Asian Pacific Journal of Tropical Medicine (2011)649-653 649

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Asian Pacific Journal of Tropical Medicine

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Document heading doi:

Phytochemical and pharmacological evaluation of prop roots of

Pandanus fascicularis Lam
Jothimani Rajeswari1*, Karthikeyan Kesavan2, Balasundaram Jayakar3
1
Srinivas College of Pharmacy, Srinivas Group of College, Mangalore, Karnataka, India
2
Department of Pharmaceutics, Institute of Technology, Banaras Hindu University, Varanasi, U.P, India
3
Vinayaka Mission’s College of Pharmacy, Salem, Tamil Nadu, India

ARTICLE INFO ABSTRACT

Article history: Objective: To evaluate the anti-inflammatory and analgesic activities of the ethanol and aqueous
Received 22 April 2011 extracts of prop roots of Pandanus fascicularis (P. fascicularis) Lam (pandanaceae). And provide
Received in revised form 30 June 2011
experimental evidence for its traditional use such as rheumatoid arthritis and spasmodic.
Accepted 15 July 2011
Methods: The anti-inflammatory activity was observed by carrageenan-induced edema of the
Available online 20 August 2011
hind paw of rats. Analgesic activities of prop roots of P. fascicularis were determined using acetic
acid induced writhing model and tail clip method in mice and rat, respectively. The ethanol
Keywords: fraction was then subjected to chromatographic analysis and a compound has been isolated
Pandanus fascicularis Lam and characterized by IR, 1H-NMR and mass spectroscopy. Results: Edema suppressant effect
Anti-inflammation of ethanol extract was found to be 37.03% inhibition whereas aqueous extract was found to be
Analgesic 63.22% inhibition after 3 h which was nearly equivalent to that of 10 mg/kg of indomethacin
Aqueous extract (67.81%). Percentage inhibition of writhing compared to control were 63.15%, 54.38%, 14.90% for
Ethanol extract aspirin, aqueous extract and ethanolic extract, respectively. Both ethanol and aqueous extracts
Writhing show significant activity against appropriate controls after 60 min of treatment on tail clip method.
Tail clip The structure of the isolated compound is may be characterized as Hepta deca-5-ene-1-ol by
analysis it’s IR, 1H-NMR and mass spectroscopy data. Conclusions: The extracts of prop roots
of P. fascicularis produce significant analgesic and anti-inflammatory activities, supporting the
traditional application of this herb in treating various diseases associated with inflammation and
pain.

in virtually all human and animal diseases.

1. Introduction Drugs which are in use presently for the management of
pain and inflammatory conditions are either narcotics or
The importance of medicinal plants in traditional non-narcotics (NSAIDS) and present well known side and
healthcare practices, providing clues to new areas of toxic effects[3,4]. On the contrary herbal medicines with good
research and in biodiversity conservation is now well absorption, less toxicity and easy availability have been
recognized. The use of traditional medicine and medicinal used since long[5]. It is therefore essential that efforts should
plants in most developing countries, as a normative basis for be made to introduce new medicinal plants to develop
the maintenance of good health has been widely observed. cheaper and effective drugs[6]. Plants represent still a large
Inflammation is the response of living tissues to injury. It untapped source of structurally novel compounds that might
involves a complex array of enzyme activation, mediator serve as lead for the development of novel drug[7].
release, extravasations of fluid, cell migration, tissue Pandanus fascicularis (P. fascicularis) Lam (syn. P.
breakdown and repair[1,2]. Inflammation has become the odoratissimus) commonly referred to as screw pines are
focus of global scientific research because of its implication palm-like evergreen trees or shrubs belong to the genus
Pandanus, order Pandanales, class Liliopsida, and division
Mangoliophyta. Pandanus comprises 500-600 species and
*Corresponding author: J. Rajeswari, Assistant Professor, Srinivas College of is distributed mainly in subtropical and tropical regions.
pharmacy Valachil, Parengipete, Mangalore, Karnataka, India.
Tel: +91 9743796028 P. fascicularis is native to South Asia and has a significant
E-mail: [email protected]; [email protected]
650 Jothimani Rajeswari et al./Asian Pacific Journal of Tropical Medicine (2011)649-653

