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South African Journal of Botany xxx (2018) xxx–xxx

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South African Journal of Botany

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An investigation of Humulus lupulus L.: Phenolic composition, antioxidant

capacity and inhibition properties of clinically important enzymes
Ş. Keskin a,⁎, Y. Şirin b, H.E. Çakir b, M. Keskin b
a
Department of Chemistry, Faculty of Science and Literature, Bilecik Şeyh Edebali University, Bilecik, Turkey
b
Department of Chemistry, Institute of Natural Science, Karadeniz Technical University, Trabzon, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Humulus lupulus L., known as hop, is used mostly in brewing industry. It is known that hops have some secondary
Received 8 February 2018 metabolites having significant biological activities. As far as we know this is the first paper reporting urease,
Received in revised form 13 April 2018 α-amylase and acetylcholinesterase inhibition activity of hop extracts. In this study, phenolic composition,
Accepted 25 April 2018
antioxidant capacities, and some clinically important enzyme inhibition properties of the methanol
Available online xxxx
extracts were investigated. Total polyphenolic content (TPC), ferric reducing antioxidant power (FRAP) and
Edited by KRR Rengasamy 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity tests were used for antioxidant activity
determinations. Inhibition activity of the extracts on three clinically important enzymes of urease, α-amylase
Keywords: and acetylcholinesterase was studied. Polyphenolic composition of the extracts was identified by using
Humulus lupulus L. RP-HPLC-UV in the presence of fourteen phenolic standards. TPC was found 7.12 ± 0.09 and 6.86 ± 0.05 mg GAE/g
Antioxidant activity in methanol extract of hop cone and leaf respectively. It was found that methanol extract of hop cones and leaves
Inhibition showed good urease (IC50 0.58 ± 0.02 and 0.87 ± 0.02 mg/mL), acetylcholinesterase (IC50 2.13 ± 0.03 and
Urease 4.18 ± 0.02 mg/mL) and α-amylase (IC50 3.92 ± 0.02 and 6.05 ± 0.03 mg/mL) inhibitory effect respectively.
Acetylcholinesterase
It can be concluded that methanol extract of H. lupulus may be an alternative for the treatment of peptic
α-Amylase
ulcer, Alzheimer's disease and diabetes mellitus.
Peptic ulcer
Alzheimer's disease © 2018 SAAB. Published by Elsevier B.V. All rights reserved.

1. Introduction formation of urinary stones and gastro-intestinal system (Upadhyay,

Humulus lupulus L. (Cannabaceae) is an industrial and medicinal
plant. In Turkey Pazaryeri, a district of Bilecik city, is the only plantation
area of the plant (Baytop, 1999). H. lupulus, a perennial and climbing
plant, is very important in the brewing industry because of its
organoleptic features. The organoleptic properties are derived from
many different bitter organic acids, essential oils, resins, and polyphenol
compounds (Cermak et al., 2015). Apart from brewing industry, hops
are used to manage anxiousness, spasms, cough, fever, inflammation
and toothache as traditional and complementary medicine since
ancient times (Liu et al., 2015; Kim et al., 2016). The plant consists of
high amount of humulones, lupulones, isohumulones and xanthohumol
and these are responsible for organoleptic characteristics and
other biological activities such as antioxidant, anti-inflammatory,
anti-microbial and anti-tumoral effects (Yamaguchi et al., 2009).
Urease (EC 3.5.1.5), a hydrolytic enzyme, catalyzes the conversion of
urea to ammonia and carbon dioxide. Bacterial ureases play important
roles in pathogenicity of some diseases such as hepatic coma, peptic
ulceration and urinary stones (Upadhyay, 2012; Horta et al., 2016).
Urease inhibitors prevent the human body from many disorders like
⁎ Corresponding author.
E-mail address: sabankeskin61@hotmail.com (Ş. Keskin).
2012). Inhibition of urease is very important to cope with Helicobacter
pylori, the bacteria which can survive in the stomach by converting
urea to ammonia and causes gastritis and ulcer (Sahin, 2015; Baltaş
et al., 2016). It was reported that many natural compounds and extracts
could inhibit infection of H. pylori and/or growth of the bacteria in terms
of urease inhibition (Kolaylı et al., 2017).
Acetylcholinesterase (EC 3.1.1.7) catalyzes hydrolysis of acetylcho-
line to choline and acetate. It is mainly found in nerve and muscle
tissues as well as central nervous and peripheral tissues (Küçükkılınç,
2014). Alzheimer is caused by the loss of some neurons in the brain
and shows indications of memory loss, dysfunction and mental retarda-
tion. It is emphasized that the deficiency of acetylcholine in the brain is
responsible for the disease. Until now, there hasn't been a definite
treatment of Alzheimer disease, but acetylcholinesterase inhibitors are
accepted as effective agents for the treatment of Alzheimer's disease
(Jazayeri et al., 2014).
α-Amylase (EC 3.2.1.1), α-1,4-glucan-4-glucanohydrolase, is an
enzyme that catalyzes the hydrolysis of the α-1,4-glycosidic linkages
in starch and related oligosaccharides. It is clearly stated in the literature
that inhibition of α-amylase coupled with α-glucosidase could consid-
erably decrease the post-prandial increase of blood glucose level
(Jayaraj et al., 2013). So inhibition of α-amylase could be a good strategy
to control blood glucose level in type 2 diabetes mellitus.

