Karthik et.

al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)

EVALUATION OF ANTI-INFLAMMATORY ACTIVITY OF CANTHIUM

PARVIFLORUM BY IN-VITRO METHOD
Kandikattu Karthik1*, Bharath Rathna Kumar P1, Venu Priya R2, Sunil Kumar K3 , Ranjith Singh.B.Rathore4
*1Department of Pharmaceutical Analysis, JNTUA - Oil Technological Research Institute, Ananthapuramu, A.P, India.
2
Department of Pharmaceutics, JNTUA - Oil Technological Research Institute, Ananthapuramu, A.P, India.
3
Department of Pharmacology, Vijaya College of Pharmacy, Hyderabad, A.P, India.
4
Department of Pharmacognosy, Vagdevi College of Pharmacy, Nellore, A.P., India.
*Corresponding author: Email: kkarthik1922@gmail.com
ABSTRACT
The present study investigates the anti-inflammatory activity of ethanolic extract of Canthium Parviflorum,
is a valuable medicinal shrubby and woody plant which has been valued for centuries in ayurvedic medicine.
Phyto-chemical analysis of Canthium parviflorum plant extracts revealed the presence of various bio-chemical
compounds such as flavonoids, glycosides, alkaloids, saponins and terpinoids. Since terpenoids and flavonoids
have remarkable anti-inflammatory activity, our present work aims at evaluating the in-vitro anti-inflammatory
activity of Canthium parviflorum by protein denaturation method. Denaturation of proteins is a well documented
cause of inflammation and rheumatoid arthritis. Several anti-inflammatory drugs have shown dose dependent
ability to inhibit thermally induced protein denaturation. Ability of CPE extract to bring down thermal
denaturation of protein is possibly a contributing factor for its anti-inflammatory activity. The data of our studies
suggests that CPE extract showed significant anti-inflammatory activity. Therefore our studies support the
isolation and use of active constituents of Canthium parviflorum in treating inflammation.
Key Words: Canthium parviflorum, flavanoids, glycosides, phytochemical analysis, inflammation.
INTRODUCTION ethanol extract of Canthium parviflorum leaves for their
anti-inflammatory activity.
Medicinal plants are believed to be an important
source of new chemical substances with potential MATERIALS AND METHODS
therapeutic effects. The research into plants with alleged
Plant material: Fresh leaves of the plant Canthium
folkloric use as anti-inflammatory agents should therefore
parviflorum were collected from Andhrapradesh, India.
be viewed as a fruitful and logical research strategy in the
The plant material was taxonomically identified by
search for new anti-inflammatory drugs. Inflammation
C.V.S.Bhaskar, Prof. in Botany, Venkatagiri Raja’s
may be potentially harmful, causing life threatening
college, SPSR Nellore, Andhra Pradesh, India. A voucher
hypersensitivity reactions and progressive organ damage
specimen has been preserved in our laboratory for future
(Robbins, 2008). NSAIDs are reported to possess
reference. The leaves were dried under shade and then
prevention of the denaturation of proteins, which act as
powdered with a mechanical grinder and stored in airtight
antigens and leads to auto-immune diseases (Insel, 1996).
container. The dried powder material of the leaves was
Canthium as herbal medicine is used for the treatment of
defatted with petroleum ether and subsequently extracted
diabetes (Ayyanar, 2008), treatment of snake bites
with ethanol in a Soxhlet apparatus. The solvent was
(Mahishi Parinitha, 2005), scabies and the ring worm
completely removed under reduced pressure and Ethanol
infection (Anitha, 2008) antioxidant and diuretic activity
extract of Canthium parviflorum leaves was obtained
(Mohideen, 2003). Canthium Parviflorum, is a valuable
(yield 18.5%). Solution of Canthium parviflorum was
medicinal shrubby and woody plant which has been
prepared freshly in distilled water and used for the studies.
valued for centuries in ayurvedic medicine. Phyto-
chemical analysis of Canthium parviflorum plant extracts Chemicals and drugs: Tris amino methane and bovine
revealed the presence of various bio-chemical compounds Serum Albumin were obtained from Himedia Lab.,
such as flavanoids, glycosides, alkaloids, saponins and Mumbai India, Ibuprfen from Sigma-Aldrich and all other
terpinoids. Different parts of Canthium parviflorum, have chemicals used were of analytical grade. Solvents were
been used traditionally for the treatment of variety of purchased from Sd-Fine Chem Ltd. All other chemicals
diseases including anaemia, toothache, cough and as a used were of analytical grade and purchased locally.
hypoglycemic agent. Roots and leaves were used to Extract Preparation: Plant parts were air dried at room
reduce swellings in inflammation. However, there is no temperature for 4 weeks to get consistent weight. The
systematic scientific report published indicating utility of dried parts were later ground to powder. 100g of dried
this plant material in the treatment of inflammation. Thus samples were extracted with hexane and ethanol using
the presence of therapeutically active flavonoids as major
constituents was the basis of selection and evaluation

