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Natural components thought to have a particular benefit nowadays. They are playing a necessary role in health attention. Sea cucumber is theorized to be a significant source of bioactive components. Moreover, Flavonoid glycosides are a group of polyphenols with different glycoside substituent that possesses diverse pharmacological activities and can help management coronavirus. So, this study aimed to extract saponin and polysaccharides from sea cucumber, also, extract both of naringin, hesperidin from orange peel and convert hesperidin to hesperetin then chemical analysis of all extracted compounds was performed to confirm their structure and evaluate their antioxidant, antibacterial and antitumor activity. The concentrations of saponin, naringen, hesperidin, hesperetin, and ascorbic acid, which scavenged 50% of DPPH radicals, were 10.50, 0.13, 0.13, 0.66, and 0.0025 mg/ml respectively. Furthermore, Cell viability of saponin, hesperidin and hespertin showed a growth inhibitory effect, IC50 28.78, 236.40 and 73.99. The obtained data indicated that sea cucumber saponin, polysaccharides, and orange peels, may provide a promising new therapeutic approach to HEPG2 cancer cells. Also, these compounds were effective antioxidant, so they may be effective and scavenge free radicals which resulted from the disease. Research Highlights a) Saponin (holothurians) and polysaccharide have been extracted from sea cucumber. b) Orange peel was treated with a different solvent to extract Naringin, Hesperidin and hespertin. c) All of the extracted compounds were chemically analyzed to confirm by chemical analysis and evaluated their antioxidant, antibacterial and antitumor activity.
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20555
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Research Article
ISSN: 2574 -1241
Isolation, Characterization and the Biological Activity
of Some Natural Components of Marine Sea Cucumber
and Orange Peel
DOI: 10.26717/BJSTR.2020.27.004463
Amira Ragab El Barky* and Tarek Mostafa Mohamed
Biochemistry Unit, Chemistry Department, Faculty of Science, Tanta University, Egypt
*Corresponding author: Amira Ragab El Barky, Biochemistry Unit, Chemistry Department, Faculty of Science, Tanta
University, Egypt
Figure 1: Graphical abstract.
ARTICLE INFO Abstract
Received: April 14, 2020
Published: April 23, 2020
Citation: Amira Ragab El B, and Tarek Mo-
stafa M. Isolation, Characterization and the
Biological Activity of Some Natural Compo-
nents of Marine Sea Cucumber and Orange
Peel. Biomed J Sci & Tech Res 27(2)-2020.
BJSTR. MS.ID.004463.
Abbreviation: FT-IR: Fourier Transforms
Infrared Spectroscopy; UV: Ultraviolet
Spectra; XRD: X-ray Powder Diffraction;
TGA: Thermal Gravimetric Analysis; DPPH:
2, 2-Diphenyl-1-Picrylhydrazyl-Hydrate;
MTT: 3-4,5-Dimethylthiazol-2-yl)-2,5-Di-
phenyltetrazolium Bromide.


of bioactive components. Moreover, Flavonoid glycosides are a group of polyphenols
with different glycoside substituent that possesses diverse pharmacological activities
and can help management coronavirus. So, this study aimed to extract saponin
and polysaccharides from sea cucumber, also, extract both of naringin, hesperidin
from orange peel and convert hesperidin to hesperetin then chemical analysis of all
          
antioxidant, antibacterial and antitumor activity. The concentrations of saponin,
naringen, hesperidin, hesperetin, and ascorbic acid, which scavenged 50% of DPPH
radicals, were 10.50, 0.13, 0.13, 0.66, and 0.0025 mg/ml respectively. Furthermore,
Cell viability of saponin, hesperidin and hespertin showed a growth inhibitory effect,
IC50 28.78, 236.40 and 73.99. The obtained data indicated that sea cucumber saponin,
polysaccharides, and orange peels, may provide a promising new therapeutic approach
to HEPG2 cancer cells. Also, these compounds were effective antioxidant, so they may
be effective and scavenge free radicals which resulted from the disease.
Keywords: Sea cucumber; Saponin; Polysaccharide; Naringin; Hesperidin; Hesperitin
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20556
Research Highlights
a) Saponin (holothurians) and polysaccharide have been
extracted from sea cucumber.
b) Orange peel was treated with a different solvent to extract
Naringin, Hesperidin and hespertin.
c) All of the extracted compounds were chemically analyzed
        
antibacterial and antitumor activity.
Introduction
       
nowadays [1]. They are playing a necessary role in health attention
and more probable to produce a pharmacologically effective
 
sea cucumber having several active components [3], distinguishing
by their nutritional value [4]. Marine invertebrates, sea cucumber
own a worthy bioactive ingredient, for instance, holothurians that
exhibited biological effectiveness and have a therapeutic effect [5].
Furthermore, it holds more than 50 forms of nutrients inclusive
amino acids, polyunsaturated fatty acids, vitamins, and trace
element and active substances such as polysaccharides, proteins
and glycosides [6]. Saponin which is a bioactive compound that
exist in large amounts in both marine sea cucumbers and sponges,
and it has a difference of biological and pharmacological effect [7],
     
agent [5,8].
Polysaccharides have numerous activities, for example, anti-
tumor, immune promoting, and antioxidant, polysaccharides
are one of the remarkable ingredients of natural compounds
[9]. Citrus juice remains are at most composed of peel, juice, and
seeds. The peel composed of bioactive compounds like those
        
       
seeds and fruit pulps, that account for 50% of the original whole
fruit mass, are a by-product of the juice, marmalade and canning
manufactures [12]. Both naringin and hesperidin that consider
        

been found in the serum of people after eating or drinking orange
and grapefruit [14]. Hesperidin considers a bioactive compound
in preventing numerous diseases, for example, lowering capillary
  