https://doi.org/10.1016/j.sajb.2018.04.017
0254-6299/© 2018 SAAB. Published by Elsevier B.V. All rights reserved.

Please cite this article as: Keskin, Ş., et al., An investigation of Humulus lupulus L.: Phenolic composition, antioxidant capacity and inhibition
properties of clinically important enzymes, South African Journal of Botany (2018), https://doi.org/10.1016/j.sajb.2018.04.017
2 Ş. Keskin et al. / South African Journal of Botany xxx (2018) xxx–xxx

The main objective of this study is to disclose the possibility of
using hops in the treatment of certain diseases such as peptic ulcer,
Alzheimer's disease (AD) and diabetes mellitus (DM). For this purpose,
methanol extract of cone and leaf parts of hop was analyzed in terms
of phenolic compositions and antioxidant properties. The inhibition of
three clinical key enzymes to clarify the potential of hop usage in
the treatment of these diseases was also carried out. The inhibition
of these enzymes with hop extracts has not been studied up to now,
for all we know this is the first paper reporting the inhibitory effect
of methanol extract of hop parts on urease, acetylcholinesterase
and α-amylase.
2. Material and methods
2.1. Materials
45 °C. The residues were re-dissolved in 2 mL of methanol and filtered
with a 0.45 μm filter. Fourteen polyphenolic standard compounds listed
in Table 1 were analyzed using HPLC (Elite LaChrom; Hitachi, Tokyo,
Japan) equipped with a reverse phase C18 column (150 mm, 4.6 mm,
5 μm; Fortis). Acetonitrile, water and acetic acid were used as mobile
phase and a programmed gradient was applied. 20 μL of samples was
injected individually at room temperature and flow rate was set as
0.75 mL/min (Can et al., 2015) HPLC-UV chromatograms of phenolic
standards was given in Fig. 1.
2.2.4. Determination of antioxidant properties
Antioxidant capacity of the samples was determined by using Ferric
reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl
(DPPH) radical scavenging activity tests respectively.

