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Evaluation of some chemical constituents, antioxidant, antibacterial and

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EVALUATION OF SOME CHEMICAL CONSTITUENTS,

ANTIOXIDANT, ANTIBACTERIAL AND ANTICANCER
ACTIVITIES OF BETA VULGARIS L. ROOT
Hossam S El-Beltagi1,2,*, Heba I Mohamed3, Basma M H Megahed4, Mohammed Gamal4, Gehan Safwat4
1
Faculty of Agriculture, Biochemistry Department, Cairo University, Giza, Cairo, Egypt
2
Cairo University, Research Park (CURP), Giza, Cairo, Egypt
3
Faculty of Education, Department of Biological and Geological Science, Ain shams University, Cairo, Egypt
4
Faculty of Biotechnology, October University for Modern Science and Art (MSA), Egypt

ABSTRACT ing salads, jam, soups and juice [7,8]. In addition,

the leaves contain a large amount of antioxidant and
Beta vulgaris is belonging to the family Che- vitamins, so it can be used as food and cooked as a
nopodiaceae and has several varieties with bulb spinach substitute [9]. Red beets have betalain
colors ranging from yellow to red. Deep red- pigments so that it has commercial and pharmaceu-
colored beet roots are the most popular for human tical uses such as natural food dye, cosmetics, drug
consumption, both cooked and raw as salad or formulations and paintings [10-12].
juice. The ethanolic extract of beetroots contains Red beets are the 10th vegetable in the world
valuable and active compounds such as carotenoids, that have antioxidants [13, 14]. These antioxidants
phenols, flavonoids, tannin, alkaloids, vitamins C, used as the scavengers of free radicals and prevent
B3, B6 and B9. Therefore, beetroot extract has the oxidative damage on proteins, DNA and lipo-
antioxidant and antimicrobial activity against gram proteins [15, 16]. The oxidative damage of macro-
positive and negative bacteria. Gram-positive bacte- molecules may lead to chronic diseases such as
ria Staphylococcus aureus and Bacillus cereus cancer, cataractogenesis, cardiovascular disease,
demonstrated higher susceptibility than Gram- neurodegenerative diseases, and stroke, which may
negative Escherichia coli and Pseudomonas typhi- prevent by the antioxidant compounds in red beets
mureum. Beta vulgaris ethanolic extract exhibit [17]. Red beets also have high concentrations of
significant anticancer activity against lung (A549) secondary metabolites (phenolic acids, flavonoids,
but slight effect against colorectal adenocarcinoma ascorbic acid) [18-20].
Caco-2 cell lines at the high concentrations of etha- The most important problems in the pro-
nolic extract (800 µg/ml). cessing of the food industry are the contamination
of microbes which affects the quality of foods and
cause economic losses [21, 22]. So that, the im-
KEYWORDS: portant strategy to overcome this problem is to use
Beta vulgaris, phenols, flavonoids, tannin, carotenoids, natural antimicrobial compounds which presented
vitamins, DPPH, antibacterial, anticancer activity. in medicinal plants and protect from fungi and
bacteria [23, 24]. Red beets used as antioxidant,
antimicrobial, anti-inflammatory, antiallergenic,
INTRODUCTION antithrombotic, antiatherogenic, cardioprotective,
and vasodilatory properties [25].
In the past few years, it's found that the use of The aim of this work is to study the chemical
synthetic drugs to protect the human from diseases composition of red beet roots and to study its effect
is unsafe to human and environment. So that, it's as antioxidant, antimicrobial and anticancer activi-
very important to use the medicinal plants which ty.
have secondary metabolites and antioxidant com-
pounds which decrease the effect of free radicals [1,
2]. MATERIALS AND METHODS
Beta vulgaris L. subsp. vulgaris is belong to
the family Chenopodiaceae (Angiosperm) [3] and it Plant Materials. The roots of Beta vulgaris
also called beetroot or garden beet [4]. Beetroot is subsp. vulgaris var. Plano (sugar beet) was collect-
annual crop, biennial herbaceous and cultivated for ed from local market in Egypt. Beta vulgaris was
their edible roots and leaves [5]. The color of beet- botanically characterized by Dr. Samah Azooz from
root is differed from yellow to red according to its Botany Department, Faculty of Agriculture, Cairo
variety. In all over the world, red beets are used in Univeristy, Egypt.
human consumption [6]. The roots are used in mak-