Hesperidin Furthermore has the ability to monitor liver cholesterol
texture by repressing the activity of 3-hydroxy-3-methylglutaryl
coenzyme A reductase [15].
Hesperidin is functionally utilized as a supplement factor in

been related to abnormal capillary leakiness as well as the ache
in the extremities, which cause pain, impairment and leg cramps.
Supplemental Hesperidin also helps in decreasing edema which
           
corona virus COVID-19 [16,17]. Naringin displayed the capacity
       
agent [19,20], anti-breast cancer agent [21], anti-allergic [22], and
hypoglycemic compounds [23]. Naringin has been considering the
anti-angiogenesis agent [24]. Also, naringin might be the active
ingredient in suppressing osteoclastogenesis and osteoclasts in
both vitro and vivo model [25]. Furthermore, naringin has the
ability to repress polymethyl methacrylate particles induced
osteolysis in vivo [25].
Materials and Methods
Saponins Extraction
Triterpenoid saponin has been extracted from marine sea
       
Zoology department, Faculty of science, Tanta University),
according to [27]. Dried body walls of marine invertebrates see
cucumber were grinding into a powder and extracted several times
with aqueous ethanol until decolorization of ethanol. The resultant
solvent was evaporated by a rotary; the remaining part was
partitioned amidst water and chloroform and left for overnight. The
top aqueous layer has been collected and treated with n-butanol,
after that, it was evaporated and concentrated in a drying oven [8].
Polysaccharides Preparation
Sea cucumber has been ground into powder; the powder has
 

trichloroacetic acid and left overnight to precipitate protein then its

(v/v) ethanol and left overnight at 4°C. The sediment which gained
via cooling centrifuge at 4000 rpm for 10 minutes [27], and then
the supernatant was waste. Also, both saponin and Polysaccharides

been ground into powder and after that it minced in boiling distal
water for polysaccharides extraction and after the end method the
ground sea cucumber has been put in aqueous ethanol for saponin
extraction, but the yield is very limited, so the best methods of the
extraction extract each of them alone.
Naringin, Hesperidin and Hesperitine Extraction
Both naringin and hesperdine have been extracted from
mature citrus orange peels, Citrus sinensis (L.) Osbeck var. Balady
(Rutaceae), was purchased from the Egyptian market and has been

(The Herbarium-TANE, Botany department, Faculty of Science,
Tanta University, Egypt, Herbarium- TANE, Index Herbariorum
New York Botanical Garden). Air dried citrus orange peels were
ground into powder. Naringin has been extracted according to
[28,29], with some modulation, 50 g of the husk powder has been

with a frequency of 40 kHz (SB-120D, Xinzhi Technology, China)
and kept for 2 hours and left overnight in the aqueous ethanol,

procedure was repeated until decolonization of the ethanol. The
         
and obtain syrup consistency. Distilled water has been added to the
obtained concentrated syrup, the mixture was agitated at 70 °C on
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20557
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
a hot plate. 10 ml of methylene chloride has been added and the
mixture left for 4 days at 25 °C to allow crystallization of naringin
in the aqueous layer.
       
Hesperidin has been extracted according to the method described
by Belboukhari et al. (2015)  
the dried citrus orange peels was soaked in the petroleum ether
after that heated to about 40°C for 1.5 h by using a hotplate, after

powder was allowed to dry at room temperature. After that, about

extract was evaporated at rotary evaporator at 70oC for 30 min until
syrup consistency was reached, the concentrated residual liquid

preserved nocturnal in cold at 4oC. The precipitated solid was the

           
material was dissolved in dimethylformamide with continuous
stirring and heating to approximately 60 °C. Then, the equivalent
quantity of distilled water was added gradually, then it was cooled
to precipitate the hesperidin and washed with little warm water.
Conversion of Hesperidin into Hesperitin:
A known weight of hesperidin which has been extracted from
citrus orange husk has been added to methanol, then concentrated
sulfuric acid has been added in a water bath, stirred and heated
about 8 hours. The homogeneous solution which obtained was
cooled, diluted by ethyl acetate and after that, it washed with

amount of acetone, and the resulting solution was added to a stirred
mixture of distilled water and acetic acid. The gained hesperetin
has been washed and cooled [30].
      -
peridin and Hesperitin:
Fourier Transforms Infrared Spectroscopy (FT-IR):
The functional groups of all extracted compounds, saponin,
Polysaccharides, naringin, hesperidin and hesperetin were
distinguished by Fourier transform infrared spectroscopy (Model-
 -1) in the
region of infrared radiation in the [Micro analytical unit, Faculty
         
