LWT - Food Science and Technology 103 (2019) 139–146

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LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Phenolic, volatile, and sensory profiles of beer enriched by macerating T

quince fruits
Pedro J. Zapataa, Alejandra Martínez-Espláa, Amadeo Gironés-Vilaplanaa, David Santos-Laxa,
Luis Noguera-Artiagab,∗, Ángel A. Carbonell-Barrachinab
a
Universidad Miguel Hernández de Elche (UMH), Escuela Politécnica Superior de Orihuela (EPSO), Departamento de Tecnología Agroalimentaria, Grupo Post-recolección
de Frutas y Hortalizas, Carretera de Beniel, km 3,2, 03312, Orihuela, Alicante, Spain
b
Universidad Miguel Hernández de Elche (UMH), Escuela Politécnica Superior de Orihuela (EPSO), Departamento de Tecnología Agroalimentaria, Grupo Calidad y
Seguridad Alimentaria (CSA), Spain

A R T I C LE I N FO A B S T R A C T

Keywords: Quince fruit (Cydonia oblonga Miller) is not very appreciated for their sensory properties for fresh consumption
Antioxidant activity but it is a fruit rich in functional compounds. The aim of this study was to take advantage of this fruit to improve
Chlorogenic acid the functional profile of one of the most popular alcoholic beverages around the world, beer. For this purpose,
Cydonia oblonga miller one quince cultivar, Vranja, and two clones ZM9 and PUM were used to brew beer and study its physico-
Descriptive sensory analysis
chemical, aromatic (headspace solid phase micro-extraction, HS-SPME), functional (phenolic compounds and
HS-SPME
antioxidant capacity) and sensory (descriptive analysis with trained panel) properties. The addition of quince
increased the total polyphenol content, the total hydroxicinnamic acids, concentration of main volatile com-
pounds related with fruity sensory descriptors, and led to higher intensities of floral and fruity sensory attributes.
Therefore, the quince beer had a better functional composition and better sensory characteristics than the
control beer.

1. Introduction appreciated for fresh consumption, but it can be consumed cooked or

processed as jam, jelly or cakes (Szychowski, Munera-Picazo, Szumny,
Beer is one of the most consumed alcoholic beverages around the Carbonell-Barrachina, & Hernández, 2014). Quince has low fat content
world, being rich in nutrients, such as carbohydrates, amino acids, and it is an important source of organic acids, sugars, crude fiber and
minerals, vitamins and phytochemicals, such as phenolic compounds. minerals, such as potassium, phosphorous and calcium (Rodríguez-
According to the Beer Purity Law from 1516, traditional German beer is Guisado et al., 2009). Several studies have showed that quince is a good
made only from barley, hop, and water, with the addition of yeast and low-cost natural source of phenolic acids and flavonoids (Legua
(Gerhäuser, 2005). et al., 2013; Wojdyło, Oszmiański, & Bielicki, 2013).
Many researches provide strong evidence for a cardio-protective Polyphenols are naturally occurring compounds found in fruits and
effect of moderate alcohol (such as beer or wine) consumption (up to 2 vegetables, and the products manufactured from these fruits and ve-
glasses a day in case of men and 1 glass a day in case of women) as well getables still contain polyphenols in significant amounts. Polyphenols
as control of lipid (cholesterol) metabolism, blood clotting and glucose are secondary metabolites of plants and may contribute to their bit-
metabolism (Giacosa et al., 2016). The relationship between alcohol terness, astringency, color, flavor, odor and oxidative stability (Pandey
consumption and cardiovascular events or all-cause mortality is de- & Rizvi, 2009). In general, antioxidants (e.g. polyphenols) reduce oxi-
scribed as a J-shaped curve, whereby maximum benefit is achieved at a dative damage to biomolecules and cells, and have been implicated in
moderate consumption level (Roerecke & Rehm, 2012; Sierksma & Kok, the pathogenesis of several human diseases and certain types of cancer
2012). These benefits are additive to the beneficial effects of other (Valko, Rhodes, Moncol, Izakovic, & Mazur, 2006).
healthy lifestyle factors such as non-smoking, physical activity and a The aim of this study was to take advantage of a fruit rich in
healthy diet (Mukamal, Chiuve, & Rimm, 2006). bioactive compounds (quince), to improve the sensory and functional
Quince fruit (Cydonia oblonga Miller) is too acid, astringent, and due profiles of a beer, to thereby, help in improving the health benefits of
to pulp hardness, bitterness, woodiness, and astringency is not very moderate consumption of this type of product.

