WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Nirmala et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 2.786

Volume 4, Issue 1, 699-703. Research Article ISSN 2278 – 4357

PHYTO CHEMICAL ANALYSIS OF PIPER BETEL LEAF EXTRACT

O. Sita kumari1, Dr. Nirmala Babu Rao2

1
Dept of Botany, RBVRR Women’s College, Hyderabad, Telangana, India
2
Department of Botany, University College for Women, Osmania University,
Hyderabad, Telangana, India.

ABSTRACT
Article Received on
11 Oct 2014, An effort has been made to find out results of phyto chemical analysis

Revised on 06 Nov 2014, made on leaf extracts of betel. Leaf Extract with ethyl Alcohol
Accepted on 02 Dec 2014 revealed the presence of tannins, anthraquinones, flavonoids, alkaloids,
Nov 2014 terpenoides, saponlns, cardiac- glycosides, glycosides, reducing sugars,
*Correspondence for phlobatanins in the betel leaf. In view of the presence of antioxidant
Author
properties of the plant, it is aimed to understand whether the plant can
Dr. Nirmala Babu Rao
be use of in fighting the malignancy and other cancer causing agents.
Department of Botany,
University College for
KEYWORDS: extracts, tannins, anthraquinones, flavonoids.
Women, Osmania
University, Hyderabad,
INTRODUCTION
Telangana, India
Piper betel Linn (Piperaceae) leaves is widely used as a post meal
mouth freshener and the crop is extensively grown in India, Sri Lanka, Malaysia, Thailand,
Taiwan and other Southeast Asian countries. Due to strong pungent aromatic flavour betel
leaves are used as masticatory by the Asian people. Its common names are betel (in English),
paan (in Indian), phlu (in Thai) and sirih (in Bahasa Indonesian). Grown abundantly in many
parts of India, betel is an evergreen dioecius herb that needs warm and moist growth
conditions for its growth (Arani Datta et al). Indian system of medicine and health has
adopted the use of betel leaves in various ways. Piper betel leaf is popular as an antiseptic and
is commonly applied on wounds and lesions for its healing effects. This particular property
has paved way for further experimental studies, which have established paan extract to have
antimicrobial and anti leshmian properties (Sarker et al., 2008). The plant has got large
number of bio molecules which show diverse pharmacological activity (S.N.Deshpande et
al). Piper betel is used to treat alcoholism, bronchitis, asthama,
leprosy and dyspepsia. Earlier, anti‐ulcerogenic activity of Piper betel was attributed to its

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Nirmala et al. World Journal of Pharmacy and Pharmaceutical Sciences

anti oxidative property. A preliminary study has reported piper betel leaves extract contains
large numbers of bioactive molecules (Devjani Chakraborty* et al). Piper betle contains a
wide variety of biologically active compounds whose concentration depends on the variety of
the plant, season and climate. Pharmacological Profile has shown antiplatelet, anti-
inflammatory effects as well as immuno modulatory, gastro protective and antidiabetic
activity (Satish A Bhalerao1* et al). Paan has been referred to in Sakta-tantra as one of the
means of achieving siddhi. It was believed that without betel chewing and offering pan to
Guru no siddhi can be gained. Tambool has also been referred to as facilitating the sadhak in
chewing dharma, yasha aisvarya, Srivairagya and mukti (D. Pradhan1* et al). It was reported
that fresh leaves contains: moisture 85.4, protein 3.1, fat 0.8, carbohydrate 6.1, fibre 2.3,
calcium 230mg, phosphorous 40mg, iron 7mg, ionisable iron 3.5mg, iodine 3.4 µ. They have
a high content of potassium nitrate (0.26-0.42%). The sugars identified in betel leaves include
glucose, fructose, maltose and sucrose. The average content of free reducing sugars in
different types of betel leaves varies from 0.38-1.46%. It also contains the enzyme like
diastase and catalase. (K.Periyanayagam1 et al). Piper betle leaves are earlier reported to
possess anticancer potential. Hence, the aqueous extract of the leaves was subjected to
cytotoxicity studies on Hep-2 cell line using Micro culture Tetrazolium and Sulphorhodamine
B assays (Chaurasia, Sundeep et al). Piper betel leaf oil can be used as an industrial raw
material for manufacturing medicines, perfumes, food additives etc. The leaves are nutritive
and contain anti carcinogens showing promise for manufacturing of a blood cancer drug
(Sengupta et al).

