Pharmacognosy Journal 5 (2013) 176e183

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Pharmacognosy Journal
journal homepage: www.elsevier.com/locate/phcgj

Original article

Pharmacognostic specications of eight cultivars of Piper betle

from eastern region of India
Atiya Akhtar Khan, Satyendra Prasad Bhatnagar, Barij Nayan Sinha, Uma Ranjan Lal*
Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi 835215, Jharkhand, India

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 22 May 2013
Accepted 17 July 2013
Available online 29 August 2013

Background: Piper betle L. is a well-known and widely used herb especially in tropical Asiatic countries,
which contains several bio-active constituents and possesses health promoting properties. The leaves of
Piper betle L. have been traditionally known for its various therapeutic uses. The pharmacognostical and
physicochemical investigations have not been carried out for eastern Piper betle cultivars.
Objective: The present investigation was therefore undertaken to determine the requisite pharmacognostic standards for their evaluation to ensure their safety and efcacy.
Materials and methods: Different Piper betle L. cultivars (Desi, Bangla, Kali, Haldia, Sanci, Meetha, Birkoli
and Magahi) were obtained from Indian States of West Bengal, Odisha, and Bihar (Table 1).
Fresh leaves were washed with water, sliced with a microtome and the transverse sections were stained
with safranin, phloroglucinol, concentrated hydrochloric acid and mounted in glycerine medium. Leaves
dried in the shade were nely powdered and passed through a sieve no.180 and a sieve no.125, separately, to obtain ne powder respectively and then subjected to microscopic examination.
Mesophyll tissue is divided up by slendest vein, which is called vein islet. The numbers of vein islet in
every epidermal cell is called Vein Islet Numbers (VIN); Vein termination number is the number of
veinlet termination per square mm of the leaf surface midway between the midrib and the margin (VTN).
Since vein islet and vein termination number is consistent and different from plant to plant, this
parameter can be used in authentication. Stomatal Index (SI) is a basilic parameter in authentication,
which can be obtained from the following formula: Stomatal Index (number of stomata/every millimeter-square)
100/(number of stomata/every millimeter-square number of epidermal cell/every
millimeter - square), which also can be used as a reliable parameter in authentication since it is
consistent in a species of plant. The results were studied using light microscope according to the usual
microscopic techniques. Different physicochemical parameters such as ash value, extractive value and
loss on drying were carried out as per WHO guidelines.
Results: In present study eight cultivars of Piper betle were compared with their distinguishing morphoanatomic details of leaf, dissociation and powders. The microscopic features were systematically
described and illustrated. The physiochemical properties such as loss on drying, total ash value, acid
insoluble ash value, water soluble ash value and extractive values were also carried out. HPTLC analysis of
petroleum ether extract and essential oil showed conrming the presence of compound in Piper betle
cultivars leaf. And detailed key authentication parameters based on these anatomic characteristics were
presented. The various morphological, microscopical, physicochemical standards can be unambiguously
used to authenticate standardization and distinguish eight cultivars of Piper betle.
Copyright 2013, Phcog.Net, Published by Reed Elsevier India Pvt. Ltd. All rights reserved.

Keywords:
Piper betle cultivars
Pharmacognosy
Microscopic
Macroscopic
Physiochemical

1. Introduction
Piper betle Linn., (Piperaceae) is an aromatic, shade loving
climber cultivated mainly in India and other tropical Asiatic

* Corresponding author.
E-mail addresses: uma@bitmesra.ac.in, urlalvv@yahoo.com (U.R. Lal).

countries (Bangladesh, Pakistan, Malaysia and Indonesia etc).1 The

betel leaf is known as Paan in India.2 There are more than 700
species of plants belonging to the genus Piper, out of which 30
species are found in India.3 It is consumed as betle quid with areca
nut (Areca catechu), katha (Acacia catechu L.), slaked lime [Ca(OH)2]
cardamom (Elettaria cardamomum), clove (Eugenia caryophyllus),
etc. which act as a breath freshener with or without tobacco.4,5 The
betle vine cultivars vary, besides in morphological characters, in

0975-3575/$ e see front matter Copyright 2013, Phcog.Net, Published by Reed Elsevier India Pvt. Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.phcgj.2013.07.002

