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IN VITRO BIO-MEDICAL STUDIES ON PSIDIUM GUAJAVA LEAVES
Taha F. Taha1, Hend A. Elakkad1, Ahmed S. H. Gendy2, Mohammed A. I. Abdelkader2
and Shaimaa Salah ELdeen Hussein3
1Department of Biochemistry, Faculty of Agriculture, Zagazig University, Egypt.
2Department of Horticulture, Faculty of Agriculture, Zagazig University, Egypt.
3Department of Pharmacology, Faculty of Pharmacy, Jouf University, Saudi Arabia.
Abstract
Distinctive parts of guava Psidium guajava L. (PG) are a great wellspring of strong and intense bio medications. The present
investigation was intended to extract and to identify the bioactive component of PG leaves, utilizing HPLC and assessed the
activity of ethanol(70%) extract against Helicobacter pyroli, tuberculosis, diabetic (Alpha-glucosidase inhibitory %), arthritic
(albumin denaturation inhibition) and aging (Anti-Collagenase) factorials. The outcomes of HPLC identification phenolic
compounds showed that ethanolic extract contained gallic acid, caffeic acid, ferulic acid, cinnamic acid and they have highest
values (4.71, 4.37, 3.82, 3.55 and 3.49µg/mg, respectively followed by resorcinol, chlorogenic, syringic acid and resormarinic
acid (3.09, 2.93, 2.85 and 2.49µg/mg, respectively), while flavonoids were quercetin, hesperetin, kaempferol, quercitrin and
rutin, where they presented 8.94, 7.61, 7.55, 7.13 and 6.37 µg/mg, respectively followed by catechin (5.12µg/mg) and apigenin
(4.83 µg/mg). The identified alkaloids were corilagin, kaempfertin, and isoquinoline and they have values 2.13, 1.89 and
1.24µg/mg, respectively. Evaluation of inhibitory effect of PGL extract stated that PGL scored the highest values for effective
concentrations (125 µg/mg) and MIC90 (26.6) comparing with activity of clarithromycin (C) (1.95 µg/mg and MIC90 (0.7),
respectively against Helicobacter pyroli activity and 7.81µg/mg and MIC90 11.94 comparing with isoniazid standard (IS) at
concentration 0.24µg/mg and MIC90 0.4 against tuberculosis activity. The IC50 of PGL against alpha-glucosidase activity
comparing with values of Acarbose (A) showed that inhibitory percentage was lower at 1000µg/mg (79.22) than the value of
acarbose (90.10) at the same concentration, while IC50 of PGL was higher (46.6) than that of acarbose (30.57). Percentage of
albumin denaturation inhibition at 1000 µg/mg of both PGL extract and DSS were 71.34 and 89.35µg/mg, respectively, while
IC50 of both were 50.26 and 15.12, respectively. Collagenase inhibition percentage of PGL recorded 82.34 but EGCGrecorded
93.24 at concentration 1000 µg/mg. The IC50 of PGL was higher (105.3) than the value of EGCG (40.3).
Key words : Anti-Helicobacter pyroli, anti-tuberculosis anti-diabetic, anti-arthritic, anti-aging and guava.
Plant Archives Vol. 19 No. 1, 2019 pp. 199-207 e-ISSN:2581-6063 (online), ISSN:0972-5210
Introduction
Guava (Psidium guajava L.), or, in other words, a
customary medicat ion is found in nations with hot
atmospheres, for example, South America, Europe, Africa,
and Asia as reported by Gutiérrez, Mitchell et al. (2008).
Leaves of P. guajava have a background marked by
use as a conventional prescription in nations, for example,
Taiwan, Japan, China and Korea as revealed by Díaz-
de-Cerio, Verardo et al. (2015).
