© The Authors, 2021, Published by the Universidad del Zulia*Corresponding author: segundo.quintana@untrm.edu.pe
Diner Mori Mestanza
1
Miguel Ángel Barrena Gurbillón
2
Segundo Manuel Oliva Cruz
2
Segundo Grimaldo Chavez Quintana
1,2*
Rev. Fac. Agron. (LUZ). 2022, 39(1): e223917
ISSN 2477-9407
DOI: https://doi.org/10.47280/RevFacAgron(LUZ).v39.n1.17
Food Technology
Associate editor: Dra. Gretty Ettiene
Keywords:
Fermentation index
Native cocoa
Fine aroma
Theobroma cacao
Response surface optimization of the cacao criollo fermentation process in the province of
Utcubamba, Amazonas-Peru
Optimización mediante supercie de respuesta del proceso de fermentación del cacao criollo en la
provincia de Utcubamba, Amazonas-Perú
Optimização através da superfície de resposta do processo de fermentação do cacau criollo na
província de Utcubamba, Amazonas-Peru
1
Universidad Nacional Toribio Rodríguez de Mendoza de
Amazonas, Instituto de Innovación y Desarrollo para el
Sector Agrario y Agroindustrial de la Región Amazonas,
Calle Higos Urco 350, Chachapoyas, Perú. CP: 01001
2
Universidad Nacional Toribio Rodríguez de Mendoza
de Amazonas, Instituto de Instituto de Investigación para
el Desarrollo de Ceja de Selva. Calle Higos Urco 350,
Chachapoyas, Perú. CP: 01001.
Received: 08-04-2021
Accepted: 15-09-2021
Published: 16-02-2022
Abstract
The fermentation process of native ne aroma cacao criollo (Theobroma
cacao) in the province of Utcubamba, Amazonas, Peru, was optimized. A
Box-Behnken statistical design was applied, whose factors were inoculum
concentration, fermentation time and inoculation sequence. With the
optimized model, a phenolic content of 12.99 mg AGE.g
-1
cocoa, a
fermentation index of 1.05, and theobromine and caffeine contents of 4.89.100
g
-1
cocoa and 1.81.100g
-1
cocoa, respectively, were obtained. Additionally,
with a panel of nine certied and accredited tasters, the basic and special
descriptive qualitative sensory attributes of the fermented and dry cocoa
beans were determined, obtaining a maximum quality score of 71.1, and the
sensory descriptors oral, fruity, nutty, sweet, bitter, acidity and adequate
astringency were identied. In the cocoa obtained with the best treatment,
64 volatile compounds from the families of aldehydes, ketones, alcohols,
esters, acids and pyrazines were identied by gas chromatography coupled
to a mass detector with solid-phase microextraction (GC-MS-SPME-HS).
In conclusion, it was possible to optimize the fermentation process of cacao
criollo to obtain cocoa with high functional and sensory properties.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2022, 39(1): e223917. January - March. ISSN 2477-9407.
2-8 |
Resumen
Se optimizó el proceso de fermentación de cacao (Theobroma
cacao) criollo nativo no de aroma en la provincia de Utcubamba,
Amazonas, Perú. Se aplicó un diseño estadístico de Box-Behnken
cuyos factores fueron la concentración de inóculo, tiempo de
fermentación y secuencia de inoculación. Con el modelo optimizado
se obtuvo un contenido fenólico de 12,99 mg AGE.g
-1
cacao,
un índice de fermentación de 1,05, contenidos de teobromina y
cafeína de 4,89.100 g
-1
cacao y 1,81.100g
-1
cacao, respectivamente.
Adicionalmente, con un panel de nueve catadores certicados y
acreditados, se determinaron los atributos sensoriales básicos y
especiales descriptivos cualitativos del cacao en grano fermentado
y seco, obteniendo una puntuación de calidad máxima de 71,1 y se
identicaron los descriptores sensoriales, oral, frutal, nuez, dulce,
amargo, acidez y astringencia adecuada. En el cacao obtenido con
el mejor tratamiento, se identicaron 64 compuestos volátiles de las
familias de aldehídos, cetonas, alcoholes, ésteres, ácidos y pirazinas,
mediante cromatografía de gases acoplada a un detector de masas con
micro extracción en fase sólida (GC-MS-SPME-HS). En conclusión,
se logró optimizar el proceso de fermentación de cacao criollo que
permite obtener cacao con elevadas propiedades funcionales y
sensoriales.
