See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/230767063

Piper betle L.: A review on its ethnobotany, phytochemistry, pharmacological profile

and profiling by new hyphenated technique DART-MS (Direct Analysis in Real Time
Mass Spectromet...

Article  in  Journal of Pharmacy Research · October 2011

CITATIONS READS

16 2,421

7 authors, including:

Kushagra Nagori Mukesh K. Singh

Rungta College of Pharmaceutical Sciences and Research Shri Rawatpura Sarkar Institute of pharmacy, kumhari, durg, (C.G.) india
16 PUBLICATIONS   70 CITATIONS    20 PUBLICATIONS   145 CITATIONS   

SEE PROFILE SEE PROFILE

Amit Alexander Tekeshwar Kumar

Rungta College of Pharmaceutical Sciences and Research Nitte University
130 PUBLICATIONS   991 CITATIONS    11 PUBLICATIONS   90 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Polysaccharide based nanoarchitect for local and systemic action View project

Drug design and its Pharmacological Screening View project

All content following this page was uploaded by Hemant Badwaik on 04 June 2014.

The user has requested enhancement of the downloaded file.

 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
Review Article Available online through
ISSN: 0974-6943 www.jpronline.info
Piper betle L.: A review on its ethnobotany, phytochemistry, pharmacological profile and profiling by
new hyphenated technique DART-MS (Direct Analysis in Real Time Mass Spectrometry).
Kushagra Nagori*, Mukesh Kumar Singh, Amit Alexander, Tekeshwar Kumar, Dhansay Dewangan, Hemant Badwaik, D.K. Tripathi
Rungta College of Pharmaceutical Sciences and Research, Bhilai 490006, Chhattisgarh, India.
Received on: 19-05-2011; Revised on: 08-06-2011; Accepted on:01-07-2011

ABSTRACT
The Piper betle plant is an evergreen and perennial creeper which is used in several traditional medicines to cure various diseases. This plant has been known
to possess antioxidant, antifungal, antiulcerogenic, antiplatelet, antidiabetic, immunomodulatory, antileishmanial, antiamoebic, anti-inflammatory, antifilarial
and antimicrobial activity. A wide range of chemical compounds in cluding chavibetol, allyl pyrocatechol, eugenol, quercetin, caryophyllene, safrole,
hydroxychavicol, a-pinene, myrcene, chavicol, Germacrene-D, a- terpineol , ß-pinene, camphene etc have been isolated from this plant. The present review
summarizes the information concerning the botany, ethnopharmacology, phytochemistry, biological activity and use of hyphenated analytical technique like
DART-MS (Direct Analysis in Real Time Mass Spectrometry) and other techniques for characterizing various compounds from P. betle plant.

Key words: P.betle, DART-MS, creeper, antioxidant, chavibetol.

INTRODUCTION
Occurrence, Botanical description and Ethnopharmacology
From the ancient time, man has been using plants as medicine. From a histori-
cal perspective, it has been evident that the fascination with plants is as old as
mankind. Herbs have provided us some of the very important life saving drugs
used in the armamentarium of modern medicine. The plant kingdom repre-
sents a rich source of organic components, many of which have been used for
medicinal & other purposes. Herbal medicines remain the major source of
health care for the world’s population 1. Currently, there is a growing interest
in plant based or herbal medicines even in the western world. In many re-
spects, the mechanism of action of the herbal drugs differs from that of the
synthetic drugs or pure compounds2. In one of the study of the World Health
Organization it is estimated that 80% of the population of developing coun-
tries relies on traditional plant based medicines for their health requirements3.
Piper betle Linn. (Betle vine) is a tropical plant closely related to the common
pepper and belongs to the family Piperaceae 4. It is extensively grown in India,
Srilanka, Malaysia, Thailand, Taiwan and other Southeast Asian countries and
has a long history of over 2000 yrs44. The stems are dichotomous, articulate,
swollen and rooted at nodes 3mm in diameter, woody and with 2.5 to 4cm long
internodes. Stem stout with pinkish-stripe along node dilated and rooting5.
The leaves are simple, spiral and exstipulate. The petiole is 5mm long, chan-
neled and pubescent. The blade is 10 x 6cm & 9.5 x 5cm, ovate to ovate –
oblong and light green below. The base of the blade is cordate and the apex is
acuminate, the secondary nerves are in three pairs. The inflorescence is an
axillary spike which is 5.5 cm long. The fruits are drupaceous, orange, and
3mm in diameter 11. The betel leaf is known as Paan (Assamese, Urdu, Hindi,
Odia, Bengali) and Bhakshyapatra, bhujangalata, bhujangavalli, divabhishta,
kalaskanda, nagavalli, nagavallika, nagini, parna, vitika (Sanskrit). Some of
the names in the regions in which it is consumed are: betel leaf vine (English),
Vetrilai (Tamil), Tamalapaku (Telugu), Vidyache pan (Marathi), Veeleyada
yele (Kannada), Vettila (Malayalam), Plu (Mon), Malus (Tetum), Maluu
(Khmer), Plue (Thai), Bulath (Sinhalese), Malu (Tokodede), Bileiy (Divehi),
Bulung samat (Kapampangan language), daun sirih (Malay language), Papulu
(Chamorro), Ikmo (Philippines) and Tr?u (Vietnamese)5,12. Ayurveda an In-
dian system of medicine has been an integral part of Indian culture and
materia medica. From the rich Indian biodiversity, it has identified various
plants/herbs that have been associated with a number of potential therapeutic
efficacies2.
PLANT PROFILE
Kingdom: Plantae
Division: magnoliphyta
Class: magnolipsida
Order: Piperales
Family: Piperaceae
Genus: Piper
Species: P. betle
*Corresponding author.
Earlier gastro protective properties of the leaf extract were reported on ex-
perimentally induced gastric lesions13. A phenolic compound hydroxychavicol
with anticarcinogenic property has also been identified in betel leaves14. Piper
betle is not recommended as a antidiabetic agent in traditional medicinal sys-
tems. However it is possible that Piper betle leaves may posses antidiabetic
properties as Piper sarmentosum and Piper longum close relatives of the
plant, have been shown to have antidiabetic properties 15, 16. The use of P. betle
L. leaves to lighten melasma may produce leukomelanosis, which manifests as
confetti-like skin depigmentation 17; also the roots (8-12g) are used in treating
rheumatism 6. The leaf produces an aromatic volatile oil containing a phenol
called chavicol which has powerful antiseptic properties. Piper betle inflores-
cence contains high levels of safrole (15mg/g wet weight) and chewing betel
quid containing Piper betle inflorescence generates a high concentration of
safrole (420 µM) 19,20. The plant possesses cardiotonic 41 wound healing3 9
chemopreventive 53 and reversible antifertility80 activities. Table 1.
Table 1 Ethnomedicinal importance of Piper betle.
Sr.No. Plant part used Ethnomedicinal Use
1. Whole Plant The liquid extract of the plant has been used traditionally in curing inflam-
mation and infection of the respiratory tract, cough, dyspnoea, indigestion,
diphtheria, hysteria as well as general and sexual debility 21 . A concoction of
indigenous Indian drugs containing P. betle dry extract was found to be an
effective long-lasting oral contraceptive 23 . Piper betle is also reported to
possess antifungal, antiseptic and anthelmintic properties to serve as a con-
traceptive for humans and to possess antihypertensive properties. 30, 31 The
users believe that chewing the ‘paan’ improves their efficiency and stamina32
. Piper betle showed hypotensive, cardiac, and respiratory depressant ef-
fects smooth and skeletal muscles relaxant actions, antimicrobial, fungicidal,
and nematocidal activity 33-35 . Medicinally P. betle is an aromatic, carmina-
tive, stimulant and astringent used as a preventive for worms and in snake bite
36
.
2. Leaves
Betel leaf has been described from ancient times as an aromatic, stimulo-
carminative (katu), astringent and aphrodisiac (kamagnisandipanam) 12 .The
leaves are credited with wound healing37, 39 . The Indian traditional system of
medicine has identified the P.betle leaves with digestive and pancreatic li-
pase stimulant activities.3 7 , 3 9 - 4 2 which has also been proved with experimen-
tal animals..43 Betel leaf is traditionally known to be useful for the treatment
of various diseases like bad breath, boils and abscesses, conjunctivitis, con-
stipation, headache, hysteria, itches, mastitis, mastoiditis, leucorrhoea, otor-
rhoea, ringworm, swelling of gum, rheumatism, abrasion, cuts and injuries
etc as folk medicine.44 Fresh juice of betle leaves is also used in many
ayurvedic preparations. 45 Leaves are also considered useful in treating bron-
chitis, difficulty in breathing and cough.37 Traditionally Piperbetle leaf extract
is reported to inhibit male reproductive competence. 46-48 Piper betelleaves
is used in eyedrops for eye injury/infection as a baby lotion for the new born,
for coughs, asthma, constipation and to arrest milk secretion.49 Essential oil
from leaves of this plant has been used for the treatment of respiratory
catarrhs and as antiseptic.5 0 . Piper betle leaves are shown to posses
hepatoprotective activity,51 antifertility on male rats48 and anti motility
effects on washed human spermatozoa 46 biological activities described for
the essential oil include antifungal, antiseptic and anthelmintic effects 80 .
In folk medicine root is known for its female contraceptive effects44 . The roots
3. Roots are used as a contraceptive and are chewed by singers to improve their voice
32

