Anti-Cancer Property Screening and Liquid ChromatographyMass Spectrometric Analysis of Areca catechu Linn. Root: An Unexplored Traditional Medicine of Kerala

Full text

Research Paper
Indian Journal of Pharmaceutical Sciences 1663
September-October 2024
Baby et al.: Anti-Cancer Property and Phytochemical Proling of Areaca catechu
The use of natural products against some prevalent ailments in human beings and household animals is a
common phenomenon in both developed and developing countries. So, documentation of this indigenous
system of knowledge is important because with the demise of the present traditional medical practitioners
this treasure house of knowledge would become extinct. Oxidative stress disorders dangerously lead to
malignant tumors, diabetes and other severe disease conditions. In the present study antioxidant property
using 2,2-diphenyl-1-picrylhydrazyl free radical scavenging assay and in vitro cytotoxicity screening
using Dalton’s lymphoma ascites cells were used to choose the bioactive solvent extract of Areca catechu
for anti-cancer screening and phytochemical profiling. In the 2,2-diphenyl-1-picrylhydrazyl free radical
scavenging assay aqueous extract was highly efficient with least half-maximal inhibitory concentration
value (53 µg/l) and in in vitro cytotoxicity screening, maximum cytotoxicity (89 %) was attained at a
concentration of 200 µg/l of ethanolic extract and minimum half-maximal inhibitory concentration
value was showed by aqueous extract (38 µg/l). So the ethanolic and aqueous extracts were selected
for the in vivo anti-cancer screening. The plant extract was found to be non-toxic in the dose of 250
mg/kg body weight of the mice. In vivo anti-cancer efficiency of the medicinal plant was screened via
ascites tumour and solid tumour models. Treatment of Areca catechu ethanolic and aqueous extracts
at different concentrations increased the survival rate of animals in ascites tumour and suppressed the
development of solid tumour. The result was compared with the commercial drug cyclophosphamide.
The aqueous extract was very efficient as an anti-tumour agent and caused 71.9 % of increase in life
span in 100 mg/kg concentration. The percentage of life span increase in cyclophosphamide treated
animals was 72.5. So, the plant extract was almost equally important to commercial drug in increasing
the rate of survival of the affected animals. The volume of solid tumour in control group on 35th d was
(4.550±0.622) mm while in case of aqueous root extract in 100 mg/kg concentration was (0.48±0.92)
mm on the same day. The value shown by the standard drug (0.643±0.111) mm was more than that
of the aqueous extract at 100 mg/kg. So, the aqueous extract of Areca catechu was subjected to high
resolution liquid chromatograph mass spectrometric analysis. 14 compounds were identified through
this analysis. That includes mitoxantrone, artemether, valporic acid, leupeptin etc., which proves the
medicinal applications of Areca catechu root.
Key words: Areca catechu, anti-oxidant property, in vitro cytotoxicity, anti-cancer property,
phytochemical profiling
Anti-Cancer Property Screening and Liquid Chromatography-
Mass Spectrometric Analysis of
Areca catechu
Linn. Root: An
Unexplored Traditional Medicine of Kerala
ALBY ALPHONS BABY*
Department of Botany, Christian College, Chengannur, Kerala 689122, India
*Address for correspondence
E-mail: albyalphonsbaby@gmail.com
Accepted 25 October 2024
Revised 27 May 2024
Received 30 September 2022
Indian J Pharm Sci 2024;86(5):1663-1670
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Human civilization begins in the forest as an integral
part of the forest ecosystem. Humans acquired unique
knowledge about various plants and animals around
them by experiences and experimentation. They
developed, maintained and preserved this knowledge
over many generations. Through years of co-evolution
and co-existence, the traditional communities are
able to identify the useful and harmful elements
around them. Traditional knowledge on health care is
very important in giving clues regarding the various
medicinally active plants, this is a very important
eld of research leading to the discovery of new

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Indian Journal of Pharmaceutical Sciences
1664 September-October 2024
bio-pharmaceuticals which are eective and less
harmful to the mankind[1].