Palabras clave: Índice de fermentación, cacao nativo, cacao criollo,
Theobroma cacao.
Resumo
O processo de fermentação do cacau criollo nativo de aroma no
(Theobroma cacao) na província de Utcubamba, Amazonas, Peru,
foi otimizado. Um projeto estatístico Box-Behnken foi aplicado
com concentração de inóculos, tempo de fermentação e seqüência
de inoculação como fatores. Com o modelo otimizado, foi obtido
um conteúdo fenólico de 12,99 mg de cacau AGE.g
-1
, um índice de
fermentação de 1,05, teobromina e cafeína de 4,89,100 g
-1
de cacau
e 1,81,100 g
-1
de cacau, respectivamente. Além disso, com um painel
de nove provadores certicados e credenciados, foram determinados
os atributos sensoriais básicos e os atributos descritivos qualitativos
especiais das amêndoas de cacau fermentadas e secas, obtendo-se
uma pontuação máxima de qualidade de 71,1 e foram identicados os
descritores sensoriais orais, frutados, frutos secos, doces, amargos,
acidez e adstringência. No cacau obtido com o melhor tratamento,
64 compostos voláteis das famílias de aldeídos, cetonas, álcoois,
ésteres, ácidos e pirazinas foram identicados por cromatograa
gasosa acoplada a um detector de massa com microextração de fase
sólida (GC-MS-SPME-HS). Em conclusão, foi possível otimizar o
processo de fermentação do cacau criollo para obter cacau com altas
propriedades funcionais e sensoriais.
Palavras-chave: Índice de fermentação, cacau nativo, cacau criollo,
Theobroma cacao.
Introduction
Among the different processes that cocoa beans must go through
(Theobroma cacao), fermentation is the rst step in the chocolate
chain (De Melo et al., 2013) this process is important and benecial
for the development of sensory characteristics. Microbial activity
in cocoa beans removes the mucilage and induces a set of internal
biochemical reactions in the cotyledon, which modify the chemical
composition of the bean and initiate the formation of aroma
precursors. During fermentation, microbial succession is produced
by the variation of temperature, pH, oxygen availability and the
compounds generated (Kongor et al., 2016), which makes the results
very heterogeneous, so the optimization of this process will enhance
and preserve the bromatological and organoleptic characteristics
of cocoa that guarantees a good quality and homogeneous product
(Sandhya et al., 2016).
According to Wacher (2011), yeasts and bacteria (lactic and acetic)
are responsible for the fermentation of cocoa pulp, which contains
carbohydrates (glucose, fructose and sucrose) at a pH between 3.3
and 4.0. They indicate that it is also necessary the presence of citric
acid, appropriate medium for the development and proliferation of
microorganisms that are essential for the transformation of cocoa
beans; on the other hand, the production of acetic acid, ethanol and
the increase of temperature, avoid the damage of the cocoa bean
preventing the enzymatic action. .
The indicator of an optimal fermentation process is the
‘‘fermentation index’’ which must be equal to or higher than one (1)
(León-Roque et al., 2016). In this sense, works have been carried
out that have sought to improve this indicator using starter cultures
that increase the efciency of fermentation. In this regard, Cempaka
et al. (2014), reported the increase of the fermentation index from
0.84 to 1.13 by initial addition of yeast, and Kresnowati et al. (2013)
observed that by employing a starter culture composed by yeast
and lactic acid bacteria, the fermentation index is improved from
0.86 to 0.95. Therefore, the objective of this work was to optimize
the fermentation process of native ne aroma cacao criollo in the
province of Utcubamba, Amazonas, Peru, by means of a response
surface.
