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Brazilian Grown Cascade Hop (Humulus lupulus

L.): LC-ESI-MS-MS and GC-MS Analysis of Chemical
Composition and Antioxidant Activity of Extracts
and Essential Oils

Aline da Rosa Almeida, Matheus Vinicius de Oliveira Brisola Maciel, Bianca

Cardoso Gasparini Gandolpho, Michelle Heck Machado, Gerson Lopes
Teixeira, Fabiano Cleber Bertoldi, Carolina Montanheiro Noronha, Luciano
Vitali, Jane Mara Block & Pedro Luiz Manique Barreto

To cite this article: Aline da Rosa Almeida, Matheus Vinicius de Oliveira Brisola Maciel, Bianca
Cardoso Gasparini Gandolpho, Michelle Heck Machado, Gerson Lopes Teixeira, Fabiano
Cleber Bertoldi, Carolina Montanheiro Noronha, Luciano Vitali, Jane Mara Block & Pedro Luiz
Manique Barreto (2021) Brazilian Grown Cascade Hop (Humulus�lupulus L.): LC-ESI-MS-
MS and GC-MS Analysis of Chemical Composition and Antioxidant Activity of Extracts and
Essential Oils , Journal of the American Society of Brewing Chemists, 79:2, 156-166, DOI:
10.1080/03610470.2020.1795586

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JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS
2021, VOL. 79, NO. 2, 156–166
https://doi.org/10.1080/03610470.2020.1795586

Brazilian Grown Cascade Hop (Humulus lupulus L.): LC-ESI-MS-MS and GC-MS
Analysis of Chemical Composition and Antioxidant Activity of Extracts and
Essential Oils
Aline da Rosa Almeidaa , Matheus Vinicius de Oliveira Brisola Maciela,b ,
Bianca Cardoso Gasparini Gandolphoa, Michelle Heck Machadoa, Gerson Lopes Teixeiraa ,
Fabiano Cleber Bertoldic , Carolina Montanheiro Noronhaa , Luciano Vitalid , Jane Mara Blocka, and
Pedro Luiz Manique Barretoa
a
Department of Food Science and Technology, Federal University of Santa Catarina, Florian
opolis, SC, Brazil; bDepartment of Biomedicine,
Catholic University Center of Santa Catarina, Joinville, SC, Brazil; cAgricultural Research and Rural Extension of Santa Catarina, Itaja
ı, SC,
Brazil; dDepartment of Chemistry, Federal University of Santa Catarina, Florian
opolis, SC, Brazil

ABSTRACT KEYWORDS
The first harvest of Humulus lupulus (var. Cascade) in Brazil in 2016 originated as an aromatic hop Antioxidant activity;
that nowadays is gaining attention from the brewery industry. However, the chemical composition Brazilian hop; essential oil;
and the properties of this hop have not been well documented to date. In this study, the chemical phenolic compounds;
terpenes
profile of the phenolic compounds and essential oils of Cascade hops produced in Brazil (BH) and
the USA (UH) was determined. The major phenolic compounds identified by LC-ESI-MS-MS in BH
and UH were protocatechuic acid, isoquercitrin, and ferulic acid (4.18–11.39; 2.44–4.90; and
1.45–4.48 mg 100 g
1, respectively). The profile of essential oils was shown to differ between the
two samples. The compounds trans-b-farnesene (20.60%), b-selinene (18.78%), myrcene (16.57%),
and a-selinene (16.22%) were the main compounds in the BH essential oil, while the essential oil
of UH showed a high content of a-humulene (35.35%), myrcene (22.35%), b-caryophyllene
(12.81%), and trans-b-farnesene (10.59%). A higher antioxidant activity (by DPPH and ABTS) was
observed for hop extracts when compared to the essential oils. The results indicated that the
Brazilian hop sample analyzed had higher radical scavenging when compared to an American hop
sample (associated with its bioactive compound composition). The Cascade hop from Brazil
appears to be a high-quality raw material showing significant potential for future industrial
applications.