H. pylori contamination has been ensnared in the
improvement of gastric malignancy, a multifactorial
malady and a main source of mortality. The hazard
factor s for gastric growth have been app eared to
incorporate natural factors and factors that impact host-
pathogen connection and in addition the mind-boggling
transaction between these variables, this meaning was
sup port ed by Zhang, Zhang et al. (2 017). Modern
lifes tyle, high feelings of anxiety, smoki ng, and
unnecessary liquor utilization, dietary lacks, and delayed
utilization of non-steroidal calming drugs (NSAIDs) are
among the most impor tant etiological ecolo gical
components as showed before by Sharifi-Rad, Fokou et
al. (2018). This bact erial contamination has been
connected to the inception of incessant gastritis that could
later prompt adenocarcinoma of the digestive tract as
stated by Sipponen and Marshall (2000). However, it may,
a few systems have been proposed to speak to t he
contribution of H. pylori disease in tumor genesis. A few
200 Taha F. Taha et al.
bacterial destructivenes s factors, for example, the
cytotoxin-related quality A (CagA) protein, present in the
DNA addition component Cag pathogenicity island
(CagPAI), were observed to be of noticeable significance
in carcinogenesisaccording to the data of Park, Forman
et al. (2018).
My cobact e r ium tub e rcu l o sis , a f acultative
int racellular organism having a place with the M.
tuberculosis complex, is the mos t vit al reason for
tuberculosis (T B) in people. In addit ion to M .
tub e rcu l o sis , differ ent individua ls f rom t he M.
tuberculosis complex that can cause tuberculosis in
people incorpor a t e Mycobact e rium bovis ,
Mycobacterium africanum, Mycobacterium microti,
and Mycobacterium canetti these reported before by
Aro, Dzoyem et al. (2015). Tuberculosis, an old yet
emerging in-divisive ailment, is one of the main sources
of human grimness and mortality as shown by Nguta,
Appiah-Opong et al. (2015). The alarming rise of multi-
tranquilize safe (MDR), widely medicate resistant (XDR)
and right now, thoroughly sedate resistant (TDR) M.
tuberculosis strains, which are hard to control with the
at present accessible fundamental enemy of tubercular
medications available and the expanded occurrence of
TB related with viral contamination such as HIV, have
as of late muddled the chemotherapeutics of tuberculosis
as stated before by Daletos, Kalscheuer et al. (2015).
Diabetes mellitus (DM), a standout amongst the most
widely recognized metabolic scatters around the world,
is expanding. In 2013, 382 million grown-ups worldwide
had diabetes, and 592 million are anticipated to be
influenced by 2035 as reported before by Guariguata,
Whiting et al. (2014). The metabolic disorder has diverse
parts, for example, stomach heftiness, impeded glucose
digestio n, dyslipidemia and hyperten sion, which
synergistically increment the peril of cardiovascular
sickness and additionally diabetes, this being obviously
associated with untimely mortality. Pre-diabetes is viewed
as a fundamental etiology of metabolic disorder, portrayed
by a mix of overabundance muscle versus fat and insulin
obstruction, and showed by hindered fasting glucose and
additionally impeded glucose resistance, along these lines
bringing about hyperglycemias revealed by Grundy (2012).
The essential focus of hyperglycemia seems, by all
accounts, to be the endothelial cells, which may actuate
endothelial brokenness and quickened atherosclerosis.
Herder, Dalmas et al. (2015) stated that these procedures
are related to the advancement of a vascular incendiary
reaction, with the inclusion of a few go-betweens, including
receptive oxygen metabolites, chemokines and master
provocative cytokines, which are unmistakably in charge
of the cardiovascular inconveniences that are the main
source of dismalness and mortality related with diabetes.
Rheumatoid arthritis (RA) is an immune system
sickness that assaults the joints of the body and causes
constant inflammation in the synovium. Inflammation in
RA joints is an exceptionally mind-boggling process and
includes the cooperation of an assortment of inflammatory
cells, auto-antibodies, and cytokines this was documented
by McInnes and Schett (2011). Conventional medications
that have been used to conquer joint inflammation and
restrain the advancement of RA include the illness altering
hostile to rheumatic medications (DMARDs) as reported
before by Conditions (2009). Traditional prescription
utilizing plant separates keeps on giving wellbeing inclusion
to more than 80% of the total populace, particularly in
the developing world. Organization (2002) recently, the
utilization of plant extracts for joint pain treatment is being
advanced in the USA, particularly after the withdrawal
of FDA-affirmed calming drugs. One of the plants that
can possibly be produced as an anti-arthritic sedate is
the guava plant (Psidium guajava). Guava contains
tannins, phenolic compounds, flavonoids, volatile oils,
sesquiterpenes and triterpenoids as investigated by Porwal
et al. (2012). Flavonoids and phenolic compounds
contained in guava have been proven to be antioxidant
and anti-inflammatory where, Barbalho et al. (2012) found
that.