presence particularly in mangrove swamps[8]. Individual 10 mL/kg by oral route was compared with single dose (5 g/
plants can reach a height of 20 meters supported by aerial kg; po.) of aqueous and ethanolic extracts of P. fascicularis.
roots. Patients and medical practitioners believe the root Access to food and water, toxic symptoms and the general
and rhizome to be effective against diabetes. The decoction behavior of mice were observed continuously for 1 h after
of the P. odorus root and rhizome has been traditionally used the treatment, intermittently for 4 h, and thereafter over a
in treating diabetic patients without much specific evidence period of 24 h. The mice were further observed for up to 14 d
and also this plant used for rheumatism and spasmodic. The following treatment for any signs of toxicity and mortality[10].
present study was deals with evaluation of anti inflammatory
and analgesic activities of the ethanol and aqueous extracts 2.5. Anti-inflammatory activity
of prop roots of P. fascicularis and isolation and structural
elucidation of ethanol extract. Twenty four albino rat each weighing 150-175 g were
used in the experiment. The animals were divided into four
groups (栺-桇) each of six animals: group (栺) received of the
2. Materials and methods negative control (caroxymethylcellulose), group (栻) received
of positive control (indomethacin 10 mg/kg) and group 栿and
2.1. Chemicals 桇 received the aqueous and ethanolic extracts at 250 mg/
kg body weight, respectively. The anti-inflammatory activity
Indomethacin was obtained from (Micro labs, Bangalore), was evaluated by the carrageenan-induced paw edema
pentazocin and aspirin were the kind gifts by APL research test in rats. The percentage of the anti-inflammatory effect
centre (Hyderabad, India). Acetic acid was purchased of ethanol and aqueous extracts were calculated by using
from Merck (Mumbai, India). Carrageenan was obtained Formula 1.
from Sigma-Aldrich Pvt. Ltd (New Delhi, India). All other
reagents were of analytical grade.
C-T 伊100
Percentage activity = (Formula 1)
2.2. Preparation of extracts C