Materials and methods
The cocoa samples were acquired in slime from the La Cruz
sector of the Cajaruro district, province of Utcubamba, Amazonas,
Peru, a native (cacao criollo) producing area of the so-called ne
aroma (common in northeastern Peru), characterized by an average
altitude of 490 m above sea level, an average temperature of 29 °C
and humidity of 61%.
The samples were transported in polypropylene bags to the
facilities of the Cooperativa Central de Productores Agropecuarios
de Amazonas (CEPROAA), where the fermentation experiment was
carried out in cubic boxes of laurel wood (Cordia alliodora), 30 cm
on each side, with 20 kg of cocoa in slime for each treatment. After
the fermentation process (according to treatments), the fermented
cocoa was dried in the sun until a moisture content of less than 7%
was obtained, then it was packaged in airtight polyethylene bags
and transferred to Laboratorio de Control de Calidad de Cacao of
Universidad Nacional Toribio Rodríguez de Mendoza de Amazonas,
for further analysis.
Obtaining the starter culture
The procedure described by Ausubel et al. (1989) was followed.
For this purpose, pure cultures (yeasts + lactic acid bacteria [BAL] +
acetic acid bacteria [BAA]) were isolated and biomass was obtained,
achieving a frank development of yeasts in 250 mL asks with liquid
YPD culture medium prepared from meat peptone (20 g.L
-1
), yeast
extract (10 g.L
-1
) and glucose/dextrose (20 g.L
-1
) in ultra-pure water
(Ausubel et al., 1989); the medium contained chloramphenicol for
bacterial growth inhibition. For lactic and acetic acid bacteria, MRS
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Mori et al. Rev. Fac. Agron. (LUZ). 2022, 39(1): e2239173-8 |
and Acetobacter broths were used. During fermentation, removal or
turning was carried out every 24 h, until completing the programmed
time according to the treatment.
Determination of the fermentation index (IF)
The IF was determined following the spectrophotometric method
described by Gourieva & Tserevitinov (1979), given by the ratio of
absorbances between 460 and 530 nm.
Determination of total phenol content
The determination of total phenols was carried out using the Folin
Ciocalteu assay, for which, a standard calibration curve was prepared
from dilutions of 0 to 16 mg.L
-1
of gallic acid (Sigma Aldrich,
Germany), following the procedure described by Pantelidis et al.
(2007). Zero point zero ve milliliters (0.05 mL) of cocoa extract and
0.45 mL of water with 2.5 mL of Folin-Ciocalteu reagent (Merck,
Germany) diluted 1:10, followed by two (2) mL of Na
2
CO
3
at 7.5 %
(m/v); then, they were completely mixed for 10 seconds in Vortex. It
was incubated in an oven at 50 ºC for ve (5) min, subsequently; the
absorbance was measured at 765 nm in a spectrophotometer (UV-
Visible S2100 UVTE, ÚNICO).
Determination of theobromine and caffeine
Theobromine and caffeine were determined by high performance
liquid chromatography (HPLC) following the method described by
Brunetto et al. (2007), using a chromatograph (Hitachi-Chromaster,
Tokyo, Japan, (LC-20AD), equipped with an autoinjector (SIL-
20A/HT), a communication module (CBM-20A) and a detector
with photodiode array (PDA, SPD-M20A), ultraviolet detection
was recorded at 278 nm. The separation was carried out on a 5
µm Supelco-LiChrospher RC C-18 column (25 cm x 4.6 mm). A
methanol/water mixture (30:70 v/v) was used as mobile phase in
isocratic mode at a ow rate of 1.0 mL.min
-1
. The identication of the
signals was performed by comparing the retention times obtained with
theobromine and caffeine standards (98 % Sigma-Aldrich, USA). For
the quantication of the concentrations of theobromine and caffeine in
the cocoa samples, a calibration curve was prepared for theobromine
and caffeine in a range from 5 to 100 mg.L
-1
, with whose areas under
the curve the respective linear function was obtained.
Determination of the pH of cocoa beans
The pH of the samples was measured following the methodology
described by Senanayake et al. (1999).