The phenolic standards of gallic acid, protocatechuic acid, 2.2.4.1. Ferric reducing antioxidant power (FRAP). The principle of this
p-hydroxybenzoic acid, vanillic acid, caffeic acid, catechin, syringic method is based on the reduction of Fe 3 + ions with in the Fe
acid, p-coumaric acid, t-cinnamic acid epicatechin, rutin, luteolin (TPTZ)3+ tripyridyltriazine complex in acidic medium to blue colored
and ferulic acid were purchased from Sigma Aldrich Chemie GmbH Fe(TPTZ)2+ complex by antioxidants (Benzie and Strain, 1999). FRAP
(Munich, Germany). Folin Ciocalteu phenol reagent and 2,4,6-Tris reagent was prepared by mixing 2.5 mL of 10 mM TPTZ dissolved in
(2-pyridyl)-s-triazine (TPTZ) were purchased from Fluka Chemie 40 mM HCL, 2.5 mL FeCl3·6H2O and 25 mL of 300 mM pH 3.60 acetate
GmbH (Switzerland). Jack bean urease, acetylcholinesterase (from buffer. A 100 μL aliquot of sample was added to 3 mL of freshly prepared
Electrophorus electricus), α-amylase (from Barley), urea, allopurinol FRAP reagent, then incubated at 37 °C for 4 min. The absorbance of
and thiourea, donepezil hydrochloride, acetylthiocholine chloride, the samples was recorded at 593 nm against distilled water blank.
5,5′-dithiobis(2-nitrobenzoic acid), glucose, 3,5-dinitrosalicylic Ferrous sulfate solution in different concentrations ranging from 100
acid, 2,2-diphenyl-1-picrylhydrazyl, methanol, glacial acetic acid, to 1000 μM was used to prepare a standard calibration curve. Trolox
acetonitrile, sodium acetate, iron(III) chloride hexahydrate (FeCl3·6H2O), was used as reference standard. Results obtained by the FRAP assay
iron(II) sulfate heptahydrate (FeSO4·7H2O) and acarbose were obtained were given as μM of ferrous equivalent Fe(II) per 100 g of sample.
from Sigma-Aldrich (St. Louis, MO, USA). All chemicals used in the study
were of analytical grade.
2.2.4.2. Free radical scavenging activity of DPPH. The radical scavenging
capacity of the methanol extracts was determined by using DPPH
2.2. Methods
assay described by Molyneux (2004). For each sample, 0.75 mL of
the extract at six different concentrations was mixed with 0.75 mL of
2.2.1. Preparation of the extracts
0.1 mM DPPH in methanol and the absorbance was recorded at
H. lupulus L. was freshly supplied by a local grower from Pazaryeri,
517 nm after incubation of 50 min at room temperature. Results were
Bilecik, Turkey in the cultivation season of 2016. Hop cones and leaves
expressed as SC50 (mg sample per mL), which corresponded to the
were air dried in a shady place. Ten grams of dried samples of hop
concentration of each sample that resulted in 50% scavenging of DPPH.
cones and leaves were separately placed in a flask with 100 mL
methanol (99%) and stirred at room temperature for 24 h, then
sonicated for 2 h with an ultrasonicator (ultrasonic Elma Schmidbauer 2.2.5. Enzyme inhibition activity of the extracts
GmbH, Germany). The mixtures were filtered with filter paper Three clinical important enzymes namely urease from Canavalia
(Whatman) and concentrated in a rotary evaporator (IKA-Werke, ensiformis (Jack bean), acetylcholinesterase from E. electricus (electric
Staufen, Germany) at 40 °C. The residues were then individually eel) and α-amylase from Barley malt (Type VIII-A) were obtained
resolved with a minimal volume of methanol and kept at 4 °C until used. commercially from Sigma Aldrich (St. Louis, MO, USA). All assays were
repeated 3 times and the results were expressed as average value
2.2.2. Determination of total polyphenolic content with ±deviations.
Determination of the total phenolic content was carried out by using
the Folin–Ciocalteu method, and gallic acid was used as standard
(Slinkard and Singleton, 1977). Shortly, 680 μL of distilled water, Table 1
RP-HPLC-UV validation parameters of phenolic compounds.
400 μL of 0.5 N Folin–Ciocalteu reagent and 20 μL of gallic acid in varied
concentrations (0.5 mg/mL–0.03125 mg/mL) were pipetted in different No Compounds R2 %RSD %RSD LODa LOQa
tubes and then the tubes were vortexed. After 3 min, 400 μL of Na2CO3 (Retention time) (Area)
solution (10%) was added into each tube and vortexed. The mixtures 1 Gallic acid 0.999 0.210 1.941 0.022 0.067
were then incubated for 2 h at room temperature. The absorbance 2 Protocatechuic acid 0.999 0.871 1.920 0.042 0.128
was recorded at 760 nm at the end of the incubation period. A standard 3 p-OH benzoic acid 0.998 0.351 3.055 0.036 0.109
4 Catechin 0.998 0.492 4.279 0.040 0.121
curve was generated and results were expressed as mg gallic acid
5 Vanillic acid 1.000 0.828 2.066 0.025 0.075
equivalents (GAE) per g of sample. 6 Caffeic acid 0.998 0.179 4.039 0.062 0.187
7 Syringic acid 1.000 0.550 0.848 0.009 0.027
2.2.3. Identification of polyphenolic compounds by HPLC 8 Epicatechin 0.999 0.429 3.819 0.030 0.090
9 p-Coumaric acid 0.999 0.204 1.562 0.010 0.030
The methanol extracts were subjected to re-extraction for HPLC
10 Ferulic acid 0.999 0.222 1.301 0.011 0.033
analysis. For this, the solvent of the extracts was removed and the 11 Rutin 1.000 0.234 3.139 0.041 0.123
residues were dissolved in 10 mL acidified (pH 2) distilled water. 12 Daidzein 0.998 0.174 1.545 0.018 0.054
Re-extraction was carried out firstly with 5 mL diethyl ether then with 13 t-Cinnamic acid 1.000 0.262 1.071 0.014 0.042
5 mL ethyl acetate for 3 times for each. Organic phases were separately 14 Luteolin 0.994 0.229 5.833 0.043 0.130