∗
Corresponding author.
E-mail address: [email protected] (L. Noguera-Artiaga).

https://doi.org/10.1016/j.lwt.2019.01.002
Received 26 September 2018; Received in revised form 29 December 2018; Accepted 2 January 2019
Available online 04 January 2019
0023-6438/ © 2019 Elsevier Ltd. All rights reserved.
P.J. Zapata et al. LWT - Food Science and Technology 103 (2019) 139–146

2. Material and methods Table 1

Quality parameters of control and new quince-beers as affected by quince
2.1. Quince fruits variety.
Parameter ANOVA† Control Vranja PUM ZM9 Pooled
In this research were used one 3 quince cultivar/clones: (i) 1 cul- Variance
tivar, Vranja, and, (ii) 2 clones, ZM9 and PUM. The reasons for selecting
pH NS 4.38 4.33 4.28 4.33 0.21
them were: (i) ZM9 clone is suitable for fresh consumption due to their
Total soluble * 5.80 a‡ 4.69 b 5.72 a 5.82 a 0.16
sensory attributes, (ii) PUM clone has high antioxidant content (suitable solids (ºBrix)
for the development of functional foods), and (iii) Vranja cultivar has Acidity NS 0.36 0.38 0.39 0.39 0.04
intermediate values of all parameters studied quality (Szychowski et al., Alcoholic grade * 5.27 b 5.62 a 5.52 a 5.51 a 0.10
(%)
2014).
Ibus bitter ** 10.96 a 8.70 b 9.52 a 8.77 b 0.06
The selected plant materials belong to the quince germplasm bank EBC (Color) * 38.50 b 37.38 b 41.38 a 38.75 b 0.87
of Universidad Miguel Hernández de Elche (Orihuela, Alicante, Spain). CIE L* * 31.64 a 30.21 b 31.52 a 31.39 a 0.33
The experiment was established according to Szychowski et al. (2014). CIE a* NS 0.68 0.52 0.60 0.56 0.08
Fruits were harvested at commercial ripening stage at first of October CIE b* NS −1.40 −1.55 −1.53 −1.48 0.09

2016, and 15 homogeneous fruits (based on color, size and absence of †

NS = not significant at p < 0.05; *, and ** significant at p < 0.05 and 0.01,
defects) were selected from each variety and used for the experiment.
respectively; ‡Values (mean of 3 replications) followed by the same letter,
within the same row, were not significantly different (p < 0.05), according to
2.2. Beer manufacturing the Tukey's multiple range test.

Prior to this study, a standard American amber ale beer was ela- (2017). The equipment consisted of a Luna C18 column (250 × 40 mm,
borated, quince was added at different weight percentages (0, 5, 10 and 5 μm particle size; Teknokroma, Barcelona, Spain) in an Agilent HPLC
15%) and it was allowed to macerate during 0, 2, 4, 6, and 8 days. The 1200 Infinity series equipped with a photodiode array detector (Agilent
main objective of this previous experience was to find the best combi- Technologies, Waldbronn, Germany) and a mass detector in series
nation (percentage of weight versus days of maceration) providing (Bruker Daltonics Ultra HCT-ESI Ion Trap, Bremen, Germany). Re-
proper beer flavor, but getting the plus of getting quince key notes and garding HPLC system, water/formic acid (99:1, v/v) and acetonitrile
no off-flavors. For this purpose, the trained descriptive sensory panel of were used as the mobile phases A and B, respectively, with a flow rate
the Agro-Food Technology Department of the Universidad Miguel of 0.8 mL/min. The linear gradient started with 1% solvent B, reaching
Hernández de Elche (UMH) was used. 15% solvent B at 15 min, 30% at 30 min, 40% at 50 min, 95% at 45 min,
After this previous analysis, the panel decided that the addition of a maintained to 50 min, and returning to 1% until 60 min. The injection
10% of quince fruit to the beer and its maceration for 10 days was the volume was 8 mL. Chromatograms were recorded at 360 and 320 nm.
best combination for the study, based on the beer, fruity, and quince-ID For capillary temperature and voltage, the ionization conditions were
flavors. 350 °C and 4 kV, respectively. The nitrogen flow rate and nebulizer
After defining the weight of fruit and maceration to be used, an pressure were 11 L/min and 65.0 psi, respectively. Collision induced
American amber ale type beer was elaborated. This beer is similar to the fragmentation experiments were performed in the ion trap using helium
typical pale ale, but with a more malty and caramelized character. This as the collision gas, with voltage ramping cycles from 0.3 up to 2.0 V.
style gives more prominence to the taste aroma and color for the special Later, an HPLC-DAD system was used for the quantification of the
malts to the detriment of the hops aroma more characteristic of the compounds with the same conditions that were used for phenolics
American styles; the color could be described as amber or even some- identification, following the chromatographic conditions described by
thing coppery. As for the bitterness, in this beer, the addition of hops Gironés-Vilaplana et al. (2013).
was reduced in the first stages of preparation to control the bitterness Individual phenolic compounds were identified by their mass in a
below levels masking other sensory attributes. Therefore, the beer had HPLC-DAD-ESI/MSn, their spectra and retention time, using analytical
only 10.9 IBUS and did not reach the range of amber ale. The color of standards (3-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, 1,3-di-
the beer was within the typical range of this style, 23.6 EBC, and also its caffeoylquinic acid and 3,5-dicaffeoylquinic acid) and using previous
alcoholic graduation, 4.7% ABV. When the beer was finished, the bibliography.
quinces (peeled and cut, were added to the beer (uniform pieces of
2 × 5 cm), and the whole mixture was left to macerate for 10 days at a
2.5. Antioxidant capacity
controlled temperature of 4 °C. After that time, the samples were bot-
tled (beer bottles of 333 mL) and were stored under refrigeration (4 °C)
The free radical scavenging activity was determined using the
for 2 weeks, until the start of the experimental analysis.
ABTS+ assay adapted to a microscale, according to Mena et al. (2011).
The antioxidant activity was evaluated by measuring the variation in
2.3. Physico-chemical parameters of beer
absorbance at 414 nm after 40 min. The assays were performed using
96-well micro plates (Nunc, Roskilde, Denmark) and an Infinite M200
The physico-chemical parameters of beer analyzed in this study
micro plate reader (Tecan, Grödig, Austria).
were pH, total soluble solids, acidity, alcoholic grade, bitter (IBUS),
color EBC (UV-2401 PC spectrophotometer, Shimadzu, Kyoto, Japan),
color CIEL*a*b* (Minolta Colorimeter CR-300, Osaka, Japan), and were 2.6. Total phenolic compounds (TPC) by the Folin-Ciocalteu's reagent
conducted using the official methods of the Analytical Division of
European Brewery Convention (European Brewery Convention, 1987). The Folin-Ciocalteu's reagent method was adapted to a microscale
assay according to Mena, Martí, Saura, Valero, and García-Viguera
2.4. Identification of phenolic compounds by HPLC-DAD-ESI/MSn and (2013). Results were expressed as mg pyrogallol per 100 g of dried
quantification by RP-HPLC-DAD weigh (dw) in quince fruits, and in mg pyrogallol per 330 mL in beers.