Preparation of samples for sensory analysis and volatile
compounds
The whole, fermented and dried beans obtained according to
the treatments were roasted in a laboratory roaster (IMSA, Peru) of
one (1) kg capacity at 120 °C for 15 min. Then, they proceeded to
shelling in a sheller (IMSA, Peru). The nibs obtained were subjected
to a conching process for 4 h at 50 °C, molded into 50 g tablets,
packed in aluminum foil and polyethylene bag with hermetic seal.
The tablets were stored frozen until further analysis.
Sensory analysis of cocoa pastes
The fermented cocoa according to treatment was dried and
processed into paste. Then, with nine (9) judges certied and
accredited by APPCacao-Sineace-Minedu-Peru, the basic and special
sensory attributes (cocoa avor, oral, fruity, nutty, sweet, bitter,
acidity and astringency) were evaluated with the ordinal scale from
1 to 100 for the total score, given by the sum of the partial scores
of each attribute, using the Tasting Card for the sensory analysis of
cocoa (USAID, Equal Exchange and TCHO, 2018).
Identication of the volatile compounds in cocoa paste
Volatile compounds were identied using gas chromatography
coupled to a mass detector and headspace-micro solid-phase
extraction (GC-MS-SPME-HS) (Rodriguez-Campos et al., 2012),
employing a divinylbenzene/carboxen/polydimethylsiloxane ber
(DVB/CAR/PDMS), 50/30 µm thick and helium as carrier gas. In
20 mL vials, 5.7 g of cocoa paste was placed, six (6) mL of ultrapure
water was added and hermetically sealed with a metallic lid and a
20 mm white silicone septa. The SPME extraction conditions were:
15 minutes of equilibrium at 50 °C, with an exposure of the ber
for 30 min at the same temperature. After the extraction time, the
ber was retracted and immediately inserted into the injection port
of the gas chromatograph-mass spectrometer (GC-MS) (Angilent
technologies, 6890N, United States), where it was kept for ve (5)
min at 250 °C temperature. Between each extraction a run of the ber
blank (clean-up) was performed for 55 min. For the identication of
the compounds, the NIST 14.L library was used.
Experimental design and statistical analysis
The Box-Behnken statistical design (Murali et al., 2000) was used
to determine the best concentration values of inoculum X1 (1.0; 1.8
and 2.6 x 10
3
UFC.mL
-1
), inoculation sequence X2 (yeasts + BAL
+ BAA at the beginning; yeasts and BAL at the beginning+ BAA
at 48 h; yeasts at the beginning + BAL at 24 h + BAA at 48 h), and
fermentation time X3 (5, 6 and 7 days), as shown in table 1. The
responses of interest were the fermentation index (IF), chemical
characteristics (total phenols, theobromine, caffeine and pH), sensory
characteristics and volatile compounds.
Table 1. Box-Behnken arrangement for optimization.
Number Variables
Experiment X1 X2 X3
1 0 - -
2 + - 0
3 0 0 0
4 0 - +
5 - 0 +
6 0 + +
7 - 0 -
8 - - 0
9 0 0 0
10 0 0 0
11 + + 0
12 - + 0
13 0 + -
14 + 0 +
15 + 0 -
+: Upper level of the variables, 0: Average level of the variable. -: Lower level of
the variables, X1: Inoculum concentration. X2: sequence of inoculation, X3: days
of fermentation
Cocoa beans in slime were placed in the fermenters and 10 mL
of the microorganisms were inoculated according to the treatments
described. To obtain the optimum fermentation parameters with cocoa
starter culture, under the Box Behnken design. A response surface
analysis was performed using Minitab 19 software. The Kruskal
Wallis non-parametric test was also applied to compare treatments
according to sensory qualication.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2022, 39(1): e223917. January - March. ISSN 2477-9407.
4-8 |
Results and discussion
Table 2 shows the average values of chemical properties of
fermented cocoa beans, in which a variation is observed due to the
set of reactions generated in the fermentation process, accentuating
some characteristic avors and aromas of the samples, as described
by Castro-Alayo et al. (2019).