Schlotmann et al. (2001) showed that the procedure
of skin aging has been isolated into two classifications:
Intrinsic and extraneous maturing Inherent skin aging or
common maturing is caused by changes in the versatility
of the skin after some time. Extraneous skin aging is
predominately a consequence of introduction to sunlight
based radiation (photoaging) as stated before by Aslam
et al. (2006). UV presentation makes physical changes
the skin because of adjus tments that happen in the
connective tissue by means of the development of lipid
peroxides, cell substance and catalysts where, Lee et al.
(2000) found that. It is a natural complex process, affected
because of the contribution of both inherent, (for example,
hereditary, hormonal and digestion changes) and outward
(especially ultraviolet A and B radiation from the sun)
factors. These fact ors prompt a decay of the skin
structure, its appearance (like wrinkles, pigmentation, and
changes in thickness of skin and so on.) and work. In
fact, the logical comprehension of the maturing procedure
empowered new test systems to be created and connected
to regular item examine. Thus, hostile to maturing property
of plant extracts would now be able to be evaluated by
restraint of particular (key) catalysts (biomarkers),
pa rticularly elas tase, hyaluronidase, an d network
In vitro Bio-Medical Studies on P. guajava Leaves 201
metalloproteinase (MMP’s), which are associated with
the biochemical procedures/ pat hway, Inhib ition of
collagenase movement assumes an essential job in
securing the uneven turn over or fast breakdown of
collagen in human aggravated or UV irradiated skin. As
of late, plants have been generally examined and found
to have hostile to collagenase and against elastase
activities as documented in the study by Meinke et al.
(2010). The utilization of cell reinforcements on the skin
is likewise an imp erative procedure to avert harm
disturbed by oxidative pressure as reported in the study
by Pinnell (2003). A high grouping of cancer prevention
agents in healthy skin details permits the infiltration of
the epidermis and dermis. The fundamental preferred
standpoint of the topical organization when contrasted
with the oral organization to treat and anticipate skin
conditions is the immediate conveyance of bioactive
substances to the objective zone, consequently wiping
out worries about foundational dissemination as stated
by Meinke et al. (2010) in their study.
The goal of thisstudy is the identification of Psidium
guajava L. leaves extract for polyphenolic, flavonoids
and alkaloids components by HPLC and assessed the
activity of ethanol 70% extract on Helicobacter pyroli,
Anti-tuberculosis, Anti-diabetic, anti-arthritic and Anti-
aging activities.
Materials and Methods
Plant material
Guava ( Psi d i um gua j a va L.) , leaves were
assembled from Salhia ranches, Sharkia, Egypt. The
leaves were dried for four days at room temperature
(20-30°C). Thereafter the dried leaves were crushed to
a fine powder.
Preparation of Psidium guajava L. leaves extract
The dried powder of leaves test was separated by
maceration with ethanol 70% (60 g dried powder into
600 ml of 70% ethanol) for 24 hr. at room temperature.
In the wake of blending, the recuperated filtrate was dried
in a rotational evaporator for 30 minutes at 40°C. The
yield of concentrates was then lyophilized and put away
at -40°C in shut compartments until required.