A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

Table 1
Collection data for samples and morphological characters of Piper betle cultivars.
Cultivars

District

State

Haldia
Bangla
Desi
Sanci

Ram Nagar
Midnapur
Midnapur
Parganas,
Halisahar
Midnapur
Machida
Nevada
Nayagarh

West
West
West
West

Meetha
Kali
Magahi
Birkoli

Bengal
Bengal
Bengal
Bengal

West Bengal
West Bengal
Bihar
Orissa

Colour

Length
(cm)

Width
(cm)

Light green
Dark green
Green
Dark green

12e14
16e19
12e14
20e23

8.0e10
14e16
5.0e7.0
15e16

Dark green
Dark green
Green to light green
Greenish yellow

14e16
17e20
7.0e9.5
9.0e11

10e11
14e17
5.5e6.5
5.0e7.0

177

Cleaning of leaf was done to study the venation pattern. Washed

leaf fragments were rst boiled in 90% alcohol for about 3e5 min to
remove chlorophyll, and then washed 2e3 times with water, then
boiled again with 10% KOH solution for 2e3 min and washed 4e5
times with water. The epidermal layer was peeled off using the help
of pointed needle and forceps and was washed in water. The margins of the cover slip were sealed with DPX (Hi-Media laboratories
Pvt Ltd, Bombay), and the slides were observed under the microscope. Stomatal index, palisade ratio, vein termination number and
vein islet number were then calculated.22

2.4. Anatomy
avour and in the aroma of the leaves and the avour and taste of
the leaves are due to the essential oil composition.6 Investigations
on the Piper betle showed a number of pharmacological properties
like anti-oxidant,7 antifungal,8 anti-microbial,9 anti-inammatory10 and radio protective11 etc. Phytochemical investigation on
leaves revealed the presence of tannins and steroidal components.12 The terpenoids and phenylpropanoids reported in Piper
betle are 1,8-cineole, cadinene, camphene, caryophyllene, limonene, pinene, chavicol, 4-allyl-pyrocatechol, carvacrol, safrole,
eugenol, and chavibetol.13,14 Phytochemical screening studies on
Piper betle leaves have revealed the presence of avonoids, saponins, mucilage, reducing sugars, carbohydrates, glycosides, protein
and amino acids and absence of alkaloids and xed oil.15 There is a
good level of traditional and experimental evidences to support
various claims and advantages of this widely available plant.
Despite its importance, only a few comparative morpho-anatomical
and physicochemical studies have been done on the leaves of Piper
betle.15e19 But our selected eastern cultivars have not been
explored. Hence the present study was undertaken to explore them
for standardization purpose. In eastern India, Piper betle L. cultivars
like Bangla, Kali, Desi, Haldia, Meetha, Sanci, Magahi and Birkoli are
widely cultivated. In present work we report macroscopic evaluation, microscopic evaluation, physicochemical parameters and
preliminary phytochemical screening for standardization and
quality assurance purposes of these cultivars from eastern region of
India.
2. Materials and methods

For microscopic studies cross section was prepared as per the

standard procedures.23 Wherever necessary, sections were stained
with safranin, phloroglucinol, concentrated hydrochloric acid and
mounted in glycerine medium.24,25 Microphotographs were taken
using Leica DME, DSC Power shot S70 DE [PICT Bridge], 7.1 megapixels, wide angle 3.6
optical zoom lens.

2.5. Powder studies

Leaves dried in the shade were nely powdered and passed
through a sieve no.180 and a sieve no.125, separately, to obtain ne
powder respectively and then subjected to microscopic examination. The sample was treated with following reagents and studied
for their components of diagnostic value, glycerine (50%) as temporary mountant phloroglucinol (2% w/v) in ethanol (90%), conc.
HCl (50%) for lignin; alcoholic ferric chloride (5% w/v) for phenolic
compounds; Iodine (I-/KI) solution (2%) for starch grains; and
ruthenium red (0.08%) in lead acetate (10%) for mucilage and
observation was recorded.26,27

2.6. Physicochemical parameters

Physicochemical values such as the percentage of ash values,
loss on drying (moisture content) and extractive values were performed according to ofcial methods prescribed (Indian Pharmacopoeia, 1996)28 and the WHO guidelines on quality control
methods for medicinal plant materials (WHO guidelines, 1992).29

2.1. Collection and authentication of plant materials

Fresh leaves of eight cultivars of Piper betle were collected from
eastern region of India (West Bengal, Bihar, and Odisha) in
November, 2010 (Table 1). Plant leaves were authenticated by the
Central National Herbarium, Botanic Garden Howrah Kolkata, India.
A voucher specimen has been deposited (CNH/10B/2011/Tech.II/
502). The plants were also authenticated by Dr. (Mrs.) M. N.
Naganandini, Department of Pharmacognosy, JSS College of Pharmacy, Mysore, India.

