Food Chemistry 342 (2021) 128304

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

Impacts of quaker beans over sensory characteristics and volatile T

composition of specialty natural coffees
Mariane Helena Sances Rabeloa, , Flávio Meira Boréma, Renato Ribeiro de Limab,
⁎

Ana Paula de Carvalho Alvesa, Ana Carla Marques Pinheiroc, Diego Egídio Ribeiroa,
Cláudia Mendes dos Santosa, Rosemary Gualberto Fonseca Alvarenga Pereirac
a
Department of Agricultural Engineering, Federal University of Lavras, POB 3037, 37.200-000 Lavras, MG, Brazil
b
Department of Statistic, Federal University of Lavras, POB 3037, 37.200-000 Lavras, MG, Brazil
c
Department of Food Science, Federal University of Lavras, POB 3037, 37.200-000 Lavras, MG, Brazil

ARTICLE INFO ABSTRACT

Keywords: The objective of this study was to evaluate the volatile composition and the sensory effect of the presence of
Quaker Quaker beans in natural specialty coffee beverage and, consequently, to confront the requirement of the
Coffee Arabica Specialty Coffee Association regarding the total absence of Quaker beans in a natural specialty coffee batch.
Roasted coffee Sensory analysis and volatile composition were performed for three different colorations of Quaker beans, added
separately to natural specialty coffee samples at seven different concentrations. Beans with color equal to or
above Agtron 82.8 negatively affected the sensory characteristics of natural specialty coffee only from the
presence of 7 Quaker beans in one cup (65 beans). Through the analysis of volatile composition, volatile
compounds formed during roasting were identified in Quaker beans from precursors present in raw immature
beans. Therefore, the color and sensory characteristics of Quaker are a consequence of the chemical composition
of an immature bean.

1. Introduction adoption of technologies for selective harvesting. However, it is

Specialty coffee has several definitions that involve both the char-
acteristics of its production and consumption chain, as well as the
physical and sensory characteristics associated with raw and roasted
beans. A coffee sample must meet several criteria to be classified and
sold as specialty coffee; one of which is not to have Quaker beans,
which are defined by the Specialty Coffee Association – SCA as im-
mature beans that do not develop during the roasting process.
Therefore, Quaker beans are considered a coffee defect and are iden-
tified only after the roasting process; in raw beans, it is not possible to
identify them. The criteria adopted for specialty coffees, including the
criterion for Quaker beans, were established by the Specialty Coffee
Association and must be followed by all countries that market specialty
coffees (Lingle, 2011a).
The higher incidence of Quaker beans is associated with changes in
beverage flavor, which is characterized as herbaceous, immature, as-
tringent, peanut, among other sensory descriptors. In turn, these sen-
sory characteristics occur due to the presence of immature beans,
commonly found in the production of natural coffees due to the lack of
common to find natural coffees sold as specialty, containing varying
amounts of Quaker beans, leading to the question whether the criterion
adopted by the Specialty Coffee Association is really valid. Moreover,
there is not a consensus on the definition of the Quaker bean based only
on the visual analysis of its color, since there are yellow beans with
different shades.
It is believed that beans that have different shades, despite having
gone through the same roasting process, have a different volatile
composition that differently affects the sensory quality of the coffee
beverage. However, there is no scientific evidence to prove the impacts
of the presence of different proportions of these beans, and their dif-
ferent shades, on the sensory quality of specialty natural coffees.
Moreover, the threshold of sensory perception regarding the presence
of Quaker beans in a specialty coffee is not yet known. This parameter is
essential to establish a tolerance for the presence of Quaker beans in a
specialty coffee sample and thus define a new criterion. It is evident
that this restrictive criterion on the commercialization of specialty
coffees with the presence of Quaker beans must be reviewed. Studies
that determine the impacts of the Quaker bean on the sensory

⁎
Corresponding author.
E-mail address: marianerabelo1@hotmail.com (M.H.S. Rabelo).