Polysaccharides, naringin, hesperidin, and hesperitin were
grounded and crushed to quite a powder with a mortar and pestle

with the help of mechanical pressure was observed at the different
coming wavelengths in FT-IR, infrared spectrum.
Determination of the Maximum Wavelength by Using
Ultraviolet Spectroscopy (UV): Maximum wavelength of each
         
     
hesperidin, and hesperetin extract were detected by Pg instruments
(UV/vis spectrometer T80, Micro analytical unit, Faculty of Science,
Tanta University, Egypt). Maximum wavelength of any component is
known as the wavelength that the component displays the farthest
absorbance. Triterpenoid marine sea cucumber saponin extract
was soluble in distilled water. Meanwhile, naringin, hesperidin,
and hesperitin were dissolved in dimethyl sulfoxide (DMSO), the
prepared extract, solutions were measured for absorbance in UV-
Visible spectrophotometer in the UV region (200 nm-800 nm) and
readings were noted down against blank. The graph was drawn
between the obtained absorbance and wavelength. The peak
obtained from the graph was taken as the most wavelength of that
compound.
X-Ray Diffraction Analysis: X-Ray Diffractometry, the patterns
of all extracted samples saponin, polysaccharide, naringin,
hesperidin, and hesperitin were determined using the X-ray
diffractometer (Siemens D5000, Germany). The investigated
          
rate average was 1°/min.
Thermal Gravimetric Analysis: Changes in the thermal
properties of saponin, polysaccharide, naringin, hesperidin,
and hesperitin were determined using [Schimadzu TG-50
 
in a previously tarred stainless-steel pan and weighted then heated
from 25 °C to 800 °C at the rate of 10 °C /min under nitrogen supply
   (Micro analytical unit of faculty of science, Tanta
University, Egypt).
Antioxidant Activity
Free Radical Scavenging of DPPH Radical: DPPH is an
antioxidant method; depend on electron-transport which offers
a violet solution in methanol [31]. Such free radical, constant at
ambient temperature, it is reduced in the existence of an antioxidant
compound, The reduction in the absorption and the changes in the
color from dark violet to light violet or yellow color of the DPPH
solution after the addition of an antioxidant has been read at
517nm., the utilize of the DPPH method supply a simple and quick
technique to estimate antioxidants activities of the compounds
under study by spectrophotometer [31]. The free radical scavenging
actions (AA) of all compounds were studied according to [32] with
  
dissolved in methanol which gives absorbance 0.90 at 517 nm [33],
25µl of the extract was added to 975µl of methanolic DPPH, the
methanolic DPPH extract mixture was shaken and put to stand in

visible spectrophotometer at 517 nm. Ascorbic acid was utilized
as a standard. The concentration of extract under study that can
diminish by 50% (IC50) amount was calculated. The free radical
scavenging action of the compound has been studied
% of Scavenging (AA) = (Abs of control Abs of the sample) / Abs of control 100 −×
% of Scavenging (AA) = (Abs of control Abs of the sample) / Abs of control 100 −×
.
Evaluation of Total Antioxidant Capacity: The total
antioxidant capacity of the extracted compounds under study
was measured as per Phosphomolybdate method according to the
proposal of [34], Phosphomolybdate method is a spectroscopic
technique that utilized to determine total antioxidant capacity, via
formation of phosphomolybdenum compound. The principle of this
assay depends on the reduction of Mo (VI) to Mo (V) by the examined
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20558
extract and next formation of a blue-green phosphate Mo (V)

was put in a glass tube and followed by 100 µ
study was added. The reaction was kept away from light and it
incubated at 95 °C for 90 min. A blank without sample was also
run. Subsequent the incubation time, the mixture has been cooled
with tap water and measured by a spectrophotometer at 765 nm.
ascorbic acid was measured as a reference.
Determination of Total Phenolic Content: The Total Phenolic
Content (TPC) has been measured according to [36], by using Folin
Ciocalteu reagent, the principle methods depend on the reduction of
tungsten and molybdenum oxides which arises a blue color, that can
be measured at 750 nm by a spectrophotometer. The concentration
of the extracted compounds under study was calculated from the
standard curve of gallic acid (0.017 - 0.1 mg/ml).
Antibacterial Activity
Saponin, naringin, hesperidin and hesperetin extract were
dissolved in distilled water and dimethyl sulfoxide respectively and
their antibacterial activities were determined using the agar well
µl of the extract under study was transferred
into each hole in the test plate Petri dish. The appearance of
a clear zone around the well in the inoculated plates is a sign of
antibacterial activities of the strains under study (all in triplicate).
The following test microorganisms were used for such purpose:
Bacteria (prokaryotic): Gram-positive cocci: Streptococcus pyogenes,
Staphylococcus aureus, and Gram-negative bacteria: Escherichia
coli, Klebsiella pneumonia, and Proteus mirabilis.
Cytotoxicity Assay
Human hepatocellular carcinoma cells (HepG2) cells and Caco-
2 cells (colon cancer cell) have been seeded in 96-well plate at a
density of 5×10, 1×10, 2×10, and 4×10 cells/well (in triplicate).
The cells have been treated with the different compounds under
the study at different concentration and measured after 22h,
post-treatment by MTT assay. Absorbance in control and treated
compounds wells have been detected by micro plate reader Elisa
[37].
Determination of Cell Viability by Using MTT Assay
Principle: Cell viability has been studied by the usage of
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye
(MTT) as the proposed method described by Oka [38]. The change
of yellow color of MTT to purple color formazan which occurs only
of the viable cells in the mitochondria that indicated the activity
of reductase enzymes. The insoluble purple formazan product is

at 630 nm using the microplate reader Elisa.
Statistical Analysis: The obtained data were statistically
analyzed by one-way analysis of variance (ANOVA) followed by the
Duncan multiple tests. All analyses were performed in triplicate and
are expressed as average as mean values ±SEM using Co Stat 6.311.