Prop roots of P. fascicularis were dried in shade and (T: Increase paw volume after ethanol and aqueous extract
powdered. The aqueous extract was prepared by cold was administered; C: Increase paw volume of control group)
maceration. The powder was soaked in equal amount of
distilled water and stirred intermittently and then left 2.6. Analgesic activity
overnight. The macerated pulp was then filtered through a
coarse sieve and the filtrate was dried at reduced pressure 2.6.1. Writhing method
in the rotor evaporator (Buchi Rotavapor R-114) and finally Twenty four albino mice each weighing 25-30 g were
freeze dried. Ethanolic extract was prepared by extracted used in the experiment. The animals were divided into four
with ethanol (95% v/v) in a soxhlet apparatus. The extract groups (栺-桇) each of six animals: group (栺) received of the
was evaporated to dryness under vacuum and dried in negative control (caroxymethyl cellulose), group (栻) received
vacuum desiccators. of positive control received pentazocine 5 mg/kg and group
栿 and 桇 received the aqueous and ethanolic extracts at
2.3. Preliminary phytochemical screening 250 mg/kg body weight, respectively. Thirty minutes after
treatment, the mice were given an intraperitoneal (ip)
The presence of various phytochemical constituents in the injection of 3% v/v acetic acid in a volume of 2 mL/kg to
extract was determined using standard screening tests[9]. induce the characteristic writhing. The number of writhes
Male albino rats (150-175 g) of Wistar strain and albino occurring between 5 and 15 min after acetic acid injection
mice (25-30 g) were obtained from the Perundurai Medical was recorded. The response of the extracts treated animals
College, Perundurai. Before and during the experiment were compared with that of control [11]. The percentage
rats were housed in polypropylene cages lined with husk in analgesic activity was calculated from Formula 2.
standard environmental conditions (temperature (25依2) 曟,
relative humidity 55%依10% and 12: 12 light: dark cycle).
The rats were fed on a standard pellet diet ad libium and Nc-Nt
Percentage protection = 伊100 (Formula 2)
had free access to water. The experiments were performed Nc
after approval of the protocol by the Institutional Animal
Ethics committee (IAEC) and were carried out in accordance (Nc is the average number of stretches of the control group;
with the current guidelines for the care of laboratory Nt is the average number of stretches of the test drug group)
animals.
2.6.2. Tail clip method
2.4. Acute toxicity studies Twenty four albino rat each weighing 150-175 g were used
in the experiment. The selected animals were divided into
The animals were randomly divided into three groups four groups (栺-桇) each of six groups: group (栺) received
(n=6). A control group having carboxymethylcellulose of the negative control (caroxymethyl cellulose), group (栻
 Jothimani Rajeswari et al./Asian Pacific Journal of Tropical Medicine (2011)649-653
651
) received of positive control received pentazocine 5 mg/ GC-Mass spectrometer spectra was recorded in SHIMADZU
kg and group 栿 and 桇 received the aqueous and ethanolic QP 50000.
extracts at 250 mg/kg body weight, respectively. A metal
artery clip was applied to the root of the mouse tail to induce 2.8. Statistical analysis
pain. A sensitivity test was carried out and animals that
did not attempt to dislodge the clip within 15 seconds were Data were presented as mean 依 SEM. Statistical differences
discarded. Analgesic activity was evaluated 0, 60, 120, 180 min between the control and treated groups were tested by one-
after oral administration of the extracts and controls. A tail way ANOVA followed by Student’s two-tailed unpaired
clip was applied and a positive analgesic response was t-test. The differences were considered to be significant at
indicated if there was no attempt to dislodge the clip within P<0.05.
5 s in any of the four consecutive trials after a time period of
2 min. The mean value was evaluated[12].
3. Results
2.7. Fractionation, isolation, purification and characterization
of compounds from the ethanol extract 3.1. Preliminary phytochemical screening

Chromatographic techniques (thin layer chromatography, Preliminary phytochemical screening revealed the
column chromatography) were used for the isolation of presence of carbohydrates, proteins, aminoacids, saponins,
compounds from the fractions. The column chromatographic tannins, phenolic compounds, alkalodies and flavonoids.
technique most commonly used for the separation of
compounds into several fractions according to the affinity or 3.2. Acute oral toxicity
solvating capacity of the compounds to the solvent used. The
study involves in fractionation and isolation of compounds The extracts of P. fascicularis were safe up to a dose of
from pharmacologically active ethanol extract. The structure 5 g/kg (po.) body weight. Over the study duration of 14 d,
of the compounds were tried to establish by spectroscopic there were no deaths recorded in the groups of mice given
methods. aqueous and ethanolic extracts of P. fascicularis. During
the observation period, P. fascicularis administration did
2.7.1. Study design not induce any variations in the general appearance or toxic
In order to carry out column chromatography, a solvent signs in the animals.
system was established by developing TLC technique. The
silica gel (100-200 mesh size) slurry was made with the 3.3. Effects of P. fascicularis on carrageenan-induced paw
solvent system established earlier. The slurry was poured edema in rats
time to time into the column very carefully and the silica gel
was allowed to settle down to from a uniform packing. Then The effect of ethanol and aqueous extract of P. fascicularis
the stop-cock of the column was opened and the excess of on carrageenan induced edema in rats is shown in Table 1.
solvent over the column head was allowed to run. The dry Edema suppressant effect of ethanol extract was found to
crude ethanol extract was mixed with small amount of silica be 37.03% inhibition whereas aqueous extract was found to
gel in a mortar to get a free flowing powder. The powdered be 63.22% inhibition after 3h which was nearly equivalent
sample was then applied carefully on the top of the prepared to that of 10 mg/kg of indomethacin (67.81%). The edema
column and successfully eluted with solvent/solvent system. suppressant effect was significant in the dose of 250 mg/
Elutes were collected in a number of conical flasks marked kg of ethanol (P<0.05) and aqueous extracts (P<0.01). P.
from fractions 1-100. Elutes were spotted successfully on fascicularis showed inhibitory effect on carrageenan induced
TLC plate and the flasks having similar spots were combined paw edema thus, exhibiting anti-inflammatory effect against
together. acute inflammation.