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TABLE 1
Microbial strains used to test the antimicrobial activities of Beta vulgaris root extract
Microbial group Indicator strain Positive control Cultivation conditions
Staphylococcus aureus (ATCC 25923) Muller-Hinton broth, 37ºC/ 24 h
Gram positive bacteria Kanamycin
Bacillus cereus (ATCC 33018) Muller-Hinton broth, 30ºC / 24 h
Escherichia coli (ATCC 8739) Muller-Hinton broth, 37ºC / 24 h
Gram negative bacteria Polymyxin
Salmonella typhimureum (ATCC 14028) Muller-Hinton broth, 37ºC / 24 h
Aspergillus niger (nrrl 326) Sabouraud dextrose broth, 25 ºC / 3days
Fungus Nystatin
Candida albicans ATCC 10231 Sabouraud dextrose broth, 25ºC / 24 h

Microbial strain. Table 1 illustrated the mi- Total carotenoid content. Total carotenoids
croorganisms which were used in this study and of red beet root were extracted using a mixture of
were obtained from the American Type Culture hexane: acetone (1:1 v/v) as described by Jeyanthi
Collection (ATCC) as well as the culture collection et al. [32]. The absorbance of carotenoid was read
of the Microbiology Lab, Cairo University Re- at 630 nm using spectrophotometer.
search Park (CURP), Faculty of Agriculture, Cairo
University. Water soluble vitamins. Sample Prepara-
tion. Water soluble vitamin were determined by
Extraction method. The roots were cleaned HPLC analysis after extraction from the sample
and washed thoroughly under tap water, and then according to Albala-Hurtado et al. [33]. Dry
the roots were freeze-dried and grinded into fine weighed 0.2 g of red beet root powder was placed
powder using an electric blender. The powder was into centrifuge tube and add 15 mL of deionized
dried in an oven at 40°C for 24 h. The fine powder water. After 15 min of ultrasonic extraction, centri-
sample (500mg) was extracted in 10 ml ethanol or fuge at 4000 rpm for 5 minutes, then quantitively
distilled H2O for 24 h using a shaker, then the ex- transfer to 25 mL volumetric flask, add water to the
tract was filtered and the samples were stored at mark. Filter through 0.2um nylon membrane before
4°C until use [26]. All analysis was done in the labs injection.
of Cairo University. Research Park (CURP), Facul-
ty of Agriculture, Cairo University, Cairo, Egypt. Instrument Conditions. Agilent 1260 infinity
HPLC Series (Agilent, USA), equipped with Qua-
Total polyphenol content. The total phenolic ternary pump, a Kinetex XB-C18 column 100 mm
content was estimated by Folin Ciocalteu method as x 4.6 mm (Phenomenex, USA), operated at 35 oC.
described by Singleton et al. [27]. The absorbance The separation is achieved using a binary linear
was measured at 765 nm using a spectrophotometer elution gradient with (A) 25 mM NaH2PO4 pH =
Thermo Scientific HERYIOS. 2.5, (B) methanol. The injected volume was 20 μL.
Detection: VWD detector set at 254 nm for ascorbic
Total flavonoid content. The flavonoids con- acids and 220nm for vitamins B3, B6, B9 and B12
tent was determined by aluminium trichloride [34].
method as described by Zhishen et al. [28]. The
absorbance was measured at 510 nm using a spec- Extraction of phenolic and flavonoid com-
trophotometer. pounds. 0.2g dry sample extracted with 20 ml
ethanol 80%, soak in brawn bottle for 24 hr at room
Total tannin contents. Tannin content in red temperature, centrifuged for 5 min, volume adjusted
beet roots was determined by using Folin-Denis to 25 ml by ethanol 80%, filtered through Whatman
reagent as described by Saxena et al. [29]. The filter paper, 10 ml of the solution evaporated to
absorbance was read at 700 nm using spectropho- dryness then dissolved in 5 ml HPLC grade metha-
tometer. nol 50%, filtered through PTFE filter with pore size
0.2 μm.
Total alkaloid contents. Alkaloids was meas-
ured according to the method described by Adham Instrument Condition for phenolic com-
[30]. pounds. Agilent 1260 infinity HPLC Series (Ag-
The percentage alkaloid was calculated as: ilent, USA), equipped with Quaternary pump, a
Percentage of total alkaloid = [Weight of residue / Zorbax Eclipse plusC18 column 100 mm x 4.6 mm
Weight of sample] ×100 i.d., (Agilent technologies, USA), operated at 30oC.
The separation is achieved using a ternary linear
Total Athocyanine content. Fresh weight of elution gradient with (A) HPLC grade water 0.2 %
Beta vulgaris root was homogenized in methanol H3PO4 (v/v), (B) methanol and (C) acetonitrile. The
containing 1% (v/v) HCl and then filtrate. The injected volume was 20 μL. Detection: VWD detec-
filtration was read at 530 and 657 nm using spec- tor set at 284 nm.
trophotometer as described by Mancinelli et al.
[31]. Instrument Condition for Flavonoids.
HPLC, Smart line, Knauer, Germany., equipped