combined and the solvents were removed in a rotary evaporator at a

Values are expressed as mg/L.

Please cite this article as: Keskin, Ş., et al., An investigation of Humulus lupulus L.: Phenolic composition, antioxidant capacity and inhibition
properties of clinically important enzymes, South African Journal of Botany (2018), https://doi.org/10.1016/j.sajb.2018.04.017
 Ş. Keskin et al. / South African Journal of Botany xxx (2018) xxx–xxx 3

Name

Area

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32

Minutes

Fig. 1. HPLC-UV chromatograms of phenolic standards. 1. Gallic acid. 2. Protocatechuic acid. 3. p-OH benzoic acid. 4. Catechin. 5. Vanillic acid. 6. Caffeic acid. 7. Syringic acid. 8. Epicatechin.
9. p-Coumaric acid. 10. Ferulic acid. 11. Rutin. 12. Daidzein 13. t-cinnamic acid. 14. Luteolin.

2.2.5.1. Determination of urease inhibition. Urease activity was deter-
mined by using the Weatherburn method. This method is based on
the determination of the amount of ammonia released from urea by
the action of urease enzyme by using the indo phenol method that has
an absorbance of 625 nm (Weatherburn, 1967). Shortly, reaction mix-
ture contained 5 μL of urea solution (100 mM), 40 μL of jack bean urease
and 5 μL of buffer (0.01 M KH2PO4, 1 mM EDTA and 0.01 M LiCl; pH 8.2).
After incubation at 35 °C 15 min, 750 μL of phenol reagent (1% w/v phe-
nol and 0.005% w/v sodium nitroprusside) was added, vortexed and
then, 750 μL of alkali reagent (0.5% w/v NaOH and 0.1% v/v NaOCl)
was added and vortexed. This mixture was incubated for 15 min more
at 35 °C and optical density was measured at 625 nm against a blank so-
lution including distilled water instead of enzyme. For the determina-
tion of the IC50 value of the extracts, activity assays were conducted at
five different extract concentrations and dose response curve was gen-
erated. Thiourea was used as standard inhibitor.
2.2.5.2. Determination of acetylcholinesterase inhibition. Acetylcholines-
terase activity was assayed by using acetylthiocholine as substrate.
Thiocholine released by the action of the enzyme reacts with Ellman's
reagent (DTNB) yielding 5-tiyo-2-nitro benzoic acid which has
maximum absorbance at 412 nm (Ellman et al., 1961). Reaction cuvette
containing 1500 μL 50 mM pH 8.0 Tris–HCl buffer, 10 μL of AChE and
20 μL DTNB (10 mM) was incubated at 37 °C for 15 min. Then 10 μL of
acetylthiocholine (200 mM) was added to the mixture and 5-tiyo-2-
nitro benzoic acid was quantified at 412 nm. For the determination of
the IC50 value of the extracts, activity assay tubes contained 50 μL
extract at five different concentrations and dose response curve
was generated. Donepezil was used as standard inhibitor.
respectively. It was reported that methanol extract of hop cones
contained 7.4 μg GAE/g total polyphenol (Maliar et al., 2017). Farcaş
et al. reported that methanol extract of hop contained 2.39 mg GAE/g
total polyphenol (Fărcaş et al., 2017). In a study carried out by Abram
