cocoa beans, so their interpretation
must be carried out within the framework of general toxicological
assessments, such as those established by the Codex Alimentarius and
the World Health Organization (Codex Alimentarius Commission.
2019; World Health Organization. 2011).
Year-on-year change in essential minerals
The increase in K in 2014 and the decrease in Ca, Mg, Fe and
Mn compared to 2013 can be explained in the framework of soil and
climatic factors. Properties such as pH, organic matter, texture and
cation exchange capacity inuence the mobility and bioavailability
of nutrients and metals in the soil (Meter et al., 2019; Barraza et al.,
2017). In addition, rainfall and water balance modify the composition
of the soil solution and therefore, the absorption of cations (Meter
et al., 2019). These patterns are comparable with reports of cocoa
in Peru and Ecuador, in which K is reported as the most abundant
macroelement, while Fe and Mn vary according to local conditions
(Arévalo-Gardini et al., 2017; Gramlich et al., 2018).
Cadmium (Cd)
Although the Cd showed a decrease between 2013 and 2014
(1.62 to 1.04 mg.kg
-1
), the dierence was not statistically signicant.
These values are in the high range of the values reported in other
producing countries such as Ecuador, Peru and Colombia, in which
concentrations usually vary between 0.5 and >1.0 mg.kg
-1
, depending
on the soil characteristics and genotype (Arévalo-Gardini et al., 2017;
Barraza et al., 2017; Meter et al., 2019).
During the study period, the European Union had already
established maximum limits of Cd in cocoa products by Regulation
(EU) 488/2014, subsequently consolidated in Regulation (EU)
2023/915. These thresholds correspond to 0.60 mg.kg
-1
for cocoa
powder and 0.80 mg.kg
-1
for chocolates with a high cocoa content
(European Union 2014; 2023). Although these limits apply to the
nal product and not to the grain, the concentrations observed in this
study conrm the need for systematic on-farm monitoring and the
application of mitigation strategies, such as liming, phosphorus and
zinc management, or agroforestry practices, which have already been
shown to be eective in other countries (Meter et al., 2019; García-
Porras et al., 2025).
It should also, be noted that the analyses were carried out on
roasted almonds. in line with a previous study (Lanza et al., 2016).
Although most, research on metals in cocoa uses raw beans. the heat
treatment applied in this study sought to simulate conditions closer
to the real processing of cocoa. This methodological dierence must
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Lanza. Rev. Fac. Agron. (LUZ). 2026, 43(1): e264313
5-6 |
be considered when comparing directly with published values for
raw grains. Consequently, the Cd levels reported in this study should
be interpreted as an indicator of potential risk for export, rather
than as a direct exceedance of regulatory limits, given that the nal
concentration in chocolate depends on multiple industrial processes:
fermentation, drying, roasting, grinding and above all, on the dilution
associated with the mixture with other ingredients (sugar, powdered
milk, cocoa butter).
Mercury (Hg). arsenic (As) and chromium (Cr)
In this study, Hg and As were detected in traces and Cr remained
stable, with no dierences between years. These ndings are
consistent with regional research in which Cd emerges as the main
contaminant of interest in Latin American cocoa, while other toxic
metals are usually at low levels (Padilla et al., 2025).
The changes observed between 2013 and 2014 can be attributed
to dierent factors that inuence the absorption and accumulation of
minerals in the grain. These assumptions are summarized in table 5.
Table 5. Climatic. edaphic and postharvest factors associated with
the year-on-year variation of minerals in cocoa beans.
Hypothesis Description References
Weather conditions
Rainfall and water balance
aect the availability of K, Ca
and Mg in the soil solution.
modifying their absorption by
the plant.
Arévalo-Gardini et al.
(2017)
Soil factors
Variations in pH, organic
matter and phosphorus
regulate the bioavailability
of Cd and its interaction with
bivalent cations (Ca²⁺, Mg²
+
).
Barraza et al. (2017);
Meter et al. (2019)
Post-harvest
practices
Fermentation and drying
processes modify the apparent
concentration of minerals and
their bioaccessibility in the
bean.
García-Porras et al.
(2025)
In summary, the interannual dierences observed in this
study can be explained mainly by the combination of climatic
conditions, soil factors and postharvest practices (table 5). These
three elements constitute the most documented mechanisms in the
literature for mineral variation in cocoa; however, other possible
determinants, such as cultivar genetics, plant-microbiota interactions,
or agricultural management practices, should not be ruled out. The
ndings presented here reinforce the need to address these factors
in an integrated manner in future studies that directly correlate soil,
climate and genotype parameters with the mineral composition of the
grain.
Nutritional implications
The high content of K and Mg conrms cocoa as an important
source of minerals, in agreement with composition data of other Latin
American cocoas (Arévalo-Gardini et al., 2017; Gramlich et al.,
2018). These minerals contribute to its nutritional value and potential
as a functional food.
Overall, the stability of the proximate composition and the
interannual dierences in minerals observed in this study are
consistent with the Latin American literature. The downward trend in
Cd, although not signicant, reinforces the importance of monitoring
by origin to comply with international regulations and maintain the
competitiveness of Venezuelan cocoa in demanding markets.
Conclusions
The proximate composition of Cata and Cuyagua cocoa beans
remained relatively stable between 2013 and 2014, except for
moisture, which was signicantly lower in 2014, which can be
attributed to climatic or post-harvest conditions.
The mineral prole showed signicant year-on-year variations:
Ca, Mg, Fe, Mn and Cu were higher in 2013, while K and Na
increased in 2014. These results suggest the combined inuence of
climatic, edaphic and postharvest factors on the mineral composition
of the grain.
Although the decrease in cadmium (Cd) was not statistically
signicant, the levels found are relevant in the face of European
regulation (Regulation (EU) 488/2014), which underscores the need
for systematic monitoring and on-farm mitigation strategies.
The high content of K and Mg conrms the importance of cocoa
as a nutritional source of essential minerals, in accordance with
reports in the literature for Latin American cocoa.
These results constitute a baseline for future studies that integrate
edaphic, climatic and genetic variables, in order to design specic
mitigation strategies by origin.
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