Betel nut (Areaca catechu (A. catechu)) from
the family Arecaceae grows in India, Malaysia,
Taiwan and many other Asian countries. Nut is
the commercially useful part and is important in
traditional medicine also. It contains alkaloids,
tannins, polyphenols and sugars[2]. Betel nut have
reported anthelmintic[3], wound healing[4], anti-
depressant[5], anti-Human Immunodeciency Virus
(HIV)[6] and anti-mycobacterial activities[7]. There
are extensive studies on the medicinal properties
and active principles in the nut and pericarp. In
traditional medicines of Kerala, A. catechu root
powder is applied for skin allergies, dierent worm
infections including ring worm infection and adding
as a component in health tonic preparations[8].
Our previous paper evaluated the anti-oxidant,
antihelmintic and anti-microbial properties of A.
catechu root crude extract[9]. The present study was
carried out to examine the anti-cancerous property
of A. catechu root and the identication of the active
principles in it using Liquid Chromatography-Mass
Spectrometric (LC-MS) analysis (g. 1).
Fig. 1: Anti-cancer property screening of A. catechu root, (I): In vitro cytotoxic property screening of A. catechu root, (a):
Ethanol extract, (b): Acetone extract and (c): Aqueous extract; (II): In vivo anti-tumor activity of A. catechu ascites tumor model, (a):
Cyclophosphamide treated group, (b): A. catechu aqueous extract treated group and (c): A. catechu ethanol extract treated group
and (III): Eect of A. catechu root extract on solid tumor induced by DLA cells (a): A. catechu aqueous extract treated mice, (b): A.
catechu ethanol extract treated mice and (c): Cyclophosphamide treated mice

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Indian Journal of Pharmaceutical Sciences 1665
September-October 2024
Braca et al.[10], was used for determining the anti-oxidant
property. Test solutions in dierent concentrations (20,
40, 60, 80 and 100) μg/l and 6.34 μM solution of DPPH
were prepared in methanol. 100 μl test solution, 100
μl DPPH and 800 μl methanol were taken in a test
tube and mixed well. Optical density of the resulting
solution was measured at 517 nm after incubation at
dark for 20 min. Methanol (900 μl) with 100 μl 6.34
μM DPPH was taken as control and methanol as blank.
The optical density was recorded and the potential of
plant extracts to scavenge the DPPH free radicals were
calculated using the formula.
Percentage of inhibition=A-B/A×100
Where, A is optical density of control and B is optical
density of sample.
Acute toxicity study:
Acute toxicity assay was performed in healthy adult
non-pregnant female Swiss albino mice (25-28 g body
weight). The mice were divided into two groups of three
each and treated with 250 mg/kg drug intraperitoneally.
The control group received 2 % carboxymethyl
cellulose suspension at the same volume.
In vitro cytotoxicity screening:
Short term cytotoxic activity of A. catechu sequential
extracts were assayed by determining the percentage
viability of the DLA cells using trypan blue exclusion
methods[11]. The cells were aspirated from the peritoneal
cavity of tumour bearing mice. The collected cells were
washed using Phosphate Buered Saline (PBS) and
checked for their viability. Dierent dilutions of the
cells were made (10-1, 10-2, 10-3) and the number of cells
in the 10-3 dilution was counted using haemocytometer
and the cell number was adjusted to 1×107 cells/ml.
This cell suspension was added to tubes containing
various concentrations of test in 1 ml PBS and the tubes
were incubated at 37° for 3 h. 100 µl of trypan blue was
added after the incubation period and the percentage of
viability were determined.
Anti-cancer eect of A. catechu on ascites tumour
bearing animals:
Ascites tumour was induced by injecting DLA cells
(1×106 cells/animal) in the peritoneal cavity of Swiss
albino mice. 36 animals get divided into six groups, each
group consist of 6 animals. Group I was maintained as
negative control (not treated with any drug). Group II-V
received 50 and 100 mg/kg body weight of aqueous and
ethanolic extracts of A. catechu (which shows highest
MATERIALS AND METHODS
Plant material:
Fresh roots of A. catechu were collected from
Mannamangalam Village of Thrissur district Kerala.
The material was authenticated by Dr. Regi Raphael
K and a voucher specimen is also deposited in the
herbarium of Botany Department, St. Mary’s College,
Thrissur, Kerala, India with voucher number SMC/
M/A-8.
Cell lines:
Daltons Lymphoma Ascites (DLA) cell-lines were
procured from Amala Cancer Research Institute,
Thrissur, Kerala, India. The mice were injected with a
suspension of cells (1×106) intra peritoneally and the
cells were aspirated from the peritoneal cavity on the
15th d.