Introduction France, and Germany) are in the northern hemisphere.

Hops (Humulus lupulus L.) are widely used in the beer
industry worldwide to impart bitterness and aroma, also
contributing to the beer’s shelf life. The production of hops
is essentially directed for brewing (98%).[1,2] However,
some reports have indicated the use of hop in medicines
for relieving mild symptoms of mental stress, insomnia,
treatment for excitability, mood disturbances (restlessness,
anxiety), and sleep disturbances.[1–3] Antifungal 4 and anti-
oxidant[5] activities of breads with added hop extracts, have
been reported in the literature. Products such as shampoos,
conditioners, and balms,[6–8] flavored waters,[9] liquor,[10]
and soda,[11] produced with hop or hop extracts, are avail-
able for sale on different websites.
The lupulin glands, produced only by female inflorescen-
ces (named strobiles or cones), are mainly responsible for
the characteristics of the hops.[2,12,13] Adequate hop growth
depends on crop location and climate conditions. Since the
hops are a cold climate crop, the largest producers (USA,
Therefore, most of the hops used in Brazil are cultivated in
the United States and Europe.[14,15] Because of their unique
aromatic characteristics, many different varieties of H. lupu-
lus may be used by breweries. The Cascade variety was
developed by the United States Department of Agriculture
(USDA) in 1956, released for cultivation in 1971, and it has
been used commercially since 1976. The Hop Growers of
America reported that from 2013 to 2017, the Cascade hop
variety occupied the major production area in the USA. A
decrease of 14.9% in the production of Cascade hop was
registered in the USA in the 2017/2018 harvest with 5257
tons, representing 8.5% of the total US production.[16]
In Brazil, after many unsuccessful attempts, the first
experiment with positive hop production was accomplished
in the municipality of S~ao Bento do Sapuca
ı (S~ao Paulo
state). This pioneering project took place at “Frutopia farm,”
and the produced hop was named the “Mantiqueira hop”
(Figure 1), which is a reference to Serra da Mantiqueira, the

CONTACT Jane Mara Block janeblock@gmail.com

Supplemental data for this article is available online at publisher’s weblink.
ß 2020 American Society of Brewing Chemists, Inc.