HPLC analysis of Psidium guajava L. leaves extract
HPLC investigat ion was perfo rmed on HPLC
(Agilent Technologies, Waldbronn, Germany) device with
a vacuum degasser, autosampler, a double pump GBC
LC 1110 and an indicator GBC UV/vis were utilized for
the chromatographic assurance. Phenolic and flavonoids
segments were dis en gaged by using KROMAS IL
segment (4.6 ×150 mm, 1.8 mm molecule estimate)
working at 25°C and a stream rate of 0.8 mL/min. what’s
more, 1 mL/min., individually) (Berek and Tarbajovská,
2002) the versatile stages utilized were methanol : water
: tetrahydrofuran with acidic corrosive (23:75:1:1%)
(Phase An) and acetonitrile (stage B) for phenolic parts
and acetonitrile: water: formic corrosive (85:14:1) and
acetonitrile (stage B) for flavonoids intensifies the
separated components were observed in grouping first
with GBC U.V/vis at 280 and 356 nm.
Conditions for alkaloids discovering HSC -18,3 µgm
particle size (50×4.6 nm I.D) column, Mobile phase :
acetonitrile, detection :UV set at 290 nm, Flow rate 1.0
ml/min, injection :20µL. The sequence of the eluted
standard was 50 µg/ml (Omer, 2013).
As ses of ant i-Helico bacter pylor i Activ ity an d
(MIC90) of PGL
Antibacterial activity of guava leaves extracts against
Helicobacter pylori was determined by micro-well
dilution methods. The inoculum of Helicobacter pylori
was prepared and the suspension to 106 CFU/mL. The
extract under investigation and the standar d drug
(clarithromycin) were prepared in dimethyl sulfoxide
(DMSO) and subsequent tow fold dilutions (1000-0.03
µg) were performed in a 96- well plate. Each well of the
microplate included 40 µl of the growth medium (brain
heart infusion (BHI) plus 10% fetal bovine serum (FBS),
10 µL of inoculum and 50 µl of the diluted extract. The
clarithromycin and DMSO are used as the positive and
negative control, respectively. The plates were incubated
at 37°C for 3 days, in 5% O2, 10% CO2 and 85% N2
atmosphere. After that, 40 µL of 3-(4,5-dimethyl-thiazol-
2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) at a final
concentration 0.5 mg/ml freshly prepared in water was
added to each well and incubated for 30 min. the change
to the purple color indicated that bacteria were biologically
active. The inhibition percentage was calculated using
the given formula:
% inhibition = Abs Control – Abs Sample × 100/Abs
Control
The concentration of samples (inhibitors) required
for 90% of inhibition (MIC 90) was determined from
corresponding dose-response curves. The MIC was taken
to the lowest concentration, where no change of color of
MT T was determined using an automatic ELISA
microplate reader at 620 nm. The MIC values were done
in triplicate.
Anti-tuberculosis activity of PGL
The anti-mycobacterial action of guava leaves extract
was assessed against Mycob acterium tuberculosis
202 Taha F. Taha et al.
(RCMB 010094-9) utilizing the microplate Alamar blue
measure (MABA) as depicted by Collins and Franzblau
(1997).
Anti- diabet ic activ ity of PGL ( α - g lucosidase
inhibitory activity)
The α-glucosidase inhibitory activity was done by
the standard s t r ategy with a minor cha nge as
recommended by Shai et al. (2011).
Anti-arthritic activity of PGL
The act ivity was per f ormed u s ing albumin
denaturation following to the method of Singh and Sharma
(2015) with few modifications.
The inhibition percentage of albumin denaturation was
calculated as follow :
% Inhibition of protein denaturation = 1-[A1/A2]100
Where, A 1 = Absorbance of control
A 2 = Absorbance of test/standard example with
albumin arrangement
The IC50 esteem was characterized as the fixation
to repress half of the protein denaturation under the
measurement conditions.
Anti-aging (Anti-Collagenase) activity of PGL
Before screening in all measures, spectra for all
concentrates were recorded on a Car y 30 0 UV-
unmistakable spectrophotometer to check for impedance
and moves in the lambda max. The method of asses was
utilized depended on spectrophotometric techniques
recommended by Thring et al. (20 09) with a few
adjustments for use in a microplate peruse.