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with binary pump, a Zorbax Eclipse plusC18 col- ried out in triplicates and the inhibition zone was
umn 150 mm x 4.6 mm i.d., (Agilent technologies, recorded as the average of the replicates± SD.
USA), operated at 35oC. Eluent: methanol: H2O In Vitro cytotoxicity assay. Human lung can-
with 0.5% H3PO4, 50:50 with flow rate 0.7 ml/min, cer (A549) and colorectal adenocarcinoma Caco-2
the injected volume was 20 μL. Detection: UV were purchased from CURP, faculty of agriculture
detector set at 273 nm and data integration by clari- at Cairo University (Egypt). Cells were maintained
tychrom@ software. This method was the modified in (DMEM) supplemented with 10% heat-
of methods Goupy et al. [35] and Mattila et al. [36] inactivated fetal bovine serum, 100 µg/ml strepto-
for fractionate the polyphenols and flavonoids, mycin and 100 unit/ml penicillin g potassium, in a
respectively. humidified 90% and 5% (V/V) CO2 atmosphere at
37ºC. The cytotoxicity of ethanolic extracts was
DPPH free Radical Scavenging activity tested by the neutral red (NR) assay as previously
(RSA). The antioxidant activity of the Beta vulgaris described [39]. Exponentially growing cells were
root extract was measured in terms of hydrogen collected using 0.25% Trypsin-EDTA and seeded
donating or radical-scavenging ability using the in 96- well plates at 20000 cells/well. After incuba-
stable DPPH method as modified by Hae-Ryong et tion (overnight), extracts were added in various
al. [37]. The reaction mixture containing 1 ml of the concentrations (10, 50, 100, 200, 400, and 800
extract at different concentrations (40, 80, 120, 150 µg/ml); 4 wells for each concentration. After treat-
μg/ml) and 1ml of DPPH (0.2mM) was vigorously ment with extracts for 24h., media were removed
shaken and incubated in darkness at room tempera- and cells were exposed to neutral red solution for 4
ture for 30 minutes. The absorbance was read at hours at 37ºC. Destin solution was used to dissolve
517nm using UV-visible spectrophotometer. Radi- the NR stained cells and color intensity was meas-
cal scavenging activity was expressed as percent of ured at 540nm microplate reader (Biotek, ELX808).
inhibition and was calculated using the following
formula:- Statistical analysis. All results were ex-
%DPPH = [ Absorbance of Control – Absorbance pressed as mean values ± standard deviation. Com-
of Sample / Absorbance of Control ] x 100 parisons were performed by analysis of variance
(ANOVA). Statistical analyses were run using SAS
Antibacterial activity. Agar disc diffusion software.
method was used to evaluate antibacterial activity
of red beet roots as describe by Bauer et al. [38].
The strains were grown on Mueller-Hinton agar RESULTS AND DISCUSSION
slants at 37°C for 24 h and checked for purity. After
the incubation, the cells were washed off the sur- Chemical constituents of red beet root. As
face of agar and suspended in sterile physiological illustrated in Table 2, the chemical constituents of
solution. The number of cells in 1 ml of suspension ethanolic extract of red beet roots contain total
for inoculation measured by McFarland nefelome- phenolic (133.5 mg /g DW), total flavonoids (1.5
ter was 5 × 107 CFU/ml. 1 ml of these suspensions mg /g DW), total tannin (5.13 mg /g DW), total
was homogenized with 9 ml of melted (45°C) alkaloid (2.1 g /100g DW), total athocyanin (63.7
Mueller-Hinton agar and poured into Petri dishes. μg/100g FW) and carotenoids (1.7 mg/100g FW).
On the surface of the agar, 5 mm diameter paper These results are similar to previous studies [40-
discs (HiMedia®, Mumbai, India) were applied and 42], who found that the main components of red
impregnated with 15 μl of samples. The plates were beet root extract are polyphenols, alkaloids, tannins,
incubated at the optimum temperature for each flavonoids, folic acid, reducing sugars and ascorbic
indicator strain (Table 1) and tested after 24, 48 and acid. In addition, folic acid and vitamins A, B, and
72 h. Growth inhibition was scored positive in the C can play important roles in brain development
presence of a detectable clear zone (ZI) around the and motor function.
disc and expressed in mm. Experiments were car-
TABLE 2
Quantitative phytochemical analysis of Beta vulgaris root
Constituents Values in ethanolic extract
Total phenolic
133.5±1.05
(mg Gallic acid /g DW)
Total flavonoid
1.54±0.047
(mg Quercetin /g DW)
Total tannin
5.13±0.085
(mg Tannic acid /g DW)
Total alkaloid
2.10±0.040
(g/100g DW)
Total athocyanin
63.73±0.032
(μg/100g FW)
Carotenoids
1.72±0.08
(mg/100g FW)
Values are mean ± SD of three replicate analyses