In ancient times flower is used as an ingredient for chewing food known as betel
4. Flower
Kushagra Nagori quid in South-East Asia .24 .
Rungta College of Pharmaceutical Sciences 5. Fruit The fruit is employed with honey as a remedy for cough 50 .
and Research, Bhilai 490006,
Chhattisgarh, India.
Tel.: + 91-09691500850, 0788-2350698
E-mail:kushagranagori13@yahoo.co.in

Journal of Pharmacy Research Vol.4.Issue 9. September 2011 2991-2997

 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
PHYTOCHEMISTRY
The chemical constituents of betel essential oil consist of mainly terpenes and
phenols53. Leaves contain protein 3.1 %, carbohydrate 6.9 %, minerals 2.3 %, and
tannins 2 %. Leaves contain bitter compounds that are about 0.7 to 2.6 %. The
nutritional composition of fresh Piper betle leaf: water (85-90%), Protein (3-
3.5%), Fat (0.4-1.0%), minerals (2.3-3.3%), Fibre (2.3%), Chlorophyll (0.01-
0.25%), Carbohydrate (0.5-6.10%), Nicotinic acid (0.63-0.89mg/100g), Vitamin
C (0.005-0.01%), Vitamin A (1.9-2.9 mg/100g), Thiamine (10-70 µg/100g), Ri-
boflavin (1.9-30 µg/100g), Tannin (0.1-1.3%), Nitrogen (2.0-7.0%), Phosphorus
(0.05-0.6%), Potassium (1.1-4.6%), Calcium (0.2-0.5%), Iron (0.005-0.007%),
Iodine (3.4 µg/100g), essential oil (0.08-0.2%, Energy (44 kcal/100g) 44. The betle
leaves have strong pungent aromatic flavor54 and the characterstic flavor of betel (3) Myrcene (C 10H 16 ) (4) Cadinene (C 15H 24)
is due to the betel phenols 55. More recent work with the leaves was found to
contain starch, diastases, sugars (0.8 to 1.8 %) and an essential oil to an extent of
4.2%, the specific gravity varies from 0.958 to 1.05712. Phytochemical investi-
gation on leaves revealed the presence of tannins and steroidal components.56.
The terpenoids include 1,8-cineole, cadinene (Fig.1.4), camphene, caryophyllene,
limonene, pinene, chavicol, allyl pyrocatechol, carvacrol (Fig.1.8), safrole, eu-
genol, and chavibetol are the major phenols found in Piper betle, there acetates
are also commonly found55,44. Eugenol was identified as the antifungal principle in
the oil.38 Some of the phytoconstituents of the plant are summarized in the Table
2.
Table. 2. Phytoconstituents of Piper betle
Active principle Plant part used Reference
(5) Camphene (C 10H 16) (6) Thujene (C 10H 16)
Chavibetol Leaves 57,22,54,58,59,60
Allyl pyrocatechol Leaves 58
Chavibetol acetate Leaves 61,62,57
Eugenol (Fig 1.1) Leaves &Flower 63,56,64
Allyl pyrocatechol Leaves 65,66,60
Quercetin Leaves &Flower 67&64
Luteolin, stearaldehyde Leaves 68, 42
a- terpineol , ß- pinene, camphene Leaves 62
Caryophyllene (Fig.1.9) leaves 57, 69,26
Safrole Leaves & Flower 63,64,70,64
Hydroxychavicol Leaves &Flower 56,71,64
ß- sitosterol Leaves & Roots 56,72,72
Diosgenin Leaves 56
?- lactone, allyl catechol , eugenol methyl ether, Leaves 73
cepharadione A, dotriacontane, triacontane
Piperine Stem 73,72
Piperlonguminine Stem 73
4-allyl resorcinol, humulene (Fig.1.17), stigmast-4-en-3,6- Roots 73 (7) Limonene (C 10H 16) (8) Carvacrol (C 10H 14O )
dione, aristololactam A-II, ß-sitosteryl palmitate, 3- ß-acetyl
ursolic acid , ursonic acid
Linalool (Fig.1.10) Leaves &Rhizome 62,22
Allyl diacetoxy benzene Leaves 38
Gallic acid, procatechuic acid, chlorogenic acid, caffeic acid, Whole plant 74
ferulic acid, ellagic acid
Isoeugenyl acetate Whole plant 69
N-Isobutyl-2E,4E-decadienamide, pipedardine, piperine Whole plant 75
4-allyl phenyl acetate ,4-allyl phenol, a- myrcene, D- Leaves 62
limonene, eucalyptol, camphor (Fig.1.15), allyl anisole, 4-allyl
phenol, ß-iso safrole (Fig.1.16), thymol (Fig.1.18), 3-allyl-6-
methoxyphenol, a-bergamotene, a-farnesene, a- muurolene, (9) Caryophyllene (C 15H 24 ) (10) Linalool (C 10H 18O )
a-bisabolene, ledol , a-cadinol, isoledene
a- tocopherol Leaves 76
Eugenyl acetate, allyl pyrocatechol diacetate, allyl Leaves 63,29
pyrocatechol monoacetate, terpinen -4-ol , thujene(Fig.1.6),
camphene (Fig.1.5), sabinene, 1,4 cineole, (Z)-ß-ocimene, (E)-
ß-ocimene, cis sabinene hydrate, fenchone, terpinolene, 2-
noanone, cis-limonene oxide, sabinene (Fig.1.14), m-cymen-
8-ol, cis-piperitol, n-decanal, thymol, 2-undecanone, iso-
ascaridole, 5-indanol, a-cubebene, a- copaene (Fig.1.10),
(E)-ß- Damascenone, vanillin , a-selinene, cuparene,
germacrene A, n-eicosane
a-pinene Leaves &Rhizome 63, 26,22
Chavicol (Fig.1.2) Leaves 63
Myrcene (Fig.1.3) Flower & Rhizome 70,63,24,22
Isoeugenol Flower 70,63,64, 62
ß- sitosteryl palmitate, dotriacontanoic acid, stearic acid, Leaves 72
cepharadione -A
(11) a- copaene (C 15H 24) (12) Germacrene D (C 15H 24 )
Methyl eugenol, flavones Flower 64
Benzene acetic acid , hexadecanoic acid, octadecanoic acid, Leaves 71
myristic acid,2,3-bis (hydroxy) propyl ester , 2 mono
palmitin, limonine (Fig.1.7), octadecanoic acid, 2,3-
bis(hydroxyl) propyl ester
Isoeugenyl acetate Leaves 69
(E)-ß-ocimene, terpinolene, allo ocimene, cymene (Fig.1.13). Leaves 27,28
ß- caryophyllene Leaves & Rhizome 77,62,22
Germacrene-D (Fig.1.12) Leaves 62