Optimization of fermentation parameters
Table 3 shows the optimized values, obtaining an IF higher
than 1, pH of 5.32, indicating an adequate fermentation, free of
undesirable compounds or acids in the cocoa beans.
Table 2. Chemical properties of fermented cocoa.
Treatments
Total phenols
(mg GAE.g
-1
)
Fermentation
index
pH
Theobromine
(g.100 g
-1
cocoa)
Caffeine
(g.100 g
-1
cocoa)
T1 15.07±0.39 0.68±0.10 5.85±0.10 5.04 2.47
T2 15.57±0.11 0.71±0.08 5.82±0.05 4.93 2.31
T3 15.46±0.42
0.90±0.42
5.78±0.10 4.58 2.04
T4 13.78±0.32 0.95±0.04 5.65±0.10 5.07 2.23
T5 20.41±0.66 0.55±0.28 5.69±0.11 5.13 2.15
T6 15.23±0.21 0.78±0.11 5.86±0.10 5.06 2.06
T7 16.93±0.17 0.99±0.09 5.59±0.03 5.12 2.25
T8 16.73±0.27 0.99±0.09 5.63±0.02 5.65 2.68
T9 15.46±0.42 0.90±0.19 5.74±0.09 4.96 2.48
T10 15.46±0.42 0.90±0.19 5.70±0.09 4.92 1.92
T11 13.93±0.86 0.97±0.02 5.80±0.04 5.81 2.35
T12 12.17±0.17 1.25±0.19 5.17±0.05 5.00 2.05
T13 17.28±0.48 0.87±0.19 5.33±0.09 6.14 2.46
T14
12.90±0.66
1.05±0.15 5.32±0.06 4.98 1.78
T15 15.37±0.41 1.18±0.09 5.27±0.10 5.65 2.40
Table 3. Response and optimized values for chemical properties
of the native ne aroma cocoa samples.
Response Optimized
<95.0
%
>95.0
%
Total phenols (mg GAE.g
-1
) 12.99 7.80 18.19
Fermentation index 1.05 0.57 1.52
pH 5.32 4.97 5.67
Theobromine concentration
(g.100 g
-1
cocoa)
4.89 4.44 5.35
Caffeine concentration (g.100 g
-1
cocoa)
1.81 1.37 2.25
The optimized model was determined: inoculum concentration
2.6 x 10
3
UFC.mL
-1
, yeast inoculation sequence and BAL at the
beginning + BAA at 48 h and fermentation time (7 days), to achieve
an adequate fermentation process (gure 1).
Figure 1. Response surface graph to optimize the fermentation
index in the fermentation process of cacao criollo.
The optimized model (gure 2), allows to obtain fermented cocoa
with low levels of total phenols (12.99 AGE.g
-1
), therefore the pastes
have low astringency (Ooi et al., 2020). Given the differences in the
results, treatments 7, 8 and 11, due to their high fermentation index
(table 2), improved microbial activity, they were able to hydrolyze
free, soluble phenolic complexes, which are easily absorbed, leading
to a decrease in seeds (Haile & Kang, 2019; Jalil & Ismail, 2008).
This has been largely attributed to oxidation of insoluble tannins
and leaching of almonds polyphenols into the surrounding pulp
and subsequent runoff on sweating (Wollgast & Anklam, 2000).
Figure 2. Response surface graph to optimize the phenolic
content in the fermentation process of cacao criollo.
The application of the microorganisms allowed to obtain a
fermentation index equal to 1.05 in seven (7) days, very consistent
results due to the obtaining of completely fermented beans with
well pronounced, light brown to dark brown striations (Ooi et al.,
2020). It should be noted that the brown color formed is due to
the oxidation reaction when catalyzing o-diphenol to o-quinone
(Hernández-Hernández et al., 2016).
The model allowed to obtain an optimized pH value of 5.32;
which indicates an adequate fermentation process, the pH of beans
leads to the generation of more free peptides and amino acids,
which would be able to form the Maillard reaction during roasting,
contributing to more preferred aroma and avor notes (Afoakwa et
al., 2008; John et al., 2019).