was used to be carried out statistical analysis and drawing the

Figure 2: FT-IR spectrums of the saponin extract.
(a) First method of preparation and
(b) The second method,
(c) FT-IR spectrum of Polysaccharide extract,
(d) Naringin extract,
(e) Hesperidin and
(f) Hespertin.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20559
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Results and Discussion
         
good source of bioactive components [39], they’re worth because
of their content of valuable ingredients, for instance, saponin
       
fruit are hesperidin, narirutin, and neohesperidin; which can be
extracted by aqueous ethanol or methanol solutions [40]. Saponin,
polysaccharide, naringen, hesperidin, and hesperitin have been
 
by several methods, for example, FT-IR of saponin (Figure 2),
    
Saponins showed characteristic bands infrared absorbance of the
hydroxyl group (OH) in 3425 and 3470 cm-1. Carbon-hydrogen
(C=H) absorption ranged from 2927cm-1. The C=C absorbance
was observed at 1406 and 1535 cm-1   
was found to be at 1639 and 1645 cm-1. Oligosaccharide linkage
absorptions to sapogenins, that is C-O-C, were evident in 1096 cm-1
region [8].
The aforementioned infrared functional group absorptions,
characteristic of saponins, have been referred to the existence of
the oleanolic acid-ester, which characterized by the C=O infrared
absorbance, these triterpenoid saponins are bidesmosides
because they own two connexions by glycones to the sapogenin
         
results that saponins are detectable in both crude aqueous and /
or alcoholic extracts by using FT-IR spectroscopy [41]. Moreover,
the FT-IR spectra of the extracted polysaccharide are displayed in
(Figure 2) and exhibit the typical signals in the range from 4000
to 400 cm. The characteristic strong broad absorption at 3431.34
cm corresponded to the O-H groups. The absorption peaks
at 2927.37 cm and 2862.68 cm indicated the aliphatic C–H
stretching vibrations. The strong band at 1639.62 cm indicated
the absorption of C=O. The results of FT-IR spectroscopy could be
         
band at 1465 and 1388 cm–1 were on behalf of the C-H deformation
vibration, the signals at 851–1245 cm
area of carbohydrates, among which the bands at 1018, 1082 cm
were the characteristic absorptions of the pyranose ring.
Typical peaks at 928cm, 837 cm, 779 cm, was assigned to

bands at 1245 cm–1     
presence of sulfate groups, a band at 1070.96 suggest the presence
           
pellets showed characteristic bands, the OH group at 3411.61 cm-1,
the C-H at 2926.49 and 2861.55 cm-1[46], the band at 1729 cm-1
which indicated presence of C=O the carbonyl stretching vibration
of the carboxyl group (COO), The infrared bands around 1626
cm-1 which indicated the C=C stretching that is attributed to the
presence of aromatic or benzene rings. The vibrational bands at
around 1457.86 cm-1 were aliphatic and aromatic (C–H) group. The
bands in the range 1360–1050 cm-1 were due to the C–O stretching
vibration of carboxylic acids and alcohols and band at 1263 and
1180 cm-1 indication of C-O-C and O–H of polysaccharides. Spectra
absorbance at the wavenumber of 900 cm-1 or less was assigned
      
cm-1 and 1080 cm-1 is characteristic to the benzene ring stretching
vibrations [48].
Figure 3: UV- spectrum of
(a) Saponin(λmax282nm),
(b) Polysaccharide(λmax262nm),
(c) Naringin(λmax215,296and396nm),
(d) Hesperidin(λmax296,312nm)and
(e) Hespertine(λmax296,313nm)
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  
a strong band of OH at 3554 and 3469 cm-1, CH (aliphatic) at 2926,
2855 cm-1, C=C (aromatic) at 1645, 1600 cm-1 and of C=O at 1735
cm-1, C-O at 1283 and 1069 cm-1. The hespertine compound which
has been obtained from hesperidin (Figure 2), The FT-IR spectrum
           -1, CH
(aromatic) at 2926 – 2861 cm-1, C=C (aromatic) at 1638, 1600, 1515
cm-1 and of C=O at 1716 cm-1, C-O at 1186cm-1 and 1089 cm-1 [5].
Utmost of the saponins compounds display a major absorption peak
in the range of 250–350 nm. Saponin extracts have max at 282 nm
[8]. Furthermore, the max of sea cucumber polysaccharide has
been displayed in (Figure 3). UV-visible spectra are mostly utilized
for the testing chromophore groups of the atom that distinguished
by a strong absorbance electronic transition. The UV spectra in the
present research showed that the maximum absorbance was at 262
nm which is matching with [49].
Moreover, The UV spectrum of naringen extract showed
maximum absorption peaks at 215.2, 296 and 396 which in
agreement with [50] they showed the absorption peaks of naringen
at 214, 283.6 and 331.1nm. Furthermore, the UV spectrum of the
hesperdine extract showed maximum absorption at 296, 312,
and 345 nm. and for hespertine at 290 nm which in accordance
with [30]. The thermal behavior of the extracted compounds was
studied by TGA in the range of 25 to 800 °C be average 10 °C min
under a nitrogen atmosphere (Figure 5). TGA of sea cucumber
saponin, the thermal decomposition occurs in two successive
steps, which indicated that saponin is a stable compound. Thermal
stability is one of the most main physicochemical properties for the
applications of the polysaccharide. The TGA analysis of isolated sea
cucumber polysaccharide was carried out and the experimental
results showed that, the degradation temperature occurs in three
successive steps.
Figure 4: XRD of the
(a) extracted Saponin and commercial reference standard saponin,
(b) Polysaccharide,
(c) Naringin,
(d) Hesperidin and
(e) Hespertin.
The results indicated that marine sea cucumber polysaccharide
extract has altitude thermal steadiness. This data suggested that
marine sea cucumber polysaccharide extract could be used for
any function and in chemical adjustment [51] Such verity points
out that, the extract has not to be submitted to the degradation
temperature to not compromise the physical integrity of the extract.
TGA analysis of naringin showed that approximately 90% weight
loss has been observed more than 200 °C, the obtained data were