Animals:
Swiss albino mice (non-pregnant females of 6-8 w age)
were purchased from Small Animal Breeding Station
(SABS), College of Veterinary and Animal Sciences,
Mannuthi, Thrissur, Kerala. The animals were kept in
well-aerated cages with controlled conditions of light
and humidity for 14 d for acclimatization. The animals
were fed with normal mouse chow (Sai Durga Food
and feeds, Banglore, India) and water ad libitum. All
experiments in the study were carried out with the prior
approval of Institutional Animal Ethics Committee
(IAEC) and were conducted as per the guidelines of
Committee for the Purpose of Control and Supervision
of Experiments on Animals (CPCSEA) constituted by
the Animal Welfare Division, Government of India.
Plant extraction and fractionation:
Washed and cleaned materials were kept in hot air oven
at 50° for 10 d. Dried material was ground into a coarse
powder using an electric blender. Sequential extracts of
varying polarity (petroleum ether, benzene, chloroform,
acetone, ethylene and water) were prepared using
column chromatography. The fractions collected were
concentrated using rotary evaporator and percentage of
yield in each solvent were calculated.
Bioactivity guided selection of the active solvent
fraction:
Anti-oxidant assay (2,2-Diphenyl-1-Picrylhydrazyl
(DPPH) free radical scavenging assay) of the serial
extracts was performed to choose the most active solvent
fraction for further studies. The procedure given by

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Indian Journal of Pharmaceutical Sciences
1666 September-October 2024
one way Analysis of Variance (ANOVA) and Duncan
Multiple Range Test (DMRT). The dierences between
the groups were considered statistically signicant at
p<0.05.
RESULTS AND DISCUSSION
Yield of the extract in dierent solvents were
determined as follows; petroleum ether-3 %, benzene-2
%, chloroform-1.5 %, acetone-2.6 %, ethanol-6
% and distilled water-18.3 %. Evaluation of the
extractive values of powdered drugs is benecial for
their evaluation especially in the cases where their
constituents cannot estimate readily. Here the aqueous
extract of A. catechu gave maximum yield.
There are several mechanisms which involved in the
antioxidant activity like free radical mediated chain
reaction termination, donation of hydrogen, hydrogen
abstraction prevention, peroxide elimination and
catalytic ion chelation. So, a single assay would not
express the antioxidant potential of a plant extract. Here
the only method, DPPH free radical scavenging activity
was used to nd out the biologically active solvent
extract for phytochemical screening.
The basic information of the ecacy of compounds in
A. catechu extracts to quench the free radicals can be
determined using DPPH free radical scavenging assay.
All the extracts screened showed scavenging of free
radicals based on their concentration (Table 1). 50 %
radical scavenging in least concentration was observed
in aqueous extract (53 µg/l). So the aqueous extract
of A. catechu was selected for anti-cancer studies and
phytochemical proling to detect the various active
principles present in it.
In the toxicity test, dose of 250 mg/kg body weight of
the mice did not cause mortality or any signs of toxicity
or change in general behaviour during the 14 d of
observation. So, it is conrmed that, the plant extract
is not toxic to animals. Table 2 shows the results of the
in vitro cytotoxicity screening of A. catechu root. Both
polar and non-polar extracts of A. catechu found to be
cytotoxic towards DLA cells. Maximum cytotoxicity
(89 %) was attained at a concentration of 200 µg/l
of ethanolic extract. Least half maximal Inhibitory
Concentration (IC50) value was showed by aqueous
extract (38 µg/l). So, ethanolic and aqueous extracts
were selected for the in vivo anticancer screening.
Animals of the control group survived only for a period
of (15±2.09) d. Treatment of A. catechu ethanolic and
aqueous extracts at dierent concentrations increased
the survival rate of animals (Table 3). One way
activity in in vitro cytotoxicity screening). Animals in
the group VI received cyclophosphamide (10 mg/kg
body weight). The drugs were given intraperitoneally
after 24 h of tumour implantation as 5 doses on alternate
days. The death of the animals due to tumour burden
was noted every day and the Percentage of Increase
in Lifespan (% ILS) was calculated using the below
mentioned formula[12].
% ILS=(T-C/C)×100
Where, T is mean survival days of treated and C is mean
survival days of control animals.