(13) Cymene (C 10H 14) (14) Sabinene (C 10H 16)

(1) Eugenol (C 10H 12O 2) (2) Chavicol (C 9H 10O )
Journal of Pharmacy Research Vol.4.Issue 9. September 2011 2991-2997
 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
(15) Camphor (C 10H 16O) (16) Safrole (C 10H 10O 2)
(17) Humulene (C 15H 24) (18) Thymol (C 10H 14O )
Fig. 1 (1-18) Chemical structures of some chemical constituents iso-
lated from various parts of Piper betle.
PHARMACOLOGICAL ACTIVITY
Antioxidant activity
Oxidative damage is an important effect of ionizing radiation on biological
membranes. It is a chain reaction. Free radicals generated from the radiolytic
decomposition of water can attack fatty acid chains of membrane lipid. A free
radical that has sufficient energy to abstract an allylic hydrogen from the
methylene carbon of polyunsaturated fatty acids can initiate the peroxidative
process. Here the presence of Piper betle leaf extract inhibited the radiation
induced lipid peroxidation process effectively. This could be attributed to its
ability to scavenge free radicals involved in initiation and propagation steps.
Oral supplementation with extract (1, 5 and 10 mg/kg) was administered daily
for 2 weeks to Swiss albino mice and the hepatic antioxidant status was analysed.
The GSH content was enhanced and no appreciable change was found in the
levels of oxidative damage in terms of lipid peroxidation. Also, the specific
activity of SOD increased in a dose dependent manner. These factors indicate
the elevation of antioxidant status in the animals. The effect on the glyox-
alase system which is considered to be activated under stress conditions was
also investigated. Our findings did not observe any significant change in gly I
and gly II activities, implying a non-stress condition after oral treatment of
the extract. The present study indicates the antioxidant activity of P. betle
leaf extract and its potential to elevate the antioxidant status 79 . Further
investigation shows that the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay of
the ethanol extracts of three varieties (Bangla, sweet, and Mysore) of Piper
betle (pan) revealed the Bangla variety to possess the best antioxidant activ-
ity that can be correlated with the total phenolic content and reducing powers
of the respective extracts. Column chromatography of the extract of the
Bangla variety led to the isolation of chavibetol (CHV), allylpyrocatechol
(APC), and their respective glucosides. The HPTLC analysis of the extracts
revealed similar chemical profiles in all three P. betle varieties, although the
concentrations of CHV and APC were significantly less in the sweet and
Mysore varieties. Among the isolated compounds, APC showed the best results
in all the in vitro experiments. It could prevent Fe (II)-induced lipid higher
than 4µg/ml 78. One study investigated that chavibetol, eugenol, methyl eu-
genol, and peroxidation (LPO) of liposomes and rat brain homogenates as well
as ç-ray-induced damage of pBR322 plasmid DNA more efficiently than CHV
2
The antioxidant activity was further shown in an aqueous IPB (inflorescence
.
Piper betle) extract and was found to be a scavenger of H2O2, superoxide
radical, and hydroxyl radical with a 50% inhibitory concentration (IC50) of
about 80, 28, and 73 µg/mL, respectively. IPB extract also prevented the
hydroxyl radical induced PUC18 plasmid DNA breaks at concentrations car-
vacrol played an important role in antioxidant effectiveness 79.
Treatment of halitosis
Piper betle L. (Piperaceae) leaves which are traditionally used in India and
China in the prevention of oral malodor was examined by bioassay-guided
fractionation to yield allylpyrocatechol (APC) as the major active principle
Journal of Pharmacy Research Vol.4.Issue 9. September 2011
which showed promising activity against obligate oral anaerobes responsible
for halitosis. The P.betle ether extract and APC could thus be used in the
prevention of halitosis in addition to the prevention of periodontal infection
caused by the colonization of oral anaerobic bacteria. The biological studies
with APC indicated that the potential to reduce methylmercaptan and hydro-
gen sulfide was mainly due to the anti-microbial activity as established using
dynamic in vitro models 65. The adhering ability of microorganisms to the
tooth surface determines the developmental stage of dental plaque. With
suitable environmental and nutrients, carcinogenic plaque will subsequently
develop. To minimize the development of carcinogenic plaque, the adhering
ability of the early settlers needs to be control problem the aqueous extract of
Piper betle is used which reduced the adherence of early plaque bacteria to an
experimental pellicle. The extracts were observed to have the ability to make
them less adherent. This could account for the significant reduction in the
hydrophobic binding capacity of the bacteria when treated with the extracts 81.
Antifungal activity
The investigation attempted to identify new antifungal agents by screening
two varieties of Piper betle (green vein and red vein) showed strong activity
against all the pathogens tested (Colletotrichum capsici, Fusarium
pallidoroseum, Botryodiplodia theobromae, Alternaria alternata, Penicil-
lium citrinum, Phomopsis caricae-papayae and Aspergillus niger) with inhi-
bition diameters significantly (P < 0.01) bigger than 2.5 mg/ml prochloraz or
10 mg/ml clotrimazole. The minimum inhibitory concentrations of the
ethanolic extracts of P. betle against these plant pathogens ranged between
0.01 mg/ml & 1 mg/m1 82.
Antileishmanial activity
An ethanolic extract of leaves of Piper betle (PB) was tested for its
antileishmanial activity that was evidenced in both promastigotes and
amastigotes, with IC50 values of 9.8 and 5.45 µg/ml, respectively; importantly,
it was accompanied by a safety index of >12-fold. This leishmanicidal activity
of PB was mediated via apoptosis as evidenced by morphological changes, loss
of mitochondrial membrane potential, in situ labeling of DNA fragments by
terminal deoxyribonucleotidyl transferase mediated deoxyuridine triphosphate
nick end labeling, and cell-cycle arrest at the sub-G0/G1 phase. Here it is
anticipated that phenolics identified in PB such as APC and chavibetol could
contribute towards the observed antileishmanial activity.68.
Radioprotective activity
The radioprotective activity of Piper betle ethanolic extract has been studied
using rat liver mitochondria and pBR 322 plasmid DNA as two model in vitro
systems. The extract effectively prevented ?-ray induced lipid peroxidation as
assessed by measuring thiobarbituric acid reactive substrates, lipid hydroper-
oxide and conjugated diene. Likewise, it prevented radiation-induced DNA
strand breaks in a concentration dependent manner. The radioprotective ac-
tivity of PE could be attributed to its hydroxyl and superoxide radicals scav-
enging property along with its lymphoproliferative activity. The radical scav-
enging capacity of PE was primarily due to its constituent phenolics, which
were isolated and identified as chavibetol and allyl pyrocatechol 58.
Antiulcerogenic activity
Pretreatment of an ethanolic extract of leaf of Piper betle Linn at a dose of
200mg/kg body weight, orally administered to rats for ten consecutive days,
was found to posseses a significant protective action against gastric lesions
induced by indomethacin. The extract pretreatment resulted in significant
increase in superoxide dismutase (SOD) and catalase (CAT) activity, increase
in mucus, hexosamine and total thiol group content, but marked reduction in
oxidatively damaged protein and peroxidised lipid levels as compared to un-
treated ulcerated control. The extract was also found to possess both superox-
ide and hydroxyl free radical scavenging action. The present observation
establish the efficacy of the extract in prevention of experimentally induced
peptic ulcer by indomethacin and antioxidant property appears to be pre-
dominantly responsible for such cytoprotective activity in the experimental
model 83. Further investigation showed the protective activity of allyl pyrocat-
echol (APC), which is the major antioxidant constituent of P.betle against the
indomethacin-induced stomach ulceration in the rat model. This was done by
treating the rat mode l with APC (2mg/kg body wt per day) and misoprostol
(1.43 µg/kg body wt per day) for 7 days and effectively heals the stomach
ulceration as revealed from the ulcer index and histopathological studies. The
healing capacities of APC and misoprostol could be attributed to their antioxi-
dant activity as well as the ability to enhance the mucin content of the gastric
tissues 66. Another investigation revealed that the healing activity of ethanolic
extracts of Piper betle against the indomethacin induced stomach ulceration
has been studied and compared with that of misoprostol. It was found that the
excellent healing activity of ethanolic extract of P. betle play a major role of
mucin protection and regeneration in the healing of non-steriodal anti-in-
flammatory drugs mediated stomach ulceration 84.
2991-2997
 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
Antiplatelet activity for all the three life-stages for water decoction was between 1.5 and 6.25 ml/
ROS (reactive oxygen species) are crucial for platelet aggregation and IPB ml. Regarding crude methanolic extract, LC100 values ranged between 7.8 and
extract has been found to inhibit the arachidonic acid (AA) induced and col- 31.25 mg/ml. Interestingly, among the different fractions, the chloroform
lagen-induced platelet aggregation, with an IC50 of 207 and 335 µg/ml, re- fraction was the most active on adult B. malayi, showing LC100 of 3.9 mg/ml,
spectively. The IPB extract also inhibited the AA-, collagen- (>100 µg/mL of followed by hexane fraction which was effective at 31.2 mg/ml; n-butanol and
IPB), and thrombin (>250 µg/mL of IPB)-induced thromboxane B2 (TXB2) aqueous fractions were ineffective 86.