The inhibition percentage was calculated according
to the following formula :
The percentage of collagenase inhibition (%)
1001
C
S
Where, ‘S’ is the corrected absorbance of the samples
containing collagenase inhibitor (the enzyme activity in
the presence of the samples), and ‘C’ is the corrected
absorbance of controls (the enzyme activity in the absence
of the samples).
The IC50 value was defined as the concentration of
the sample to inhibit 50% of collagenase under the assay
conditions.
Statistical analysis
The results are conveyed as means ± SD (Standard
Deviation). All tests were performed in triplicate and
reiterated no under three times. Quantifiable complexity
between bundles was controlled by one-way examination
of progres s (ANOVA). Ap-es t eem < 0.05 was
considered statistically significant.
Results
Identification and quantification of different phenolic
acids, flavonoids and alkaloids in Psidium guajava
L. (PGL) µg/mg leaves ethanol 70% extract
The HPLC chromatogram of ethanol 70% extract
of Psidium guaj ava L demonstrated that identified
phenolic comp ounds were gallic acid , resorcinol,
chlorogenic acid, caffic acid, coumaric acid, ferulic acid,
cinnamic acid, resormarinic acid and syringic acid as
showed in (table 1) among them gallic acid, caffic acid,
coumaric acid, ferulic acid and cinnamic acid have highest
concentration 4.71, 4.73, 3.82, 3.55 and 3.49 µg/mg,
resp ectively followed by resorcino l (3.09µg/mg),
chlorogenic acid (2.93µg/mg) and syringic acid (2.85 µg/
mg). Res u lts in table 1 revea led tha t flavon oids
compounds were 7 identified compounds catchin,
kaempferol, rutin, quercetin, hesperetin, apigenin and
quercitrin among them quercetin, hesperetin, kaempferol,
quercitrin and rutin recorded highest concentration (8.94,
7.61, 7.55, 7.13 and 6.37 µg/mg), respectively followed
by catchin (5.12 µg/mg) and apigenin (4.83 µg/mg). Also,
table 1 results showed three compounds of alkaloids were
identified, which a r e ka emp f ertin (1.89µg/mg) ,
isoquinoline (1.24µg/mg) where corilagin had the highest
concentration (2.13µg/mg) . It can be noticed that
flavonoids presented the highest fraction of PGL ethanolic
70% extract, especially quercetin (8.94µg/mg) which
known as a high antioxidant agent.
Anti-Helicobacter pylori activity (% Helicobacter
pylori inhibitory) of Psidium guajava L. (PGL) µg/
mg leaf ethanol 70% extract
The (% Helicobacter pylori inhibitory) extend from
21.34 ±1.2 at concentration 0.24µg/mL to 100 ±2.5% at
concentration 125 µg/mL for guava leaves ethanol extract
and 81.35± 1.5 to 100±0.00% for clarithromycin at
concentrates extended from 0.24 to 1.94 µg/mL. MIC90
were 26.6 and 0.7 for guava extract and clarithromycin,
respectively (tables 2).
Anti-tuberculosis activity (TB) of Psidium guajava
L. (PGL) µg/mg leaves ethanol 70% extract
The Anti-mycobacterial action of Psidium guajava
L. extract was given in (table 3). The outcomes indicated
most activity against the M. tuberculosis (TB Inhibitory
%) 100±0.00, at concentration 7.81µg/mL of PGL, but
of IS was 100 inhibitory percentage at concentration 0.24
µg/mL. The MIC90 value of PGL was 11.94, while it
was 0.4 of IS. These results were illustrated in table 3.
Anti-diabetic activity (Alpha-glucosidase inhibitory
%) of Psidi um guajav a L. (PGL) µg/mg leaves
ethanol 70% extract
The Psidium guajava ethanol 70% ex t ract
uncovered a noteworthy inhibitory activity of the alpha-
glucosidase enzyme. The percentage inhibition at 7.81 –
1000 µg/mL convergences of Psidium guajava separate
demonstrated a dosage subordinate increment in rate
hindrance. The rate restraint changed from 29.53±1.3-
79.22 ±1.5% for the most astounding focus to the least
fixation this was close to Acarbose (An) as standard the
Table 1 :HPLC analysis data of phenolic, flavonoid and
alkaloids compounds in Psidium guajava L. (PGL)
µg/mg leaf ethanolic 70% extracts.