Phenolic compounds were identified using the method and chro- 2.7. Volatile composition
matographic conditions previously described by Gironés-Vilaplana,
Villaño, Moreno, and García-Viguera (2013) and Martínez-Esplá et al. The method selected to study the volatile composition of beer

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Table 2
Phenolic acids identified in the three varieties of quince fruits by HPLC-DAD-ESI/MSn.
Retention time (min) Identity HPLC-DAD [M-H]-(m/z) HPLC-ESI(−) MSn (m/z)
λ (nm) (% base peak)

16.7 3-O-caffeoylquinic acid (neochlorogenic acid) 240, 304, 323 353 MS2[353]: 191 (100), 179 (44), 135 (11)
MS3[353 → 191]: 127 (100), 173 (88)
17.9 3-O-coumaroylquinic acid 246, 307, 327 337 MS2[337]: 163 (100)
20.8 5-O-caffeoylquinic acid (chlorogenic acid) 240, 305, 322 353 MS2[353]: 191 (100), 179 (2)
MS3[353 → 191]: 127 (100), 173 (79)
24.4 5-O-coumaroylquinic acid 245, 305, 326 337 MS2[337]: 191 (100)
31.5 3,5-di-O-caffeoylquinic acid 244, 304, 325 515 MS2[515]: 353 (100), 191 (16)
MS3[515 → 353]: 191 (100), 179 (15)