which indicated the decomposition of naringen was found to be
around 250 °C. On the other hand, hesperdine decomposition steps
   
266 and 500 °C and from 500 to 800 °C which can be attributed
to the decomposition of hesperidin Cao et al. (2018) and the
thermogram of hespetine extract show the thermal decomposition
of hesperetin occur of about 44.19 % and 19.9% which has been
°C and from 550 to 800°C.
The XRD analysis was performed to determine the crystalline
nature of the extracted compounds and to provide the qualitative
information of different elements in these compounds. The XRD
spectrum of both saponin sea cucumber extract and standard
saponin (Figure 4) displayed a distinct diffraction peak at 2
values of 31.94° and 45.7° and for standard at 12.7, 16.56,1
9.67, 20.13, 21.41, 23.89, 27.73, 31.86,37.63, 38.73 and 40.9°
respectively, which indicated that sea cucumber saponin is a highly
crystalline compound. On the other hand, the XRD of sea cucumber
polysaccharide has been displayed in (Figure 5), Showed a distinct
diffraction peaks at 2 values of 31.83°, 45.79 and 56.44 which in
agreement with [42], polysaccharide being semi-crystal and did
not have periodical structure, could only show a diffuse region
corresponding to the maximum value of the diffraction when
         
even unable to make a judgment on the chemical composition of
polysaccharide [42].
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Figure 5: TGA analysis of
(a) Saponin,
(b) Polysaccharide,
(c) Naringen,
(d) Hesperidin and
(e) Hespertin.
Moreover, the X-ray of naringin showed that naringin was
      
peaks in the diffractogram which in accordance to [52], there are
        
angles of 10.87º, 14.26º, 18.48º, 21.31 and º35.98 were detected,
         
XRD results of hesperidin extract (Figures 6-9) there are six of the
most prominent peaks from hesperidin diffractogram at angles of
8.6°, 12.28°, 13.72°, 15.69°, 16.32 and 21.53° were detected which
        
XRD of hesperetin showe that hespertine have 10 of the most
prominent peaks from hesperetin diffractogram at angles of 8.63°,
11.69°, 12.27°, 13.67°, 15.57, 16.32, 20.70, 21.44, 23.43 and 29.12
which showed the crystalline form. Free radicals are thought to
have a remarkable part in numerous diseases.
Figure 6: Percentage of DPPH radical scavenging activity of
(a) ascorbic acid,
(b) Saponin and
(c) Naringen, Hesperidin and hespertin.
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20562
Figure 7:
(a) Total antioxidant activity of ascorbic acid,
(b) Saponin,
(c) Naringen, Hesperidin and hespertin
Figure 8: Standard curve of gallic acid.
It should be studied to measure them and display the oxidative
damage that they cause [54]. Vitamin C is a potent antioxidant, that
can scavenge singlet oxygen, superoxide, and hydroxyl radicals
have a positive effect of a scavenger of free radicals [55]. The DPPH
method was utilized to detect the ability of the extract under study
in scavenging free radical [56]. In the DPPH radical scavenging
method, antioxidant compounds combine with DPPH and convert
it pale violet. The grade of change dark violet color to pale violet or
yellow denotes the ability of the compound to scavenge free radical
[57]. It has been shown that all extracted compounds under study
can effectively scavenge DPPH, the scavenging reaction between
DPPH and antioxidant compounds (H-A) is due to the capacity of
the extracted compounds to change DPPH color as a stable free
radical. For example, saponin extract IC50% was 10.50 mg/ml
which in agreement with [58].
Saponin may be depending on their structure in eliminating
free radical as it contains a number of the hydroxyl group (OH) in
its structure. DPPH is vastly utilized to estimate the free radical
scavenging of different antioxidant materials and polyhydroxy
aromatic components [59]. Naringen orange peels extract
which scavenged 50% of DPPH radical was 0.13 mg/ml which
in agreement with [60]. Also, Hesperidin extract showed potent
scavenging activates of free radical which scavenged 50% by 0.13
mg/ml. The antioxidant properties of hesperidin result from their
chemical structure, hydroxyl and methoxy system, the mutual
          