PGL
Concentration
(µg/mg)
Gallic acid 4.71
Resorcinol 3.09
Chlorogenic acid 2.93
Caffeic acid 4.37
Coumaric acid 3.82
Ferulic acid 3.55
Cinnamic acid 3.49
Resormarinic acid 2.49
Syringic acid 2.85
Catechin 5.12
Kaempferol 7.55
Rutin 6.37
Quercetin 8.94
Hesperetin 7.61
Apigenin 4.83
Quercitrin 7.13
Kaempfertin 1.89
Isoquinoline 1.24
Corilagin 2.13
Phenolic compounds
Flavonoid compounds
Alkaloids
compounds
Table 2 :Inhibitory percentage and MIC90 of PGL ethanolic
extract comparing with values of clarithromycin (C)
against Helicobacter pylori activity.
C PGL
Concentration (µg/mL)
Helicoba cter pylori
inhibitory (%)
125 100±0.00 100 ±2.5
62.5 100±0.00 93.25 ±0.58
31.25 100±0.00 82.34 ±0.72
15.63 100±0.00 73.14 ±0.63
7.81 100±0.00 66.32 ±1.2
3.9 100±0.00 60.38 ±1.3
1.95 100±0.00 54.28 ±1.2
0.98 92.45±1.2 49.31 ±2.5
0.48 87.65±0.58 32.25 ±0.63
0.24 81.35±1.5 21.34 ±1.2
00±0.00 0±0.00
*MIC90 0.7 26.6
Table 3 :Inhibitory percentage and MIC90 of PGL ethanolic
extract comparing with values of isoniazid standard
(IS) against anti-tuberculosis activity (TB)
IS PGL
Concentration (µg/mL)
TB Inhibitory (%)
31.25 100 ±0.00 100±0.00
15.63 100 ±0.00 100±0.00
7.81 100 ±0.00 100±0.00
3.9 100 ±0.00 93.25 ±2.5
1.95 100 ±0.00 86.34 ±1.5
0.98 100 ±0.00 71.28 ±0.58
0.48 100 ±0.00 59.38 ±0.63
0.24 100 ±0.00 38.14 ±2.1
0.12 92.35 ±0.58 21.07 ±1.5
0.06 86.34±1.5 10.69 ±0.36
00 ±0.00 0 ±0.00
MIC90 0.4 11.94
Table 4 :Inhibitory percentage and IC50 of PGL ethanolic
extr act comparing with values of Acarbose (A)
against alpha -glucosidase activity (as diabe tic
parameter).
A PGL
Concentration (µg/mL)
Alpha-glucosidase inhibitory
%
1000 90.10 ±0.58 79.22 ±1.5
500 86.34 ±1.20 74.51±2.1
250 71.34 ±1.50 68.72±1.2
125 63.42 ±2.10 62.41±0.58
62.5 60.14 ±0.72 58.41±1.3
31.25 50.31 ±1.50 41.85±0.72
15.63 43.28 ±1.20 36.13±2.1
7.81 32.15 ±0.58 29.53±1.3
00 ±0.00 0 ±0.00
*IC50 30.57 46.6
In vitro Bio-Medical Studies on P. guajava Leaves 203
outcomes were gone from 32.15 ±0.58 to 90.10 ±0.58 at
same concentrates (table 4).
Inhibition of protein denaturation (%) by Psidium
gua j a v a L. (PGL) µg /mg leaves et hano l 70 %
extract
The consequences of in vitro denaturation of egg
albumin of P. guajava L. condensed in (table 5) guava
leaves extract demonstrated an inhibition in protein
denaturation with an IC50 estimation of 50.26 μg/mL (IC50
estimation of Diclofenac sodium is 15.12 μg/mL). As
appeared, P. guajava L. had demonstrated hindrance of
protein denaturation closest to Diclofenac sodium
showing that P. guaj ava L. is an intense enemy of
ligament operator. The greatest level of hindrance was
communicated at 1000 μg/mL.