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The presence of the secondary metabolites in DW), ferulic acid (0.68 mg/100g DW), o-Coumaric
red beet root has contributed to its medicinal value acid (1.31 mg/100g DW) and cinnamic acid (0.6
as well as physiological activity [43]. Phytochemi- mg/100g DW). These results are similar to Vulić et
cal components are responsible for both pharmaco- al. [53] who reported that beetroot contain ferulic,
logical and toxic activities in plants [44]. They are vanillic, p-hydroxybenzoic, caffeic and protocate-
used for therapeutic purposes to cure various dis- chuic acids.
eases and to heal injuries [45]. For instance, flavo- In addition, the ethanolic extract of red beet
noids have been shown to have antibacterial, anti- root contains a number of flavonoids compounds
inflammatory, anti-allergic, antiviral, antineoplastic such as myricetin (19.3 mg/100g DW), neringenin
and antioxidant, which act as free radical scavenger (19.9 mg/100g DW), kaempferol (3.0 mg/100g
and metal chelators [46, 47]. Alkaloids contribute DW) and apigenin (2.56 mg/100g DW). Similar
to plant species fitness of survival and have phar- results reported by Pyo et al. [54] recognized the
macological effects and are used as medication and following: catechin (6.7 mg/100 g FW), myricetin
recreational drugs [48]. They protect the plants (2.2 mg/100 g FW), quercetin (7.5 mg/100 g FW)
against infection with insects by the production of and kaempferol (9.2 mg/100 g FW).
the bitter taste that repels insects from feeding on Also, Ben Haj Koubaier et al. [55] found that
plant leaves. Tannins may provide protection the presence of five phenolic acids (ferulic, vanillic,
against microbial degradation of dietary proteins in syringic, ellagic, and caffeic), three flavonoids
the rumen [49]. In addition, carotenoids have pro- (quercetin, kampferol, and myricetin) for roots of
tective effects against several diseases such as can- red beet by using Liquid chromatography–mass
cer, coronary heart disease, inflammatory reactions, spectrometry. These flavonoids act as antioxidation,
and age-related macular degeneration [50] and act antiinflammation and inhibition of tumor prolifera-
as antioxidant [51]. tion [56].