Figure 1. Brazilian Cascade hops: (a) frozen hops, (b) lyophilized hops, (c) width, and (d) length of a hop cone.
region where it was cultivated. Production of around
2000 kg of hops of the Cascade variety was harvested in
2016. According to the farmers, the current production
comes from 950 plants that produce about 800 kg per year.
The Brazilian hop was collected and bagged as cones and is
used only by the Baden brewery in the production of a spe-
cial beer named Baden M€arzen. This beer style, originating
in Bavaria, is currently produced with 100% Brazilian hops.
This innovation in the Brazilian brewery may encourage the
production of hops, as it was the first successful attempt to
produce this raw material on a large scale without using
artificial techniques in Brazil.
The chemical composition of hops includes a wide variety
of phenolic compounds such as catechin, epicatechin, ferulic,
protocatechuic, 4-hydroxybenzoic, vanillic, gallic, q-couma-
ric, trans-cinnamic, gentisic, caoric, and ellagic acids, rutin,
morin, quercetin, syringe, coumarin, naringenin, and prenyl-
flavonoids (e.g., xanthohumol).[2,12,17–19] In addition, hops
also present essential oils, rich in terpenes such as myrcene,
a-humulene, trans-b-farnesene, and b-caryophyllene.[12,17,20]
The concentration of phenolic compounds, mainly flavo-
noids, are suggested to present beneficial effects on human
health and disease prevention.[17] This information high-
lights the importance of evaluating the composition of hops
and its essential oils, also verifying which compounds may
influence hop antioxidant properties.
Thus, this study aimed at assessing the chemical compos-
ition of the Brazilian Cascade hop and its essential oil, as
well as the antioxidant activity.
Material and methods
Material
Hop (Humulus lupulus L.) samples of the Cascade variety
from Brazil (BH) and the USA (UH) were donated by Brasil
Kirin (Itu, Brazil) and Brewtime (S~ao Jos
e dos Campos,
Brazil), respectively. The BH samples were cultivated
between October 2015, and April 2016 in the municipality
of S~ao Bento do Sapuca
ı (Brazil; 22 370 32.300 S 45 340 54.000 W)
and were used in the form of cones. The UH samples from
the 2016 harvest were supplied in the form of pellets. Hop
samples were dried using an LJJ02 freeze-dryer (JJ
Cientifica, S~ao Carlos, Brazil) for 24 h; then, they were
milled in a Q298A21 knife mill (Quimis, Diadema, Brazil)
JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 157
and stored at
20  C in polyethylene plastic bags for further
analysis. All standards used were purchased from Sigma
Aldrich (St. Louis, USA), and the reagents were analytical
grade. All analyses were performed in triplicate.
Physicochemical analysis of hop samples
Moisture (constant weight at 105  C), protein content
(Kjeldahl method), lipids (Soxhlet with petroleum ether),
ash (incineration at 550  C), crude fiber, and carbohydrate
by difference (n ¼ 3) were performed according to AOAC
Official Methods.[21]
Identification and quantification of phenolic compounds
of hops
The content of phenolic compounds from hop samples was
evaluated as described by Schulz et al.[22] Briefly, 1 g of
freeze-dried BH or UH was hydrolyzed using 10 mL of
hydrochloric acid. After cooling at room temperature, the
pH of the solution was adjusted to 2 using 6 mol L
1
NaOH. Then, a partition extraction (three times) was per-
formed with 10 mL of ethyl ether (25 ± 2  C). The obtained
fractions were centrifuged at 3000 g for 10 min, and the solv-
ent was evaporated using a rotary evaporator Fisatom 802
(S~ao Paulo, Brazil) at 50 ± 2 C under vacuum. The aliquot
obtained was resuspended with 1 mL of methanol (HPLC
grade). The identification and quantification of phenolic
compounds were carried out using an Agilent LC system
(Waldbronn, Germany). The separation was performed in a
SynergiTM Polar-RP 80 A column (4.0 lm particle size,
150 mm, 2.0 mm internal diameter, Phenomenex, USA).
Liquid chromatographic analysis was carried out using a
mobile phase gradient consisting of (A) 95% methanol in
water and (B) 0.1% formic acid in water, using a flow rate
of 250 lL min
1. The separation proceeded at 30  C using
segmented gradient elution as follows: 0–5 min, 10% A;
5–7 min, 90% A; 7–10 min, 90% A; 10–17 min, 10% A. The
HPLC system was coupled to a mass spectrometry system
consisting of a hybrid triple quadrupole/linear ion trap mass
spectrometer Q Trap 3200 (Applied Biosystems/MDS Sciex,
Concord, Canada). The experiments were performed using
the TurboIonSprayTM source (electrospray-ESI) in negative
ion mode. The capillary needle was maintained at
4500 V.
The MS-MS parameters were curtain gas, 400  C; gas
158 A. da ROSA ALMEIDA ET AL.
Table 1. Physicochemical composition of Cascade hops from Brazil (BH) and
the USA (UH) on a dry basis.
Sample