Anti-aging (Collagenase inhibitory %) of Psidium
gua j a v a L. (PGL) µg /mg leaves et hano l 70 %
extract
The incubation of the enzyme with the concentrate
fundamentally inhibited the protein. A grouping of 1000
µg/mL extract indicated 82.34 ±1.2% catalyst hindrance,
while bring down convergences of 7.81, 15.63, 31.25
and 62.5 µg/mL caused 6.31±0.72%, 16.31 ±1.3%, 22.14
±0.58% and 41.14 ±1.2% protein enzyme inhibition,
res pectively. These outcomes were compared with
Epigallocatechin gallate the outcome at high concentrate
1000 µg/mL was 93.24±1.78% though bring down
groupings of 7.81, 15.63, 31.25 and 62.5 µg/mL standard
caused 19.34 ±2.5%, 32.55 ±0.58%, 48.69 ±0.63% and
53.21±1.2% enzyme inhibition, respectively (table 6).
Discussion
Guava leaves extracts are a potential wellspring of
common cell reinforcements as showed by res ults
obtained by Ojan and Nihorimbere (2004), who stated
that these cell reinforcement properties are related to its
phenolic compounds, for example, ferulic acid, quercetin,
guavin B, ascorbic acid, Gallic corrosive and caffeic acid
these results were supported by Thaipong et al. (2005).
The phenolic profile of Spanish guava leaves has as of
late been accounted for with an appraisal of the grouping
of various flavonoids. Among these, flavonols and flavan-
3-ols were the real subclasses found in the Andalusia
guava leaves as stated by Díaz-de-Cerio et al. (2016),
who revealed thatmain phenolic segments beforehand
recognized in the product of Psidium guaja va are
glycosides of myricetin, quercetin, kaempferol, luteolin,
apigenin and isorhamnetin, a B-type proanthocyanidin
(gallocatechin-catechin) and two benzophenone mixes
stated recently by Rojas-Garbanzo et al. (2017). Results
in this study were in harmony with those reported above
attributed to the highest effect of these compounds as
an antioxidant agent.
Guava leaves extract give has well extreme against
Helicobacter pylori and as Anti-tuberculosis. Anti H.
pylori Potency of Psidium guajava L. leaves was IZD
= 33.0 ± 2.3 mm (240 µg/circle) as reported by Salehi et
al. (2018). T he numbers of synergetic interactions
between PGL against M. tuberculosis are few. In this
way, more examination is required into the system of
activity of flavonoids. Tulin (2015) demonstrated that
quercetin is a piece of the 3-hydroxy gathering, which
enhances antimycobacterial activities and synergistic
cooperation. Kaempferol likewise has a functioning
Table 5 :Inhibitory percentage and IC50 of PGL ethanolic
extract comparing with values of diclofenac sodium
standard (DSS) drug against protein denaturation.
DSS PGL
Concentration (µg/mL)
Inhibition of protein
denaturation (%)
1000 89.35 ±0.58 71.34 ± 1.5
500 84.12 ±1.2 62.58 ± 2.1
250 76.52 ±0.63 60.14 ± 1.5
125 70.14 ±0.58 58.34 ± 1.2
62.5 68.28 ±0.63 51.39 ±2.1
31.25 59.14 ±1.2 47.84 ±1.5
15.63 51.21 ±0.58 31.24 ±1.3
7.81 31.12 ±1.2 16.85 ±1.5
00 0
*IC50 15.12 50.26
Table 6 :Inhibitory percentage and IC50 of PGL ethanolic
extract comparing with values of Epigallocatechin
gallate EGCG against aging marker (Collagenase
activity as aging parameter).