et al. it was reported that hop leaf extract had 15 mg GAE/g total polyphe-
nol (Abram et al., 2015). It was stated that total polyphenolic content of
hop could be affected by many factors including, growing area, weather
condition, extraction methods and the solvent used (Abram et al., 2015).
3.2. Determination of phenolic components by HPLC
Fourteen individual phenolic compounds of the methanol samples
were analyzed by using HPLC, the results were summarized in Table 2.
It was found that ten phenolic compounds were detected in the hop
cones, and eight phenolic compounds were identified in the leaves
respectively. Luteolin, rutin, ferulic acid and catechin were the most
abundant phenolic compounds detected in the hop cone extracts, and
rutin, epicatechin, vanillic acid and daidzein were the most abundant
phenolic compounds detected in the hop leaf extract as well. Similar
results were reported in different studies. p-Coumaric acid, rutin, caffeic
acid and catechin were reported to be found in ethanol extract of hop
cones (Abram et al., 2015). Inui et al. (2017) stated that phenolic
composition of hop could differ depending on the hop cultivars, harvest
time and harvest year.
3.3. Determination of antioxidant properties
FRAP and DPPH methods were frequently used antioxidant tests,
mostly natural extracts. The results were summarized in Table 3.

2.2.5.3. Determination of α-amylase inhibition. α-Amylase activity was Table 2

assayed according to the modified DNS method described by Bernfeld Phenolic content of methanol extract of Humulus lupulus L.
(1955). Reaction mixture containing 300 μL of 1% soluble starch and Phenolic standard H. lupulus L. cone extract H. lupulus L. leaf extract
300 μL of enzyme solution was incubated for 30 min at 35 °C. Equal (μg/g sample) (μg/g sample)
volume of DNS reagent was added into tubes and kept in a boiling Gallic acid nd nd
water bath to quantify generated reducing sugars as glucose equivalent. Protocatechuic acid 5.71 ± 0.02 nd
All characterization assays were performed in triplicate. IC50 value of the p-OH benzoic acid nd 2.12 ± 0.07
extracts was determined at five different extract concentrations at Catechin 369.44 ± 0.09 nd
Vanillic acid nd 102.54 ± 0.12
standard assay condition and dose response curve was generated.
Caffeic acid 20.98 ± 0.01 nd
Acarbose was used as reference inhibitor. Syringic acid 43.32 ± 0.02 nd
Epicatechin nd 825.57 ± 0.04
3. Results and discussion p-Coumaric acid 50.24 ± 0.03 45.23 ± 0.02
Ferulic acid 472.61 ± 0.05 25.85 ± 0.01
Rutin 903.48 ± 0.04 1338.94 ± 0.05
3.1. Determination of total polyphenolic content Daidzein 92.40 ± 0.01 120.95 ± 0.02
t-Cinnamic acid 14.84 ± 0.02 49.13 ± 0.03
Total phenolic content of the methanol extracts was determined. Luteolin 1261.14 ± 0.02 nd
It was found that hop cones and hop leaves contained 7.12 ± Results are the mean value of three parallel experiments with ±standard deviations.
0.09 mg GAE/g and 6.86 ± 0.05 mg GAE/g phenolic compounds nd: not determined.