2.7. Preliminary phytochemical screening

The shade dried and coarsely powdered leaves were extracted
successively with different solvent by using soxhlet apparatus and
analysed using simple chemical tests for preliminary phytochemical screening of various groups of phytoconstituents such as alkaloids, avonoids, phenolic acids, sterols, cardiac glycosides,
tannins and so on by using standard procedure.30,24

2.2. Macroscopic analysis

2.8. HPTLC studies

Details macroscopic and taxonomic features of the leaves viz.

size, shape, colour, surface, venation, apex, margin, base, lamina
and texture were studied according to the standard procedures
mentioned in standard textbooks.20,21

Qualitative densitometric analyses (HPTLC) was performed for

the development of characteristic ngerprinting prole for petroleum ether (60 e80 ) extract and essential oil of leaf of Piper betle
cultivars. 10 ml of the sample solution (10 mg/mL for petroleum
ether extract; 5% solution in petroleum ether for essential oil) was
applied and plates were developed in solvent system [(hexane:
ethyl acetate; 8.8:1.2) for petroleum ether extract, (toluene: ethyl
acetate; 7:3)] for essential oil. Developed plates were scanned at
various wavelengths.

2.3. Leaf constants

Fresh leaves were washed and small fragments of leaves were
taken from the middle region of the lamina of mature leaves.

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A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

3. Results and discussion

3.3. Anatomy

3.1. Macroscopic characters

The comparative anatomy of eight cultivars of Piper betle

showed structural similarities (Fig. 2). All the cultivars of Piper betle
leaves shows thick cuticle on the upper epidermis and thin on the
lower epidermis. Piper betle leaves showed barrel shaped four
layered upper and two layered lower epidermis (8m
4m) (Fig. 2a, g
and i). Sclerenchymatous cells for support present above the
schizogenous secretory canal (Fig. 2a and b). The Schizogenous
secretory canal on the adaxial surface was present in all the cultivars of the species (Fig. 2a). Distinct phloem tissue can be seen on
the ventral surface and well developed xylem tissue towards
the dorsal surface of the midrib. Xylem consist of tracheids

The detailed macroscopical description was studied and shown

in (Table 1) (Fig. 1).

3.2. Leaf constants

Quantitative microscopy is an important tool for identifying
characteristic of leaf, stomatal number, vein islet and vein termination numbers are present in (Table 2) and (Fig. 1a, b and d).

Fig. 1. Different Piper betel L. cultivars, Adaxial view and leaf constant measurement photographs; A) Desi; B) Bangla; C) Kali; D) Haldia; E) Sanci; F) Meetha; G) Birkoli and H) Magahi;
a,b) Apostomatic and hypostomatic tetra-cyticstomatal complex, face view of the adaxial and abaxial sides of the epidermis (40
), epidermal cells, subsidiary cells; c,e) Rosette type
crystal, Balloon like epidermal cells; d) Vein termination, vein islet and f) Conjugation tube like structure, glandular trichomes. Abbreviations: ST) Stomata; SBC) Subsidiary cells; EC)
Epidermal cells; RC) Rosette crystals; BE) Balloon like epidermal cells; VT) Vein termination; VI) Vein islet; COS) Conjugation tube like structure and GT) Glandular trichome.