samples was headspace solid phase micro-extraction (HS-SPME). For Range Test (Tukey's test), using IBM SPSS statistics 21 software (SPSS
analysis of volatiles, 15 mL of beer and 1.5 g of NaCl were mixed and Inc., Chicago, IL, USA). Additionally, data of the relationship among
equilibrated for 10 min at 40 °C. The volatile compounds were adsorbed physico-chemical parameters, phenolic acids content, sensory profile,
on a DVB/CAR/PDMS fiber of a length of 2 cm (50 min at 40 °C). and quince beers were subjected to principal component analysis using
Similar extraction procedure has been successfully used in pomegranate a XLSTAT software (version 2014.1).
wine (Andreu-Sevilla, Mena, Martí, García Viguera, & Carbonell-
Barrachina, 2013) and fruit liquors (Gironés-Vilaplana, Calín-Sánchez,
3. Results and discussion
Moreno, Carbonell-Barrachina, & García-Viguera, 2015). The equip-
ment consisted of a Shimadzu chromatograph GC-17A (Shimadzu
3.1. Quality parameters of the beers
Corporation, Kyoto, Japan), a Shimadzu mass spectrometer detector
GC-MS QP-5050A, and a SupraWax-280 column (Teknokroma S. Co.
The results obtained after the analysis of the main physico-chemical
Ltd., Barcelona, Spain). Analyses were carried out using helium as
parameters of the beers are shown in Table 1. The addition of quince to
carrier gas at a flow rate of 0.8 mL/min, in a split ratio of 1:10, and the
beer did not have a statistically significant effect on pH, acidity, or on
following program: (a) initial temperature 40 °C; (b) rate of 5.0 °C/min
the color coordinates CIE a* and CIE b*. However, it had significant
up to 100 °C; (c) rate of 4 °C/min up to 220 °C and hold for 1 min; (d)
effects on the total amount of soluble solids, alcohol content, bitterness,
rate of 20 °C/min to 260 °C and hold for 5 min. Injector and detector
IBU color and the color coordinate CIE L*.
temperatures were held at 230 and 260 °C, respectively. Experiments
The addition of quince to beer slightly increased the alcohol content
were run in triplicate, and the relative abundance of each compound
(Table 1). The control sample was the one with the lowest alcohol
was expressed as the percentage (%) of the total arbitrary area units.
concentration (5.27%), while the samples ZM9, PUM and Vranja had
concentrations of 5.51, 5.52 and 5.62%, respectively. This slight var-
2.8. Sensory evaluation with trained panel iation could be due to the fact that during the time of maceration a
second fermentation occurred due to the sugar contribution of the
To describe the quince beers a sensory evaluation with trained panel fruits.
was carry out. Ten panelists, aged between 22 and 49 years, 50% fe- On the other hand, as it can see in Table 1, the beer obtained by
male, associated with the Agro-Food Technology Department of the maceration with quince of the variety Vranja had a concentration of
Universidad Miguel Hernández de Elche (Orihuela, Alicante, Spain), soluble solids of 4.69 ºBrix, while the rest of the samples under study
with more than 500 h of testing experience with fruits, vegetables, meat had a slightly higher concentration (average of 5.78 ºBrix). The same
and beverages, were selected to participate in this study (Andreu- happened with the CIE L* color coordinate, in which the Vranja sample
Sevilla et al., 2013; Gironés-Vilaplana et al., 2015). had the lowest value than the rest of the samples (∼1 unit). It is de-
Panelists discussed the main attributes of beers involved in the vi- monstrated that differences less than 2 units in the CIEL*a*b* system
sual, olfactory, and gustative phases, during 2 preliminary orientation are imperceptible for the human eye (Galindo et al., 2015); thus, the
sessions, each lasting 60 min. For the evaluation, samples were served difference in lightness values among the samples did not imply any
in glasses, odor-free, and covered by watch glasses. Samples were significant change. In the case of the color obtained by EBC, significant
served at a controlled temperature of 20 ± 2 °C in a testing room with differences were also found, although in this case it was the PUM
a combination of natural and fluorescent light. sample the one having the highest value as compared to the other
To quantify the intensity of the beers attributes, the panel used a samples (PUM = ∼41, Control, Vranja, and ZM9 = ∼38).
numerical scale from 0 to 10, with 0.5 increments; where 0 represented Finally, in the case of the bitterness of the samples, the beers ob-
no perceptible intensity, and 10 extremely strong intensity. The panel tained with the quinces of the Vranja and ZM9 varieties had slightly
after orientation sessions, agreed to evaluate the following attributes: lower values (8.70 and 8.77, respectively), than the control sample
(i) appearance: color and foam persistence; (ii) flavor: malt-biscuit, grain (10.96). In contrast, the sample obtained with the variety PUM
cereal, toasted, caramel, nuts, bitter almond, hop, floral, pine resin, (Ibus = 9.52) caused no change in bitterness relative to the control
fruity (citrus, tropical, pome fruit, and berries), herbal, cooked vege- sample (Table 1).
table (DMS, dimethyl sulfide), yeast, earthy-leather, alcoholic, sweet,
sour, salty, bitter, astringent, heat, spicy, body, and aftertaste. Three 1
3.2. Phenolic compounds: hydroxycinnamic acids
h-sessions were held for the samples evaluation, and all 4 beers were
tested in triplicate.
Phenolic compounds of quince fruit and peel have been extensively
studied (Carbonell-Barrachina, Szychowski, Vásquez, Hernández, &
2.9. Statistical analysis Wojdyło, 2015; Fattouch et al., 2007; Silva et al., 2005). The HPLC-
DAD-ESI/MSn analysis of the hydromethanolic extracts of quince fruits
The data presented in this study are the mean values of, at least, 3 revealed a wide range of different phenolic acids (Table 2). 5-O-caf-
replicates and were expressed as the mean ± standard deviation. All feoylquinic acid (chlorogenic acid) and 3-O-caffeoylquinic acid (neo-
the data were subjected to analysis of variance (ANOVA) and a Multiple chlorogenic acid) were identified and differentiated by MSn, having

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All the parameters measured in the quince fruits are expressed as units/100 g of dry weight. †NS = not significant at p < 0.05; *, **, and *** significant at p < 0.05, 0.01, and 0.001, respectively; ‡Values (mean of 3
both of them only displayed a MS2 base peak m/z of 191 with a sec-

15.90 ± 1.15 a
0.724 ± 0.011
0.51 ± 0.05 a
0.65 ± 0.06 b
0.30 ± 0.01 b
3.24 ± 0.21 b
ondary ion at m/z 179. These two acids, chlorogenic and neo-
chlorogenic acids, were also confirmed by the addition of external
Hydroxicinnamic acids (individual and total quantities in mg/100 g), TEAC (mmol Trolox/100 g) and total phenolic contents (TPC in mg/100 g) of different varieties of quince fruits and new designed beers.