1
Parameter (g 100g ) BH
Protein 12.20 ± 0.15a
Moisture 9.03 ± 0.12a
Lipids 7.95 ± 0.07a
Ash 6.81 ± 0.21a
Crude fiber 22.20 ± 0.54a
Carbohydrates 54.01 ± 0.89a
Differences between samples are indicated by lowercase letters (t-test, p < 0.05).
1.45 psi; gas 2.45 psi; and CAD gas, medium. Other parame-
ters for DP - Declustering Potential; EP - Entrance Potential;
CEP - Collision Energy Potential; CE - Collision Energy; CXP
- Collision Cell Exit Potential of the 46 phenolic compounds
tested are listed in the supplementary material (Table S1).
The software Analyst (version 1.5.1) was used for the
LC–ESI-MS-MS system control and data analysis.
Obtaining the extracts by ultrasound-assisted extraction
The extracts used in the antioxidant activity were obtained
according to Almeida et al.[23] One gram of sample (BH or
UH) was suspended in 34 mL of an aqueous solution con-
taining 49% (v/v) of ethanol, at 52  C. Ultrasound-assisted
extraction was performed in a Q5.9137A ultrasonic bath
(Ultronique, S~ao Paulo, Brazil) with 5.7 L of working volume
(internal dimensions of 30  15  15 cm) at 37 kHz for
30 min. After the extraction, the samples were centrifuged
for 25 min at 1956 g in a Z200A Hermle centrifuge
(Gosheim, Germany). The supernatant was collected and fil-
tered using a Whatman No 1 filter paper and flushed with
nitrogen. The extracts of Brazilian hop (BHE) and USA hop
(UHE) were then stored in amber bottles under refrigeration
(6 ± 2  C) until further analysis.
Obtaining essential oils from the hops
The essential oils were recovered by steam distillation with a
Clevenger apparatus using 50 g of BH or UH. Briefly, the
sample was transferred to a flask containing 1500 mL of dis-
tilled water. Then it was heated at the boiling point (100  C)
for 30 min and cooled to 75  C. After 7 h, the essential oils
(BHO, and UHO for Cascade hop from Brazil or USA,
respectively) were collected and stored in 2 mL centrifuge
tubes covered with aluminum foil to protect them from light
and stored under refrigeration (5  C).[24]
Chemical profile of essential oils
The chemical profile of essential oils was obtained using a
GCMS - QP2010 gas chromatograph coupled to a mass
spectrometry detector Shimadzu, model LC-20AT (Tokyo,
Japan), using a ZB-5MS capillary column (30 m  0.25 mm
 0.25 mm, Phenomenex, Torrance, U.S.A.). The sample
injection was 1 mL, the injector temperature was 250  C, and
the carrier gas was helium at the flow of 1.0 mL/min. An
isotherm of 4 min at 60  C was used, and then the
temperature was increased (6  C/min) to 210  C and kept in
isotherm for 6 min. The quantification of each component
was made by the normalization of the percentage of the peaks
UH in the total ion chromatogram (TIC). The total area was the
16.55 ± 0.10b sum of all areas of the eluted peaks (100%). Identification by
8.63 ± 0.06b GC-MS was based on the comparison of mass spectra with
15.41 ± 0.43b
10.31 ± 0.09b GC-MS NIST data libraries, and by comparing the retention
14.68 ± 0.67b rates calculated with those found in the literature (database –
51.25 ± 0.67b
WebNIST, GMD). The retention indices were calculated
according to van Den Dool and Kratz.[25]
Antioxidant assays
DPPH radical inhibition assays
DPPH (2,2-diphenyl-1-picrylhydrazyl) assay was carried out
as described by Brand-Williams et al.,[26] with modifica-
tions.[27] Briefly, 0.6 mmol L
1 of DPPH was dissolved in
50 mL ethanol. The DPPH solution was diluted with ethanol
in order to achieve an absorbance value between 0.500 and
0.600 at 515 nm. Then 100 lL of the stock solutions of each
extract was added in 2.9 mL of the adjusted DPPH solution.
After 30 min, the absorbance at 515 nm was determined
using a BEL-PHOTONICS 2000 spectrophotometer (S~ao
Paulo, Brazil). Trolox was used as a standard. The analysis
was performed in triplicate, and the results were expressed
as mmol Trolox equivalent antioxidant capacity mL
1 (mmol
TEAC mL
1).
ABTSþ radical inhibition assays
The ABTS [2,2-azino-bis(3-ethyl benzothiazoline-6-sulphonic
acid)] assay was carried out according to Re et al.[28] with
modifications. Briefly, 10 mL of ABTS solution (7 mM) was
mixed with 10 mL potassium persulfate (2.45 mM). The solu-
tion was homogenized and stored in an amber flask in the
darkness for 16 h. The ABTS solution was diluted with etha-
nol to achieve an absorbance value of 0.700 ± 0.02 at 734 nm.
Then 60 lL of extract stock solution was added in 2.84 mL of
adjusted ABTS solution, and after 6 min, the absorbance at
734 nm was measured in a spectrophotometer. Trolox was
used as the standard. The analysis was performed in triplicate,
and the results were expressed as mmol Trolox equivalent
antioxidant capacity mL
1 (mmol TEAC mL
1).
Statistical analysis
The obtained data from triplicate determinations were
expressed as mean and standard deviations. The comparison
of means was performed by the Students t-test at 5% signifi-
cance level using Statistica 7.0 (StatSoft, USA).
Results and discussion
Physicochemical analysis of hops samples
Table 1 shows the physicochemical composition of BH and
UH expressed on a dry basis. The results showed statistical
differences (p > 0.05) on all parameters evaluated among
samples. Cascade hops evaluated herein presented significant
 JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 159