HPLC of soluble vitamins. The results in Ta- TABLE 4

ble 3 reported that the ethanolic extract of red beet- HPLC analysis of phenolic and flavonoid
root contains vitamin C (26.2 mg/100g DW), vita- compounds of Beta vulgaris root
min B3 (1.67 mg/100g DW), vitamin B6 (6.17 Phenolic compounds Conc. mg/100g DW
mg/100g DW) and vitamin B9 (2.60 mg/100g DW). Gallic acid 11.01
These results are similar to Odoh and Okoro [41] Catechol 7.38
who found that beetroot contains significant p-Comuaric acid 0.74
amounts of vitamins, especially vitamin C (4.36 Ferulic acid 0.68
mg/100 g). o-Coumaric acid 1.31
Cinnamic acid 0.60
The results showed that red beet root has a
flavonoid compounds
high concentration of ascorbic acid. This vitamin
plays an important role in human nutrition, includ- Myricetin 19.25
Neringenin 19.92
ing growth and maintenance of tissues, the produc-
Kaempferol 3.02
tion of neurotransmitters, hormones and immune Apigenin 2.65
system responses. Vitamin C is an important anti-
oxidant and reduces the adverse effects of reactive
Antioxidant activity of red beet root. The ef-
oxygen species which caused damage to macromol-
fect of antioxidants on DPPH radical scavenging
ecules such as lipids, DNA and proteins, which are
was thought to result from their hydrogen donating
related to cardiovascular disease, cancer and neuro-
ability. DPPH is a stable free radical and accepts an
degenerative diseases [52].
electron or hydrogen radical to become a stable
diamagnetic molecule. The reduction capability of
TABLE 3
DPPH radicals was determined by the decrease in
Water soluble vitamins contents of
its absorbance at 517 nm induced by antioxidants. It
Beta vulgaris root
is visually noticeable as a discoloration from purple
Values
Vitamin contents to yellow. The scavenging of DPPH radicals in-
(mg/100g DW)
creased with increasing extract concentration from
Vitamin C (Ascorbic acid) 26.23±0.32
Vitamin B3 (Niacin) 1.67±0.05
40, 80, 120 and 150 μg /mL (Table 5). The IC50
Vitamin B6 (Pyridoxine) 6.173±0.16 value of ethanolic extract of beet root was 55.82 μg
Vitamin B9 (Folic acid) 2.60±0.08 /mL concentration. IC50 value indicate the concen-
Values are mean ± SD of three replicate analyses tration of the test sample required to inhibit 50% of
the free radicals. The IC50 value is a parameter
HPLC of phenolic compounds. Data in Ta- widely used to measure the free radical scavenging
ble 4 showed that the ethanolic extract of red beet activity [57]; a smaller IC50 value corresponds to a
root contains a number of phenolic compounds higher antioxidant activity.
such as gallic acid (11 mg/100g DW), catechol (7.4 Enzymatic and nonenzymatic antioxidants are
mg/100g DW), p-Comuaric acid (0.74 mg/100g molecules that have the ability to scavenge free