Vranja
standards. Differences on MSn were also used to differentiate 3-O-
coumaroylquinic acid and 5-O-coumaroylquinic acid, with MS2 of 163
and 191, respectively. Furthermore, 3,5-di-O-caffeoylquinic acid was
also identified, and was also corroborated with the use of an external

17.55 ± 1.18 a
0.733 ± 0.008
0.97 ± 0.09 a

3.56 ± 0.20 a
0.46 ± 0.03 b

0.30 ± 0.01 b

standard.
The phenolic acid with the highest differences among quince vari-
eties was neochlorogenic acid, with Vranja having the highest quantity
ZM9

of this acid (Table 3); the high differences in this phenolic acid among
varieties was also reflected in the differences in the total content of the
hydroxycinnamic acids. On the other hand, the differences of the con-
16.74 ± 1.12 a
0.722 ± 0.012

tents of chlorogenic acid and 3,5-di-O-caffeoylquinic acid among vari-

0.94 ± 0.08 a
0.32 ± 0.01 a
3.46 ± 0.24 a
0.39 ± 0.02 c

eties were smaller than that of neochlorogenic acids; however, there

were statistically significant differences and ZM9 and PUM had the
PUM

highest contents of chlorogenic acid, while ZM9 had the highest content
of 3,5-di-O-caffeoylquinic acid.
Regarding to newly designed quince beers, the same phenolic acids
replications) followed by the same letter, within the same row, were not significantly different (p < 0.05), according to the Tukey's multiple range test.

(neochlorogenic, chlorogenic and 3,5-dicaffeoylquinic) than in quince

13.47 ± 0.85 b
0.715 ± 0.010
0.11 ± 0.02 c
0.26 ± 0.02 c
2.55 ± 0.11 c
0.00 ± 0.02 d

fruits were detected; besides, 3-O-coumaroylquinic acid and 5-O-cou-

maroylquinic acid were also found, but only in trace amounts.
Control
Beers

However, the contents found in beer were more than one order of
magnitude smaller than those initially found in the quince fruits; for
example, the mean total content of hydroxycinnamic acids (mean of all
treatments) were 180 mg/100 g and 3.2 mg/100 mL in quinces and
294.97 ± 18.80 c
202.80 ± 3.14 a
106.64 ± 2.65 b

beers, respectively.
77.06 ± 0.36 a

3.36 ± 0.07 b

7.93 ± 0.92 b

It is important to comment that although control beers displayed the

same phenolic profile, the phenolic acid contents were significantly
Vranja

lower than those found in the new quince-beers (Table 3); this ex-
perimental finding clearly demonstrated that the hydroxycinnamic
acids come from the maceration of the quince fruits in the beer. No
neochlorogenic acid was detected in control beers, but there was a
543.90 ± 24.31 a
111.55 ± 0.12 a

178.85 ± 4.01 b

significant positive correlation (R2 = 0.828, p < 0.05) between the

12.75 ± 0.85 a
51.40 ± 0.62 b

4.21 ± 0.12 a

content of this compound in the beers and in the quinces (Vranja >
ZM9 > PUM). In the case of chlorogenic and 3,5-di-O-caffeoylquinic
acids, the control beer had low but measurable contents of these two
ZM9

compounds, and the newly designed quince beers had significantly

higher contents, due to quince contribution, and mainly in the PUM
beer. Finally, is important to note that although Vranja fruits had the
412.62 ± 15.42 b
102.29 ± 0.21 c

157.59 ± 0.54 c
‡

highest contents of both neochlorogenic acid and total hydro-

11.46 ± 1.05 a
41.02 ± 0.12 c

4.11 ± 0.02 a

xycinnamic acids, this fact was not fully reflected in the Vranja-beer
which had the highest content of neochlorogenic acid, but the lowest
Fruits

total content of hydroxycinnamic acids of all the quince-beers.

PUM

All these phenolic compounds, identified and quantified in quince

beers in higher quantities than in the control one, have been linked to
beneficial effects in various human diseases, particularly cardiovascular
Beers

diseases and cancer (El-Seedi et al., 2012). Moreover, these phenolic

NS
**

**
*
*
*

acids have been recently reported as a potential class of natural com-

pounds for the management of lipid metabolism and obesity (Alam
ANOVA†

Fruits

et al., 2016), and as neuroprotective agents (Villareal et al., 2016).