Table 2. Phenolic profile of Cascade hops produced in Brazil (BH) and in the USA (UH).
Phenolic compounds (mg 100 g
1) Chemical structure BH UH
4-Aminobenzoic acid 0.19 ± 0.03a 0.12 ± 0.01b

Salicylic acid 0.04 ± 0.00a 0.01 ± 0.00b

Cinnamic acid <LOQ <LOQ

p-Anisic acid 0.06 ± 0.00 <LOQ

Mandelic acid 0.19 ± 0.01 <LOQ

Vanillin 0.08 ± 0.00a 0.03 ± 0.00b

4-(Hydroxymethyl)benzoic acid <LOQ –

Protocatechuic acid 11.39 ± 1.04a 4.18 ± 0.17b

p-Coumaric acid 0.84 ± 0.04a 0.19 ± 0.01b

Vanillic acid 0.42 ± 0.03a 0.06 ± 0.00b

(continued)
160 A. da ROSA ALMEIDA ET AL.

Table 2. Continued.
Phenolic compounds (mg 100 g
1) Chemical structure BH UH
Gallic acid 0.18 ± 0.01a 0.18 ± 0.01a

Caffeic acid 1.38 ± 0.04a 1.37 ± 0.10a

Syringaldehyde <LOQ <LOQ

Ferulic acid 4.48 ± 0.08a 1.45 ± 0.62b

Syringic acid 0.31 ± 0.00a 0.08 ± 0.01b

Sinapic acid – 0.01 ± 0.00

Apigenin 0.01 ± 0.00a 0.01 ± 0.00a

Galangin <LOQ <LOQ

Naringenin <LOQ –

Aromadendrin 0.04 ± 0.00 <LOQ

(continued)
 JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 161

Table 2. Continued.
Phenolic compounds (mg 100 g
1) Chemical structure BH UH
Catechin 0.09 ± 0.02b 0.25 ± 0.07b

Epicatechin – 0.01 ± 0.00

Quercetin 3.00 ± 1.16a 1.51 ± 0.08b

Taxifolin 0.03 ± 0.00 –

Chlorogenic acid 0.80 ± 0.09b 1.05 ± 0.13b

Isoquercitrin 4.90 ± 0.28a 2.44 ± 0.16b

Rutin 1.18 ± 0.00a 1.06 ± 0.15a

LOQ ¼ limit of quantification. Significant differences among samples (t-test, p < 0.05) are indicated by lowercase letters.