Theobromine and caffeine concentrations
The optimal values of theobromine and caffeine were 4.89 and
1.81 g.100 g
-1
cocoa respectively; values that indicate the intensity
of bitterness in the cocoa paste (Brunetto et al., 2007), these
compounds are associated with the index fermentation of cocoa
beans (Cardoso et al., 2020).
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Mori et al. Rev. Fac. Agron. (LUZ). 2022, 39(1): e2239175-8 |
Sensory analysis of native ne cacao criollo paste with
fermented aroma with different treatments
Figure 3 shows the results obtained in the sensory analysis of
the quality attribute of native ne aroma cocoa.
Figure 3. Sensory analysis of the quality attribute of native ne
aroma cocoa.
Four treatments (T11, T14, T15 and T2) formed the best scored
group (close to 70) in terms of quality of the cocoa paste obtained
per treatment, as shown in gure 3a.
The same treatments that allowed obtaining higher quality
were those that obtained the best aroma score, as shown in gure
3b. Average scores of seven (7) for treatments 11, 14 and 15,
evidence that the fermentation was adequate and that the volatile
compounds precursors of the aroma were enhanced. Fermentation
treatments 11, 14, 15, 2 and 9, obtained the best scores in sensory
acidity. The judges revealed a citric acidity, this could be due to
4-methylpentanoic and beta-myrcene acids, identied in T14, the
treatment with the highest sensory acceptance (p<0.05), as can be
seen in Figure 3c, which confer an acidity similar to a sweet orange,
generated and transferred to the cotyledons during the fermentation
process, being this type of acidity benecial for special cocoa
samples (Utrilla-Vázquez et al., 2020). The same treatments had
the highest scores in avor and aftertaste (gures 3d and 3e).
Identication of the volatile compounds of native ne aroma
cocoa
The volatile compounds precursors of aroma were identied in
treatment 14 (table 4), due to it was the best treatment. Among the
identied volatile compounds, aldehydes, ketones, alcohols, esters,
acids and pyrazines, mainly responsible for special aromas and
avors, such as Linalool, 2-Nonanol (oral, menthol, cinnamon,
citrus and fruit aromas); 3-ethyl-2,5-dimethyl pyrazine, tetramethyl
pyrazine, 2-ethyl-6-methyl pyrazine, trimethyl pyrazine,
2,3,5-trimethyl-6-ethylpyrazine (aromas of roasted cocoa, sweet
panela, sweet caramel) (Utrilla-vázquez et al. , 2020). Figure 4
shows chromatogram of volatile compounds identied by GC-MS-
SPME-HS.
Table 4. Volatile compounds identied in fermented samples of native ne aroma cacao criollo.
Compound name Group Chemical formula TR Compound aroma description*.
Dimethyl ether Ether C
2
H
6
O 4.855 Fruits
Acetone
Aldehydes and ketones
C
3
H
6
O 5.475 Fruits
2-Methylpropanal C
4
H
8
O 7.096 Sweet
Acetic acid Acid C
2
H
4
O
2
7.625 Bitter, vinegar
2,3-Butanedione
Ketones
C
4
H
6
O
2
7.943 Buttery
2-Butanone C
4
H
8
O 8.194 Sweet
Butanimidamide C
4
H
10
N
2
10.688 …
2-Methylbutanal Aldehydes and ketones C
5
H
10
O 11.108 Malt, chocolate
2,3-Pentanedione Ketones C
5
H
8
O
2
12.345 Toasted almond, cocoa, yogurt, nuts
Pentanal Aldehydes and ketones C
5
H
10
O 12.541 Fruits
Acetoin Aldehydes and ketones C
4
H
8
O
2
12.982 Butter
3-(1methylethyl)oxoethane
2-Methylpropanoic acid
C
6
H
12
O 14.283 ...
Acid C
4
H
8
O
2
14.515 ...
2,3-Butanediol Alcohol C
4
H
10
O
2
16.446 ...