***

***
***
**

**
*

3.3. Antioxidant activity (TEAC) and total phenolic content (TPC) of

quince fruits and newly designed quince beers
¥
Total hydroxycinnamic acids (mg/100 g)
3,5-Dicaffeoylquinic acid (mg/100 g) ¥

The free radical scavenging ability of numerous vegetal and food

¥

samples are usually studied using a moderately stable nitrogen-centered

Neochlorogenic acid (mg/100 g)
Chlorogenic acid (mg/100 g) ¥

radical species, ABTS+ radical (Oboh & Ademosun, 2012). With respect
TEAC (mmol Trolox/100 g) ¥
TPC (mg pyrogallol/100 g) ¥

to the quince fruits, PUM and ZM9 exhibited similar antioxidant ca-
pacities, and were significantly higher than that of the Vranja fruits
(Table 3), due to phenolic acid content, mainly chlorogenic acid.
Nevertheless, the contribution of fruits activity in the antioxidant ac-
tivity of new beers was low, because the newly designed quince beers
Parameter

had similar values of ABTS+ to those of the control beers. These ABTS+
Table 3

results of beers were significantly correlated with chlorogenic acid

content (R2 = 0.737, p < 0.05) and with total hydroxycinnamic acids
¥

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Table 4
Relative abundance (% of total area) of volatile compounds, retention time, RT (min) and retention indexes, RI [Exp. = experimental and Lit. = literature (NIST
National Institute of Standards and Technology, 2018)] identified in control and quince beers as affected by quince variety.
Volatile Compound Sensory descriptors RT (min) RI ANOVA† Control Vranja PUM ZM9 Pooled Variance

Exp. Lit. (%)

Ethyl acetate Anise, pineapple 3.13 874 886 NS 0.76 b‡ 0.77 b 1.00 a 1.04 a 0.05
Ethyl butanoate Banana, pineapple 3.89 921 926 NS 0.16 0.17 0.21 0.23 0.05
Ethyl 2-methylbutanoate Apple, plum 6.10 1028 1034 NS 0.01 0.07 0.08 0.09 0.03
Ethyl isovalerate Apple, fruity 6.49 1043 1037 NS 0.01 0.04 0.05 0.04 0.02
Isoamyl acetate Banana, pear 6.90 1058 1054 * 1.54 b 1.88 b 2.32 a 2.26 a 0.34
3-Methyl-1-butanol Oily 8.33 1111 1118 NS 3.64 3.16 3.88 3.85 0.25
Ethyl hexanoate Apple, pineapple 11.11 1204 1208 ** 3.24 b 4.45 a 4.67 a 4.22 a 0.32
Hexyl acetate Cherry, floral 12.93 1265 1262 NS 0.10 0.13 0.13 0.11 0.08
Amyl isovalerate Sweet, mango 13.61 1287 1303 NS 0.03 0.04 0.07 0.10 0.05
3-Methyl-2-buten-1-ol Green, fruity 14.53 1318 1320 NS 0.04 0.05 0.04 0.03 0.02
Ethyl heptanoate Berry, peach 14.76 1326 1327 NS 0.20 0.31 0.35 0.30 0.06
Ethyl 2-hexanoate Apple, banana 15.10 1337 1336 NS 0.02 0.05 0.02 0.02 0.02
1-Hexanol Green, herbaceous 15.33 1345 1344 NS 0.04 0.06 0.06 0.06 0.02
Heptyl acetate Apricot, oily 15.95 1366 1370 NS 0.13 0.16 0.15 0.12 0.03
Methyl octanoate Fruity, citrus 16.35 1380 1387 NS 0.07 0.12 0.16 0.12 0.02
Nonanal Lemon, oily 16.53 1386 1389 * 0.34 b 0.30 b 0.64 a 0.81 a 0.10
Ethyl octanoate Apricot, floral 17.87 1431 1430 ** 41.8 c 45.5 a 43.7 b 41.7 c 0.86
1-Heptanol Apple, apricot 18.34 1446 1452 NS 0.09 0.16 0.22 0.23 0.04
Theaspirane A Raspberry, grape 19.71 1492 1491 ** 0.10 b 1.03 a 0.27 b 0.37 b 0.12
Theaspirane B Raspberry, grape 20.87 1531 1523 ** 0.30 b 1.44 a 0.36 b 0.45 b 0.21
Linalool Floral, citrus 21.13 1539 1542 NS 0.67 0.55 0.63 0.66 0.13
1-Octanol Citrus, woody 21.34 1548 1553 NS 0.20 0.14 0.16 0.30 0.04
Ethyl decanoate Grape, pear 23.91 1633 1635 *** 7.46 a 7.34 a 1.53 c 3.57 b 0.67
Isoamyl octanoate Green, fruity 24.50 1653 1652 ** 0.69 a 0.37 b 0.37 b 0.16 c 0.11
Methyl benzoate Fruity 24.70 1659 1651 * 0.24 b 0.47 a 0.57 a 0.43 a 0.08
(Z)-Ethyl 9-decenoate Fruity, fatty 25.44 1685 1691 * 6.79 a 5.22 b 6.39 a 6.77 a 0.51
1-Decanol Fatty, waxy 27.45 1754 1763 NS 0.15 0.12 0.10 0.14 0.02
Citronellol Geranium, rose 27.54 1757 1755 NS 0.15 0.18 0.17 0.18 0.03
2-Phenylethyl acetate Melon, pineapple 28.94 1806 1811 NS 1.97 1.75 2.01 2.14 0.23
Ethyl dodecanoate Fruity, floral 29.82 1838 1848 *** 3.79 a 2.89 b 0.82 c 1.36 c 0.31
Hexanoic acid Cheese, fatty 30.12 1848 1850 NS 1.09 0.83 1.20 1.04 0.30
Phenylethyl alcohol Honey, rose 31.51 1898 1902 *** 10.8 b 11.0 b 16.7 a 15.2 a 0.41
4-Ethyl guaiacol Smoky, meaty 34.66 2015 2024 ** 1.32 a 0.33 b 0.14 b 0.15 b 0.07
Ethyl tetradecanoate Waxy, soapy 35.35 2041 2049 NS 0.33 0.19 0.16 0.10 0.08
Octanoic acid Oily, cheese 35.74 2064 2068 ** 10.6 a 7.9 b 10.4 a 11.2 a 0.36
Ethyl hexadecanoate Waxy 40.46 2248 2255 NS 0.34 0.16 0.15 0.10 0.10
Decanoic acid Fatty, citrus 41.19 2278 2270 * 0.72 a 0.64 a 0.20 b 0.34 b 0.11