https://doi.org/10.1016/j.foodchem.2020.128304
Received 9 November 2019; Received in revised form 21 August 2020; Accepted 30 September 2020
Available online 06 October 2020
0308-8146/ © 2020 Elsevier Ltd. All rights reserved.
M.H.S. Rabelo, et al.
characteristics of a specialty coffee will support the Specialty Coffee
Association to define new criteria for the presence of Quaker beans.
Thus, it would enable the commercialization of natural coffees as being
specialty, even with the presence of Quaker beans, opening the market
to countries that mostly produce natural coffees, providing greater
added value to the product.
In view of the importance of determining the sensory, physical and
chemical characteristics of a Quaker bean, besides the amount of
Quaker beans present in a sample that is capable of negatively influ-
encing the quality of the coffee beverage, the objective of this research
was to evaluate the impacts of the presence of Quaker beans on the
sensory characteristics and volatile composition of specialty natural
coffee. In this context, some strategies were adopted, such as the defi-
nition of color references to classify a bean as Quaker; determination of
the effects on sensory characteristics and the threshold of sensory
perception that the presence of Quaker beans causes in a specialty
natural coffee; determination of volatile compounds present in Quaker
beans responsible for causing changes in sensory characteristics and,
finally, the tolerance of Quaker beans in a specialty natural coffee
sample.
2. Material and methods
2.1. Raw material
To carry out this research, a commercial batch of natural coffee
(Coffea arabica L.) was randomly selected, originating from the
Mantiqueira de Minas region (Minas Gerais, Brazil) and supplied by the
company CarmoCoffees.
From the commercial batch, two sub-batches were formed. The first
sub-batch was used to comprise the Control treatment and was formed
by flat-shaped beans, the size was approximately of 6.35 mm (beans
classified in relation to size through a 16/64 in. round hole sieve) and
free from defective beans such as black, black-green, immature, sour,
insect-damaged, broken and badly formed. This sub-batch was classi-
fied as a specialty coffee, with a sensory score greater than 80 points,
according to the criteria described by the Specialty Coffee Association –
SCA (Lingle, 2011b). The second sub-batch, used to comprise the other
treatments, was formed by beans classified as immature defects, se-
lected automatically by an optical electronic selector (Sortex B Color-
Vision, Buhler). The aforementioned defects are described by Norma-
tive Instruction No. 8 of the Ministry of Agriculture, Livestock and
Supply of June 11, 2003 (BRASIL, 2003).
2.2. Roasting process
For the roasting process of the sub-batches of natural coffee used in
this research, a temperature versus time roasting curve was developed.
This roasting curve was developed with the main objective of stan-
dardizing the roasting process for both sub-batches. The first sub-batch
of natural coffee beans was used to develop the roast curve, as it was
homogeneous between the beans and represented the Control treat-
ment. In addition, due to the presence of immature beans in the second
sub-batch, which does not occur in the first sub-batch, the determina-
tion of the roasting end point based on the color of the beans became
unviable.
For the development of the roasting curve (available in the
Supplementary Material – Fig. 1) the parameters total roasting time and
final color of the beans were defined following the guidelines of the SCA
Protocols – Cupping Specialty Coffee (Lingle, 2011b). Thus, the total
roasting time was defined as 12 min and Agtron disc # 55 (SCA Roast
Classification Color System – Disks) the final color of the beans. The
roasting process was performed in an Atilla roaster model Gold Plus,
with a standardized initial roasting temperature of 150 °C, and this
value was defined based on pre-tests.
Four roasting batches were performed for each sub-batch, with each
2
Food Chemistry 342 (2021) 128304
batch representing a replication. Sensory analysis was performed in 3
replications, and volatile composition analysis was performed in 4 re-
plications.
2.3. Experimental design
The roasted beans of the second sub-batch were divided into three
groups of beans with a yellowish shade. For the definition of these
groups, the beans of the first sub-batch were used as a reference for
standard coloring (Agtron Scale − 53.7); the group of beans with the
most discrepant coloring of the standard was then defined. Based on the
two coloring extremes, the existence of two distinct groups was ob-
served, one with a coloring that approached the lighter end and another
that approached the standard coloring. At first, the three groups of
beans were named Color 1, Color 2 and Color 3, with Color 3 being the
most yellowish/light extreme.
To evaluate the sensory effect and the threshold of sensory per-
ception of Quaker beans of different shades present in a specialty nat-
ural coffee, three experiments were set, each experiment corresponding
to a group of beans (Color 1, Color 2 and Color 3). In these experiments,
each group of beans was added to the Control coffee at different con-
centrations, each concentration being a treatment of the experiment.
Each treatment was represented by a cup, containing 65 beans, which
correspond to approximately 8.25 g. In each experiment, the beans of
each group were inserted into the Control coffee at 7 different con-
centrations, defined through preliminary tests. Each of the concentra-
tions was tasted by the panelist in 3 replications, and each replication
represented a roasting batch.
For the Color 1 group, the 7 concentrations used were 0, 5, 10, 15,
20, 25 and 30 beans in a total of 65 beans per cup. For the Color 2 and
Color 3 groups, the 7 concentrations used were 0, 1, 4, 7, 10, 13 and 16
beans in a total of 65 beans per cup. In addition, it should be noted that
the concentrations referring to zero beans of the groups Color 1, Color 2
and Color 3 correspond to Control coffee, that is, Control coffee is the
specialty natural coffee used as a reference to measure the effect of the
inclusion of Quaker beans different shades.
In the case of bean color analysis and its volatile composition, the
Control, Color 1, Color 2 and Color 3 groups were analyzed individually
in 4 replications, with each replication representing a roasting batch.
2.4. Analysis of bean color