content of total carbohydrates ranging between 49.38 and
64.01 g 100 g
1 for the samples obtained from the USA and
Brazil, respectively. The BH showed an approximately 34%
higher crude fiber content and half of the lipid content dis-
played by the UH sample. The content of protein in the
hops samples varied from 12.20 to 16.55 g 100 g
1, while ash
ranged from 6.81 to 10.31 g 100 g
1. The moisture content
in the BH and UH was lower than 10%.
Patzak, et al.[29] reported that the physicochemical com-
position of hops can be affected by environmental aspects
such as the type of soil where it grows, the geographical
location, year of harvest and intrinsic differences between
the varieties. In the present study, the main variations
observed in the composition between the samples were
probably due to the use of the whole BH sample in natura,
which contained, a few, numbers of branches and leaves
that are sources of fiber, carbohydrates and humidity. On
the other hand, the UH sample was a commercial product
in the form of pellets that undergoes a process combining
humidity, heat, and pressure, thus facilitating lipid exposure
and extraction from the matrix.[30,31]
Identification and quantification of phenolic compounds
of BH and UH hops
Table 2 shows the identification and quantification of the
phenolic compounds found in the hops. Among the 46
162 A. da ROSA ALMEIDA ET AL.

Table 3. Chemical composition of the essential oils of hops of the Cascade variety from Brazil (BHO) and the USA (UHO).
Compounds Chemical structure RT RI RI-lit BHO (%) UHO (%)
Myrcene 7.79; 7.82 989; 990 992 16.57 22.35

Isobutyric acid 8.57 1017 1020 – 0.83

Unidentified – 10.95 1101 – – 0.41

Geraniol 14.85 1252 1253 – 1.13

2-Undecanone 15.86 1293 1295 4.27 –

a-Ylangene 17.84 1379 1376 – 0.92

b-Caryophyllene 18.83; 18.85 1424; 1425 1427 5.24 12.81

Bergamottin 19.08; 19.10 1436; 1437 1436 0.88 0.51

a-Aromadendrene 19.24 1443 1441 2.37 –

trans-b-Farnesene 19.47; 19.48 1454; 1455 1457 20.60 10.59

a-Humulene 19.60; 19.62 1460; 1461 1460 0.32 35.35

Geranyl propionate 19.80 1470 1475 – 0.60

Unidentified – 19.97 1477 – 1.81 –

c-Muurolene 20.00 1479 1480 – 1.46

Unidentified – 20.28 1492 – – 0.33

(continued)
 JOURNAL OF THE AMERICAN SOCIETY OF BREWING CHEMISTS 163

Table 3. Continued.
Compounds Chemical structure RT RI RI-lit BHO (%) UHO (%)
b-Selinene 20.32; 20.33 1494; 1495 1495 18.78 2.69

a-Selinene 20.46; 20.47 1501 1501 16.22 2.93

Unidentified 20.62 1509 – – 1.91

c-Cadinene 20.81 1518 1520 – 1.36

d-Cadinene 20.90 1522 1524 0.46 2.11

Unidentified – 20.95 1525 – 0.52 –

Unidentified – 21.01 1528 – – 0.45
Unidentified – 21.31 1543 – 2.82 –
Selina-3.7(11)-diene 21.40 1547 1546 3.09 –

Unidentified – 21.76 1565 – 0.90 –

Unidentified – 22.30 1592 – 0.43 –
Unidentified – 22.79 1616 – – 0.77
Unidentified – 23.05 1630 – 0.29 –
Unidentified – 23.64 1661 – 0.49
Unidentified – 23.65 1662 – 1.97 –
Unidentified – 23.70 1665 – 1.78 –
Unidentified – 23.85 1672 – 0.68 –
Total identified – – – – 88.80 95.64
Total unidentified  – – – – 11.20 4.36
RT ¼ Retention time (min). RI ¼ Retention index. RI-lit ¼ Retention index of the literature (NIST, 2018). TR and IRc separated by semicolons are values for BHO
and UHO, respectively. Components that diverge from interpretation by mass spectra or with a retention index not consistent with the values found in
the literature.