production by more than 90%. However the effect of thrombin-induced ag-
gregation was not significant. These results indicated that aqueous compo- Anti-inflammatory activity
nents of IPB are potential ROS scavengers and may prevent the platelet The study reported that the crude leaf powder suspension of P. betle was
aggregation possibly via scavenging ROS or inhibition of TXB2 production 78. evaluated for acute and chronic anti-inflammatory study at a dose of 300 mg/
kg. Diclofenac sodium was used as the standard drug. Carrageenan and dextran
Immunomodulatory activity models were studied for acute inflammation while cotton pellet induced granu-
Many of the disorders today are based on the imbalances of immunological loma was used for chronic inflammation study. ANOVA followed by Dunnett’s
processes. This necessitates the search for newer and safer immunomodulators. test were employed for statistical analysis. The study indicates that the P.betle
With this objective a study was conducted to explore the immunomodulatory exhibit significant anti inflammatory activity 85.
activity of the methanolic extract of Piper betle L. The MPb consists of
mixture of phenols, flavonoids, tannins and polysaccharides. Both in vitro as Anti-amoebic & anti-giardial activity
well as in vivo evaluation was carried out. The effects of MPb on lymphocyte The anti-amoebic activity of P.betle was studied with chloroform extract of
proliferation, interferon-C receptors and the production of nitric oxide were P.betle leaf which was commonly used by AIDS patients in southern Thailand.
measured in vitro. Further, the extract at different dose levels was studied in It was screened at a concentration of 1,000 µg/ml, against Entamoeba
vivo for the humoral and cellular immune responses on mice immunized with histolytica strain HTH-56:MUTM and strain HM1:IMSS growing in vitro.
sheep red blood cells.the result showed that P.betle significantly suppressed The extract was incubated with 2×105E. histolytica trophozoites/ml of me-
phytohaemaglutinin stimulated peripheral blood lymphocyte proliferation in dium at 37°C under anaerobic conditions for 24 h. The cultures were examined
a dose-dependent manner. The decrease in antibody titre and increased sup- with an inverted microscope and scored (1-4) according to the appearance and
pression of inflammation suggests possible immunosuppressive effect of ex- numbers of the trophozoites the extract caused inhibition and retested using
tract on cellular and humoral response in mice. The study indicated the poten- the same conditions but with concentrations that ranged from 31.25 to 1,000
tial of MPb as a novel candidate for immunosuppressive activity. The same µg/ml using E.histolytica strain HM1:IMSS, and the IC50 values for each ex-
could be further evaluated for its anticancer activity or as a potential candi- tract was calculated. The chloroform extract of P.betle with IC50 value of 55.2
date in the treatment of autoimmune disorders such as rheumatoid arthritis, µg/ml was classified as active i.e. with an IC50 of less than 100 µg/ml. The IC50
systemic lupus erythomatous or emphysema 56. of a standard drug metronidazole was 1.1 µg/ml 87. Also the study investigated
the anti-giardial activity of chloroform extract of P.betle leaf which was
Antimicrobial activity commonly used as self medication by AIDS patients in southern Thailand. The
The methanolic and aqueous extracts showed strong activity against the yeasts: standard drug metronidazole and plant extract was incubated with 2 x 105
C. albicans, and M. pachydermatis. The crude essential oil exhibited a broad- trophozoites of Giardia intestinalis per millilitre of growth medium in 96-well
spectrum strong antimicrobial activity against all test organisms. The stron- tissue culture plates under anaerobic conditions for 24 h. The chloroform
gest activity was observed against C. albicans, followed by S. aureus and M. extract of P.betle was shown to be active i.e. with an IC50 of 100 < µg/ml. the
pachydermatis62. Further an investigation reported the antimicrobial activity results shows that the MIC & IC50 values of P.betle leaf have potential use
of crude aqueous extract of P. betle on Streptococcus mutans. Transmission against infection caused by G. intestinalis 88.
electron microscopy (TEM) was used to determine the effect of the extract
on the ultrastructure of S. mutans. The study showed cells with nucleoid ANALYTICAL TECHNIQUES USED IN IDENTIFICATION OF CHEMI-
material coagulated in to thick electron dense filaments and destruction of the CAL CONSTITUENTS OF P. BETLE
plasma cell membrane and inner cell wall. The effect is more significant with Profiling of Piper betle Linn. by Direct Analysis in Real Time Mass
the higher concentration of the extract. The effect of crude extract on the Spectrometric Technique.
ultra structure of S. mutans observed in this study could be due to the fatty Identification and quantitation of bioactive components from raw materials
acids and hydroxyl fatty acid ester components present. The hydrophobic and processed products has been routinely performed by HPLC-PAD, HPLC-
parts of the compound may enable them to partition the lipids of the bacterial MS,GC-MS, etc. Although such methods have been successfully adopted to
cell membrane, thereby disturbing the structures and rendering them more various fields of natural products and crude herbal drugs, there is an increasing
permeable. When the membrane is more permeable, other components present requirement for more efficient and prompt analytical techniques in order to
in the extract could make its way in to the bacterium and coagulate the manage vast numbers of samples. The quick and efficient analysis and real
nucleoid while maintaining the cell intact. Analysis of the effect on the acid time detection of bioactive components in raw materials and processed prod-
producing propertieswas analysed by pH drop assay. The extract was found to ucts would be one of the most important steps in facilitating the successful
significantly reduce acid producing properties of the bacteria 71. Antibacterial development of products containing botanical drugs and functional foods. In
activity of P.betle was checked against 15 clinically important bacterial strains order to evaluate the extent of the advantage offered by DART-MS in provid-
it shows moderate activity against B.cereus. Here cefotaxime (100µg/disc) was ing real time analysis of natural products from raw materials, a pilot study was
used as a standard. From results it can be said that P. betle can be explored performed with the well known anti-oxidant botanical drug Piper betle 94 . The
further as they are able to resist bacterial growth up to some extent 85. new technique referred to as Direct Analysis in Real Time has been coupled to
the AcuuTOF-LC atmospheric pressure ionization mass spectrometer to per-
Mosquito larvicidal and Tyrosinase inhibition activity mit high-resolution, exact mass measurements of gases, liquids, and solids 96.
While mosquito larvicidal activity was carried out with the essential oil and Using a helium plasma DART ionizes atmospheric water and generates water
methanolic and aqueous extracts of P. betle. The essential oil exhibited the clusters which in turn ionize the sample held in the gas stream. The resulting
larvicidal activity significantly with 2 h and 24 h, LD50 value of 86 and 48 spectra are relatively clean and simple. As DART ion source can ionize mol-
ppm, respectively. The methanolic extract of P. betle showed larvicidal activ- ecules directly from the surface plant products, which can be analyzed directly
ity with 2 h and 24h with LD 50 value of 153 and 125 ppm, respectively, without sample preparation 97, 98, 99. The eight varieties of Piper betle leaves
whereas the aqueous extract showed slight activity. The study suggested that were analysed by DART-MS named Bangla, Desawari, Deshi, Jaleshar green,
the essential oil and methanolic extract of P. betle is a potential natural Jaleshar white, Kalkatiya, Mahoba and Saufia. The results were shown in the
mosquito larvicide 62. Inhibition of mushroom tyrosinase by the essential oil, Table 3.
methanolic and aqueous extracts of P. betle was evaluated. The essential oil
exhibited concentration-dependent inhibition of mushroom tyrosinase, giv- Table .3. Exact mass data from the DART mass spectra of Piper betle leaves
ing an IC50 value of 126 ppm. The essential oil was fractionated into two Molecular Measured Calculated Molecular Error Remarks
fractions. The fraction I showed a strong inhibition of tyrosinase activity weight mass mass formula (mmu)
concentration-dependently, at an IC50 value of 115 ppm. The presence of the
134 135.08128 135.08099 C9 H11 O 0.29 Chavicol
hydroxyl group at the 4 position of the aromatic ring (4-allylphenol, eugenol) 150 151.07640 151.07590 C9 H10 O2 0.50 Allylpyrocatechol
in the essential oil, may play an important role in the inhibition of tyrosinase 164 165.09242 165.09155 C10 H13 O2 0.87 Chavibetol
62 165 166.08864 166.08680 C9 H12 NO2 0.84 Phenyl alanine
.
174 175.07764 175.07590 C11 H11 O2 1.74 Unknown
176 177.09109 177.09155 C11 H13 O2 -0.46 Chavicol acetate
Antifilarial activity 192 193.08772 193.08647 C11 H13 O3 1.25 Allylpyrocatechol
Both water decoction and crude methanol extract of BM in general appeared acetate
206 207.10140 207.10212 C12 H15 O3 -0.72 Chavibetol acetate
to possess good antifilarial efficacy against all the life-stages of B. malayi, 251 252.12287 252.12358 C13 H18 NO4 -0.71 Unknown
viz. adult worms, vector derived infective larvae and microfilariae. The LC100
Journal of Pharmacy Research Vol.4.Issue 9. September 2011 2991-2997
 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
Table No. 4 Chemical constituents obtained by different analytical techniques in P.betle.
P. betle varieties Part used Identification method used Active principle Reference