2.7.2. Analysis of fraction F2 3.4. Effects of P. fascicularis on acetic acid-induced writhing

The fraction F2 containing 20-36 conical flasks having reflex in mice
similar spots on TLC plate were combined. Then the
fractions were subjected to TLC by using petroleum ether: The analgesic activity of P. fascicularis was evaluated by
benzene (50: 50) as a solvent system. The expected bands using the writhing test. Oral administration both aqueous
were separated off and eluted with petroleum ether 100%, (P<0.01) and ethanolic (P<0.05) extracts of P. fascicularis
petroleum ether: benzene (50: 50), yield of 150 mg obtained. (250 mg/kg), 30 min before the acid injection, produced a
The compound was obtained as dull white sticky amorphous significant inhibition of acetic acid-induced abdominal
waxy solid. The fraction was characterized by spectroscopy constrictions in mice (Table 1). Aspirin (300 mg/kg), a
techniques like Perkin-Elmer Vector 22 model FT-IR standard NSAID used as positive control, also produced
Spectrophotometer (Nujol),1H-NMR spectra were recorded in significant inhibition of acetic acid-induced writhing
a BRUKER DPX-200 MHz using TMS as internal standard and response ( P < 0 . 01 ). Percentage inhibition of writhing
652 Jothimani Rajeswari et al./Asian Pacific Journal of Tropical Medicine (2011)649-653

Table 1
Effect of ethanol and aqueous extracts of P. fascicularis Lam, on carrageenan induced rat paw edema and acetic acid induced writhing in mice.
Sl. no Design of treatment Dose (mg/kg) Increase in paw edema at the end of 3 h No. of writhing for 5 min
1 Control - 0.87依0.03 57.00依2.24
2 Indomethacin 10 0.28依0.01** -
3 Aspirin 300 - 21.00依1.12**
4 Ethanolic extract 250 0.60依0.01* 48.50依2.10*
5 Aqueous extract 250 0.32依0.02** 26.00依1.76**
P<0.05, **P<0.01 compared to control (n=6); Students t-test.
*

Table 2
Effect of ethanol and aqueous extracts of P. fascicularis Lam, on rat by tail clip method.
Pain reaction
Sl. no Design of treatment Dose (mg/kg)
0 min 60 min 120 min 180 min
1 Control - 3.00依0.10 3.20依0.16 3.50依0.18 3.50依0.13
2 Pentazocine 5 4.00依0.24 14.00依1.74* 9.50依1.42* 7.90依0.84*
3 Ethanolic extract 250 3.20依0.12 8.50依0.84 *
8.00依0.74 6.40依1.10**
4 Aqueous extract 250 5.00依0.16 13.50依1.24* 9.00依1.17 7.40依0.62*
P<0.05, **P<0.01 compared to control (n=6); Students t-test.
*