The color of the whole beans was measured using the SCA Roast
Color Classification System, using the M−Basic AGTRON Scale col-
orimeter. In addition, the beans color was also measured by the CIEL * c
* h° color space using the Konica Minolta CR-300 colorimeter.
2.5. Sensory evaluation
The sensory evaluation was carried out according to the re-
commendations of SCA Protocols – Cupping Specialty Coffee (Lingle,
2011b), carried out by five panelists certified as Q-graders. A sensory
evaluation form (Supplementary Material – Fig. 2) was developed to
show a better description of the sensory attributes of coffee and meet
the objectives of this study. The form was based on the Check All That
Apply sensory methodology – CATA (Adams, Williams, Lancaster, &
Foley, 2007) and the Intensity Scale methodology.
2.6. Analysis of volatile composition
For the analysis of volatile composition, the beans of each group
(Color 1, Color 2 and Color 3) and the Control coffee were analyzed
separately, that is, only beans from the same group. The beans from
each group and from the Control coffee were ground separately in a
analytical grinder (IKA, model A11), after grinding, 2 g of sample were
placed in vials for analysis. During grinding process, liquid nitrogen was
M.H.S. Rabelo, et al.
used to preserve the samples. Each group of beans was analyzed in four
replications, with each replication representing a roasting batch.
The volatile compounds were extracted using the static headspace,
as described by Maeztu et al. (2001), with adaptations. The GC–MS
model QP − 2010 SE (Shimadzu) equipped with an NST-100 column
(30 m × 0.25 mm × 0.25 μm) that has a polyethylene glycol phase
similar to a Carbowax ® column. The vials containing the samples were
placed in the equipment and remained in it for 30 min until reaching
equilibrium at 70° C. The volatile phase was injected into the gas
chromatograph (GC) with subsequent detection using a mass spectro-
meter (MS), and the injector temperature was adjusted to 220 °C. He-
lium gas, used as a carrier gas for the volatile phase, was maintained
throughout the analysis at a flow rate of 1 mL min−1. The heating of the
oven was programmed as follows: for 6 min, the temperature of the
oven was maintained at 25 °C, right after the temperature was heated
up to 70 °C at a rate of 10 °C min−1, until 95 °C at 5 °C min−1, until
115 °C to 10 °C min−1, up to 170 °C to 5 °C min−1 and, finally, to 215 °C
at 40 °C min−1. Therefore, the total chromatographic running time was
35 min.
Data analysis was performed using the GCMSsolution software
(version 4.4, Shimadzu Corporation, Japan) and the NIST NIST/EPA/
NIH 2014 database. Chemical identification of each compound was
performed by comparing the MS spectra with the dataset. The results
were expressed in relative percentage area that corresponds to the peak
area of each identified compound, calculated from the chromatogram
total area.
2.7. Statistical analysis
2.7.1. Analysis of bean color
Each group of Quaker beans (Color 1, Color 2 and Color 3) and
Control coffee had their color evaluated separately, totaling four
treatments. To check the difference in color between treatments, the
colorimetric parameters Agtron, L *, c * and h° were subjected to
analysis of variance (ANOVA) and, when significant differences were
identified in the F test, the Scott-Knott test was applied, using the
SISVAR software (SISVAR, version 5.3). A randomized block design
(DBC) with four blocks was used, each block corresponding to a
roasting batch.
2.7.2. Sensory analysis
Three experiments were carried out, set in a randomized block de-
sign (RBD), with three blocks. In each experiment, a group of Quaker
beans was evaluated, inserted at different concentrations in a Control
coffee. Each concentration studied is characterized by one treatment;
thus, seven treatments were studied for each experiment.
The results of the sensory analysis were submitted to multivariate
analysis, through principal component analysis (PCA), using the CHE-
MOFACE software (CHEMOFACE, version 1.64). In this multivariate
analysis, the data used were self-scaled and a matrix [m × n] was
constructed with the sensory scores of the “n” attributes evaluated for
the “m” samples evaluated. In addition, in order to improve the un-
derstanding of the effect of treatments on the sensory attributes studied,
the results of the sensory attributes obtained through sensory analysis
(Global Score, Sweetness, Acidity, Body, Astringency, Bitterness and
Finalization) were submitted to analysis of variance (ANOVA) and,
when significant differences in the F test were detected, the Dunnett
test was applied, at 5% significance, using the Statistical Analysis
System software (SAS®, version 9.4, USA).
As it is a frequency data, the terms describing the Aroma and Flavor
attributes were submitted to the analysis of generalized linear models,
considering the Bernoulli distribution and logistic link function. The
purpose of this analysis was to observe the probability of sensory per-
ception of certain sensory descriptors of Aroma and Flavor with the
increase in the concentration of the number of Quaker beans in the
Control coffee.
3
Food Chemistry 342 (2021) 128304
Finally, the hypergeometric distribution was used to determine the
probability of a cup (8.25 g) containing a certain number of Quaker
beans, considering different concentrations in the sample (100 g). The
different concentrations corresponded to the presence of 0 to 100
Quaker beans in a 100 g roasted coffee sample.
2.7.3. Volatile compounds
For the evaluation of volatile composition, an experiment was car-
ried out using a randomized block design (RBD) with four blocks, each
block corresponding to a roasting batch. In this experiment, each group
of Quaker beans (Color 1, Color 2 and Color 3) and Control coffee were
evaluated individually, totaling four treatments.
The relative percentage areas of volatile compounds found in the
treatments were subjected to analysis of variance (ANOVA) and, when
significant differences were identified in the F test, the Scott-Knott test
was applied, using the SISVAR software (SISVAR, version 5.3). For a
better understanding of the relationship between the treatments and
volatile composition, principal component analysis (PCA) was per-
formed, using the CHEMOFACE statistical software (CHEMOFACE,
version 1.64). In this multivariate analysis, the data used were self-
scaled and a matrix [m × n] was constructed with the relative areas of
the “n” chromatographic peaks identified for the “m” samples eval-
uated.
