tylophora indica
TRANSCRIPT
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___________________________________________Research Paper
Qualitative Studies of Bioactive Compounds in Leaf of T ylophora
indica (Burm. F.) M err. Sanjeet kumar*
Department of Life Science, Regional Institute of Education (NCERT), Bhubaneswar, Orissa, India.
_____________________________________________________________________________________________
ABSTRACT
Tylophora indica (Burm. F.) Merr. is a common wild climber of Odisha, known as “Sawasmari”, belongs to family
Asclepidaceae.The leaves are used as traditional medicine in asthma or other respiratory problems among rural
and tribal community of Odisha.The qualitative phytochemical screening and Thin Layer Chromatography(TLC)
were used to investigate the bioactive compounds. Presence of alkaloids in different solvents revealed the potent
medicinal properties of Swasmari. Presence of tannin, saponin and terpenoid in aqueous extract revealed more
pharmacological activity.
Key Words: Tylophora indica, Medicinal value, TLC, Phytochemical screening.
INTRODUCTION
Human beings cannot survive on this earth for long
without the plant because its product and their active
constituent played an important role in day to day
life. Plant materials remain important resources to
combat serious diseases in the world. The traditional
medical methods, especially the use of medicinalplants, still play a vital role to cover the basic health
needs in the developing countries like India. The
medicinal value of these plants lies in some chemical
active substances that produce a definitephysiological action on the human body. The most
important of these bioactive constituents of plants are
alkaloids, tannin, flavnoid and phenolic
compounds1.Therefore over the last few years,
researchers have aimed at identifying and validating
plant-derived substances for the treatment of various
diseases commonly observed among rural and tribal
community. Interestingly it is estimated that morethan 25 % of the modern medicines are directly or
indirectly derived from plants. It is worth mentioning
that Indian medicinal plants are considered as a vastsource of several pharmacological principles and
compounds that are commonly used as homeremedies against multiple ailments
2.The concept of
developing drugs from plants used in indigenous
medical system is much older, while in some cases
direct link between a local and biomedical use exists,
in other cases the relationship is more
________________________________________
*Address for correspondence:
E-mail: [email protected]
complex3.Tylophora indica is a perennial, much
branched, pubescent twining or climbing herbs grow
up to 1.6 meter, leaves ovate-oblong to elliptic-
oblong,3.2-9.8 cm long and 1.2-7.4 cm wide,
yellowish sap belongs to family asclepidaceae, found
in most forest tracts of Odisha and indigenous to
India. It is mainly used for bronchitis and bronchial
asthma as a traditional medicine among the rural andtribal people of Odisha. Its jaundice
4, inflammation
5,
antitumer6, immunomodulatory
7, antioxidant
8, anti-
asthmatic9, smooth muscle relaxant
10,
antihistaminic10, hypotensive, analgesic,anticonvulsant
10and antirheumatic
10activities are
reported. The leaves are used as diaphoretic, emetic
and expectorant11
. Also the anticancer activity of
tylophorin has been reviewed12.
Phytochemical
literature reveals the presence of tylophorin, type of
alkaloids, tylophorinidine, tylophorinine,tyloindicine-
A,D,F,G,H, and I, 14-hydroxy tylophorine, stigma
sterol, octa cosanyl, skimmianide, tannins, saponinsand flavonoids in Tylophora indica
13, 14,15,16,17,18. In
Odisha it is found up to 1000 meters high such as
Niyamgiri hills, Simlipal Biosphere Reserve forests,Karlapat sanctuary, Gandhmardan hills etc. Kol tribal
communities of Simlipal biosphere reserve forest useit in respiratory problems.
MATERIAL AND METHODS
1. Collection of plant materials The leaves of T.indica were collected inOctober 2010, from Germplasm
conservation garden of NBPGR Base Centre
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Cuttack and authenticated by Dr. R.C.
Misra, Sr.Sc. NPGR Base Centre, Central
Rice Research Institute, Cuttack.
2. Preparation of Plant extract
The collected leaves were first washed in tap
water and dried than cut in to small piecesand again washed in distill water. Then
leaves were left for air drying under shade.
After air drying the leaves were grinded in
mechanical grinder and powderd material
was kept in airtight container for further use.The sample powder was extracted at a
solute-solvent ratio of 1:10 by percolation
method. Crude extract were kept in small
bottles for phytochemical screening.
3. Phytochemical Investigation
Qualitative analysis of bioactive compounds
were carried out following the methodsprovided by Harborne
19(1973), Trease and
Evans20
(1989) and Kumar et al (2007)21
.
4. Preparative TLCGlass plate was cleaned with ethyl acetate
then left for 15 min. 3 gm of silica gel wastaken in a beaker and 15 ml water was
added. Then slurry was poured over the
glass plate and was allowed to dry, than
heated by hot plate for few minute to
activation of TLC plate.