A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

179

Fig. 2. Transverse section of leaf of different Piper betle cultivars; A) Magahi; B) Meetha; C) Sanci; D) Haldia; E) Kali; F) Bilkori; G) Bangla and H) Desi. Enlarged view of transverse
section of leaf of Piper betle gure 2G): a, b) Showing the presence of SC e Sclerenchymatous cells; SZ e Schizogenous secretory canal; T e Trichomes; UE e Upper epidermal cells
(Two to three layer); c) Showing the presence of tracheoid idioblasts like structure; MXY e Metaxylem; PXY e Protoxylem; PH e Phloem with SBS e Sclerenchymatous bundle
sheath; CVB e Collateral vascular bundle and V e Vessels; d) UPP e Upper palisade parenchyma, LPP e Lower palisade parenchyma; OC e Oil cell; e) SPF e Sclerenchymatous
pericycle bre; f) OC e Oil reserves/oil cells, CO e Collenchymatous cells, PO e Prism of calcium oxalate; g) LE e Lower epidermal cells (side view), TT e Tector trichomes; h) PC/G e
Polygonal Parenchymatous cells or ground tissue with oil cell; and i) LE e Lower epidermal cells (one to two layer), ground tissue/Parenchymatous cells.

(3.96e10.45 m), xylem parenchyma, protoxylem, and metaxylem

towards lower periphery. One conjoint collateral vascular bundle is
encircled with parenchymatous cells with sclerenchymatous
bundle sheath (Fig. 2c). Tracheoid idioblasts like structure have

been noticed from the vessels elements of midrib and petiole

(Fig. 2c). It is also characterized by presence of polygonal parenchymatous cells with oil cell was observed (Fig. 2h). The leaves are
hypostomatic, tetra-cyticstomatal complexes are common which is

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A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

Fig. 3. Powder characters showing the presence of ST) Stomata (40
, 100
); OC/OR/SC) Oil cells/oil reserves/ secretary cells; ROC) Ruptured oil cells; LEC) Lower epidermal cells;
UEC) Upper epidermal cells; VE) Spiral vessel element; SVF) Spiral vessel bre; PO) Prism of calcium oxalate and TOC) Translucent oil cells. Multicellular glandular trichomes (40
)
of Piper betle L. cultivars a) Meetha; b) Magahi c, d, e) Sanci, f,g) Desi; h,i) Bangla; j,k) Bilkori; l,m) Kali and n) Haldia. Uniseriate (r, v) and multicellular tector trichomes (o, p, q, s, t, u).

the characteristic feature of the Piperaceae.31,32 Hypostomatic

tetra-cytic stomata in lower epidermis and apostomatic in upper
epidermis of Piper betle leaves (Fig. 1a and b). Some times stomatal
complexes are cyclocytic but twin stomata and just aposed type are

also seen (Fig. 1c 10
) and (Fig. 1e 100
). Hypostomatic, tetracyticstomatal complexes in lower epidermis and apostomatic in
upper with heterogenous mesophyll, were previously reported.
They have a multiple epidermis in both the adaxial and abaxial

A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

Table 2
Leaf constant of leaves of Piper betle cultivars {Mean  SD}.
Cultivars

Stomatal no.

Magahi
Haldia
Bangla
Desi
Sanci
Birkoli
Meetha
Kali

19
16
9.0
12
17
12
17
10










1.6a
2.4
1.6
1.8
1.8
1.8
1.2
1.2

Stomatal index
27
21
25
20
29
24
19
28










3.8
2.1
2.0
0.9
1.4
1.4
1.4
1.4

Vein termination no.

11
12
15
10
13
11
20
14










1.9
3.0
3.9
1.4
1.9
3.2
2.2
2.1

Vein islet no.

6.0
5.0
7.0
6.0
5.0
6.0
6.0
7.0










1.1
1.0
1.0
0.5
0.9
0.8
1.1
1.1

Each value is the mean  standard deviation of triplicate determinations.

Table 3
Moisture & Ash values of leaves of Piper betle cultivars {Mean  SD}.
Cultivars Moisture content Total ash Water soluble ash Acid insoluble ash
Magahi
Haldia
Bangla
Desi
Sanci
Birkoli
Meetha
Kali
a

3.1  0.1a
4.4  0.2
6.0  0.3
11.0  0.1
5.5  0.3
7.0  0.1
8.0  0.2
8.5  0.3

3.0  0.1
2.0  0.1
2.8  0.1
3.2  0.1
3.6  0.3
2.3  0.1
2.8  0.3
2.9  0.2

1.3  0.1
1.7  0.15
1.0  0.1
1.2  0.15
1.1  0.15
1.4  0.15
1.9  0.1
1.3  0.1

0.2  0.05
0.8  0.1
0.3  0.1
0.7  0.1
0.5  0.2
0.1  0.05
0.5  0.15
0.4  0.1

Each value is the mean  standard deviation of triplicate determinations.