3. Results and discussion
3.1. Coloring reference for Quaker beans
The color of roasted beans is one of the parameters used to measure
the roasting degree of a coffee. The SCA Protocols – Cupping Specialty
Coffee standardized the roasting degree for the evaluation of a coffee
sample and adopted the Agtron System – SCA Roast Color Classification
System to measure the color of the beans (Lingle, 2011b).
The beans that presented yellowish tones that differ from the de-
fined pattern for roasting, in most cases, are considered Quaker beans
whereas, in the scientific literature, reference values have not yet been
described to consider a bean as Quaker. Therefore, in this research,
color references for Quaker beans were proposed based on the Agtron
Scale – SCA Roast Color Classification System and the CIEL*c*h°
System. Table 1 presents the color of the Control coffee and the pro-
posed colorations of the beans of groups Color 1, Color 2 and Color 3,
measured by the Agtron colorimetric system and the CIEL*c*h° uni-
versal colorimetric system. The result of the analysis of variance, also
presented in Table 1, showed that all colorimetric parameters used
indicated significant differences between treatments.
As can be seen in Table 1, the beans that make up Color 1 group
showed a color that was called Brownish Quaker, the beans that make
up Color 2 group showed a color that was called Brown Yellowish
Quaker and the beans that make up Color 3 group were named Yel-
lowish Quaker. In the Agtron Scale, the bean color varied between 61.2
and 95.35; the higher the Agtron value, the more yellow the bean color
Table 1
Evaluation of the color of roasted coffee beans with different shades by the
Agtron and CIE L* c* h° systems for treatment composition.
Colorimetric parameters Treatments
Control Color 1 Color 2 Color 3
Agtron* 53.7d
61.2c
82.8b
95.35a
CIEL*c*h° L** 21.3c 23.9b 28.1a 29.15a
c** 16.3d 18.6c 22.2b 26.36a
h°* 59,9c 61.2b 61.6b 63.83a
*significantly different by the F test. Mean values within the same line followed
by different lowercase superscript letters differ significantly (p < 0.05) by the
Scott-Knott’s test.
M.H.S. Rabelo, et al.
Fig. 1. Principal Component Analysis used to assess the sensory attributes as a function of each color group. Graphics: A – Color 1, B – Color 2, C – Color 3.
is and, the lower the Agtron value, the more brown the bean color is. In
the CIEL*c*h° system, regardless of the group, the beans showed a color
between yellow and red, with an angle around h° = 62°. Chromaticity
(c*) was more intense for Color 3 group. In addition, the beans of Color
3 group had a lighter luminosity (L*) than Color 1 group. Therefore,
luminosity (L*) and chromaticity (c*) were the coordinates that showed
the greatest variation. This variation was expected, since the coordinate
luminosity (L *) represents a gray scale, and chromaticity (c*) re-
presents color saturation.
3.2. Evaluation of the sensory attributes of specialty natural coffee
Through the results of the sensory analysis, the relationship be-
tween the increase in the concentration of Quaker beans in each group
with the sensory attributes of the beverage of specialty natural coffee
was evaluated using the principal component analysis (PCA), for each
group of beans (Fig. 1).
In Fig. 1, it is observed that the vectors represent the sensory at-
tributes and the points represent the concentrations of Quaker beans. In
this sense, for Color 1 group, it was found that there is a separation of
4
Food Chemistry 342 (2021) 128304
sensory attributes by the first principal coordinate. Bitterness, as-
tringency and body are located on the positive part of PC1, where the
concentrations of 15, 20 and 30 beans are also found. However, overall
score and aftertaste, sweetness and acidity are located on the negative
side of PC1, along with concentrations of 0, 5, 10 and 25 beans. It was
expected that there would be a clear separation between the highest
and lowest bean concentrations; however, it did not occur, since the
concentration of 25 beans was grouped with the Control treatment and
the lowest concentrations of beans. Thus, an analysis of variance
(Supplementary Material – Table 1) was performed in order to verify
whether there is a significant difference between treatments. It was
observed that the presence of beans from Color 1 group at different
concentrations in specialty natural coffee did not significantly alter the
final score and none of the sensory attributes evaluated. Therefore,
beans in Color 1 group, with Agtron 61.2 coloring, are not to be con-
sidered Quaker beans.
For Color 2 group (Fig. 1B), the first pricipal component (PC1) ac-
counted for most data variability. Bitterness and astringency, which are
unwanted sensory attributes in the coffee beverage, are located in the
same quadrant of concentrations 7, 10, 13 and 16 beans. In turn,
M.H.S. Rabelo, et al.
Fig. 2. Curves of perception probability for aroma and flavor for each category of Quaker beans. Graphics: A – Aroma/Color 1, B – Aroma/Color 2, C – Aroma/Color
3, D – Flavor/Color 1, E – Flavor/Color 2, F – Flavor/Color 3.
concentrations 0, 1 and 4 are in the opposite quadrant, together with
the final score and the sensory attributes acidity, sweetness and com-
pletion. From this result, it is possible to affirm that, from 7 beans in a
cup, the beverage of specialty natural coffee is changed, causing bit-
terness and astringency. The results of the variance analysis of Color 2
bean group corroborated the results presented in the PCA and showed
that the presence of 16 beans in a cup caused significant changes in
bitterness and astringency. In addition, the natural coffee beverage is
no longer considered specialty at a concentration of 7 beans in Color 2
group; from this concentration of beans, the overall score differed sta-
tistically from the Control and showed a value below 80 points.
Color 3 group (Fig. 1C) showed similar results to Color 2 group.
Bitterness and astringency were located in a quadrant opposite to that
of the final score and the attributes sweetness, acidity, body and finish,
5
Food Chemistry 342 (2021) 128304
being separated by the first principal component, which accounted for
90.46% of data variability. Regarding the amount of beans present in
the beverage of specialty natural coffee, the concentrations 13 and 16
beans were located in the same quadrant of bitterness and astringency
and opposite to the others. The second principal component (PC2) ac-
counted for only 5.34% of data variability in Color 3 bean group, but it