MATERIALS & METHODS

Plant material was collected from the college campus. Chemicals such as wagnes
reagent,chloroform,2% H2SO4, Concentrated sulphuric Acid,10%Lead acetate, Benedict’s
reagent, 0.1% ferric chloride, Fehling’s solution, dilute NaOH,2% HCL, 10% Ammonia,
10%HCL, distilled water, Ethyl Alcohol are provided by the management of the college.

Preparation of solutions: a) Fehling’s solution:- A mixture of equal volume of copper

sulphate, sodium potassium tartar ate and sodium hydroxide is prepared in a beaker.
b)Wagner’s Reagent:- Mixing 2gm of Iodine, 6gm of potassium iodide in 100ml of water.

Collection of sample: Healthy leaves of betel were taken and washed under running water to
remove the dust and other external pollutants. The plant leaves were air dried for few days
(normally 15 to 21days).

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Nirmala et al. World Journal of Pharmacy and Pharmaceutical Sciences

Grinding the sample: The dried leaves are grinded to a fine powder in a mixer and the
powder is collected in clean polythene bags.

Preparation of plant extract with ethyl Alcohol: Taken 10 gms of leaf powder and added
50ml of ethyl alcohol stirred it constantly for 30 minutes and the solution was kept at room
temperature for 24 hours (minimum) and then filtered. The filtered solution is again filtered
with watman filter paper No.3 and then it was stored at 4 degrees centigrade (in a freezer)
until use.

TESTS AND RESULTS

1) Phyto chemical screening: Chemical test is carried out on the ethyl alcohol extract of
piper betel using standard procedures to identify the constituents.

2) Procedure for alkaloids: 2ml of extract is taken and added 2ml of wagner’s reagent a
brownish precipitate indicate the presence of alkaloids.

3) Cardiac glycosides: 2ml of extract is dissolved with 2ml of chloroform and concentrated
sulphuric acid is carefully added to form a layer. Deep reddish brown colour at the inter
face of steroid ring indicates the presence of cardiac glycosides.

4) Flavonoids: 2ml of extract is treated with 2 ml of 10%lead acetate. Yellowish green

colour indicates the presence of flavonoids.

5) Saponins: 2ml of extract is dissolved with 2ml of Benedicts reagent. Blue black ppt
indicates the presence of saponins.

6) Tanins: 2ml of extract is treated with 0.1% of ferric chloride. Brownish green indicates
the presence of tannins.

7) Terplenoides: (Salkowski test) 2ml of extract is dissolved with 2ml of chloroform and
concentrated sulphuric acid is carefully added to form a layer. A reddish brown colour
indicates the presence of terpenoids.

8) Anthraquinones: 1ml of extract is boiled with 10% HCL for few minutes in a water
bath. It is filtered and allowed to cool. Equal volume of CHCl3 is added to the filtrate few
drops of 10% Ammonia is added to the mixture and heat. Formation of rose pink colour
indicates the presence of anthraquinones.

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Nirmala et al. World Journal of Pharmacy and Pharmaceutical Sciences

9) Glycosides: The extract is hydrolysed with HCL solution and neutralised with NaoH
solution. A few drops of Fehlings solution A&B are added red precipitate indicates the
presence of glycosides.

10) Reducing sugars: The extract is shaken with distilled water and filtered. The filtrate is
boiled with drops of Fehling’s solution A&B for few minutes. An orange red precipitate
indicate the presence of reducing sugars.

11) Phlobatanins: The extract is dissolved in distilled water and filtered. The filtrate is boiled
with 2% HCL solution. Red precipitate shows the presence of phlobatanins.