Table 4
Extractive values of leaves of Piper betle cultivars with successive extraction.
(Soxhlation) {% (w/w)  SD}.
Cultivars

Pet-ether
(60e80  C)

Magahi
Haldia
Bangla
Desi
Sanci
Bilkori
Meetha
Kali

2.8 
3.0 
4.6 
3.2 
2.0 
2.8 
4.2 
4.9 

0.1a
0.26
0.15
0.15
0.15
0.3
0.15
0.4

Ethyl acetate
3.5
4.4
2.2
15.4
1.0
7.0
6.1
4.4










0.15
0.17
0.15
0.25
0.15
0.15
0.15
0.15

Chloroform
4.0
2.1
13
2.0
5.2
5.6
3.7
2.6










0.15
0.15
0.1
0.1
0.4
0.4
0.15
0.1

Methanol
2.1
9.4
12.4
8.1
14.5
7.5
2.5
12.4










0.1
0.20
0.25
0.15
0.35
0.30
0.11
0.26

Each value is the mean  standard deviation of triplicate determinations.

surfaces with secretory glandular trichomes and tector trichomes

respectively15,18,19 Conjugation tube like structures is present between the twin stomata in Sanci cultivar (Fig. 1f). The apical cell of
trichome is slightly pointed or clavately shaped are multicellular
and uniseriate and are more restricted in the midrib and petiole
region (Fig. 2b, g and i; Fig. 3oev). Cultivar Magahi leaves has more
stomatal number (Table 2) and trichome frequency (Fig. 2A)

181

frequency than the other cultivars. All the cultivars of the species
studied have a heterogenous mesophyll with a single palisade parenchyma layer (Fig. 2d) and three to four compactly arranged
spongy parenchyma is also observed. The margin is similar in all the
cultivars studied where the sub-epidermal cell layer is collenchymatous at the midrib (Fig. 2f). Calcium oxalate crystals are mostly
prismatic types (Fig. 2f and 3L) and oil reserves (Fig. 2d, f and h;
Fig. 3C, D, F, I and K) are observed in the cortical collenchymas and
in parenchyma regions of the petiole.

3.4. Powder study

The most common features of powder of leaves show presence
of glandular trichomes which have uniseriate and multicellular
apical cell and a short pedicel (Fig. 3aen) are found in the sub
epidermal cells on both sides. Non-glandular trichomes (tector
trichomes) are multicellular and uniseriate are more restricted at
the midrib and petiole regions (Fig. 3oev). Oil reserves {yellow
(Fig. 3I, F), reddish yellow (Fig. 3K) and red in colour (Fig. 3C, D)
found in the cortical collenchyma and in parenchyma regions of the
petiole. The powder study of leaves also shows the presence of
ruptured oil cells (Fig. 3E), epidermal cells (Fig. 3F, H), xylem vessel
element (Fig. 3J), stomata (Fig. 3A and B), spiral xylem bers
(Fig. 3G) and prismatic calcium oxalate monocrystals with a pyramidal base occur in parenchymatous and collenchymatous petiole
cells. (Figs. 2f and 3L). Balloon like epidermal cells and rosette type
crystals are observed (Fig. 1C). These characteristics are observed
and are reported for the rst time in these cultivars.

3.5. Physicochemical parameters

Physicochemical parameters like percentage of moisture content, total ash, acid insoluble ash, water soluble ash values were
determined. Ash values of a drug give an idea of the earthy matter
or the inorganic composition and other impurities present along
with the drug. Piper betle cultivars showed lower content of water
soluble and acid insoluble ash followed by total ash. Materials
contain free water, the relative amount being dependent upon
the physical and chemical properties of the material. The drying
process stops the degradation processes caused by yeasts or enzymes, prevents the development of microorganisms and oxidation and hydrolysis reactions. The moisture content in fresh plants
leaves varies from 60% to 80%. The drying process reduces
the moisture content to 5%e12%. In Desi cultivar moisture content
was high 11.0  0.1 followed by other cultivars (Table 3). The
extractive values are primarily useful for the determination of
exhausted or adulterated drug. The water soluble extractive was
low. (Table 4).