made an important contribution, in which the concentrations located in
the negative quadrant of PC1 were separated by PC2 in three different
regions. The first region consists of concentrations 1 and 4 beans; this
region is completely opposed to astringency and bitterness. The second
region comprises the treatment corresponding to the Control bean
group, which is aligned with the overall score, finish and sweetness,
with sweetness being the most striking characteristic of this coffee. In
the third region, there are concentrations of 7 and 10 beans which,
M.H.S. Rabelo, et al.
despite being very close to the treatment corresponding to the Control
bean group, suffer great influence from bitterness. When verifying if
there are significant differences between bean concentrations for Color
3 group through analysis of variance, it was observed that the presence
of beans in this group significantly affected the final score and the at-
tributes sweetness, acidity, astringency and finish. The beverage of
specialty natural coffee presented less sweetness in relation to the
treatment corresponding to the group of Control beans from the pre-
sence of 13 beans. The attributes acidity, astringency and finish differed
from the Control from 16 beans. The final score of the beverage differed
from the Control treatment, based on the concentration of 7 beans and
is no longer considered specialty.
In this research, it could be observed that the presence of Quaker
beans in all experiments affected two attributes of great importance for
the quality description of a specialty coffee. The first was sweetness
which, according to the International Coffee Organization Organizacion
Internacional Del Café (1991), is one of the most desirable flavor
characteristics in the beverage. Furthermore, compounds related to the
perception of sweetness participate in important chemical reactions,
which occur during roasting, such as the Maillard reaction and/or
caramelization, originating compounds responsible for the formation of
beverage color, flavor and aroma (Borém, Ribeiro, Pereira, Da Rosa, &
Morais, 2006). The second attribute affected was acidity, an important
organoleptic parameter in coffee, which may be pleasant or not, de-
pending on the nature of the predominant acid. A pleasant acidity
contributes to the liveliness of the coffee, increases the perception of
sweetness and gives it a characteristic of fresh fruit. However, excessive
acidity can be unpleasant and indicate an unusual characteristic of a
coffee (Illy & Viani, 2005; Lingle, 2011b; Malta, 2011; America, 2009).
Based on the observed results, it is possible to affirm that, from the
presence of 7 beans for both Color 2 and Color 3 groups, the sensory
characteristics of the beverage of a specialty natural coffee are nega-
tively affected, causing greater bitterness and astringency and affecting
the overall score, making the coffee no longer classified as a specialty
coffee.
3.3. Analysis of aroma and flavor of specialty natural coffee
Aroma is the first attribute to be perceived when drinking coffee;
the second attribute is flavor, which is closely linked to aroma. When
they are positive, aroma and flavor contribute to the complexity of the
beverage of a specialty coffee. However, defective beans can negatively
affect the aroma and flavor of the beverage (Toci & Farah, 2014).
Through the results of the sensory analysis, it was determined that the
presence of beans from groups Color 1, Color 2 and Color 3 in the
specialty natural coffee affected the aroma and flavor of the beverage,
with its predominant sensory perception being described as “green/
immature”. The aroma and flavor of the beverage of specialty natural
coffee free from Quaker beans, that is, the treatment of the Control bean
group, have been described as “chocolate” and “caramel”, which are
positive sensory descriptors for a specialty coffee.
Fig. 2 shows regression models that were developed to analyze the
probability of sensory perception of aroma and flavor as a function of
the increase in the concentration of beans in each group in specialty
natural coffee. The adjusted regression equations, which define the
probability of perception of the descriptors “Green/Immature”, “Cho-
colate” and “Caramel” for Aroma and Flavor are included in the
Supplementary Material, due to the presence of Quaker beans in each
group.
In Fig. 2, it can be seen that the presence of beans from Color 1
group in specialty natural coffee provided the sensory perception of the
“chocolate” and “green/immature” aromas. At lower concentrations, a
“chocolate” aroma was predominant and, at higher concentrations, the
perception of “green/immature” notes, predominated. However, the
probability of perceiving the “green/immature” aroma in none of the
studied concentrations is greater than the probability of perceiving the
6
Food Chemistry 342 (2021) 128304
“chocolate” aroma. In other words, although “green/immature” notes
were present, they did not harm the beverage of specialty natural
coffee, as “chocolate” notes were prevalent at all concentrations stu-
died. Regarding flavor, when evaluating the influence of Color 1 group
on specialty natural coffee, the beans belonging to this group provided
a slight “green/immature” flavor. The probability of detecting this de-
scriptor at the maximum concentration studied was 0.32 (Fig. 2D). This
result was very similar to that found for aroma, which confirms the low
sensory perception and influence of this group of beans in the beverage
of a specialty natural coffee.
In Color 2 bean group, the sensory notes of “chocolate”, “caramel”
and “green/immature” were detected in the aroma and flavor of spe-
cialty natural coffee, shown in Fig. 2B and E, respectively. At low
concentrations of beans from Color 2 group in specialty natural coffee,
the sensory perception of “chocolate” and “caramel” was predominant
for both aroma and flavor. With the increase in the concentration of
beans in this group, the sensory perception of “green/immature” be-
came more evident. From the concentration of 14 beans, the aroma of
“green/immature” became more evident and overcame the probability
of perceiving the aromas of “chocolate” and “caramel”. The same oc-
curred with flavor, but from the presence of 8 beans in Color 2 group.
For flavor and aroma of Color 3 group, shown respectively in Fig. 2C
and F, the inclusion of beans from this group in specialty natural coffee
resulted in the perception of “green/immature” notes. Notes of “cho-
