Estimation of flavonoid lantana camara linn verbenaceae

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* Corresponding author: Sorabh kumar agrawal
E-mail address: sorabh44@gmail.com
Available online at www.ijrpp.com
Print ISSN: 2278 – 2648
Online ISSN: 2278 - 2656 IJRPP | Volume 2 | Issue 1 | 2013 Research article
Anti Cancer activity of Cinnamomum Malabatrum against Dalton’s ascitic
lymphoma
*Sorabh kumar agrawal, R.C.Chipa, K.C.Samanta Suresh.
Gyan Vihar School Of Pharmacy, Jaipur, Rajasthan, India.
ABSTRACT
The aim of the research is to find out new anti cancer drugs from indigenous plant which are potent, nontoxic or
minimal toxic and to investigate the anti cancer activity of Cinnamomum Malabatrum. The powder of Cinnamomum
Malabatrum was successively Extracted with Petroleum Ether, Chloroform, Acetone, Ethyl alcohol and Aqueous
.The preliminary phytochemical test were done and the LD50 values for both alcohol and aqueous extract
determined. The anti cancer activity of the alcoholic (625 mg / kg. p.o.) and aqueous Extract (500 mg / kg. P.O.)
were assessed in Dalton’s Ascitic Lymphoma induced cancer.
KEY WORDS: Cinnamomum malabatrum, Delton,
s ascitic lymphoma.
INTRODUCTION
Herbal medicine is the oldest form of healthcare
known to Mankind herbs has been used by all
cultures throughout history. It was an integral part of
the development of modern civilization. Primitive
men observed and appreciated the great diversity of
plants available to him. The Cinnamomum
Malabatrum is moderate ever green tree, bark smooth
or slightly longitudinal cracked light brown, leaves
are opposite or sub-opposite, elliptic to oblong,
glabrous, pink when young, 3-nerved from close
above the base almost to the apex. Flowers long, pale
yellowish, fruits ellipsoid. Cinnamomum Malabatrum
leaves contain chemical composition of cinnamic
aldehyde, euganol, β-caryophyllene, benzaldehyde,
camphor, cadinene, α-terpinol, limonene, geraniol,
euganol acetate, ocimene, γ-terpinene, benzyl
cinnamate, β-phellandrene and benzyl acetate. The
oil from bark contains cinnamaldehydes (70-85%) as
a major constituents. The plant also contain 3,4’,5,7-
tetra hydroxyl flavones, 3,3’,4’,5,7-pentahydroxy
flavones, kaempferol-3-O-sophoroside and quercetin
3-O- rutin. The plant has been traditionally used as
stimulant, carminative, haemostatic, astringent,
diaphoretic, deobstruent and galactogogue. The bark
also prescribed in bowl complaints such as dyspepsia,
flatulence, diarrhea and vomiting. The leaves are
carminatives and are used in colic and rheumatism.
They are sweetish, heating, useful in vata, scabies,
disease of the anus and rectum, tridosha, piles and
heart troubles. The dried buds are used with various
combinations in cough and urinary disease. The plant
is also used for treatment of some tumors. Hence
present study was aimed to investigate the anti cancer
activity of plant extract of Cinnamomum Malabatrum
on DAL induced cancer in Swiss Albino Mice.
MATERIAL AND METHODS
The bark of Cinnamomum Malabatrum were
collected from ABS Botanical Conservation,
International Journal of Research in
Pharmacology & Pharmacotherapeutics