RESULT AND DISCUSSION
The present study has authenticated the medicinal
values of T.indica. Preliminary qualitative analysis of
bioactive compounds revealed the presence of alkaloids in methanol extract and shown spot on TLCafter applying dragendorff’s reagent. The presence of
saponin indicated in methanol and aqueous while
terpenoids and tannin present only in Aqueous extract
(Table 4). Okwu and Okwu22
(2004) reported that
tannins have astringent properties, healing of wounds
and inflamed mucous membranes. Tannins arereported to exhibit antiviral, antibacterial, anti-tumor
activities and was also reported that certain tannins
are also able to inhibit HIV25
replication selectively
and is also used as anti-diuretic26
. The presence of
tannin in aqueous extract (Table 4) is likely to be
responsible for the free radical scavenging effectswhich provide potent medicinal value of Tylophora
indica as an antioxidant medicinal plant. Alkaloids
shown only in methanol extract but on TLC in
acetone and chloroform extract (Table 2 & 4)
indicated the analgesic, anti-spasmodic andbactericidal effects
23.Alkaloids and saponin are
known to be effective for the treatment of syphilis
and other venereal disease27
. Oliver-Bever24
(1986)
and Okwu23
(2004) had earlier reported that saponins
have antibiotic properties and so help the body to
fight infections and microbial invasion. Also, it is
used as a mild detergent and in intracellular
histochemistry staining to allow antibody access to
intracellular proteins, also reported in hyperchloles-
trolaemia, hyperglycaemia, antioxidant, anticancer,anti- inflammatory and weight loss and have anti-
fungal properties25.
CONCLUSION
It is concluded that T.indica is a plant with a variety
of ethnic medicinal uses, presence of bioactive
compounds such as alkaloids, tannin, saponin and
terpenoids shown the potential of biological and
pharmacological action of this plant. Present study
might be useful to supplement scientific information
in regard to its medicinal value andphytocompounds.Further more scientific studies
needed to know its biological action on disease.
ACKNOWLEDGEMENT
Author gratefully acknowledge Sr. Scientist R.C.
Misra, OIC Dr. D. R. Pani, NBPGR Base Centre
Cuttack and Director, Biotechnotric Pvt Ltd.
Bhubaneswar, for their valuable suggestion,
identification and provide lab facilities to complete
this work successfully.
Table 1: Identification of spot on TLC plate and
determination of Rf Values
Extract Mobile phaseRf
valuesMethyl
alcoholChloroform: Methyl alcohol = 9:1 0.825
Acetone Chloroform: Methyl alcohol = 9:1 0.900
Benzene Chloroform: Methyl alcohol = 9:1 0.723
Acetic
acidChloroform: Methyl alcohol = 9:1 0.800
Aqueous Chloroform: Methyl alcohol = 9:1 0.676
Hexane Chloroform: Methyl alcohol = 9:1 0.380
00.10.20.30.40.50.6
0.70.80.91
Chlorof orm:Methyl
alcohol=9:1
Chlorof orm:Methyl
alcohol=9:1
Chlorof orm:Methyl
alcohol=9:1
Chlorof orm:Methyl
alcohol=9:1
Chlorof orm:Methyl
alcohol=9:1
Chlorof orm:Methyl
alcohol=9:1
Rf =distancetraveledby
thespotf rom
theoriginline(cm)
/ distan
cetraveledby
thesloventf ront(cm)
Organic solvents
Variations of Rf values in different organic solvents
Rf values
Fig. 1: Variations of Rf values in different solvents
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Table 2: Identification of spot of
Alkaloids on TLC Plate
(1 = Present & 0= absent)Solvents Alkaloids
Methyl
alcoholPresent
Hexane Absent
Acetone PresentChloroform Present
Benzene Absent
Acetic acid Absent
Aqueous Absent
Variations of Alkaloid in different organic solvents
0
0.20.4
0.6
0.8
1
1.2
M e t h y
l a l c o
h o
H e x
a n e
A c e t o n
e
C h l o r o
f o r m
B e n z e
n e
A c e t i c
a c i d
A q u e
o u s
solvents
P r e s e n t a n d
A
b s e n t a s
1
&
0
Alkaloids
Fig. 2: Variations of Alkaloid in different organic
solvent
Table 3: Colour variations of different solvent
extractSolvent Colour appearance (Naked eye)
Methyl alcohol Light green
Hexane Yellow with green florescence
Acetone Deep green
ChloroformCreamy white with brown layer on upper
part
Benzene Light yellowish brown
Acetic acid Brown
Aqueous Light brown
Table 4: Qualitative analysis of bioactive
compounds of leaf of Tylophopra indica.(1 = Present & 0= absent)
1 2
31
2
3
1 2 3
1
2 31 2
3
0
0.2
0.4
0.6
0.8
1
1.2
Solvents
Variations of Bioactivecompounds in different solv
Fig. 3: Variability of Bioactive compounds of leaf
of Tylophora indica using different solvents
Leaves and stem of Tylophora indica
Leaves of Tylophora indica in Simlipal Biosphere
reserve forests,Orissa.
Bioactive
compound
Methanol
extract
Hexane
extract
Aqueous
extract
Tannins 0 0 1
Saponin 1 0 1
Flavonoids 0 0 0
Alkaloids 1 0 0
Terpenoids 0 0 1
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