International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

___________________________________________Research Paper

Qualitative Studies of Bioactive Compounds in Leaf of Tylophora indica (Burm. F.) Merr.
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 medicinal plants, 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 definite physiological 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 more than 25 % of the modern medicines are directly or indirectly derived from plants. It is worth mentioning that Indian medicinal plants are considered as a vast source of several pharmacological principles and compounds that are commonly used as home remedies against multiple ailments2.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: sanjeet.biotech@gmail.com complex3.Tylophora indica is a perennial, much branched, pubescent twining or climbing herbs grow up to 1.6 meter, leaves ovate-oblong to ellipticoblong,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 and tribal people of Odisha. Its jaundice4, inflammation5, antitumer6, immunomodulatory7, antioxidant8, antiasthmatic9, smooth muscle relaxant10, 10 antihistaminic , hypotensive, analgesic, anticonvulsant10 and antirheumatic10 activities 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,tyloindicineA,D,F,G,H, and I, 14-hydroxy tylophorine, stigma sterol, octa cosanyl, skimmianide, tannins, saponins and flavonoids in Tylophora indica13, 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 use it in respiratory problems. MATERIAL AND METHODS 1. Collection of plant materials The leaves of T.indica were collected in October 2010, from Germplasm conservation garden of NBPGR Base Centre

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International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

2.

3.

4.

Cuttack and authenticated by Dr. R.C. Misra, Sr.Sc. NPGR Base Centre, Central Rice Research Institute, Cuttack. Preparation of Plant extract The collected leaves were first washed in tap water and dried than cut in to small pieces and 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. Phytochemical Investigation Qualitative analysis of bioactive compounds were carried out following the methods provided by Harborne19 (1973), Trease and Evans20 (1989) and Kumar et al (2007)21. Preparative TLC Glass plate was cleaned with ethyl acetate then left for 15 min. 3 gm of silica gel was taken 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.

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 hyperchlolestrolaemia, hyperglycaemia, antioxidant, anticancer, anti- inflammatory and weight loss and have antifungal 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 and phytocompounds.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 Methyl alcohol Acetone Benzene Acetic acid Aqueous Hexane Mobile phase Chloroform: Methyl alcohol = 9:1 Chloroform: Methyl alcohol = 9:1 Chloroform: Methyl alcohol = 9:1 Chloroform: Methyl alcohol = 9:1 Chloroform: Methyl alcohol = 9:1 Chloroform: Methyl alcohol = 9:1 Rf values 0.825 0.900 0.723 0.800 0.676 0.380

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 TLC after applying dragendorffs 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 are reported 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 effects which 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 and bactericidal effects23. 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

Variations of Rf values in different organic solvents

y mb o d r e f l t e o v p r s a ) e t m h e c t c ( y n t b a n t o i r d s f e d l / t e v ) n e a mv r c o t ( s l e e c n e i n l h a n t t s i i g d i r = o f e Rh t

1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0

R values f

l y h t 1 e : M9 : = ml r o o f h o o c r l o l a h C

l y h t 1 e: M9 : = ml r oo f h o oc r l o l a h C

l y h t 1 e : M9 : = ml r o o f h o o c r l o l a h C

l y h t 1 e : M9 : = ml r o o f h o o c r l o l a h C

l y h t 1 e: M9 : = ml r oo f h o oc r l o l a h C

l y h t 1 e: M9 : = ml r oo f h o oc r l o l a h C

Organic solvents

Fig. 1: Variations of Rf values in different solvents

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International Journal of Research in Pharmaceutical and Biomedical Sciences

ISSN: 2229-3701

Table 2: Identification of spot of Alkaloids on TLC Plate (1 = Present & 0= absent)

Solvents Methyl alcohol Hexane Acetone Chloroform Benzene Acetic acid Aqueous Alkaloids Present Absent Present Present Absent Absent Absent

Variations of Bioactive compounds in different solvents

1.2 1 0.8 1 3

t 0.6 n e v 0.4 l
Fig. 3: Variability of Bioactive compounds of leaf of Tylophora indica using different solvents

Variations of Alkaloid in different organic solvents

P r esent a nd A bse nt a s 1 & 0
1.2 1 0.8 0.6 0.4 0.2 0
c oh ol He xa n A ce e to n e Ch l o ro fo r Be m nz A ce e n e t ic ac i d Aq ue o us

S 0.2 0

Alkaloids

Me

th y l al

solvents

Fig. 2: Variations of Alkaloid in different organic solvent Leaves and stem of Tylophora indica Table 3: Colour variations of different solvent extract
Solvent Methyl alcohol Hexane Acetone Chloroform Benzene Acetic acid Aqueous Colour appearance (Naked eye) Light green Yellow with green florescence Deep green Creamy white with brown layer on upper part Light yellowish brown Brown Light brown

Table 4: Qualitative analysis of bioactive compounds of leaf of Tylophopra indica. (1 = Present & 0= absent)
Bioactive compound Tannins Saponin Flavonoids Alkaloids Terpenoids Methanol extract 0 1 0 1 0 Hexane extract 0 0 0 0 0 Aqueous extract 1 1 0 0 1

Leaves of Tylophora indica in Simlipal Biosphere reserve forests,Orissa.

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Orange Spot after applying dragndroff reagent on methanolic extract

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