Cyclobutanol, 2-ethyl C
6
H
12
O 17.664 …
Hexamethyl cyclotrisiloxane C
6
H
18
O
3
Si
3
18.061 …
3-Furaldehyde Aldehydes and ketones C
5
H
4
O
2
19.34 Toasted almond
Butanoic acid, 2-methyl Acid C
5
H
10
O
2
19.477 …
3-Furanmethanol Alcohol C
5
H
6
O
2
20.166 Mint
2-Heptanone Aldehydes and ketones C
7
H
14
O 22.094 Fruit, plantain
Styrene C
8
H
8
22.656 Sweet, cinnamon, coffee.
1-(2-Furanyl) ethanone C
6
H
6
O
2
23.214 …
*(Utrilla-vázquez et al., 2020).
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Rev. Fac. Agron. (LUZ). 2022, 39(1): e223917. January - March. ISSN 2477-9407.6-8 |
Table 4. Volatile compounds identied in fermented samples of native ne aroma cacao criollo (continuation).
Compound name Group Chemical formula TR Compound aroma description*.
2,5- Dimethylpyrazine
Pyrazines
C
6
H
8
N
2
23.401 Caramel
2,3- Dimethylpyrazine C
6
H
8
N
2
23.708 Roasted cocoa
4-Methylpentanoic acid C
6
H
12
O
2
23.950 Fruity, pineapple
2-Furancarboxaldehyde, 5-methyl C
6
H
6
O
2
25.692 …
Octametil ciclotetrasiloxano C
8
H
24
O
4
Si
4
25.884 …
Benzaldehyde Aldehydes and ketones C
7
H
6
O 26.094 Pleasant almonds
Beta-Myrcene C
10
H
16
26.965 Citrus
Benzonitrile Nitriles C
7
H
5
N 27.061 Almonds
2-Ethyl-6-methylpyrazine,
Pyrazines
C
7
H
10
N
2
27.435 Sweet caramel
Trimethylpyrazine C
7
H
10
N
2
27.617 Roasted cocoa
2-Ethenyl-6- methylpyrazine C
7
H
8
N
2
28.385 Sweet caramel
Acetate 2-heptanol C
9
H
18
O
2
28.845 …
3,6,6- trimethylbicyclo (3.1.1)
hept-2-ene
C
10
H
16
28.975 …
Benzyl alcohol Alcohol C
7
H
8
O 29.129 Sweet, owery
1-Methyl-5-(1-methylethenyl)