Please cite this article as: Keskin, Ş., et al., An investigation of Humulus lupulus L.: Phenolic composition, antioxidant capacity and inhibition
properties of clinically important enzymes, South African Journal of Botany (2018), https://doi.org/10.1016/j.sajb.2018.04.017
4 Ş. Keskin et al. / South African Journal of Botany xxx (2018) xxx–xxx
Table 3
Antioxidant properties of methanol extract of Humulus lupulus L.
Antioxidant test H. lupulus L. H. lupulus L.
Cone extract Leaf extract
FRAP μM FeSO4·7H2O/g sample 926.54 ± 0.02 873.71 ± 0.07
DPPH SC50 (mg/mL) 0.78 ± 0.01 0.99 ± 0.01
Results are the mean value of three parallel experiments with ±standard deviations.
It was found that methanol extract of the hop cones and the leaves
exhibited a good ferric reducing inhibition activity as 926.54 ± 0.02
and 873.71 ± 0.07 μM FeSO4·7H2O/g sample. Moreover methanol ex-
tract of the hop cones and the leaves showed good radical scavenging
activity with the SC50 value of 0.78 ± 0.01 and 0.99 ± 0.01 mg/mL,
respectively.
It was reported that methanol extract of hop leaves had much lower
DPPH scavenging activity (IC50 0.02 mg/mL) than methanol extract of
hop cones (IC50 0.01 mg/mL) (Abram et al., 2015). It was stated that
methanol extract of wild hop shoots exhibited 0.5 mg trolox equiva-
lent/g sample (Maietti et al., 2017). Mocan et al. studied the Morina
persica L. by preparing water, methanol and acetone extract and they
reported that methanol extract showed both the highest DPPH radical
scavenging activity as 48.09 ± 0.86 mg trolox equivalent/g extract and
highest ferric reducing activity as 82.85 ± 2.53 mg trolox equivalent/g
extract (Mocan et al., 2016). Higher antioxidant capacity of hop cone
extract may be the result of higher polyphenolic content of the hop
cone extract.
3.4. Enzyme inhibition activity of the extracts
Inhibition effect of the extracts on urease, acetylcholinesterase and
α-amylase was firstly investigated. The results were expressed as IC50
and summarized in Table 4. It was found that methanol extract of hop
cones and leaves was a much better inhibitor than thiourea, standard
inhibitor of urease. It was also found that methanol extract of the hop
cone showed better inhibition activity on all tested enzymes than meth-
anol extract of the hop leaf. It is clear from the results that the extracts
have good inhibition activity on acetylcholinesterase and α-amylase
enzymes as well. High inhibitory effect of extracts could be explained
by its polyphenolic composition as the extracts are rich in phenolic
acid and catechins. The inhibitory activity of phenolic acids and
catechins extracted from different sources was reported previously
(Hara and Honda, 1990; Nagai et al., 2004; Kolaylı et al., 2016).
There has been a visible increase in researches of the plant
metabolites exhibiting interesting biological properties. It is known
that phenolic compounds are secondary metabolites of plants and
they exhibit broad spectrum of biological activity. These compounds
are not required in daily diet for the human body to function appropri-
ately, but they enhance resistance to various diseases and infections
(Kolaylı et al., 2011; Żołnierczyk et al., 2015). Over the past 20 years,
the tendency towards the use of natural remedies has increased because
of the side effects of synthetic medicines and the resistance of disease
factors to these medicines. It is stated that treatment of peptic ulcer
caused by H. pylori depends on urease inhibition as the bacteria can
survive in the acidic environment of the stomach by converting urea
to ammonia (Altındiş and Özdemir, 2003). Peptic ulcer caused by
Table 4
IC50 values of the extracts.
H. lupulus L. H. lupulus L. Standard inhibitor
Cone extract Leaf extract IC50 (mg/mL)
IC50 (mg/mL) IC50 (mg/mL)
Urease 0.58 ± 0.02 0.87 ± 0.02 (Thiourea) 12.01
Acetylcholinesterase 2.13 ± 0.03 4.18 ± 0.02 (Donepezil) 0.03
α-Amylase 3.92 ± 0.02 6.05 ± 0.03 (Acarbose) 1.26