†
NS = not significant at p < 0.05; *, **, and *** significant at p < 0.05, 0.01, and 0.001, respectively; ‡Values (mean of 3 replications) followed by the same letter,
within the same row, were not significantly different (p < 0.05), according to the Tukey's multiple range test; t.i.: Tentatively identified; na: not available.

content (R2 = 0.733, p < 0.05).

Although these phenolic acids have been described as chain-
breaking antioxidants that probably act as radical scavengers (Cos
et al., 2002), the similar values of the ABTS+ activity in all beers
(control and quince) demonstrated that other non-identified com-
pounds were also responsible, at least in part, for this antioxidant ac-
tivity (Fattouch et al., 2007). Furthermore, the possible pro-oxidant
effects of hydroxycinnamic acids previously reported (Razzaghi-Asl,
Garrido, Khazraei, Borges, & Firuzi, 2013) can be also responsible in
part for the similar antioxidant activities of beers.
The total phenolic contents, TPC, of quince fruits showed that ZM9
fruits were the richest ones, followed by PUM and Vranja (Table 3). This
same trend was observed for TPC in the quince beers, with the ZM9-
beer having significantly higher TPC than Vranja and control beers. The
TPC values were positively correlated with ABTS+ data (R2 = 0.880,
p < 0.005 for quince fruits, and R2 = 0.827, p < 0.01 for quince
beers), demonstrating the key, but not exclusive, role of these phyto-
chemicals on the antioxidant activity of the quince fruits and of the new
quince beers.
Nonetheless, TPC values must be interpreted with caution because
Fig. 1. Relative abundance (% ± SE) of the main chemical families in control
and quince beers. Values (mean of 3 replications) followed by the same letter,
the Folin Ciocalteau's reagent can react, not only with phenolics, but
within the same group, were not significantly different (p < 0.05), according also with a variety of non-phenolic reducing compounds, including
to the Tukey's multiple range test. amino acids (tryptophan), certain purines, hydrazine, hydroxylamine,
tertiary aliphatic amines, and other inorganic and organic reducing
agents, leading to an overestimation of the phenolics content.

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Table 5 Moreover, different phenolics may have various responses to the Folin-
Descriptive sensory analysis of control and quince beers as affected by quince Ciocalteu's reagent, presenting lower absorption resulting in under-
variety. The intensity scale used range from 0 (no intensity) to 10 (high in- estimations of compounds too (Ikawa, Schaper, Dollard, & Sasner,
tensity). 2003). Thus, these results should be evaluated together with those
Sensory Attribute ANOVA† Control Vranja PUM ZM9 Pooled obtained by the analysis of phenolic acids by HPLC-DAD-ESI/MSn.
Variance