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radicals before they damage the cells. Antioxidants rium causing food poisoning. On the other hand, a
can be endogenous or obtained exogenously, either weak antimicrobial activity was found against Sal-
through diet or by dietary supplements [58]. Scien- monella typhimureum (ZI = 7.11 mm). The extract
tific evidence suggests that antioxidant compounds showed no effect on the fungus used (Aspergillus
in food play an important role as a health protecting niger and Candida albicans). In general, red beet
factor. Beet root ranks among the top ten most extracts have exhibited antibacterial activity against
powerful vegetables with respect to its antioxidant a wide range of gram-positive bacteria and gram-
capacity ascribed with a total phenol content of 50– negative bacteria; however, no inhibitory activity
60 μmol/g dry weight [13]. Kähkönen et al. [14] was found against the fungi and yeasts studied [60-
reported that beet root contains considerable 63].
amount of phenolic acids such as ferulic, protocate- In terms of antimicrobial potential, gram-
chuic, vanillic, p-coumaric, phydroxybenxoic, and positive bacteria (Bacillus, Micrococcus, Staphylo-
syringic acids [59]. coccus, and Streptococcus) have been found more
susceptible to red beets than gram-negative (Esche-
TABLE 5 richia coli and Pseudomonas aeruginosa) [62-63,
Antioxidant activity of Beta vulgaris root against 53]. The red beetroot extract contains a high
DPPH method. amount of phenolic compounds which may cause
Conc. (μg/ml) DPPH % in ethanolic extraxt the disrupting of the cell wall structure of gram-
40 49.20 positive bacteria [64-66]. The inhibitory effect of
80 50.638 ethanolic extract on gram-negative bacteria is at-
120 52.763 tribute to their outer membrane, consisting of dou-
150 70.351 ble-layered, highly hydrophilic lipopolysaccharide
IC50 (μg/ml) 55.823 molecules, and unique periplasmic space [67-68].
These bacteria caused the infectious diseases
Phenolic compounds present in red beet de- on human health and also affect on food safety. The
crease oxidative damage of lipids improves antioxi- phytochemicals derived from red beetroot have
dant status in humans, scavenges free radicals, been widely exploited for their colorant properties;
exhibits inflammatory effect, anticancer property however, there is much potential for utilizing their
and reduces the risk of chronic illnesses such as antimicrobial properties, particularly in food-related
cancer and cardiovascular diseases [18]. In addi- applications and consumer products targeting hu-
tion, the antioxidant properties of phenolic com- man health and beauty. For example, red beet com-
pounds are mainly because of their redox potential, pounds could be incorporated into products such as
which allows them to act as reducing agents, hy- active antimicrobial food packaging [69], cosmetic
drogen donators, metal chelators and singlet oxygen products [70-72] and also in active drug formula-
quenchers. tions for the treatment against disease in the form of
supplements, topical sprays, and ointments.
Antimicrobial activity of red beet root. The
agar diffusion method used to evaluate the antibac- Anticancer activity of red beetroot. Data in
terial and antifungal activity of ethanolic extract of Table 7, showed that the cytotoxic activity of red
red beetroot by using selected gram-positive, gram- beetroot as an anticancer agent (towards to lung)
negative bacteria and fungus. The diameter of the and IC50 dose. The percentage of lung cancer cell
inhibition zone (ZI) is shown in Table 6. line (A549) viability was decreased with increasing
The data indicate that the extract exhibited the the concentrations of the methanolic extract of red
activity against the investigated food pathogens. beetroot (Figure 1). On the other hand, the viabitity
Gram positive bacteria Staphylococcus aureus and of colorectal adenocarcinoma Caco-2 is not affected
Bacillus cereus demonstrated higher susceptibility by all concentrations of red beet root except the
than Gram-negative Escherichia coli and Salmonel- high concentrations (800 μg/ml) which showed
la typhimureum. The extract showed antibacterial slight decrease in the viability of Caco-2 cell line.
activity against Staphylococcus aureus (ZI = 12.5 Cancer is often associated with increased risk of
mm), one of the most common gram-positive bacte- death and the toxic side effects caused by the mod-
ern medicine.

TABLE 6
Antibacterial activities of Beta vulgaris root against selected bacterial strains and fungus.
Inhibition zone (mm)*
Samples Gram positive bacteria Gram negative bacteria Fungus
S. aureus B. cereus E. coli S. typhimureum A. niger C. albicans
Beta vulgaris root in
12.54±0.35 9.25±0.16 8.37±0.21 7.11±0.0 - -
ethanolic extract
Values are mean ± SD of three replicate analyses, *Well size = 5 mm

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TABLE 7
Anticancer activities of Beta vulgaris root.
Lung cell line Colorectal adenocarcinoma
Concentrations (A549) Caco-2
(μg/ml)
Viability %
10 100 100
50 99.8 100
100 88.6 100
200 86.4 100
400 82.8 100
800 78.0 95.5
IC50 (μg/ml) 17800 4831

FIGURE 1
Morphological observation of cancer cell lines (A549) by 40X magnification power.

Many cancer patients seek alternative and presence of phenolic groups, flavonoids, betaxan-
complementary methods of treatment such as usage thins and betacyanins [74]. Polyphenolic com-
of phytomedicine. Natural dietary agents have pounds might inhibit cancer cells by xenobiotic
drawn a great deal of attention because of their metabolizing enzymes that alter metabolic activa-
potential to suppress cancers and to reduce the risk tion of potential carcinogens, while some flavo-
of cancer development by decreasing oxidative noids could also alter hormone production and
stress, which plays a significant role in the patho- inhibit aromatase to prevent the cancer cells [76,
genesis and pathophysiological process of cancer 77]. The mechanism of action of anticancer activity
[73]. Previous studies have shown that beetroot has of phenols could be by disturbing the cellular divi-
an excellent antioxidant property which can con- sion during mitosis at the telophase stage. It was
tribute to the anticancer activity [74-75]. Previous also reported that phenols reduce the amount of
phytochemical studies of Beta vulgaris indicate the

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cellular protein and mitotic index and colony for- side effects. In vitro investigations in the present
mation during cell proliferation of cancer cells [78]. study provide substantial evidence that beetroot
peel; an inedible waste product is a potent source of
antioxidant, antimicrobial agent and anticancer
activity thereby indicating its use as a value-added
component for functional.

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