Indian (Saufia, Bangla, Leaves DART-MS Chavicol, allylpyrocatechol, chavibetol, 55

Desawari, J.Green, J.White, Phenyl alanine, chavicol acetate, chavibetol acetate,
Kalkatiya, Mahoba, Deshi Paan ) allylpyrocatechol acetate, allylpyrocatechol diacetate.
Vietnamese Rhizome oil Combination of capillary GC, a- cadinol, d- cadinene. 22
GC-MS & 13 C-NMR.
Leaves GC & GC-MS Isoeugenol and isoeugenyl acetate 89
Taiwanese Flowers GC/MS Safrole (28%), & myrcene (26%) 24
HPLC and TLC Safrole, hydroxychavicol, eugenol, isoeugenol, eugenol methyl ether.
S. marcescens treated betlevine Leaves and roots HPLC Gallic,, protocatechuic, chlorogenic, caffeic, ferulic, ellagic acid. 74
P.betle Herba Whole plant TLC, HPLC-DAD, GC-MS Piperdardine, piperine. 75
—— Leaves GLC Safrole, eugenol, allyl pyrocatechol diacetate, methyl chavicol,terpineol. 91
Betle inflorescence Water extract HPLC Safrole (78.9%), eugenol (6.2%) 92
Srilankan Leaves GC-MS Safrole, allyl pyrocatechol diacetate, eugenol & eugenol acetate 63
Indian (South) Leaves GC-FID, GC-MS, olfatometry. Chavibetol, eugenol, allyl pyro catechol derivatives. 93
Indian Leaves GC, GC-MS ß - caryophyllene (less than 26.5% ) 77
Indian (Bangla) Leaves Column Chromatography Chavibetol, allyl pyrocatechol 60
Malasian Leaves GC-MS Hydroxychavicol,hexadecanamide,2-monopalmitin, hexadecanoic acid, 2,3- 71
bis(hydroxy)propyl ester, myristic acid, 2,3-bis(hydroxy)propyl ester, benzeneacetic
acid, alpha-hydroxy phenyl.