compared to control were 63.15%, 54.38%, 14.9% for aspirin, and 4.44. The spectrum shows δ value 2.01 for terminal
aqueous extract and ethanolic extract, respectively. methyl group, δ: 3.3 for six protons due to three methylene
groups directly attached of the OH group resonated at
3.5. Effects of P. fascicularis on tail clip method in rat δ: 2.25 as a broad singlet. All other methylene protons
resonated at δ: 1.25 and it also indicates the presence of
The analgesic effect of ethanol and aqueous extracts of hydroxyl protons. This spectrum also indicates the presence
P. fascicularis was evidently effective by giving the dose of long chain aliphatic nature of the compound.
of 250 mg/kg body weight and their activity exhibited was Mass spectrum of the compound shows the following
roughly equivalent to the standard drug of pentazocine fragmentation pattern: 256 (M+), 236, 201, 183, 167, 153, 137,
5 mg/kg (Table 2). Both ethanol and aqueous extracts show 153, 97, 83, 69, 61 and 43. The mass spectrum showed it to
significant activity (P<0.01) against appropriate controls after be a straight chain compound (Base peak at m/z 43). The
60 min of treatment. Between these two extracts, aqueous values indicate the presence of methylene groups and long
extracts shows maximum activity when compared to ethanol chain nature of the compound. Formation of fragments at m/
extract. z 83 and 167 confirm the position of double bond at C-5.
On the basis of the above data the isolated compound may
3.6. Isolation and identification of the active compound be characterized as Hepta deca-5-ene-1-ol. Structure: H3C
- (CH2)10 CH=CH (CH2)3 CH2OH. Molecular formula: C17H34O.
The ethanol extract has been selected for isolation of Molecular weight: 254.
the available active constituents, which has the polarity
in between the acetone and aqueous. The purity of
isolated compound was checked by TLC using different 4. Discussion
solvent system. Rf value of the isolated compound 0.62
(benzene: methanol = 90 : 10 ). The isolated compound The anti-inflammatory and analgesic effects of the ethanol
was characterized by its physical, chemical as well as and aqueous extracts of P. fascicularis were investigated
spectrometric analysis. It is dull white sticky amorphous in this study. Inflammation has different phases the first
waxy crystalline compound soluble in pet ether, benzene phase is caused by an increase in vascular permeability,
and chloroform. The melting point is 58-60 曟 (uncorrected). the second one by infiltrate of leucocytes and the third one
IR data of the isolated compound shows the intense peaks at by granuloma formation. We determined anti-inflammatory
the following frequency: 3 784 cm-1, 3 340 cm-1, 2 919 cm-1, activity by using inhibition of carrageenan- induced
2 852 cm , 1 667 cm , 1 621 cm , 1 457cm , 1 374 cm ,
-1 -1 -1 -1 -1
inflammation which is one of the most feasible methods
1 052 cm , 923 cm , 852 cm and 722 cm . The peaks at
-1 -1 -1 -1
to screen anti-inflammatory agents. The development of
2 919 cm , 282 cm and 1 457 cm show the paraffinic
-1 -1 -1
carrageenan-induced edema is bi-phasic the first phase
nature of the compound. Peak at 3 340 cm-1 indicates the is attributed to the release of histamine, serotonin and
presence of hydroxyl group and peaks at 1 667 cm-1 shows kinins and the second phase is related to the release of
the presence of double bond. Band at 722 cm-1 shows its prostaglandins and bradykinins[13].
long chain nature. We observed that aqueous extract of P. fascicularis
1
H-NMR Spectrum of the isolated compound shows the δ at the given dose possess significant inhibition against
values at the following ppm: 0. 87, 1.25, 2.01, 2.52, 3.37, 3.75 carrageenan-induced paw edema in rats whereas ethanol
 Jothimani Rajeswari et al./Asian Pacific Journal of Tropical Medicine (2011)649-653
653

extract exhibited feeble effect in the treatment of acute

inflammation and pain. This response tendency of the Acknowledgements
extract in carrageenan-induced paw edema revealed good
peripheral anti-inflammatory properties of the extract. The The authors wish to acknowledge Mr. G.V.S. Murthy, Joint
anti inflammatory effect of aqueous extract may be due to Di-rector, Scientist, C-I/C, Botanical survey of India, Tamil
the presence of flavonoids. Flavonoids are known to inhibit Nadu Agricultural University Campus, Coimbatore for
the enzyme prostaglandin synthetase, more specifically the identification and authentication of P. Fascicularis.
endoperoxidase and reported to produce anti-inflammatory
effects. Since, prostaglandins are also involved in the
pain perception, inhibition of their synthesis might be the References
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Conflict of interest statement 2004; 92: 317-324.

We declare that we have no conflict of interest.