C ring, and arrangement of the hydroxyl group and double bond
[61]. On the other hand, hespertine scavenged DPPH radical with
IC50 % equal to 0.66 mg/ml. Moreover, total antioxidant capacity is
determined through phosphomolybdenum complex formation [34]

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20563
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Figure 9: Antitumor activity of
(a) saponin,
(b) Hesperidin and
(c) Hespertin.
The results also showed that the total antioxidant capacity
of all compounds under the study increased with the increase of
its concentration. The extracted compounds have the ability to
scavenge free radicals and could act as a strong free radical inhibitor
or scavenger due to its chemical structure and its ability to donate
electrons, which is in accordance with [62]. The total phenolic
content was determined using the folin ciocalteu reagent which
gives a blue color to the solution; this indicated that orange peels

compounds from orange peels (Table 1). The hespertine extract
showed a small clear zone around the well in the inoculated plates
of Staphylococcus aureus which is an indication of antibacterial
activities of the strains under this study this is may be due to the
hesperidin hydrolysis and the change of their structure. MTT assay
was determined to investigate the biological activity of all extracted
compounds under study.
Table 1: Concentration of phenolic compounds of naringen,
hesperidin and hesperitin.
Compound Name Weight per volume
mg/ml
Total phenolic
contents mg/ml
Naringen 0.465 0.111±0.0075a
hesperidin 0.55 0.079±0.001b
hesperitin 0.50 0.057±0.001c
MTT assay showed that saponin, hesperidin, and hespertin on

that reduces cell viability by 50%. The mechanism of the extracted
components as antioxidants may be due to inhibit the peroxidation
of linoleic acid induced by Fe2+ and auto-oxidation in membranes
cerebral and inhibiting the production of reactive oxygen species
including hydroxyl radicals and nitric oxide [63] Saponins displayed
anticancer properties by attaching various cancer-related proteins
and Pathways [64]. So, marine-derived natural products such as

Conclusion

by FT-IR, UV, XRD and TGA analysis. The obtained data indicated
that sea cucumber saponin, polysaccharides, and orange peels,
Naringin, Hesperidin, and Hesperitin may provide a promising
new therapeutic approach to HEPG2 cancer cells. In addition, these
compounds were an effective antioxidant, so they may be effective
and scavenge free radicals which resulted from the disease and may
manage corona virus.
Acknowledgement
          
   
Science Tanta University, for her continuous help and cooperation
in the interpretation of the TGA and to Dr. Thanaa Mahmoud Ali
       
University) for her continuous help and cooperation.


References
1.          
   Citrus junos Sieb ex Tanaka) peels: a response
surface methodology study. J Food Meas Charact 11: 364-379.
2. Vandavasi SR, Ramaiah M, Gopal PN (2015) In vitro standardization of
Dendrobium normale falc. For free radical
scavenging activity. Journal of Pharmacognosy and Phytochemistry 3(5):
107-111.
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20564
3.               
polysaccharides from sea cucumber, Acaudina molpadioidea, in cecal
ligation and puncture-induced sepsis. CURRENT topics in nutraceutical
research 11(1-2): 29-34.
4.               
dyslipidemic effects of polysaccharidic extract from sea cucumber
processing liquor. Electronic Journal of Biotechnology 28: 1-6.
5. El Barky AR, Ali EMM, Mohamed TM (2017) Marine Sea Cucumber
Saponins and Diabetes. Austin Pancreat Disord 1(1): 1-7.
6. 
and free radical scavenging activities of a sulphated polysaccharide
extracted from abalone gonad (Haliotis Discus Hannai Ino). Food Chem
121: 712-718.
7.     
-mixturein crude extracts from leaves of Acanthopanax senticosus harms
by saponin structural correlation and mass spectrometry. Analytica
Chimica Acta 57(1-2): 198-203.
8. El Barky AR, Hussein SA, Alm Eldeen AA, Hafez YA, Tarek M, et al. (2016)
Anti-diabetic activity of Holothuria thomasi saponin. Biomedicine &
Pharmacotherapy 84: 1472-1487.
9.             
polysaccharide from Phellinus linteus induces G2/M phase arrest and

181.
10. Suetsugu T, Iwai H, Tanaka M, Hoshino M, Quitain A, et al. (2013)
Extraction of Citrus Flavonoids from Peel of Citrus Junos Using
Supercritical Carbon Dioxide with Polar Solvent. Chemical Engineering
and Science 1(4): 87-90.
11. Benavente Garcia O, Castillo J (2008) Update on uses and properties of
      
        
6185-6205.
12.         
Characterization. In: Encyclopedia of Food Sciences and Nutrition.
   
6000.
13. Peterson JJ, Dwyer JT, Beecher GR, Bhagwat SA, Gebhardt SE, et al. (2006)
Flavanones in oranges, tangerines (mandarins), tangors, and tangelos: a
compilation and review of the data from the analytical literature. J Food
Compost Anal 19: S66-S73.
14. 
(2006) Determination of naringin and hesperidin in citrus fruit by high-
performance liquid chromatography. The antioxidant potential of citrus
fruit. acta chromatographica 17: 108.
15. Horcajada MN, Habauzit V, Trzeciakiewicz A, Morand C, Gil Izquierdo
A, et al. (1985) Hesperidin inhibits ovariectomized-induced osteopenia
and shows differential effects on bone mass and strength in young and
adult intact rats. J Appl Physiol 104(3): 648-654.
16. Meneguzzo F, Ciriminna R, Zabini F, Pagliaro M (2020) Accelerated
production of hesperidin-rich citrus pectin from waste citrus peel for
prevention and therapy of COVID-19. Preprints.
17.           
      
reveals velpatasvir, ledipasvir, and other drug repurposing candidates.
F1000Research 9: 129.
18.            