Anti-cancer eect of A. catechu on solid tumour
bearing animals:
Animals were divided into 6 groups and each group
carried 6 mice. Viable DLA cells aspirated from the
peritoneal cavity of ascites tumor bearing mice in the
concentration 1×106 cells in 0.1 ml PBS were transplanted
into the right hind limb of mice. 50 mg/kg and 100 mg/
kg drugs (aqueous and ethanolic extracts of A. catechu
root) were administered intraperitoneally after tumour
transplantation and continued for 10 consecutive
days. Control group received only DLA cell line and
standard group were treated with cyclophosphamide in
the concentration 10 mg/kg. The development of solid
tumor in each group was determined by measuring
the diameter of tumour in perpendicular planes using
vernier calipers in for every 7 h. The tumour volume
was calculated using the formula[12]
V=4/3πr12r2
Where, r1 is the minor radius and r2 is the major radius.
LC-MS analysis:
LC-MS system facilitates the analysis of samples,
which have diculty to analyze traditionally. Even
though the technique Gas Chromatography-Mass
Spectrometry (GC-MS) is powerful as an analytical
agent, many compounds are impossible to analyze
with GC-MS. LC-MS is suitable for the analysis
of large, ionic, thermally unstable and non-volatile
compounds. A mass spectrometer combined with a
LC can detect masses characteristic of a compound
or a class of compounds. The system can selectively
detect compounds of interest in a complex matrix, thus
making it easy to nd out and identify.
Statistical analysis:
The numerical data obtained were statistically analyzed
and expressed as mean±Standard Deviation (SD). The
signicant levels of comparison were analyzed using

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Indian Journal of Pharmaceutical Sciences 1667
September-October 2024
dierent; both aqueous and ethanolic plant extracts are
equally signicant in action to the commercial drug
cyclophosphamide. And the number of days survived in
these groups is signicantly higher than the treatments
in group 1, 2 and 4. Treatment groups 2 and 4 shows no
signicant dierence in the number of days survived.
Number of days survived is signicantly lower in the
rst group compared to all other groups.
ANOVA was carried out for comparing number of days
survived among dierent treatment groups. F-value
was found to be signicant at 0.01 level as the p<0.01.
This shows that there exists signicant dierence in
the number of days survived among dierent treatment
groups. DMRT was carried out as post hoc analysis
to nd out which of the groups are homogeneous and
which of them are signicantly dierent. Results shows
that number of days survived are not signicantly
S. No. Concentration
(µg/l)
Percentage of inhibition
Petroleum
ether Benzene Chloroform Acetone Ethanol Distilled water
1 20 14.5±2.1 18.6±0.67 5.03±4.2 6.9±1.89 24.6±0.72 12.6±0.51
2 40 24.6±1.5 25.5±0.92 17±1.15 18.2±2 35.4±0.8 33±0.92
3 60 39.6±0.57 32.8±2.42 26.3±0.38 27.6±0.56 49.1±1.10 60.5±1.49
4 80 47.6±1.15 48±2.3 48.6±1.56 49.4±1.08 72.2±2.1 76.46±1.8
5 100 59.5±0.7 61.8±0.72 67.6±1.72 63±1 85.5±1.05 92.8±1.13
IC50 84±0.84 83±1.59 82±0.98 81±1.05 61±1.15 53±0.87
Note: Values expressed as mean±SD
TABLE 1: DPPH FREE RADICAL SCAVENGING PROPERTY OF A. Catechu ROOT
S. No. Concentration
(µg/l)
Percentage of inhibition
Petroleum
ether Benzene Chloroform Acetone Ethanol Distilled water
1 10 8±0.67 11±0.62 10±0.65 9±0.61 10±0.58 9±0.48
2 20 20±0.87 19±0.83 31±0.78 37±0.76 30±0.67 21±0.73
3 50 42±1.63 40±0.93 44±0.97 54±1.23 57±0.69 36±0.98
4 100 61±0.72 49±1.23 61±0.69 68±0.87 63±1.67 59±01.46
5 200 84±0.48 61±1.56 79±0.88 82±0.58 89±0.95 76±1.93
IC50 value 40±0.93 41±0.97 45±0.73 56±0.97 55±0.83 38±1.12
Note: Values expressed as mean±SD
TABLE 2: IN VITRO CYTOTOXIC PROPERTY SCREENING OF A. catechu ROOT
TABLE 3: IN VIVO ANTI-TUMOUR ACTIVITY OF A. catechu: ASCITES TUMOUR MODEL
S. No. Treatment Number of days survived Percentage increase in life span
1 DLA cells alone 15.0±2.09c-
2 DLA+A.catechu ethanolic extract 21.7±3.01b28.75
(50 mg/kg body weight)
3 DLA+A.catechu ethanolic extract 26.9±2.80a66.25
(100 mg/kg body weight)
4 DLA+A.catechu aqueous extract 23.8±2.14b42.5
(50 mg/kg body weight)
5 DLA+A.catechu aqueous extract 26.5±1.07a71.9

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Indian Journal of Pharmaceutical Sciences
1668 September-October 2024
ethanolic and aqueous extracts of A. catechu eectively
increased the life span of aected mice. Highest activity
was observed in aqueous extract.