The mass spectrometer used was a JMS-100 TLC (AccuTof) atmospheric pressure
ionization time-of-flight mass spectrometer (Jeol, Tokyo, Japan) fitted with a
DART ion source. The mass spectrometer was operated in positive-ion mode with
a resolving power of 6000 (full-width at half-maximum).The orifice 1 potential
was set to 28 V, resulting in minimal fragmentation. The ring lens and orifice 2
potentials were set to 13 and 5 V, respectively. Orifice 1 was set to a temperature
of 100°C. The RF ion guide potential was 300 V. The DART ion source was
operated with helium gas flowing at approximately 4.0 L/min. The gas heater was
set to 300°C. The potential on the discharge needle electrode of the DART source
was set to 3000 V; electrode 1was 100 V and the gridwas at 250 V. Freshly cut
pieces of betel leafwere positioned in the gap between the DART source and mass
spectrometer for measurements. Data acquisition was from m/z 10 to 1050. Exact
mass calibration was accomplished by including a mass spectrum of neat polyeth-
ylene (PEG) glycol (1:1 mixture PEG 200 and PEG 600) in the data file. m-
Nitrobenzyl alcohol was also used for calibration. The mass calibration was accu-
rate to within_0.002 u. Using the Mass Center software, the elemental composi-
tion could be determined on selected peaks. PCA analysis was carried out using
Minitab 14 statistical analysis software (Trial Version) 55.
APPLICATION OF DART – MS
DART has been interfaced to mass spectrometry for the analysis of counterfeit
antimalarials100,101 formulated products102 bioanalytical samples103 chemical reac-
tions104 metabolic stability 105 fatty acid methyl esters106 flavors and fragrances97.
Some chemical constituents obtained by different analytical techniques in P.betle
are shown in Table 4.
CONCLUSION
The scientific research on P. betle suggests a huge biological potential of this
plant. It is strongly believed that detailed information as presented in this
review on the phytochemical, analytical techniques and various biological
properties of the extracts might provide detailed evidence for the use of this
plant in different medicines. The phytochemical variations and efficacy of
the medicinal values of P.betle is dependent on geographical locations and
seasons. Betle quid are very commonly used by local people of India. Betel
quid generally consists of betel leaf (from the Piper betle L. vine), areca nut
(from the Areca catechu tree), catechu (a tannin-rich powder) and slaked lime
(calcium hydroxide), to which tobacco is often added. There have been many
reports that chewing betel quid causes oral cancer. There is a demand to
standardize the toxic properties of Betle quid and their detailed clinical trials.
After proper processing, identification and removal of the harmful properties
of quid, they may be utilized to prepare a good, nourishing and Ayurvedic
medicine. At the same time, the organic and aqueous extract of P.betle could
be further exploited in the future as a source of useful phytochemical com-
pounds for the pharmaceutical industry.
REFERENCES
1. Mohammad Y, Mohammad I, Antiasthmatic herbal drugs: a review, International
Journal of Pharmacy and Pharmaceutical Sciences, 2(3), 2010 28-29.
2. Jitesh S, Birija S, Soumyaditya M, Gamre S, Chattopadhyay S, et al, Antioxidant
Activity of Piper betel Leaf Extract and Its Constituents. Journal of Agricultural &
Food Chemistry, 54, 2006, 9046?-9054.
3. Patra A, Jha S, Murthy P.N, Phytochemical & pharmacological potential of
Hygrophila spinosa T. Anders, Pharmaconosy review 3(6), 2009, 330-341.
4. Gunther E, The Essential Oils, (Van Nostrand: New York), 5, 1952, 160-161.
5. Chaveerach A, Mokkamul P, Sudmoon R, Tanee T, Ethnobotany of genus Piper
(Piperaceae) in Thailand. Ethnobotany Research and Applications 4, 2006, 1547-
3465.
6. Dan N.V, Medicinal plants in Vietnam, WHO Regional Publication, Western
Pacific Series, 1990, 3.
7. Tobacco Habits Other than Smoking; Betel Quid and Areca-nut Chewing and Some
Related Nitrosamines. IARC Monographs on the Evaluation of the Carcinogenic
Risk of Chemicals to Humans, (IARC Press: Lyon) 1985, 37.
8. Sankaranarayanan R, Oral cancer in India: An epidemiological and clinical
review, Oral Surgery, Oral Medicine, Oral Pathology 69, 1990, 325–330.
9. Moore S R, Johnson N.W, Pierce A.M, and Wilson D.E., The epidemiology of mouth
cancer: a review of global incidence, Oral Diseases, 6, 2000, 65–74.
10. Avon S.L, Oral mucosal lesions associated with use of quid, Journal of the Canadian
Dental Association 70(4), 2004, 244–248.
11. Wiart C, Medicinal plants of Asia & Pacific, (Pub-CRC Press) Taylor & Francis
Group, 2006, 25-26.
12. Chopra R.N, Chopra I C, Indigenous Drugs of India. 2 nd Edition, (Pub- Academic
Publishers) 1958, 372.
13. Majumdar B, Ray Chaudhuri S.G, Ray A, Bandyopadhyay S.K, Effect of ethanol
extract of Piper betle Linn leaf on healing of NSAID-induced experimental ulcer: a
novel role of free radical scavenging action. Indian J. Exp. Biol,. 41, 2003, 311-315.
14. Verma A, Kumar N, Ranade S.A, Genetic diversity amongst landraces of a dioecious
vegetatively propagated plant, betelvine (Piper betle L.) J. Biosci,. 29(3) 2004, 319-
328.
15. Peungvicha P, Thirawarapan S.S, Temsirikkul R, Watanabe H, Prasain J.K, Kadota S,
Hypoglycaemic effect of the water extract of Piper sarmentosum in rats. Journal of
Ethnopharmacology, 60, 1998, 27–32.
16. Purohit A, Daradka H.M.M, Anti-diabetic efficacy of Piper longum fruit (50% etha-
nol extract) on alloxan induced diabetic rats, Journal of the Diabetic Association of
India, 38, 1999, 22–23.
17. Bour-Jr Wang, Yue-Liang Guo, How-Ran Guo, Ho-Yuan Chang, Piper betle L. inflo-
rescence causes allergic contact dermatitis of the hands during betle quid assembly.
Contact dermatitis 58, 2008, 368–370.
18. Yu-Ting Chung, Chiu-Lan Chen, Cheng-ChungWu, Shan-An Chan, Chin-Wen Chi,
Tsung-Yun Liu, Safrole-DNA adduct in hepatocellular carcinoma associated with
betel quid chewing, Toxicology Letters 183, 2008, 21-27.
19. Hwang L.S, Wang C.K, Shen M.J, Kao L.S, Phenolic compounds of Piper betle
flower as flavoring and neuronal activity modulating agents, Occurrence and Chem-
istry, American Chemical Society, Washington DC, 1992, 200–213.
20. Wang C.K, Hwang L.S, Phenolic compounds of betel quid chewing juice. Food
Sci,. 20, 1993, 458–471.
21. Gilani A.H, Aziz N, Khurram I.M, Rao Z.A, Ali N.K, The Presence of Cholinomimetic
and Calcium Channel Antagonist Constituents in Piper betle Linn. Phytotherapy
Research 14, 2000, 436–442.
22. Thanh Le, Dung N.X, Casanova J, Leclercq P.A, Combination of capillary GC, GC/
MS & 13 C-NMR for the characterization of the rhizome oil of Piper betle L. (Piperaceae)
from Vietnam. Spectroscopy 13, 1996/1997, 131-136.
23. Das P.C, Patent G.B, 1445599 760811, Chemical Abstracts 86, 1976, 21786.
24. Hwang L.C, Wang C.K, Sheu M.J, Kao L.S, ACS Symp. Ser. 1992, 506:200.
25. Sharma M.L, Rawat A.K.S, Balasubramanyam V.R, Singh A, Indian Perfumer 27(2),
1983, 91.
26. Sharma M.L, Rawat A.K.S, Balasubramanyam V.R, Indian Perfumer 31(4), 1987,
319.
27. Rawat A.K.S, Tripathi R.D, Khan A.J, Balasubramanyam V.R, Biochem. Syst. Ecol.
17(1), 1989, 35.
28. Balasubramanyam V.R, Rawat A.K.S, J.plant Crops, 18(2), 1990, 78.
29. Sothy N, Lo V.N, Dung N.X, Vietnamese J of Pharmacy 5, 1989, 8.
30. Parmar V.S, Jain S.C, Bisht K.S, Phytochemistry 46, 1997, 597-673.
31. Philip H.E, William S.B, Evangeline J.F, J.Agric. Food Chem. 32, 1984, 1254-
1256.
32. Usmanghani K, Saeed A, Alam M. T, Piper betle In, Indusyunic Medicine, (Univer-
sity of Karachi Press), Karachi. 1997, 340-341.
33. Evans P. H, Bowers W. S, Funk E. J, Identification of Fungicidal and Nematocidal
Components in the Leaves of Piper Betel (Piperaceae).Journal of Food Agricultural
and Chemistry 32, 1984, 1254-1256.