681-688.
19. Jain M, Parmar HS (2011) Evaluation of antioxidative and anti-
 

20.             
   
cigarette smoke-exposed rats. J Med Food 15(10): 894-900.
21.    
cell cycle arrest and apoptosis in human breast cancer MCF-7 cells. Arch
Pharm Res 34(12): 2125-2130.
22.           
         
glycosides on chemical substance induced dermatitis in mice. J Nat Med
63(4): 443-450.
23. Badea M, Olar R, Uivarosi V, Marinesu D, Aldea V (2012) Synthesis and
 
as potential insulin-mimetic agents. J Therm Anal Calorim 107: 279-285.
24.      
improves functional recovery by Increasing BDNF and VEGF expression,
inhibiting neuronal apoptosis after spinal cord injury. Neurochem Res
37(8): 1615-1623.
25. 
induced osteolysis with naringin. Int Orthop 37(1): 137-143.
26.               
cucumber alleviate orotic acid-induced fatty liver in rats via PPARa and

27.            
Sources on the Production and Carbohydrate Composition of
Exopolysaccharide by Submerged Culture of Pleurotus citrinopileatus.
Food and Dr Analysis16: 61-67.
28.     
method for the large scale isolation of naringin from pomelo (Citrus
grandis) peel. International Journal of Food Science and Technology 44:
1737-1742.
29. Tang D, Zhu C, Zhong S, Zhou MD (2011) Extraction of naringin from
pomelo peels as dihydrochalcone’s precursor. J Sep Sci 34(1): 113-117.
30.         
   Citrus sinensis Peels. Der
Pharma Chemica 7(2): 1-4.
31.           
capacity assays. J Agric Food Chem 53(6): 1841-1856.
32.     
phenolic compounds from sage ( ). J Agric Food Chem
46(12): 4869-4873.
33. Zengin G, Aktumsek A, Guler GO, Cakmak YS, Yildiztugay E (2011)
Antioxidant properties of methanolic extract and fatty acid composition
of Centaurea urvillei       
123-132.
34. Prieto P, Pineda M, Aguilar M (1999) Spectrophotometric quantitation
of antioxidant capacity through the formation of a phosphomolybdenum
   
Biochem 269(2): 337-341.
35. Singh S, Singh RP (2008) In vitro methods of assay of antioxidants: An
overview. Food reviews international 24(4): 392-415.
36.            in vitro
         
Phoenix dactylifera L. (Ajwa and Zahedi Dates). Int J Adv Sci Technol
35(35): 2319-2682.
37.           In vitro anti-tumor
activity of isorhamnetin isolated from Hippophae rhamnoides L. against

38. 
MTT assay adapted for primary cultured hepatocytes: application
to proliferation and cytotoxic assays. Bioscience Biotechnology and
Biochemistry 56(9): 1472-1473.
Volume 27- Issue 2 DOI: 10.26717/BJSTR.2020.27.004463
20565
Copyright@ Amira Ragab El Barky | Biomed J Sci & Tech Res | BJSTR. MS.ID.004463.
39.         
involved in UVA-induced autolysis in sea cucumber Stichopus japonicus.
J Photochem Photobiol B 158: 130-135.
40. Ma Y, Ye X, Fang Z, Chen JC, Xu GH, et al. (2008) Phenolic Compounds and
Antioxidant Activity of Extracts from Ultrasonic Treatment of Satsuma
Mandarin (Citrus unshiu Marc.) Peels. Journal of Agricultural and Food
Chemistry 56(14): 5682-5690.
41. Almutairi MS, Ali M (2014) Direct detection of saponins in crude extracts
of soapnuts by FTIR. Natural Product Research 29(13): 1271-1275.
42. 
       Flammulina velutipes
polysaccharide with Zn. Food and agricultural immunology 28(1): 162-
177.
43. 
of a neutral extracellular glucan from Lactobacillus reuteri 
Carbohydrate Polymers 106: 384-392.
44.               
conformation, and immunomodulatory activity of the polysaccharide
      
Polymers 150: 149-158.
45.      
from Panax japonicus CA Meyer and their antioxidant activities.
Carbohydr Polym 101: 386-391.
46.          
scavenging capacity of naringin extracted from Citrus uranium peel
against free radicals. Prospect 14(2): 31-35.
47.           
In Vitro  
Activity of Citrus Fruits Extracts from Aceh, Indonesia. Antioxidants
6(1): 11.
48.     
molecularly imprinted naringin prepared via reverse atom transfer
radical polymerization with excellent recognition ability in a pure
aqueous phase. RSC Adv 7: 28082-2809.
49.           
Characterization of Extracellular Polysaccharides Produced by
Cyanobacterium Arthrospira platensis. Biotechnology and Bioprocess
Engineering 14: 27-31.
50.              
Determination of Flavonoids in Different Parts of Citrus reticulata