colate” and “caramel” were also perceived for aroma and, for flavor,
only notes of “chocolate”. Notes of “green/immature” were perceived
more than notes of “chocolate” and “caramel” in aroma with the pre-
sence of a smaller number of beans than in the other groups, from the
presence of 8 beans in Color 3 group. For flavor, from 7 beans, the notes
of “green/immature” were more perceived than the notes of “choco-
late”.
Therefore, it was observed that, only from the presence of 7 Quaker
beans, with a color equal to or greater than Agtron 82.2, in a cup of
specialty natural coffee, there were changes in the sensory attributes
capable of being perceived by a highly trained sensory panel. Thus, the
threshold of sensory perception for the presence of Quaker beans, with
a color equal to or greater than Agtron 82.2 in specialty natural coffees,
is 7 beans in a cup.
3.4. Hypergeometric distribution
The beans from Color 2 and Color 3 groups present in a specialty
natural coffee caused noticeable sensory differences in the beverage
only from the concentration of 7 beans, that is, up to 4 Quaker beans
per cup, it was not enough to change the sensory characteristics of the
natural coffee beverage classified as specialty. This result represents a
change in tolerance in relation to the presence of Quaker beans, unlike
the criterion that was established by the Specialty Coffee Association
(Lingle, 2011a) for batches of specialty coffees, in which a sample of
100 g of roasted coffee cannot contain Quaker beans.
Thus, considering that a tasting cup contains 8.25 g of roasted
coffee, there was a need to convert the tolerance of 4 Quaker beans per
cup to a sample of 100 g of roasted coffee. This conversion was not
carried out directly, as it must be taken into account that, when making
a random 8.25 g roasted coffee intake, more than 4 Quaker beans may
be present. Therefore, it is necessary to consider the probability that
more than 4 Quaker beans are present in a cup of coffee (8.25 g) to
establish a maximum number of Quaker beans for a 100 g sample of
roasted coffee.
Consequently, assuming a maximum tolerance of four Quaker beans
present in a cup, the hypergeometric distribution was applied in order
to define the probability of a cup containing more than four Quaker
beans in a random 8.25 g shot, considering 100 g samples containing
quantities of Quaker beans ranging from 0 to 100 beans (Fig. 3). Ac-
cording to the hypergeometric distribution shown in Fig. 3, it can be
observed that, if a 10% probability of a cup containing more than 4
M.H.S. Rabelo, et al.
Fig. 3. Probability of a cup containing more than four Quaker beans, depending on the number of Quaker beans present in a sample (100 g), through Hypergeometric
Distribution.
Quaker beans is considered, the 100 g sample can contain a maximum
of 20 Quaker beans. Similarly, considering a 1% probability that a cup
contains more than 4 Quaker beans, it can contain up to 12 Quaker
beans in 100 g of roasted coffee.
Therefore, the results showed that the presence of more than four
Quaker beans in a single cup is capable of altering the sensory char-
acteristics of the beverage of a specialty natural coffee and the prob-
ability of more than four beans in a cup depends on the initial con-
centration of Quaker beans in the 100 g sample. Thus, these results may
support the Specialty Coffee Association to establish new criteria for the
marketing of specialty coffee batches, with regard to the tolerance of
the presence of Quaker beans.
3.5. Volatile composition
The sugars, amino acids and proteins present in raw coffee beans are
responsible for most of the beverage characteristic flavor. During the
coffee roasting process, through the Maillard reaction, caramelization
and pyrolysis, these sugars, amino acids and proteins give rise to nu-
merous volatile compounds, such as furans, pyrazines, ketones, pyr-
roles, pyridines, alcohols and aldehydes.
The correlation between volatile compounds and bean groups was
carried out through Principal Component Analysis, shown in Fig. 4. The
first principal component (PC1) was responsible for 91.71% of data
variability and the second pricipal component (PC2) accounted for for
6.95%. In Fig. 4, it is possible to observe that the bean groups Control
and Color 1 were grouped in the negative part of the first principal
component (PC1) and showed greater chemical affinity with the com-
pounds of the classes; furan, ketone, aldehyde, pyrrole, phenol, acid,
amide and amine. The groups Color 2 and Color 3 were grouped in the
positive part of the first principal component (PC1) and had greater
affinity with pyrazine, aldehyde, alcohol, pyrrole, pyridine and thia-
zole. In the evaluation of the volatile composition of beans from groups
Color 1, Color 2, Color 3 and Control, thirty-six volatile compounds
were identified (Table 2). The main classes of compounds, in order of
abundance, were: furans (10 compounds), pyrazines (8), ketones (5),
aldehydes (3), alcohols (2), pyrroles (2), phenols (1), pyridines (1),
acids (1), thiazole (1), amide (1) and amine (1), in accordance with the
literature (Flament, 2002). Among all the volatile compounds shown in
Table 2, 27.8% belong to the Furan group and 22.2% to the Pyrazine
7
Food Chemistry 342 (2021) 128304
group. According to several researchers (Akiyama et al., 2005; Maeztu
et al., 2001; Sanz, Czerny, Cid, & Schieberle, 2002; Toci & Farah, 2008;
Yeretzian, Jordan, & Lindinger, 2003), not only are Furans and Pyr-
azines the main and most abundant compounds, but also the main
classes of volatile compounds that contribute to the characteristic
aroma of roasted coffee.
Furans are an important class of volatile compounds in roasted
coffee, both in terms of number of compounds and contribution of
aroma to the beverage. More than 140 furan compounds have already
been identified in coffee. This class, in general, was very important to
confirm the existence of differences between treatments. The largest
relative area of these compounds was observed in the beans of the
Control group, followed by Color 1, Color 2 and Color 3 groups. The
result obtained from the analysis of variance showed that all com-
pounds of the Furan group indicated a significant difference between
treatments (Table 2). For some compounds, such as furfuryl alcohol, the
difference between the Control bean group and Color 1 group was not
identified, confirming the result presented in the Principal Component
Analysis.
Toci and Farah (2014) studied defective beans called PVA (black,