The result of the HR LC-MS of aqueous extract
revealed the presence of valuable compounds with
proved medicinal properties in the root of A. catechu.
Mitoxantrone, which is available in the trade name
Novantrone® is used in the treatment of certain cancers,
mostly in metastatic breast cancers, non-Hodgkin’s
lymphoma and acute myeloid leukemia. It is a type II
topoisomerase inhibitor, it disrupts Deoxyribonucleic
Acid (DNA) synthesis and repair in both healthy and
cancer cells by intercalation between DNA bases[16].
Artemether is a medication used in the treatment of
malaria. In the case of severe malaria, artemether is
given in its injectable form[17]. Valproic acid is used as
the medication in the treatments of epilepsy, migraine
headache and bipolar disorders. Due to the broad
spectrum action of valproate, it is used in the treatments
of anticonvulsant activity, as a rstline treatment in
tonic clonic seizures, mycoclonic seizures and absence
seizures. It also used as second line treatment for
infantile spasms and partial seizures[18].
Leupeptin, it is a well-known antioxidant and anti-
inammatory agent widely used in the medical eld[19].
The present study reveals that, this plant is a reservoir
of several medicinally active compounds. So, there
is no doubt that this plant is very promising as a
traditional medicinal plant. Proteases are enzymes that
play a crucial role in the regulation of various cellular
processes, including cell growth, dierentiation and
apoptosis (programmed cell death). Leupeptin is a
protease inhibitor that has been studied for its potential
role in cancer therapy. Dysregulation of proteases has
been implicated in cancer development and progression.
Leupeptin inhibits several proteases, including serine,
cysteine and thiol proteases, thereby aecting multiple
pathways involved in cancer[20]. Potential mechanisms
through which leupeptin may exert its anti-cancer eects
are apoptosis induction, cell cycle arrest, angiogenesis
inhibition and metastasis suppression[21].
Development of solid tumor found to be reduced in
the A. catechu root treated groups when compared
to the control group from 21st d of observation. The
volume of solid tumour in control group on 35th d was
(4.550±0.622) mm while in case of aqueous root extract
in 100 mg/kg was (0.48±0.92) mm on the same day.
The value shown by the standard drug (0.643±0.111)
was more than that of the aqueous extract at 100 mg/kg.
The result was presented in the Table 4.
From these results it is clear that the plant extracts are
highly ecient as anti-tumour agents, the percentage
of increase in life span is increasing with the increase
in concentration of the plant extract in ascites tumour
and solid tumor development is signicantly reduced
by the plant drug. Aqueous extract of A. catechu, which
shows maximum anti-cancer property were given for
High Resolution LC-MS (HR LC-MS). 59 compounds
were detected through the analysis; from the list
database formula dierence in the range of -10 to +10
is considered as signicant. 14 compounds are present
in signicant amount which is given in the Table 5.
Medicinal plants are nature’s gift to human beings
to lead a healthy, disease free life. Most of these
plants used today are believed to be much safer and
proved as elixir in the treatment of various ailments.
Plant derived compounds have played an important
role in the development of several clinically useful
anticancer agents[13]. Oxidative stress induced by an
imbalance between production of reactive oxygen
species and antioxidants are associated with pathogenic
disease conditions like carcinogenesis[14]. So, radical
scavenging activity is very important in the searching
of natural sources of cancer drugs.