EGCG PGL
Concentration (µg/mL)
Collagenase inhibition %
1000 93.24 ±1.78 82.34 ±1.2
500 84.22 ±1.3 73.25 ±1.2
250 69.35 ±0.58 66.17 ±0.63
125 58.98 ±1.5 53.97 ±2.5
62.5 53.21 ±1.2 41.14 ±1.2
31.25 48.69 ±0.63 22.14 ±0.58
15.63 32.55 ±0.58 16.31 ±1.3
7.81 19.34 ±2.5 6.31 ±0.72
00 ±0.00 0 ±0.00
*IC50 40.3 105.3
204 Taha F. Taha et al.
compound required beside the hydroxyl gatherings in ring
B, giving potentiation activity of flavonoids.
Guava leaves extract indicates anti-diabetic action,
the in vitro activities of α-glucosidase proteins in guava
extrac t was a polymerized polyphenol. Moreover,
polysaccharides from guava leave additionally displayed
α-glucosidase hindranceas revealed by Zhang et al.
(2016).
The IC50 of guava leaves extract to inhibition of
protein denaturation was near to diclofenac sodium
standard (DSS). The provocative reaction happens when
cells and body tissues are harmed by natural, chemical,
or physical upgrades, for example, microorganisms, injury,
poisons, or warmth. It is a standout among the most
imperative safeguard components, or, in other words,
evacu atio n of the ha r mful imp rovements and
commencemen t of the recup era t ing p rocedure.
Macrophages are entered players in different fiery
ill nesses and in the resistant reaction where they
discharge pro inflammatory middle people and proteins,
including interleukin-6 (IL-6), tumor corruption factor-a
(TNF-a), cyclooxygenase-2 (COX-2) and inducible nitric
oxide synthase (iNOS) as stated by Grip et al. (2003).
Plant extracts have been utilized as a wellspring of
pharmaceuticals for a wide assortment of a human issue.
Homegrown and characteristic items have as of late
gotten expanded consideration in view of their organic
and pharmacological activities as stated by Eze et al.
(2017). Psidium guajava L. leaves have been utilized
for the treatment of stiffness, fever, joint pain and other
provocative conditions as found (Kim et al., 2015) in
their study. Quercetin may add to mitigating action as
mentioned by Metwally et al. (2010). Flavonoids have
natural activities, for example, antioxidant, anti-aging,
hostil e to cancer - caus ing, mit i gat ing, agains t
atherosclerosis, cardiovascular insurance and change of
endothelial capacity, the hindrance for angiogenesis and
cell multiplication activities as stated by Rahman et al.
(2015) in their study. Phenolic mixes add to pain relieving,
calming, against microbial, hepatoprotective and cancer
prevention agent activities (Choudhury et al., 2012).
The inhibitory effects on collagenase act ivities
showed that 82.34±1.2% ethanol PGL extract ata
concent ration of 1000μg/mL pos ses sed moderate
inhib ition comp a r a b le to that of 9 3 .2 4 ±1.78%
Epigallocatechin gallate at the same concentration. The
effect of PGL as anti-aging may be due to its polyphenolic
components. Sin and Kim (2005) inspected the inhibitory
activities of different flavonoids, including the flavanones,
flavones/isoflavones and flavonols, on collagenase from
Clostridium histolyticum to build up their helpful potential
against skin aggravation and photoaging. As a rule, the
flavonols were more grounded inhibitors than the flavones/
isoflavones, and this showed the significance of the C-3
hydroxyl substitution. Quercetin was the most dynamic
flavonoid the IC50 was 286 microM. This work needs to
study the action of guava leaves extract on therapeutic
activities under-cons idered es pecially Heli cobacter
pyroli, anti-tuberculosis and anti-aging.
Conclusion
In conclusion, Psidium guajava L. leaves as cheap
raw material contains high amount from Gallic acid,
Quercetin and Corilagin according to HPLC analysis these
compounds play a great role as antioxidants activities.
The ethanol 70% extract of the plant gave high possess
inhibition for Helicobacter pyroli, Anti-tuberculosis, Anti-
diab etic, anti-arthritic and Anti-aging, PGL will be
accepted to be more adequate by the human body, while
reducing the risk of using synergistic drugs.
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