green and burnt) and observed that, in general, defective roasted beans
contained lower concentrations of total furans compared to healthy
mature beans (control). This fact is probably a reflection of the initial
composition, that is, of the precursors present in raw beans. According
to Trugo (2004), furans are mainly derived from carbohydrate de-
gradation (Amadori rearrangement). Flament (2002) identified three
volatile compounds, belonging to the furan group, in the thermal de-
gradation products of carbohydrates and concluded that the volatile
compound furfural has sweet notes, aroma of caramel, cinnamon and
almond, and the compounds 2-methylfuran and furfuryl alcohol have a
fruity and caramel aroma, respectively. In another study, Toci and
Farah (2010) investigated the initial composition of PVA and control
beans, and it was observed that raw control beans had a higher content
of sucrose, compared to defective beans, with the lowest levels being
observed in black beans, followed by burnt and, lastly, immature. Im-
mature beans are chemically very similar to ripe ones, once they ori-
ginate from immature fruits, differing only by their degree of ripeness.
Regarding the Pyrazine group, which represented the second largest
group of volatile compounds identified in this study (Table 2), not all
compounds effectively contributed to the separation of treatments. Toci
M.H.S. Rabelo, et al.
and Farah (2014) observed higher levels of heterocyclic compounds,
such as pyrazine, in defective roasted beans, especially in immature
beans. Baltes and Bochmann (1987), in their model study, observed that
the relationship between the content of amino acids and sugars in a
reaction system determines the concentration of pyrazines during
roasting. A greater amount of amino acids in relation to the sugar
content provided an increase in the formation of heterocyclic com-
pounds such as pyrazine. Vasconcelos, Franca, Glória, and Mendonça
(2007) observed higher concentrations of free amino acids in immature
beans compared to mature beans. Regarding the carbohydrate content,
Mazzafera (1999) observed a lower sucrose content in immature beans,
in comparison with mature beans, corroborating the study by Toci and
Farah (2010).
As shown in Table 2, methylpyrazine was the compound of the
Pyrazine group that had the peak with the largest relative area, be-
coming important in separating the treatments Color 2 and Color 3. The
formation of this compound can occur by different routes, for example,
in the reaction of asparagine with sugars, such as glucose, fructose,
sucrose and arabinose (Odell, 1974) cited by Flament, 2002). It can also
be formed by condensation (involving Strecker degradation of an amino
acid) of pyruvaldehyde with glyoxal (Wang, Kato, & Fujimaki, 1969)
cited by Flament, 2002). The flavor is characterized by notes of nut,
roasted, green and bitter almond (Flament, 2002).
In Table 2, it was possible to observe that pyridine was the com-
pound that presented the peak with the largest relative area among all
compounds and significantly contributed to the separation of treat-
ments. This compound belongs to the class of Pyridines which, in
general, provides burnt, roasted and green notes, in addition to as-
tringency and bitterness (Maga, 1981). This volatile compound, found
mainly in roasted coffees, represents 25% of the pyrolysis products of a
trigonelinamonohydrate sample. Pyridine can also be formed from
other precursors, being found in model reactions between glucose and
8
Food Chemistry 342 (2021) 128304
Fig. 4. Principal Component Analysis of
Volatile compounds present in the beans of
groups Color 1, Color 2, Color 3 and Control.
Legend: 1 - Furfural, 2 - Furfuryl formate, 3 -
2-Methylfuran, 4 - 2-Vinylfuran, 5 - 2-Furfuryl
alcohol, 6 - 2-(Methoxymethyl)-furan, 7 - 2,5-
Dimethylfuran, 8 - Furfurylacetate, 9 - 5-
Methylfurfural, 10 - 2-Acetylfuran, 11 -
Pyrazine, 12 - Methylpyrazine, 13 - 2,5-
Dimethylpyrazine, 14 - 2,6-Dimethylpyrazine,
15 - Ethylpyrazine, 16 - 2,3-
Dimethylpyrazine, 17 - 2-Ethyl-6-methylpyr-
azine, 18 - 2-Ethyl-3-methylpyrazine, 19 - 5-
Ethyl-4-methyl-3-heptanone, 20 - 2,3-
Pentanedione, 21 - 3,4-Hexanedione, 22 - 4-
Hydroxy-3-hexanone, 23 - 2-Hydroxy-3-pen-
tanone, 24 - Butanal, 25 - 2-Methylbutanal, 26
- 3-Methylbutanal, 27 - 1-Pentanol, 28 - 3-
Methyl-3-buten-1-ol, 29 - 1-Methyl-1H-pyr-
role-2-carboxaldehyde, 30 - 1-Methyl-1H-
pyrrole, 31 - 3-Methyl-Phenol, 32 - Pyridine,
33 - 4-Hydroxy-butanoic acid, 34 - 4-
Methylthiazole, 35 - Methyl-urea, 36 - 5-
Nonylamine.
amino acids (Flament, 2002). Pyridine has a pungent, penetrating and
diffusive odor, also described as nauseating but, with various dilutions,
it becomes hot, burnt and roasted (Flament, 2002).
2-Methylbutanal, another compound that influenced the char-
acterization of treatments, is a product of isoleucine pyrolysis, but it can
also be generated by the Maillard reaction. This volatile compound
provides the malt flavor present in coffee (Flament, 2002). Guyot,
Petnga, and Vincent (1988) observed a decrease in the global content of
free amino acids with an increase in the maturity of coffee fruits. In
fruits classified as immature, the content of serine, aminobutyric acid,
valine, leucine, isoleucine and methionine was clearly more relevant
than in samples of ripe parchment fruits. Thus, this study helps justify
the high content of 2-methylbutanal present in Quaker beans.
Mature beans had a higher sucrose content than those immature,
which probably causes a greater formation of ketones during the
roasting process. Ketones are the result of carbohydrate pyrolysis,
which produces more complex ketone compounds, its main formation
route (Flament, 2002; Trugo, 2004). This fact corroborates the result
shown in Table 2, with the beans of the Control group having a higher
relative percentage.
4. Conclusion
With the objective of verifying which quantity of Quaker beans is
able to negatively influence the beverage of a specialty natural coffee, it
was possible to conclude that, for the beans of groups Color 2 and Color
3, there was a difference in sensory perception only from the con-
centration of 7 Quaker beans in a cup (65 beans). That is, only with the
presence of 10.8% of Quaker beans in a tasting cup, the beverage of a
specialty natural coffee is negatively altered. Quaker beans caused
greater astringency and bitterness to the beverage of specialty natural
coffee, causing natural coffee to stop being classified as a specialty
M.H.S. Rabelo, et al. Food Chemistry 342 (2021) 128304