Array Detection. Molecules 15(8): 5378-5388.
51. Xu C, Yang C, Mao D (2014) Fraction and chemical analysis of antioxidant
      
Pharmacogn Mag 10(37): 66-69.
52.           
as a shell material for encapsulation of naringin: Production and
physicochemical characterization. Journal of Food Engineering 161: 68-
74.
53. Varghese JJ, Mallya R (2015) Formulation development and evaluation
        
Pharmaceutical Research 4(08): 1149-1170.
54.         
oxidative damage in vivo and in cell culture: how should you do it and
what do the results mean. Br J Pharmacol 142(2): 231-255.
55.     
nickel induced hepatic lipid peroxidation in rats. J Basic Clin Physiol
Pharmacol 12(3): 187-195.
56.          
Phenolic Content of Selected Tropical Fruits from Malaysia, Extracted
with Different Solvents. Food Chemistry 115(3): 785-788.
57.           
Citronellal and Crude Extracts of Cymbopogon citratus by 3 Different
Methods. Pharmacology & Pharmacy 5(4): 395-400.
58. MO N, AOT A (2017) Antioxidant and Inhibitory Effects of Saponin
Extracts from Dianthus basuticus
in Type 2 Diabetes In vitro. Pharmacogn Mag 13(52): 576-582.
59.           
Non-reductive scavenging of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) by
peroxyradical: a useful method for quantitative analysis of peroxyradical.
Chem Pharm Bull 53(6): 714-716.
60.           
induced toxicity in rats: an in vivo and in vitro study. International
journal of pharmaceutical sciences and research. IJPSR 2(1): 137-144.
61.          
deprived rats. J Nutr 130(7): 1766-1771.
62.       
Property of Hesperidin. Int J Chem Stud 1(4): 2321-4902.
63. 
against peroxynitrite. Free Radic Res 38(7): 761-769.
64.  
Medicines as Anticancer Agents. Molecules 21(10): 1326.
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ISSN: 2574-1241
DOI: 10.26717/BJSTR.2020.27.004463
Amira Ragab El Barky. Biomed J Sci & Tech Res
... Hesperidin consists of aglycone, hesperetin, and the sugar rutinoside [2]. Both hesperidin and hesperetin exhibit various biological activities including anti-inflammatory, antibacterial, and antitumor properties, as well as the potential to reduce capillary permeability [3,4]. Furthermore, hesperidin has been shown to have an inhibitory effect against SARS-CoV-2, the causative agent of COVID-19 [4]. ...
... Both hesperidin and hesperetin exhibit various biological activities including anti-inflammatory, antibacterial, and antitumor properties, as well as the potential to reduce capillary permeability [3,4]. Furthermore, hesperidin has been shown to have an inhibitory effect against SARS-CoV-2, the causative agent of COVID-19 [4]. ...
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Numerous epidemiological studies have reported that particulate matter 2.5 (PM2.5) causes skin aging and skin inflammation and impairs skin homeostasis. Hesperidin, a bioflavonoid that is abundant in citrus species, reportedly has anti-inflammatory properties. In this study, we evaluated the cytoprotective effect of hesperidin against PM2.5-mediated damage in a human skin cell line (HaCaT). Hesperidin reduced PM2.5-induced intracellular reactive oxygen species (ROS) generation and oxidative cellular/organelle damage. PM2.5 increased the proportion of acridine orange-positive cells, levels of autophagy-related proteins, beclin-1 and microtubule-associated protein light chain 3, and apoptosis-related proteins, B-cell lymphoma-2-associated X protein, cleaved caspase-3, and cleaved caspase-9. However, hesperidin ameliorated PM2.5-induced autophagy and apoptosis. PM2.5 promoted cellular apoptosis via mitogen-activated protein kinase (MAPK) activation by promoting the phosphorylation of extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38. The MAPK inhibitors U0126, SP600125, and SB203580 along with hesperidin exerted a protective effect against PM2.5-induced cellular apoptosis. Furthermore, hesperidin restored PM2.5- mediated reduction in cell viability via Akt activation; this was also confirmed using LY294002 (a phosphoinositide 3-kinase inhibitor). Overall, hesperidin shows therapeutic potential against PM2.5-induced skin damage by mitigating excessive ROS accumulation, autophagy, and apoptosis.
... Flavonoid ingredient component as hesperidin and naringin possesses a diverse pharmacological activity [29], they can ameliorate the elevated levels of blood glucose and glycosylated hemoglobin (Hb A1C) [30], and also can manage coronavirus [29]. Moreover, vitamin C as lemon, orange or guava can increase immunity, manage a variety of viral infections and can kill the host of coronavirus [31], furthermore, Ginger was effective in blocking viral attachment and internalization and can prevent plaque-forming which results in respiratory syncytial virus infection. ...
... Flavonoid ingredient component as hesperidin and naringin possesses a diverse pharmacological activity [29], they can ameliorate the elevated levels of blood glucose and glycosylated hemoglobin (Hb A1C) [30], and also can manage coronavirus [29]. Moreover, vitamin C as lemon, orange or guava can increase immunity, manage a variety of viral infections and can kill the host of coronavirus [31], furthermore, Ginger was effective in blocking viral attachment and internalization and can prevent plaque-forming which results in respiratory syncytial virus infection. ...
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