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Research and Training Center, Kaaripatti, Tamilnadu
in month of July,2009. The plant was identified and
authenticated by Dr.A.BALASUBRAMANIAN. The
bark part of Cinnamomum Malabatrum was shaded
dried at room temperature and then powdered with a
mechanical grinder. The powder was passed through
sieve no.40 and stored in an air tight container for
further use .The solvent used Petroleum ether (60-80
c), Chloroform, Acetone, Alcohol 95% w/v and
Distilled water for decoction. The Petroleum ether
extract was obtained using Soxhlet apparatus.
Alcoholic extract was obtained with ethyl alcohol
95% w/v for 18 hrs using Soxhlet apparatus. The
aqueous extract was prepared with the remaining
mass by maceration process for 7 days. The extract
was dried at 55 ºC in a water bath. The Percentage
yield of Petroleum ether, Chloroform, Acetone,
alcoholic and aqueous extract were 1.92%
,1.68%,5.16%,3.89%and 12.2% respectively.(Table
no.1)
Phytochemical Screening
The preliminary phytochemical screening of
Alcoholic and aqueous extract of Cinnamomum
Malabatrum were carried out as described by
khandelwal. (Table no.2)
Animals
Male & Female Swiss albino mice weighing between
20-25 gm were used for this study . The animals
were obtained from animal house of Sri
Venkateshwara Enterprises, Bangalore, India. On
arrival the animal were placed randomly and
allocated to treatment groups in propylene cages with
paddy husk as bedding. Animals were housed at a
temperature of 24± 2ºC and relative humidity 45-
55% with 12:12 hours light/dark cycle. The animals
had free assist to food and water . The animals were
habituated to laboratory conditions for 48 hours prior
to the experimental protocol to minimize any non-
specific stress. All the experimental process and
protocols used in this study were reviewed by the
institutional animal ethical committee (Pharmacology
project no. 02/2010) and were in according with the
guideline of the CPCSEA.
Acute Toxicity Study
Acute oral toxicity of alcoholic and aqueous extract
were determined using adult Swiss albino mice of
both sexes. The control animals received normal
saline orally. The Alcoholic extract of Cinnamomum
malabatrum was administered once orally at various
dose levels (6000 to 6400 mg/kg body wt. to group of
6 mice of both sexes about equal in number which
have been fasting overnight (about 18h.). The
aqueous extract of Cinnamomum malabatrum was
administered orally at various dose levels (4400 to
5200 mg/kg body wt) to other group of animals. The
animals were observed continuously for 2 hours and
then occasionally for further 4 hours and finally
overnight mortality recorded.
Experimental Protocol
Test compound The Alcoholic & Aqueous extract
of Cinnamomum malabatrum (625mg/kg & 500
mg/kg body weight) and standard drug 5-fluorouracil
(20 mg/kg i.p) were used. The following chemicals
were obtained from the indicated commercial.
Experimental Setup
Swiss albino Mice (20-25 gm) used in the present
studies was procured from listed suppliers of Sri
Venkateshwara Enterprises, Bangalore, India. The
animals were fed with standard pellet diet (Hindustan
lever Ltd. Bangalore) and water ab libitum. All the
animals were acclimatized for a week before use. The
alcoholic & aqueous extract of Cinnamomum
malabatrum was dissolved in normal saline.
The rats were divided into five groups of 6
animals in each
Group І : Normal Control
Control animal were received normal saline
Group ІІ : Tumor control
Animals were inoculated with 1X106 cells per
mouse intraperitoneal
Group III : Standard Group
Animals were injected 5-fluorouracil (20mg/kg)
intraperitoneal along with DAL cells (1X106 ) cells
per mouse treatment.
Group IV : Test group- I
Animals were treated with alcoholic extract of
Cinnamomum malabatrum 625mg/kg orally along
with DAL cells (1X106 cells /mouse)
Group V : Test group -II
Animals were treated with aqueous extract of
Cinnamomum malabatrum 500 mg/kg orally with
DAL cells (1X106 cells/mouse) treatment.

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EVALUATION PARAMETERS
Effect on Solid Tumor Volume by Simultaneous
Method
To determine the effect of alcoholic and aqueous
extract of Cinnamomum malabatrum on solid tumor,
all the animals were injected with 1x106
cells/mouse
in phosphate buffered saline into the right hind limb
of all the animals simultaneously. All the treatments
were started from the next day of inoculation and
were continued for five alternative days. The
measurement of tumor radii was taken from 11th
day
of tumor induction and was repeated on every 5th
day
for a period of 30 days. The volume of tumor mass
was calculated from the formula V= 4/3π r2
where ‘r’
is mean of r1 and r2 which are the two independent
radii of the tumor mass (Table no.3).
Effect On Peritoneal Cells Counts in Mice
One group of animals was treated with aqueous
extract at a dose of 500mg/kg/day, p.o. of stem bark
of Cinnamomum malabatrum once for a single day
and the second group received the same treatment for
two consecutive days. Similar treatment was given to
other group with Alcoholic extract at a dose of 625
mg/kg/day/p.o. the untreated first group was used as
control. Peritoneal exudates of alcoholic and aqueous
extract groups were collected after 24 hr and 48 hr of
treatment by repeated intraperitoneal wash with
normal saline (0.9% w/v) and the cells were counted
in each of the treated groups under WBC newbauer’s
chamber and compared with those of normal control
(Table no.4) .
Effect On Body Weight(21-22)
Four groups of six mice each were transplanted
intraperitoneal with 1x106
Dalton’s ascetic tumor
cells. After 24h, the first and second group were
orally treated with Alcoholic and aqueous extract of
Cinnamomum malabatrum. The third group, serving
as the control, received normal saline (0.9% w/v).
Treatments were continued for 9 days. Body weights
were recorded every 5th
day till 40 days of treatments
or till the death of the animal (Table No.5).
RESULTS AND DISCUSSION (23-24)
Preliminary phytochemical studies show the presence
of Flavonoids, Fixed oil, Amino acids, Tannins and
Phytosterols according to Table no.2. The Stem bark
extract of Cinnamomum malabatrum was found to be
minimal toxic. Treatment of mice with DAL
produces cancer. DAL is one of the most common to
produce tumor then present study demonstrated as
aqueous extract exhibited significantly dose
depended anti cancer activity. Alcoholic and
aqueous extract shows significant decrease in Solid
tumor volume, Increase in Peritoneal cell count and
body weight compare to tumor control group. It can
be concluded that Cinnamomum malabatrum stem
bark extracts posses a protective effect against DAL
induced cancer in mice.
Table No.1 Data Showing the Extractive Values of Cinnamomum malabatrum
Plant
name
Part used
Method of
extraction
Extracts Extractive Values (w/w)
Cinnamomum
malabatrum
Stem bark
Continuous
hot
percolation
Petroleum Ether
1.92%
Chloroform
1.68%
Acetone
5.16%
Alcoholic
3.89%
Aqueous
6.67%