phenolic compounds evaluated (Supplementary Material Table
S1), 25 were identified, and 20 were quantified in the BH. In
the UH, 24 compounds were identified, and 18 phenolic com-
pounds were quantified. Significant differences (p < 0.05) were
observed in the content of some phenolics among BH and
UH samples. The results showed that the major phenolic com-
pounds found in the samples were protocatechuic acid, fol-
lowed by isoquercitrin, ferulic acid, caffeic acid, and rutin. The
compounds 4-(hydroxymethyl) benzoic acid, naringenin, and
taxifolin were found in BH. These results showed that the
evaluated Cascade hops are matrices rich in bioactive com-
pounds with similar chemical composition.
Chemical composition of essentials oils
The essential oil in hops has been widely studied, and its
composition shows a wide variety of compounds such as
myrcene, linalool, a-humulene, b-caryophyllene, undecanone-
2, geranyl acetate, b-selinene, and a-selinene. The essential oil
profile of hops has been related to its high antioxidant and
antimicrobial activities.[32–34]
Steam distillation provided a 1.25 and 3.85% extraction yield
for Brazilian and USA hops, respectively. Table 3 shows the
main compounds found in BHO and UHO. Trans-b-farnesene
(20.60%) followed by b-selinene (18.78%), myrcene (16.57%),
a-selinene (16.22%), b-caryophyllene (5.24%) and 2-Undecanone
(4.27%) were the major compounds identified in BHO. On the
other hand, UHO showed a composition rich in a-humulene
(35.35%) myrcene (22.35%), b-caryophyllene (12.81%), and
trans-b-farnesene (10.59%). The composition of essential oil, as
well as the content of trans-b-farnesene in Brazilian Cascade
hops, has not been previously reported in the literature.
The trans-b-farnesene is a highly valued compound in
aromatic hops, and its content in the Cascade variety usually
164 A. da ROSA ALMEIDA ET AL.
Table 4. Antioxidant activity of extracts and essential oils from Cascade
hops samples.
Sample ABTS (lmol TEAC mL
1 ) DPPH (lmol TEAC mL
1 )
BHE 8632.23 ± 10.29a 8272.40 ± 206.06a
UHE 7924.56 ± 7.31b 7585.52 ± 65.79b
BHO 7073.95 ± 3.28c 3297.88 ± 59.37c
UHO 3904.82 ± 5.57d 1425.09 ± 29.68d
TEAC, Trolox equivalent antioxidant activity; BHE, Brazilian hop extract; UHE,
USA hop extract; BHO, Brazilian hop essential oil; UHO, USA hop essential
oil. Equal letters in the same line mean no significant difference
(t-test, p < 0.05).
ranges between 3.0 and 7.0%. However, some varieties of
hops may not synthesize it.[35] This compound may serve as
a fingerprint for the Brazilian Cascade hop, as it appears as
the major compound in the essential oil. The content of
myrcene in the samples is in agreement with previous
reports that showed a range of 7.4 to 22% in wild accessions
and over 30% in commercial hop varieties.[5] Other constit-
uents reported in BHO and UHO include b-caryophyllene
and a-humulene. These compounds have been related to the
quality of aromatic hops and may vary among different spe-
cies and different regions.[20]
It has been reported that a-humulene is one of the most
common terpenes found in the essential oil of many vari-
eties of hop. The differences in the composition of the
studied samples may be related to different growing condi-
tions such as soil and climatic conditions of the region
where they were produced. In addition, differences may be
related to the type of sample studied (cones and pellets).
Mongelli et al.[20] reported 39 compounds in cone hops of
the Cascade variety from the USA. In their study, the main
compounds found were myrcene (47.13%), a-humulene
(15.83%), and b-caryophyllene (5.95%). A similar content of
b-caryophyllene was verified in the BHO sample.
DPPH and ABTS antioxidant activity
Table 4 shows the antioxidant activity for BHO and UHO
and for its ethanolic extracts (BHE and UHE). The data
showed that the extracts presented higher antioxidant activ-
ity than the essential oils. This result may be associated with
the hop’s composition, which is rich in phenolic compounds