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Karthik et.al Indian Journal of Research in Pharmacy and Biotechnology
Soxhlet apparatus. The extracts were concentrated under
reduced pressure by using rota vapour.
Phytochemical analysis: Phytochemical analysis was
carried out for the presence of various phytochemical
Table.1.Phyto chemical Screening of Canthium parviflorum extracts
Test Hexane Extract
Alkaloids
Saponins
Tanins
Phlobatanins
Steroids
Flavonoids
Anthraquinones
Glycosides
Reducing sugars
Terpenoids
+ve sign indicates the presence and –ve sign indicates the absence
In-vitro anti-inflammatory activity:
Antidenaturation activity: A solution of 0.2% w/v of
BSA was prepared in a Tris Buffer Saline and pH was
adjusted to 6.8 using glacial acetic acid stock solutions of
10,000µg/ml of all the extracts were prepared by using
ethanol as a solvent. From these stock solutions 4 different
concentrations of 1,100, 200 and 500µg/ml were prepared
by using ethanol as a solvent. 50µl (0.05ml) of each
extract was transferred to Eppendorf tubes using 1ml
micropipette. 5ml of 0.2% w/v BSA was added to the
entire above Eppendorf tubes. The control consists of 5ml
of 0.2% w/v BSA solution with 50µl ethanol. The
standard consists of 100µg/ml of Ibuprofen in ethanol
with 5ml 0.2% w/v BSA solution. The test tubes were
heated at 72ºC for 5 minutes and then cooled for 10
minutes. The absorbance of these solutions was
determined by using a UV-VIS Double beam
spectrophotometer (ELICO SL 244) at a wavelength of
660nm. Each experiment was carried out in triplicate and
the mean absorbance was recorded. The percentage of
inhibition of precipitation was determined on a percentage
basis relative to control using the formula.
Percentage of inhibition of denaturation =
absorbance of control  absorbance of extract
×100
absorbance of control
RESULTS AND DISCUSSION
Anti-inflammatory properties - Inhibition of albumin
denaturation: Denaturation of proteins is a well
documented cause of inflammation. As part of the
investigation on the mechanism of the antiinflammation
activity, ability of different solvent plant extract protein
denaturation was studied. It was effective in inhibiting
IJRPB 1(5) www.ijrpb.com
ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
constituents i.e. saponins, steroids, phenol, alkaloids and
tannins, flavonoids, glycosides, etc. Phytochemical
screening was performed by employing standard screening
tests and the results are represented in table1.
Ethanolic Extract
+ +
+ +
+ +
+ +
- -
+ +
- -
+ +
- +
+ +
heat induced albumin denaturation (Table 2). Maximum
percentage of inhibition 67.5 was observed from Ethanol
extract followed by hexane (56.4). All the solvent extracts
inhibited the albumin denaturation, the ethanol extract
stood first compared to hexane extracts. Ibuprofen, a
standard anti-inflammatory drug showed the maximum
inhibition 71.89% at the concentration of 100 μg/ml.
Inflammation is the response of living tissues to injury. It
involves a complex array of enzyme activation, mediator
release, extravasations of fluid, cell migration, tissue
breakdown and repair (Vane, 1995). Denaturation of
proteins is a well documented cause of inflammation and
rheumatoid arthritis (Mizushima, 1966). Several anti-
inflammatory drugs have shown dose dependent ability to
inhibit thermally induced protein denaturation (Grant,
1970). Ability of Canthium parviflorum extract to bring
down thermal denaturation of protein is possibly a
contributing factor for its anti-inflammatory activity. The
anti-inflammatory activity of Canthium parviflorum
extract found may be due to the presence of
therapeutically active flavonoids. The therapeutic
applications of flavonoids on inflammation have
previously been reported (Middleton, 2000;
Havsteen,2002). The data of our studies suggests that
Canthium parviflorum showed significant anti-
inflammatory activity by in vitro methods tested. Further
studies involving the isolation and purification of the
chemical constituents of the plant and the investigations in
the biochemical pathways may result in the development
of a potent anti-inflammatory agent with a low toxicity
and better therapeutic index.
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Karthik et.al Indian Journal of Research in Pharmacy and Biotechnology
Table.2.Anti denaturation of BSA in presence of Canthium parviflorum extracts
Type of sample Concentration(μg/ml)
Ethanolic
Ethanolic
Ethanolic
Ethanolic
Hexane
Hexane
Hexane
Hexane
Ibuprofen
CONCLUSION
Canthium parviflorum is a valuable medicinal
shrubby and woody plant which has been valued for
centuries in ayurvedic medicine. Phyto-chemical analysis
of Canthium parviflorum plant extracts revealed the
presence of various bio-chemical compounds such as
flavonoids, glycosides, alkaloids, saponins and terpenoids.
Since glycosides and flavonoids have remarkable anti-
inflammatory activity, our present work aims at evaluating
the in-vitro anti-inflammatory activity of Canthium
parviflorum by protein denaturation method. Denaturation
of proteins is a well documented cause of inflammation
and rheumatoid arthritis. The data of our studies suggests
that Canthium parviflorum extract showed significant anti-
inflammatory activity. Therefore our studies support the
isolation and use of active constituents of Canthium
parviflorum in treating inflammation.
ACKNOWLEDGEMENT
The authors are thankful to Prof.Dr.N.Devanna,
Director, Oil Technological Research Institute, JNTUA,
Ananthapuramu for the encouragement and facilities
provided to carry out this research work.
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