Table 2 Quaker bean are compounds formed during the roasting process from
Relative percentage area (%) of volatile compounds extracted from each precursors present in raw beans from immature harvested fruits.
treatment. Finally, based on the resultd of this research, the Specialty Coffee
Volatile compounds Treatment (%) Association – SCA criterion as to the presence of Quaker beans can be
revised to a maximum tolerance of 12 Quaker beans in 100 g of natural
Control Color 1 Color 2 Color 3 specialty coffee, considering 1% probability of sensory perception.
Furans
1 Furfural* 6.486d 5.385c 3.233b 1.776a CRediT authorship contribution statement
2 Furfuryl formate* 0.420d 0.352c 0.187b 0.078ª
3 2-Methylfuran* 11.368d 10.253c 6.903b 4.513ª Mariane Helena Sances Rabelo: Conceptualization, Methodology,
4 2-Vinylfuran* 0.269b 0.268b 0.186ª 0.142ª
Formal analysis, Investigation, Data curation, Writing - original draft,
5 Furfuryl alcohol* 20.356c 19.743c 16.049b 12.494ª
6 2-(Methoxymethyl)-furan* 0.190c 0.179c 0.127b 0.092ª Writing - review & editing, Visualization, Project administration. Flávio
7 2.5-Dimethylfuran* 0.703d 0.616c 0.356b 0.221ª Meira Borém: Conceptualization, Methodology, Investigation,
8 Furfuryl acetate* 1.867c 1.829c 0.880b 0.442ª Resources, Writing - original draft, Writing - review & editing,
9 5-Methylfurfural* 3.011d 2.307c 1.147b 0.624ª Supervision, Project administration. Renato Ribeiro de Lima:
10 2-Acetylfuran* 0.633d 0.479c 0.235b 0.133ª
Conceptualization, Methodology, Formal analysis, Data curation. Ana
Pyrazines Paula de Carvalho Alves: Methodology, Formal analysis,
11 Pyrazine* 1.933a 2.335b 3.454c 3.904d
Investigation, Data curation, Writing - original draft, Writing - review &
12 Methylpyrazine* 9.321ª 11.282b 13.367c 14.514d
13 2.5-Dimethylpyrazine* 0.671ª 0.855b 1.040c 1.067c editing. Ana Carla Marque Pinheiro: Conceptualization,
14 2.6-Dimethylpyrazine* 0.940ª 1.185b 1.371c 1.286c Methodology. Diego Egídio Ribeiro: Methodology, Investigation.
15 Ethylpyrazine* 0.878ª 1.087ª 1.524b 1.725b Claudia Mendes Santos: Methodology, Investigation, Data curation,
16 2.3-Dimethylpyrazine* 0.231ª 0.294b 0.393c 0.545d Writing - original draft, Writing - review & editing. Rosemary
17 2-Ethyl-6-methylpyrazine* 0.180ª 0.263b 0.413c 0.485d
18 2-Ethyl-3-methylpyrazine* 0.154ª 0.277b 0.510c 0.602d
Gualberto Fonseca Alvarenga Pereira: Conceptualization,
Methodology, Resources, Project administration.
Ketones
19 5-Ethyl-4-methyl-3-heptanone* 0.401b 0.415b 0.374b 0.287ª
20 2.3-Pentanedione* 3.577d 2.581c 1.955b 1.160ª Declaration of Competing Interest
21 3.4-Hexanedione* 0.200d 0.156c 0.072b 0.046ª
22 4-Hydroxy-3-hexanone* 0.447d 0.387c 0.213b 0.095ª
The authors declare that they have no known competing financial
23 2-Hydroxy-3-pentanone* 0.212d 0.162c 0.087b 0.031ª
interests or personal relationships that could have appeared to influ-
Aldehydes
ence the work reported in this paper.
24 Butanal* 1.171c 1.379d 1.014b 0.840ª
25 2-Methylbutanal* 8.030ª 8.088ª 14.369b 18.473c
26 3-Methylbutanal* 2.091d 1.851c 1.688b 1.288ª Acknowledgments
Alcohols
27 1-Pentanol* 0.085ª 0.108ª 0.311b 0.422c The authors thank the National Council of Scientific and
28 3-Methyl-3-Buten-1-ol* 0.090ª 0.090ª 0.185b 0.226c Technological Development (CNPq), the Foundation for Research
Pyrroles Support of the State of Minas Gerais (FAPEMIG), the Coordination for
29 1-Methyl-1H-Pyrrole-2- 0.271 0.211 0.203 0.231 the Improvement of Higher Education Personnel (CAPES), the National
Carboxaldehyde Science and Technology Institute of Coffee (INCT Café). The authors
a b c
30 1-Methyl-1H-Pyrrole* 1.402 1.597 2.436 3.141d
would like, in particular, to thank CARMOCOFFEES for donating the
Phenol coffees used in experiment. Finally, to thank to the large team of stu-
31 3-Methylphenol* 0.365c 0.372c 0.172b 0.077a dents from LPPA and Polo de Excelencia do Café/UFLA who voluntarily
Pyridine helped in the data collection.
32 Pyridine* 17.551a 19.548b 22.571c 27.141d

Acid Appendix A. Supplementary data

33 4-Hydroxy-Butanoic-acid 0.274 0.318 0.278 0.260

Thiazole Supplementary data to this article can be found online at https://

34 4-Methylthiazole* 0.052a 0.063a 0.089b 0.126c doi.org/10.1016/j.foodchem.2020.128304.
Amida
35 Methyl-urea* 1.694d 1.502c 1.108b 0.693a References
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