12) Result: By this analysis we can conclude that Piper betel consists of Tannins,
Anthraquinones, Flavanoids, Alkaloids, Terpenoids, Saponins, Caediac glycosides,
Glycosides, Reducing sugars.

Table showing results of Phyto Chemical Analysis with Ethanol Extract

Sl. No Phytochemicals Piper betel Betel leaf
1 Tanins Positive
2 Anthraquinones Positive
3 Flavanoides Positive
4 Alkaloides Positive
5 Terpenoids Positive
6 Saponins Positive
7 Cardiac glycosides Positive
8 Glycosides Positive
9 Reducing Sugars Positive
10 Phlobatanins Positive

Human usage and Disease Management

a) Alkaloids are anaesthetic agents and found in medicinal plants.
b) Cardiacglycosides are strong and direct action on the heart, kidneys and used for diuretic
and lowering the blood pressure.
c) Flavoinoids are anti oxidant elements strengthen capillary volves of heart and improve
blood circulation.
d) Saponins are useful as expectorants, anti bacterial and anti viral.
e) Tanins draw the tissue closure together and fights infection.
f) Terpenoids are useful as aromatic agents and used for flavouring the food.
g) Anthraquinins are having laxative properties and useful to large intestine.

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DISCUSSION AND CONCLUSION

Being a commercial crop the growth of the plant has been limited to a few varieties. Research
is to be done on the lost variants which have more medicinal properties. Wild varieties such
as to be nurtured and developed for disease management.

REFERENCES
1. Arani Datta, Shreya Ghoshdastidar and Mukesh Singh*; Antimicrobial Property of Piper
betel Leaf against Clinical Isolates of Bacteria; Journal of Pharma Sciences and Research
(IJPSR), 2011; 2(3): 104-109.
2. Sarkar A, Sen R, Saha P, Ganguly S, Mandal G, Chatterjee M (2008). An ethanolic
extract of leaves of Piper betle (Paan) Linn mediates its antileishmanial activity via
apoptosis. Parasitol. Res, 2008; 102(6): 1249-55.
3. S.N. Deshpande, D.G. Kadam; Gcms Analysis and Antibacterial Activity of Piper
Betle(Linn) Leaves Against Streptococcus Mutans; Asian J Pharm Clin Res, 2013; 6: 5.
4. Devjani Chakraborty*, Barkha Shah; Antimicrobial, Anti-Oxidative And Anti-Hemolytic
Activity Of Piper Betel Leaf Extracts; International Journal of Pharmacy and
Pharmaceutical Sciences; 2011; 3: 3.
5. Satish A Bhalerao1*, Deepa R Verma2, Rohan V Gavankar2, Nikhil C Teli2, Yatin Y
Rane3,Vinodkumar S Didwana2 and Ashwin Trikannad2; Phytochemistry,
Pharmacological profile and Therapeutic uses of Piper Betle Linn. Research and Reviews:
Journal of Pharmacognosy and Phytochemistry, 2013; 1.
6. D. Pradhan1*, Dr. K. A. Suri2 , Dr. D. K. Pradhan3 and P. Biswasroy 4; Golden Heart of
the Nature: Piper betle L; Journal of Pharmacognosy and Phytochemistry; 2013; 1: 6.
7. K.Periyanayagam1, M.Jagadeesan2 , S.Kavimani3 , T.Vetriselvan4; Pharmacognostical
and Phyto-physicochemical profile of the leaves of Piper betle L. var Pachaikodi
(Piperaceae) - Valuable assessment of its quality; Asian Pacific Journal of Tropical
Biomedicine, 2012.
8. Chaurasia, Sundeep; Kulkarni, Giriraj Tirupatirao; Shetty, Laxmi Narayan; Mishra,
Brahmeshwar; Phytochemical Studies and In vitro Cytotoxicity Screening of Piper betle
Leaves Extracts; Journal of Pharmacy Research, Nov 2011; 4(11): 4187.
9. Sengupta, Rupa; Banik, Jayanta K; A Review On Betel Leaf (Pan); International Journal
of Pharmaceutical Sciences & Research, Dec 2013; 4(12): 4519.

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