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Table No.2 Data showing the Preliminary Phytochemical Screening for extracts of
Cinnamomum malabatrum
Phytochemical
constituents
Petroleum
ether
Chloroform Acetone Alcohol (95%) Aqueous
ALKALOIDS - - - - -
FLAVANOIDS - - + + +
CARBOHYDRATE - - - + +
SAPONINS - - - + +
TRITERPENS + + -
STEROLS + - -
TANNINS - - + + +
GLYCOSIDES - - - - -
+ VE = Present -VE = Absent
Table No.3 Data showing the Effect of Cinnamomum malabatrum on Solid tumor volume
(Mean± SEM n=6 in each group) in rats
*P<0.01 Values are represented as mean ± S.E.M (n=6)
One-way ANOVA followed by Student-Newman-Keuls post test (P< 0.001) is used.
a-vs group I and b-vs group II.
Groups Treatment Solid Tumor Volume (ml)
15th
days 20th
days 25th
days 30th
days
I Tumor Control 7.56 ± 0.22 8.16 ± 0.25 8.96 ± 0.25 11.16 ± 0.33
II Standard 4.56 ± 0.22* 4.16 ± 0.21* 4.16 ± 0.25* 3.16 ± 0.21*
III Test Group- I 6.16 ± 0.25* 6.66 ± 0.22* 7.16 ± 0.22* 7.6 ± 0.25*
IV Test Group – II 5.56 ± 0.22* 6.33 ± 0.21* 7.33 ± 0.21* 8.1 ± 0.34*

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Table No.4 Data showing the Effect of Cinnamomum malabatrum on Peritoneal Cell Count
(Mean± SEM n=6 in each group) in rats
Values are represented as mean ± S.E.M (n=6)
One-way ANOVA followed by Student-Newman-Keuls post test (P< 0.001) is used.
a-vs group I and b-vs group II.
Table No.5 Data showing the Effect of Cinnamomum malabatrum on Body Weight
(Mean± SEM n=6 in each group) in rats
One-way ANOVA followed by Student-Newman-Keuls post test (P< 0.001) is used.
a-vs group I and b-vs group II.
ACKNOWLEDGEMENT
Authors are thankful to Management and Staff for
providing the necessary facilities to conduct this
study and are thankful to Ms.Divya for help
pertaining to complete this study. Authors are also
thankful to all persons whose help in this work.
Groups Treatment Peritoneal cell count (1X106
)
I
Normal Control 5.08 ± 0.01
II Tumor Control 3.21 ± 0.07 ***
III Standard 10.21 ± 0.02 ***
IV Test Group –I 9.48 ± 0.13**
V Test Group- II 14.21 ± 0.12
S.N
o.
Group Dose 7th
Day 14th
Day 21st
Day 28th
Day 35th
Day
I Normal
control
- 22.56 ± 0.77 23 ± 0.52 24.16 ± 0.52 27.66 ± 0.66 31.83 ± 0.83
II Tumor
control
- 27.86 ± 0.22* 40.56 ± 0.22* 50.16 ± 0.65* - -
III Standard 20mg/kg/
day
24.36 ± 0.62 24.5 ± 0.32** 26.56 ± 0.6** 29.33 ± 0.6 33.46 ± 0.47
IV Test Group
–I
625
mg/kg/day
25.05 ± 0.25 30.56 ± 0.33** 32.6 ± 0.66** 34.56 ± 0.32** 37.34 ± 0.33
V Test Group
–II
500
mg/kg/day
24.36 ± 0.33* 29.76 ± 0.76** 30.5 ± 0.82** 31 ± 0.25 35.83 ± 0.30

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