Journal of Pharmacy Research Vol.4.Issue 9. September 2011 2991-2997

 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
34. Bangar G.P, Rao R.E, Varma K.C, Antimicrobial activity of leaves and oil of Piper
betle. Ind. J. Pharm. 28, 1966, 327.
35. Ali S.M, Mehta R.K, Preliminary pharmacological and anthelmintic studies of the
essential oil of Piper betle,. Ind. J. Pharm. 32, 1970, 132.
36. The Wealth of India, The dictionary of Indian raw materials and industrial products.
Raw material, revised ed. New Delhi, (Publication and information directorate, CSIR),
1992, 5.
37. Mula S, Banerjee D, Patro B.S, Barik A, Bandopadhayay S.K, Chattopadhyay S,
Inhibitory property of the Piper betle phenolics against photosensitization induced
biological damages, Bioorganic and medicinal chemistry 16, 2008, 2932-2938.
38. Dubey P, Tripathi S.C, Z. Pflanzenkrankh. Pflanzenschutz 94(3), 1987, 235.
39. Santhanam G, Nagarajan S, Wound healing activity of Curcuma aromatica and Piper
betle, Fitoterapia 61, 1990, 458-459.
40. Chatterjee A, Pakrashi S.C, Treatise of Indian Medicinal Plants, (CSIR Publication
New Delhi), 1995, 26.
41. Deshpande S.M, Upadhyay R.R, Singh R.P, Chemical study of Piper betel leaves,
Curr. Sci. 39, 1970, 372.
42. Rawat,AKS, Tripathi RD, Khan AJ, Balasubrahmanyam V.R, Biochem. Syst. Ecol.
17(1), 1989, 35.
43. Prabhu M.S, Patel K, Saraawathi G, Srinivasan K, Effect of orally administered betel
leaf (Piper betel leaf Linn.) on digestive enzymes of pancreas and intestinal mucosa
and on bile production in rats, Indian J. Exp. Biol. 33, 1995, 752-756.
44. Guha P.,Betel Leaf: The Neglected Green Gold of India,. J. Hum. Ecol. 19(2),
2006, 87-93.
45. Sharma R.N, Contribution of Ayurveda towards management of bronchial asthma,
Sachitra Ayurveda 44, 1991, 349-352.
46. Ratnasooriya W.D, Jayawardena K.G.I, Premkumara G.A.S, Antimotility effect of Piper
betle (L) leaf extract on washed human spermatozoa, Journal of Natonal Science Council
of Srilanka, 18, 1990, 53-60.
47. Ratnasooriya W.D, Premakumara G.A.S, Piper betle leaves impairs masculine sexual
behavior of rats, Medical Science Research, 24, 1996, 303-306.
48. Ratnasooriya W.D, Premakumara G.A.S, Piper betle leaves reversibly inhibits fertil-
ity of male rats, Vidyodaya Journal of Science 7, 1997, 15-21.
49. Burkill I.H, A Dictionary of economic products of the Malay Peninsula, (Ministry of
Agriculture and Cooperatives, Kuala Lumpur, Malaysia), 1966.
50. Chandra T, Sadique J, Somasundaram S, Effect of Elipta alba on inflammation and liver
injury, Fitoterapia 58, 1987, 23-31.
51. Saravanan R, Prakasam A, Ramesh B, Pugalendi K.V, Influence of Piper betle on
hepatic marker enzymes and tissue antioxidant status in ethanol-treated wistar rats,
Journal of Medicinal Food, 5, 2002, 197–204.
52. Santhakumari P, Prakasam A, Pugalendi K.V, Modulation of oxidative stress param-
eters by treatment with Piper betle leaf in streptozotocin induced diabetic rats. In-
dian Journal of Pharmacology 35, 2003, 373-378.
53. Azuine M.A, Amonkar A.J, Bhide S.V, Chemopreventive efficacy of betel leaf extract
and its constituents on 7, 12-dimethyl benza(a)anthracene induced carcinogenesis
and their effect on drug detoxification system in mouse skin. Indian J Exp Biol 29,
1991, 346–51.
54. Bandyopadhyay S, Pal B, Bhattacharya Ray M, Roy K.C, Anti-leishmanial activity
of betel leaf extract, United States Patent Application Publication 2002, Pub. No.:
US2002/0150636 A1.
55. Bajpai V, Sharma D, Kumar B, and Madhusudanan K.P, Profiling of Piper betle Linn.
Cultivars by direct analysis in real time mass spectrometric technique, Biomedical
Chromatography published online 28 june 2010, DOI: 10.1002/bmc.1437.
56. Kanjwani D.G, Marathe T.P, Chiplunkar S.V, Sathaye S. S, Evaluation of
Immunomodulatory Activity of Methanolic Extract of Piper betel. Scandinavian
Journal of Immunology 67, 2008, 589–593.
57. Suppakul P, Ead N.S, Phoopuritham P, Antimicrobial and antioxidant activities
of betel oil. Nat. Sci. 40, 2006, 91-100.
58. Bhattacharya S, Subramanian M, Bauri A.K, Kamat J.P, Radioprotective property of
the ethanolic extract of Piper betel leaf. J. Radiat. Res. 46, 2005, 165-171.
59. Ganguly S, Mula S, Chattopadhyay S, Chatterjee M, An ethanol extract of Piper betle
Linn. mediates its anti-inflammatory activity via down-regulation of nitric oxide. J
Pharm Pharmacol. 59, 2007, 711–718.
60. Rathee J, Patro S, Birija S, Mula S, Gamre S, Chattopadhyay S, Journal of Agricultural
and food chemistry 2006, 54(24).
61. Arambewela L.S.R, Arawwawala L.D, Ratnasooriya W.P, Antidiabetic activities of
aqueous & ethanolic extracts of Piper betle leaves in rats Journal of Ethopharmacology
102, 2005, 239-245.
62. Row L.C.M, Ho J.C, The Antimicrobial Activity, Mosquito Larvicidal Activity,
Antioxidant Property and Tyrosinase Inhibition of Piper betle, J. Chin. Chem. Soc.
56 (3), 2009, 1-6.
63. Mohottalage S, Tabacchi R, Guerin P.M, Components from Srilankan Piper betle L.
leaf oil & their analogues showing toxicity against the housefly, Musca domestica.
Flavour & Fragr. J, 22, 2007, 130-138.
64. Lin M.H, Chou F.P, Huang H.P, Hsu J.D, Chou M.Y, Wang C.J, The tumor promoting
effect of lime-piper betel quid in JB6 cells, Food and chemical toxicology 41, 2003,
1463-1471.
65. Ramji N, Iyer R, Chandrasekaran S, Phenolic antibacterials from Piper betle in the
prevention of halitosis, Journal of Ethnopharmacology 23, 2002, 149-152.
Journal of Pharmacy Research Vol.4.Issue 9. September 2011
66. Bhattacharya S, Banerjee D, Bauri A.K, Chattopadhyay S, Bandyopadhyay S.K,
Healing property of the Piper betel phenol, allylpyrocatechol against indometha-
cin-induced stomach ulceration and mechanism of action World J Gastroenterol
13(27), 2007, 3705-3713.
67. Tasdemir D, Kaiser M, Brun R, Yardley V, Schmidt T.J, Tosun F, Ruedi P,
Antitrypanosomal and antileishmanial activities of flavonoids and their analogues:
in vitro, in vivo, structure activity relationship, and quantitative structure-activity
relationship studies, Antimicrob. Agents Chemother. 50, 2006, 1352–1364.
68. Sarkar A, Sen R, Saha P, Ganguly S, Mandal G, Chatterjee M, An ethanolic extract of
leaves of Piper betle (Paan) Linn mediates its antileishmanial activity via apoptosis.
Parasitol. Res 102, 2008, 1249-1255.
69. Luu H.V, Hoc T.C, Xuat Ban B.Y.T, Chemical chemical constituents of essential oil of
Piper nigrum L. and the essential oil of Piper betle L. in Nghe An province. 11, 2003,
15-17.
70. Hwang L.S, Wang C.K, Sheu M.J, Kao L.S, ACS Syp. Ser. (Phenol. Compd. Food Eff.
Health I) 506, 1992, 200-213 (Chem. Abstr. 1993; 118: 58461x).