such as flavonoids and phenolic acids (Tables 2 and 3).[18]
Krofta et al.[36] described that during the processing of
hop to produce pellets, the raw material is exposed to tem-
peratures of 50–60  C and airflow. Because this process causes
slight damage to the polyphenols, minimal losses (<5%) on
the antioxidant activity of hops were reported. Therefore, the
differences observed for antioxidant activity of BH and UH
may be attributed to the higher concentration of the phenolic
compounds in BH (Table 2), which directly affected the anti-
oxidant power. The high concentration of chlorogenic acid in
the USA hop (UH), which is one of the most thermally
unstable phenolic compounds, suggests that pelletization may
keep the polyphenols in the pellets. However, studies compar-
ing Brazilian and American hops both in the form of pellets
is still necessary to confirm this hypothesis.[1,36]
A comparison between the results for the antioxidant
activity reported in the literature for hops is difficult because
of the different methods used and the wide variety of hops
available. For instance, the inhibition of the DPPH radical
in hop extracts between 70 and 80% has been reported in
the literature for the Galena variety, and Saaz and Spalter
Select varieties,[19,36] while Kowalczyk et al.[1] reported an
EC50 ranging from 1.0 at 1.5 mg mL
1 in hop extracts of the
Magnum and Marynka variety.
This is the first report showing the antioxidant activity of
the Brazilian Cascade essential oils. The study showed that
both BHO and UHO have high antioxidant activity (Table
4), with the Brazilian sample showing almost twice the value
found in the USA sample, suggesting that the composition
of BHO has a higher impact on the scavenging of radicals.
Conclusion
In this initial study, the content of phenolic compounds
determined in Cascade hops and the extracts from Brazil
was higher when compared to the USA sample tested. The
major compounds in BH and UH were protocatechuic acid,
isoquercitrin, ferulic acid, quercetin, caffeic acid, and rutin.
The essential oil profile for the Brazilian Cascade hop pre-
sented a high content of trans-b-farnesene, b-selinene, myr-
cene, a-selinene, b-caryophyllene, and 2-Undecanone. When
compared to the essential oil, extracts from hops showed
higher antioxidant activity due to their higher content of
phenolic compounds, as confirmed by the liquid chromatog-
raphy. Additional studies will be required to investigate
Brazilian grown hops, but initial results appear promising.
Acknowledgments
The authors would like to thank Coordination for the Improvement of
Higher Education Personnel - CAPES (grants 88882.316463/2019-01,
88882.316461/2019-01, 88882.344940/2019-01, and 1744193) for the
scholarships of Gerson Lopes Teixeira, Carolina Montanheiro
Noronha, Michelle Heck Machado, and Aline da Rosa Almeida,
respectively. J. M. Block thanks CNPq for the productivity grants
(311070/2018-3). The authors also are grateful to Brasil Kirin and
Brewtime for donating the samples.
Conflict of interest
The authors have no conflict of interest to disclose.
Authors’ contributions
A. R. Almeida, M. V. O. B. Maciel, and P. L. M. Barreto designed the
study. A. R. Almeida conducted the experiments, collected data, and
discussed the results. M. H. Machado, F. C. Bertoldi, L. Vitali, and B.
G. Gandolpho helped to conduct the experiments and collect data. A.
R. Almeida and C. M. Noronha drafted the manuscript. G. L. Teixeira
and J. M. Block edited the manuscript.
ORCID
Aline da Rosa Almeida http://orcid.org/0000-0003-4750-6319
Matheus Vinicius de Oliveira Brisola Maciel http://orcid.org/0000-
0001-8702-1063
Gerson Lopes Teixeira http://orcid.org/0000-0002-3442-3525
Fabiano Cleber Bertoldi http://orcid.org/0000-0003-4750-5164

Carolina Montanheiro Noronha http://orcid.org/0000-0003-
2068-7439
Luciano Vitali http://orcid.org/0000-0002-1664-6346
Pedro Luiz Manique Barreto http://orcid.org/0000-0001-8716-8962
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