Results are the mean value of three parallel experiments with ±standard deviations.
Please cite this article as: Keskin, Ş., et al., An investigation of Humulus lupulus L.: Phenolic composition, antioxidant capacity and inhibition
properties of clinically important enzymes, South African Journal of Botany (2018), https://doi.org/10.1016/j.sajb.2018.04.017
H. pylori is a disease that could not be cured completely even by using
three or more drug combinations (Özden, 2016). It was reported in a
study that fresh hops showed inhibitory effect on the growth of
H. pylori isolated from the patients suffering from gastritis (Cermak
et al., 2015). In the present paper, urease inhibition was reported
for the first time by methanol extract of the hop cones and leaves.
Urease inhibitory activity of methanol extract of Hibiscus schizopetalus
with an IC50 value of 80.1 ± 0.87 μg/mL was reported (Zahid et al.,
2014). Methanol and water extracts of fifteen medicinal plants, except
H. lupulus, were tested by Mahernia et al. for jack bean urease inhibitory
activity and it was reported that they showed different inhibitory
activity with IC50 values ranging from as high as 409.90 μg/mL to as
little as 36.17 μg/mL (Mahernia et al., 2015). It was reported that
methanol and water extracts of Aphloia theiformis showed good
urease inhibition activity with IC50 values as 148.80 ± 6.46 μg/mL and
228.57 ± 3.56 μg/mL respectively (Picot et al., 2017).
As mentioned before there has not been a certain cure for Alzheimer
disease and acetylcholinesterase inhibitors have been considered to be
effective in prevention of the disease (Jazayeri et al., 2014). In this
paper, we report the acetylcholinesterase inhibition by methanol
extract of the hop cones and leaves for the first time. It was reported
that methanol extract of Rhizophora mucronata leaf exhibited AChE in-
hibitory activity with an IC50 value of 59.31 ± 0.35 μg/mL (Suganthy
and Devi, 2016). It was reported that methanol extract of seven
medicinal plants parts (root, stem or rhizome) exhibited AChE
inhibitory activity with IC50 values ranging from 16.74 to 73.69 μg/mL
(Vinutha et al., 2008).
As the inhibition of α-amylase along with the α-glucosidase is an
approach for the treatment of diabetes mellitus, many medical plant
extracts have been screened for α-amylase and/or α-glucosidase
inhibitory activity. It was reported that water, ethanol and hexane
extracts of Eugenia dysenterica leaf at 1 mg/mL concentration caused
93.08, 98.95 and 78.01 percentage inhibition for α-amylase activity
respectively (deSouza et al., 2012). It was reported that ethanolic
extract of Andrographis paniculata showed a weak alpha-amylase
inhibitory activity (IC50 = 50.9 ± 0.17 mg/mL) for porcine pancreatic
amylase (Subramanian et al., 2008). It was reported that grape seed
(IC50 = 8.7 ± 0.8 μg/mL), green tea (IC50 = 34.9 ± 0.9 μg/mL),
and white tea (IC50 = 378 ± 134 μg/mL) extracts showed different
inhibition properties for human saliva α-amylase (Yılmazer-Musa
et al., 2012).
4. Conclusion
In this paper we reported the phenolic composition, antioxidant ac-
tivity of methanol extract of the hop cones and the leaves. The inhibition
of some medically important enzymes by the extracts was reported for
the first time as well. Our results clearly showed that even though the
total polyphenolic content of the leaf extract was less than that of
cone extract, leaf of H. lupulus L. could be used for phenolic source as
food additive or preservative. Our results obtained for enzyme inhibi-
tions are quite compatible with the findings reported in literature.
Methanol extract of hop cones showed higher antioxidant capacity
and higher enzyme inhibition properties, because of its higher phenolic
composition. By comparing with the given literature data, it could be
concluded that the hop cones and leaves might be useful for the
treatment of many diseases such as peptic ulcer, Alzheimer disease
and diabetes mellitus.
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