Cytotoxicity is one of the chemotherapeutic targets of
antitumor drugs[15]. Most of the clinically proved anti-
tumour agents possess signicant cytotoxic activity
in cell culture systems. The cytotoxic activity of A.
catechu root extracts against DLA cell lines partially
explains its signicant anti-tumour activity. The drug
shows toxicity towards the tumour cell line and not toxic
to normal cells. The anti-cancer activity was evaluated
using ascites tumour and solid tumour models. Both
(100 mg/kg body weight)
6 DLA+cyclophosphamide (10 mg/kg) 26.7±2.07a71.5
F 22.58**
p <0.001
Note: **Signicant at 0.01 level and means having same letter as superscript are homogeneous

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Indian Journal of Pharmaceutical Sciences 1669
September-October 2024
Tumor volume in
treated group
Day of determining tumour volume
1st 7th 14th 21st 28th 35th
Control 0.440±0.0317 0.643±0.20 0.956±0.16 2.972±0.452 3.790±0.62 4.550±0.622
Standard
cyclophosphamide
(10 mg/kg)
0.442±0.092 0.635±0.87 0.982±1.45 1.742±0.172 1.122±0.227 0.643±0.111
Ethanol extract (50
mg/kg) 0.456±0.072 0.67±0.87 0.964±0.15 1.84±0.98 1.359±0.31 0.816±0.13
Ethanol extract
(100 mg/kg) 0.426±0.03 0.643±0.201 0.953±0.86 1.615±0.12 1.18 ±1.79 0.71 ±0.69
Aqueous extract (50
mg/kg) 0.450±0.02 0.643±0.20 0.984±0.236 1.64±0.68 1.359±0.3 0.88± 0.85
Aqueous extract
(100 mg/kg) 0.439±0.04 0.621±0.068 0.956±0.168 1.554±0.49 1.036±0.48 0.48 ±0.92
Note: Values expressed as mean of tumor volume in cm±SD
TABLE 4: EFFECT OF A. catechu ROOT EXTRACT ON SOLID TUMOUR INDUCED BY DLA CELLS
S. No. RT Mass Name Formula DBDIFF (parts per million) HITS database
1 0.145 426.2983 Leupeptin C20H38N6O4-6.62 15
2 0.147 218.1145 3-hydroxysebacic acid C10H18O54.4 2
3 0.153 737.5105
1-heptadecanoyl-2-(9Z-
tetradecenoyl)-sn-glycero-3-
phosphoserine
C37H74N2O10P -3.27 5
4 0.153 616.413 6'-hydroxysiphonaxanthin C40H56O5-0.35 4
5 0.154 196.1093 4-(2-hydroxypropoxy)-3,5-dimethyl-
Phenol C11H16O33.54 11
6 0.154 513.2771 Sulfolithocholylglycine C26H43NO7S -2.03 3
7 0.155 567.2872 Dihydrodeoxystreptomycin C21H41N7O11 -1.45 1
8 0.156 639.3083 Protorifamycin I C35H45NO10 -6.2 1
9 0.168 296.1823 Farnesylthioacetic acid C17H28O2S -4.31 10
10 0.17 466.3841 Ergosterol acetate C32H50O2-6.53 3
11 0.177 320.1461 Valproic acid glucuronide C14H24O83.09 15
12 0.177 444.2033 Mitoxantrone C22H28N4O6-5.44 1
13 0.177 298.1769 Artemether C16H26O53.74 6
Note: RT: Retention Time
TABLE 5: COMPOUNDS PRESENT IN THE A. catechu ROOT AQUEOUS EXTRACT
In conclusion the results of the in vitro cytotoxicity
screening and anti-tumour studies of A. catechu shows
that, it can act as a source of active compounds for
the preparation of anti-cancer drugs. The presence of
various secondary metabolites like alkaloids, saponins,
phenols, steroids and avonoids provides some
scientic evidence for the biological activities and also
account for the pharmacological uses. LCMS analysis
of the aqueous extract shows the presence of large
number of compounds with proved medicinal uses.
So this unravelled medicinal plant will be a prominent
contributor of medicinal compounds in the near future.
Acknowledgements:
The authors are thankful to the Department of Botany,
St. Mary’s College Thrissur and Amala Cancer
Research Centre, Thrissur, Kerala, India for providing
the laboratory facilities to carrying out the research.

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Indian Journal of Pharmaceutical Sciences
1670 September-October 2024
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Conict of interest:
The authors declared no conict of interests.
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