71. Nalina T, Rahim Z.H.A, The crude aqueous extract of Piper betle L. & its antibacterial
effects towards Streptococcus mutans. American Journal of Biotechnology & Bio-
chemistry 3(1), 2007, 10-15.
72. Parmar V.S, Jain S.C, Gupta S, Sharma N.K, Wengel J, Polyphenols & alkaloids from
Piper species, Phytochemistry 49(4), 1998, 1069-1078.
73. Ghosh K, Bhattacharya T.K, Chemical constituents of Piper betle Linn. (Piperaceae)
roots. Molecules, 10, 2005, 798-802.
74. Lavania M, Chauhan P.S, Chauhan S.V.S, Singh H.B, Nautiyal C.S, Induction of Plant
defence enzymes and phenolics by treatment with plant growth promoting
rhizobacteria Serratia marcescens NBRI1213, Current microbiology 52, 2006, 363-
368.
75. Stohr J.R, Xiao P.G, Bauer R, Constituents of Chinese Piper species & their inhibi-
tory activity on prostaglandin & leukotriene biosynthesis in vitro, Journal of
Ethnopharmacology 75, 2001, 133-139.
76. Bhattacharya S, Mula S, Gamre S, Kamat J.P, Bandyopadhyay S.K, Chattopadhyay S,
Inhibitory property of Piper betel extract against photosensitization induced dam-
ages to lipids & proteins, Food Chemistry, 100, 2007, 1474-1480.
77. Chieng J.C, The essential oil composition of the leaves of five Piper species for taxo-
nomic purposes, ACGC Chemical Research Communication, 19, 2005, 6-12.
78. Lei D, Chan C.P, Wang T.M, Lin B.R, Chang M.C, Wang Y.J, Antioxidant & antiplatelet
effects of aqueous inflorescence Piper betle extracts, Agric. Food Chem. 51, 2003,
2083-2088.
79. Dorman H.J.D, Figueiredo A.C, Barroso J.G, Deans S.G, In vitro evaluation of anti-
oxidant activity of essential oils and their components, Flavour Fragr. J, 15, 12-16.
80. Sarkar M, Gangopadhyay P, Basak B, The reversible antifertility activity of Piper
betel Linn. on Swiss albino male mice, Contraception 62, 2000, 271–274.
81. Razak F.A, Othman R.Y, Rahim Z.H.A, The effect of Piper betle & Psidium guajava
extracts on the cell surface hydrophobicity of selected early settlers of dental plague,
Journal of Oral Science, 48(2), 2006, 71-75.
82. Mohamed S, Saka S, Sharkawy S.H, Ali A.M, Muid S, Antimycotic screening of 58
Malaysian plants against plant pathogens, Pestic. Sci. 47, 1996, 259-264.
83. Majumdar B, Chaudhari S.R, Ray A, Bandyopadhyay S.K, Potent antiulcerogenic
activity of ethanol of leaf of Piper betle Linn by antioxidative mechanism, Indian
Journal of Clinical Biochemistry, 17(1), 2002, 49-57.
84. Bhattacharya S, Chaudhuri S.R, Chattopadhyay S, Bandyopadhyay, Healing proper-
ties of some Indian medicinal plants against indomethacin-induced gastric ulcer-
ation of rats, J. Clin. Biochem. Nutr., 41, 2007, 106-114.
85. Vaghasiya Y, Nair R, Chanda S, Investigation of some piper Species for Antibacterial
and anti-inflammatory property. International journal of Pharmacology, 3(5), 2007,
400-405.
86. Nath V, Singh A.P, Landrace/gender-based differences in phenol and thiocyanate
contents and biological activity in Piper betle L, Current Science, 91(6), 2006, 746-
749.
87. Sawangjaroen N, Phongpaichit S, Subhadhirasakul S, Visutthi M, Srisuwan N,
Thammapalerd N, The anti amoebic activity of some medicinal plants used by AIDS
patients in southern Thailand, Parasito. Res. 98, 2006, 588-592.
88. Sawangjaroen N, Subhadhirasakul S, Phongpaichit S, Siripanth C, Jamjaroen K,
Sawangjaroen K, The in vitro anti-giardial activity of extracts from plants that are
used for self-medication by AIDS patients in southern Thailand, Parasito. Res. 95,
2005, 17-21.
89. Dung T.L, Xuan N, Lu Van, Piet A, Chemical composition of the leaf oil from Piper
betle L., cultivated in Vietnam, Journal of Essential oil-Bearing Plants 5(1), 2002,
38-42.
90. Hwang L.C, Wang, C.K, Sheu M.J, Kao L.S, ACS Symp. Ser., 5(6), 1992, 200.
91. Kumarasinghe S.P.W, Karunaweera N.D, Arambewela L.S.R, Ihalamulla R.L, Larvi-
cidal effects of mineral turpentine, low aromatic white spirits, aqueous extracts of
Cassia alata & aqueous extracts, ethanolic extracts & essential oil of betel leaf (Piper
Betle) on Chrysomya megacephala. International Journal of Dermatology, 41, 2002,
877-880.
92. Chen S.J, Wu B.N, Yeh J.L, Lo Y.C, Chen I.S, Chen I.J, C-fibre-evoked autonomic
cardiovascular effects after injection of Piper betle inflorescence extracts, J. of
Ethnopharmacology, 45, 1995, 183-188.
93. Leopold J, Christiane P, Gerhard B, Wilhelm F, Muhammed A.M, Analysis of the
2991-2997
 Kushagra Nagori et al. / Journal of Pharmacy Research 2011,4(9),2991-2997
essential oil of Piper betle leaves from South-India using GC/FID, GC/MS and olfac-
tometry, Sci. Pharm. 67(4), 1999, 305-311.
94. Chattopadhyay I, Kaushik B, Bandyopadhyay U, Banerjee R.K, Turmeric and
curcumin: Biological actions and medicinal applications, Current Science 87(1),
2004, 44-53.
95. Cody R.B, Laramee J.A, Nilles J.M, Durst H.D, Direct Analysis in real time (DART)
mass spectrometry, JEOL News 40(1), 2005, 8-12.
96. Cody R.B, Laramee J.A, Durst H.D, Anal. Chem, 77(8), 2005, 2297-2302.
97. Haefliger O.P, and Jeckelmann N, Direct mass spectrometric analysis of flavors and
fragrances in real applications using DART, Rapid Communications in Mass Spec-
trometry, 21(8), 2007, 1361-1366.
98. Madhusudanan K.P, Banerjee S, Suman P.S, Khanuja S.P.S, Chattopadhay S.K, Analy-
sis of hairy root culture of Rauvolfia serpentina using direct analysis in real-time
mass spectrometric technique, Biomedical Chromatography 22, 2008, 596–600.
99. Kim H.J, Jang Y.P, Direct analysis of curcumin in turmeric by DART-MS, Phytochemi-
cal Analysis, 20, 2009, 372–377.
100 .Nyadong L, Hampton C, Leung H, Newton P, Green M, de Jesus V, Fernandez F.M,
Presented at the 54 th meeting of the American society for Mass Spectrometry, Seattle,
WA, May 28 –June 1, 2006.
101 .Fernandez F.M, Cody R.B, Green M.D, Hampton C.Y, McGready R, Sengaloundeth
S, White N.J, Newton P.N, Chem. Med. Chem., 1, 2006, 702-705.
102 .Marcus A, New A, Presented at 54th meeting of the American society for Mass Spec-
trometry Seattle, WA, May 28 –June 1, 2006, poster.
103 .Wu J.T, Musselman B, Presented at the 54th meeting of the American Society of Mass
Spectrometry Seattle, WA, May 28 –June 1, 2006.
104 .Gomez M, Presented at the 54th meeting of the American Society of Mass Spectrom-
etry Seattle, WA, May 28 –June 1, 2006.
105 .Lochansky M.I, Gomez M.A, Williams J.D, Johnson R.L, Miller L.A, Presented at
54 th meeting of the American society for Mass Spectrometry Seattle, WA, May 28 –
June 1, 2006, poster.
106 .Pierce C.Y, Barr J.R, Cody R.B, Massung R.F, Woolfitt A.R, Moura H, Thompson
H.A, Fernandez F.M, Chem Commun,. 8, 2007, 807-809.

Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.4.Issue 9. September 2011 2991-2997

View publication stats