Visual
3.4. Volatile composition
Color *** 5.2 bc‡ 6.1 a 4.8 c 5.5 b 0.2
Foam persistence NS 4.5 4.8 4.8 5.0 0.3
Flavor Thirty-seven compounds were identified in the volatile profile of
Malta-biscuit NS 2.2 2.1 2.0 1.9 0.2 quince beers (Table 4). The most abundant compound were ethyl oc-
Grain cereal NS 2.9 2.6 2.8 2.5 0.3 tanoate, with a ∼43% of total area of identified compounds [this
Toasted NS 5.7 4.6 5.3 4.9 0.4
Caramel ** 5.4 a 4.4 b 4.5 b 3.7 c 0.3
compound is product of fermentation by Saccharomyces yeast (Saerens,
Nuts NS 0.3 0.1 0.1 0.1 0.2 Delvaux, Verstrepen, & Thevelein, 2010)], followed by phenylethyl
Bitter almond * 0.3 ab 0.1 b 1.0 a 0.5 ab 0.3 alcohol (∼13%), octanoic acid (∼10%), and ethyl 9-decenoate (∼6%).
Hop * 1.3 ab 0.9 b 1.5 ab 1.8 a 0.3 All of these compounds are typical in the volatile composition of any
Floral *** 0.3 c 2.4 a 1.3 b 1.5 ab 0.3
beer (Alvim, de Cássia Oliveira Gomes, Garcia, de Lourdes Almeida
Pine resin NS 0.0 0.0 0.0 0.0 0.0
Fruity * 0.4 b 1.7 a 1.6 a 1.0 ab 0.4 Vieira, & de Resende Machado, 2017; Olaniran, Maharaj, & Pillay,
Citrus NS 0.8 1.2 0.7 1.2 0.2 2011; Verstrepen et al., 2003).
Tropical NS 0.0 0.0 0.0 0.3 0.2 In general, esters with 21 compounds, were the most abundant
Pome fruit * 0.0 b 0.9 a 1.3 a 0.7 a 0.2 chemical group (∼68%), followed by alcohols with 7 compounds
Berries NS 0.0 0.0 0.0 0.0 0.0
Herbal NS 0.7 0.8 1.2 1.0 0.2
(∼18%), acids with 3 compounds (∼12%), monoterpenoids, and nor-
Cooked vegetable NS 0.0 0.0 0.0 0.0 0.0 isoprenoids with 2 compounds (∼1%), and aldehydes and phenolic
Yeast NS 2.6 1.9 1.8 2.2 0.3 compounds with 1 compound (∼1%) (Fig. 1).
Earthy-leather NS 1.5 1.0 0.8 0.9 0.3 Significant differences were found in 15 out the total 37 compounds
Alcoholic NS 1.0 1.0 1.1 1.1 0.1
reported. Quince beers had higher concentrations of some volatile
Sweet NS 2.3 2.8 2.5 2.3 0.3
Sour NS 0.8 0.8 0.8 0.8 0.1 compounds, such as methyl benzoate or ethyl hexanoate (compounds
Salty NS 0.0 0.0 0.0 0.0 0.0 related with fruity descriptors) than control beers. Otherwise, quince
Bitter ** 2.0 b 2.0 b 2.4 a 2.1 ab 0.2 beers had lowest concentrations of 4-ethyl guaiacol, ethyl dodecanoate,
Astringent NS 0.7 0.6 0.8 0.7 0.2 and isoamyl octanoate, which are compounds sensory related with
Heat NS 0.8 0.8 0.8 0.8 0.1
Spicy NS 1.4 1.3 1.3 1.3 0.1
smoky, floral, and green notes, respectively (Table 4).
Body * 2.0 b 2.7 ab 3.3 a 3.4 a 0.2 If the quince samples are compared, ZM9 and PUM had higher
Aftertaste NS 2.1 1.7 2.3 2.2 0.3 concentration of compounds related with fruity, sweet, and citric sen-
sory descriptors than Vranja sample (such as ethyl acetate, isoamyl
†
NS = not significant at p < 0.05; *, **, and *** significant at p < 0.05, 0.01, acetate, nonanal, or phenylethyl alcohol), but Vranja had highest con-
and 0.001, respectively. ‡Values (mean of 3 replications) followed by the same
centration of the main compound found, ethyl octanoate, that is related
letter, within the same row, were not significantly different (p < 0.05), ac-
with pear and pineapple sensory descriptors (Table 4). In global, Vranja
cording to the Tukey's multiple range test.
had highest concentration of esters than ZM9 and PUM, compounds

Fig. 2. Principal component analysis (PCA) scores plot showing the relationship among physico-chemical parameters, phenolic acids content, sensory profile, and
quince beers.

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NIST National Institute of Standards and Technology. (2018) Retrieved May 2018, from
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Luis Noguera-Artiaga was funded by an FPU grant from the Spanish Spanish quince tree clones (Cydonia oblonga Miller). Scientia Horticulturae, 122(3),
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