Table 5
Behaviour of powdered leaves on treatment with different chemical reagents.
Reagents

Bangla

Magahi

Haldia

Desi

Sanci

Bilkori

Meetha

Kali

1N NaOH (aq.)
1N NaOH (alk.)
H2SO4 (dil.)
HCl (dil.)
HCl (Conc.)
Acetic acid
5% Fecl3
5% Iodine
Ammonia
1N HNO3

Brown
Dark brown
Yellow
Buff colour
Dark brown
Olive green
Brownish yellow
Maroon red
Dark brown
Yellow

Brown
Dark brown
Pale yellow
Light buff
Brown
Light green
Yellow
Maroon red
Dark brown
Dark yellow

Brown
Dark brown
Brown
Light buff
Dark brown
Dark brown
Yellow
Blood red
Brownish green
Brick red

Brown
Dark brown
Light brown
Buff colour
Dark brown
Olive green
Brownish yellow
Maroon red
Dark brown
Yellow

Brown
Dark brown
Dirty white
Blackish colour
Straw colour
Light green
Yellowish brown
Dark reddish
Brown
Yellow

Brown
Dark brown
Pale
Buff colour
Yellow
Light green
Yellow
Maroon
Brownish green
Yellow

Brown
Dark brown
Buff colour
Buff colour
Brownish Green
Light green
Yellow
Blood red
Light brown
Yellow

Brown
Dark brown
Buff
Off white
Dark green
Green
Yellowish green
Maroon red
Yellow
Orange

Abbreviations: aq) Aqueous; alk) Alkaline; 1N) Normal; dil) Dilute; Conc.) Concentrated.

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A.A. Khan et al. / Pharmacognosy Journal 5 (2013) 176e183

Fig. 4. Qualitative HPTLC ngerprint proles of Piper betle cultivars A) Petroleum ether extract (at 200 nm) B) Essential oils (250 nm).

3.6. Preliminary phytochemical screening and HPTLC studies

References

Behaviour of powdered leaves of Piper betle cultivars on treatment with different chemical reagents are furnished in Table 5.
Preliminary phytochemical screening of methanolic extract
revealed the presence of carbohydrates, protein, amino acids, triterpenoids, steroids, glycosides, avonoids and absence of alkaloids, xed oils, saponins. HPTLC studies performed on petroleum
ether extract and essential oil shows differences in track prole of
all the cultivars (Fig. 4). Further there is need to explore the
chemical differences in the volatile oil content as well as in other
extracts. Chemical Markers for each cultivar would help in their
differentiation quickly.
In brief the leaves of Piper betle cultivars are hypostomatic, tetracytic (Fig. 1a, b) stomatal complex which seems to be characteristic
of Piperaceae family. Oil cells occur in the parenchyma, collenchyma and vascular tissues of the petiole and midrib. In the blade,
they can appear in the mesophyll or sub epidermis. The oil secretory cells are common in Piperaceae.32 Prismatic calcium oxalate
monocrystals with a pyramidal base occur in parenchymatous and
collenchymatous petiole cells.33 Tracheoid idioblasts like structure
were previously reported.34 Conjugation tube like structure was
present between twin stomata in Sanci cultivar (Fig. 1F). Schizogenous oil cavity was observed with secretory epithelial layer at the
midrib and in petiole35 (Fig. 2a). According to Cristina, translucent
secretions are found in newly sectioned leaf of Piper species36
(Fig. 3D).
As there is no pharmacognostical work on record of this traditionally much valued drug, the present work was taken up with a
view to lay down standards, which could be useful to detect the
authenticity of this medicinally useful plant. Macro and micro
morphological standards discussed here can be considered as
identifying parameters to authenticate the drug.

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Conicts of interest
All authors have none to declare.

Acknowledgement
This work was supported by grant from the CSIR, (Council of
Scientic and Industrial Research), Government of India, New
Delhi. (09/554 (0030)/2011 EMR-1). The authors are very much
thankful to Dr. S. Jha (Professor) and Dr. K. Jayaram (Associate
Professor), Dept. of Pharmaceutical Sciences, Birla Institute of
Technology, Mesra, Ranchi, Jharkhand, India, for valuable support
during research work.

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