cyclohexene
C
10
H
16
29.199 Essential oils, citrus
1,3,3-Trimethyl,
tricyclo[2.2.1.0(2,6)]heptane
C
10
H
16
29.509 Essential oils, citrus
Benzeneacetaldehyde Aldehydes and ketones C
8
H
8
O 29.738 Almond, cherry, strawberry
1-(1h-pyrrol-2-yl) ethanone, C
6
H
7
NO 30.161 …
3-Methyl 2-cyclohexen-1-one C
7
H
10
O 30.325 …
Acetophenone Aldehydes and ketones C
8
H
8
O 30.783 Sweet
3-Ethyl-2,5- dimethylpyrazine
Pyrazines
C
8
H
12
N
2
30.913 Roasted cocoa,
Tetramethylpyrazine C
8
H
12
N
2
31.267 roasted, chocolate
2-Methoxyphenol C
7
H
8
O
2
31.507 Sweet
Ethyl 2-(5-methyl-5-vinyl
tetrahydrofuran-2-yl)propan
-2-yl carbonate
C
13
H
22
O
4
31.575 …
2-Nonanol
Alcohol
C
9
H
20
O 31.715 Fruits, roses
Linalool C
10
H
18
O 31.810 Floral, mentholated, cinnamon, citrus
Nonanal Aldehyde C
9
H
18
O 31.984 Rose, orange, citrus
Phenylethyl alcohol Alcohol C
8
H
10
O 32.718 Floral
2,3-Dimethyl 2,4,6-Octatriene C
10
H
16
33.007 Citrus, essential oils
4h-Pyran-4-one, 2,3-dihydro-3,5-
dihydroxy-6-methyl
C
6
H
8
O
4
33.918 …
2,3,5- Trimethyl-6-
ethylpyrazine
Pyrazines C
9
H
14
N
2
34.204 Sweet caramel
Ethyl bezoate Ester C
9
H
10
O
2
35.026 Sweet, fruit, cherry, grapes.
(3R,6S)-2,2,6- Trimethyl-6-
vinyltetrahydro-2H-pyran-3-ol
Alcohol C
10
H
18
O
2
35.155 …
Octanoic acid, ethyl ester Ester C
10
H
20
O
2
35.468 Orange aroma, citrus
Dodecane C
12
H2
6
35.81 …
Benzeneacetic Acid
Ethyl ester
C
10
H
12
O
2
37.666 …
Acetic acid, phenylethyl ester C
10
H
12
O
2
38.167 …
Benzeneacetaldehyde, alpha.-
ethylidene C
10
H
10
O 38.957 Green, fresh
*(Utrilla-vázquez et al., 2020).
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Mori et al. Rev. Fac. Agron. (LUZ). 2022, 39(1): e2239177-8 |
Compound name Group Chemical formula TR Compound aroma description*.
Cyclotetradecane
C
14
H2
8
42.865 ...
1-Butanol, 3-methyl-, benzoate
C
12
H
16
O
2
43.289 …
5-Methyl-2-phenyl-2-hexenal
C
13
H
16
O 46.478 Cocoa
1H-2-Benzopyran-1-one,
3,4-dihydro- 8-hydroxy-3-methyl
C
10
H
10
O
3
48.988 …
*(Utrilla-vázquez et al., 2020).
Table 4. Volatile compounds identied in fermented samples of native ne aroma cacao criollo (continuation).
Figure 4. Chromatogram of volatile organic compounds
precursors of the aroma of the T14 treatment.
Sixty-four volatile compounds were identied: 14 aldehydes and
ketones; 7 pyrazines, 7 alcoholic compounds, 4 esters, 4 organic acids
and other compounds, which confer the special notes to the cocoa
samples (Rodríguez-Campos et al., 2012). At the end of fermentation,
aldehydes and ketones represent the highest percentage of the total
content of volatile compounds, which confer the pleasant notes of
almond, butter or oral; pyrazines and alcohols are among the most
important avor groups of cocoa as they confer notes of chocolate,
roasted coffee, fruity, oral, menthol, cinnamon, citrus and especially
the Linalool volatile compound present in samples of cocoa from the
Amazon region. Similar compounds were found in samples of Criollo
and Trinitario cocoa grown in Chiapas, Mexico (Utrilla-Vázquez et
al., 2020).
Conclusions
This study allowed to optimize the fermentation process of T.
cacao native ne aroma, nding the optimal model. Emphasizing the
importance of the fermentation process presented in the particular
characteristics of the chemical composition and volatile compounds
of native ne aroma cocoa from the Amazon region.
The microorganisms added in the fermentation process allowed
accelerating a set of chemical reactions that give rise to the formation
of avor precursors and the formation of a qualitatively and
quantitatively very important aroma precursor fraction, which was
evidenced by the volatile compounds identied.
Regarding to the sensory analysis, a nal maximum quality
score of 71.1 was obtained, inuencing the astringency due to the
concentration of phenols, the bitterness due to the theobromine and
caffeine content, and the special avors and aromas due to the volatile
compounds, especially from the aldehyde, ketone, ester, pyrazine and
alcohol families, which confer these special notes to cocoa, allowing
to have extraordinary samples.
Funding source
Instituto de Investigación para el Desarrollo Sustentable Ceja de
Selva (INDES-CES) de la Universidad Nacional Toribio Rodríguez
de Mendoza de Amazonas, through CINCACAO project /
Contrato N° 026-2016-FONDECYT: Círculo de Investigación para
la Innovación y el Fortalecimiento de la Cadena de Valor del Cacao
Nativo Fino de Aroma en la Zona Nor Oriental del Perú, fully
financed the research
work.
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