ArticlePDF Available

Antioxidant activity of methanol extract of pandanus fascicularis Lam

Authors:

Abstract

Plants are widely used in traditional and folklore medicine. Use of medicinal plants and their products are almost doubled over the last decade in developing countries and the present trend of wide spread interest in alternative therapies is well known. Pandanus fascicularis is used in traditional systems of medicine to treat varying conditions like rheumatism, fever, headache, earache and also used as antispasmodic. No evidence of scientific study is available on this plant. So the present study is aimed at investigating the invitro antioxidant activity of methanolic extract of Pandanus fascicularis. Present study is aimed at investigating the invitro antioxidant activity of methanolic extracts of leaves of Pandanus fascicularis(MEPF) by four different invitro methods. The lipid peroxidation was assayed by estimating the thiobarbituric acid reactive substances (TBARS) in different concentrations of MEPF on normal rat liver homogenates. The reduced glutathione (GSH) was assayed in liver homogenates of different concentrations of MEPF using the method of Ellman et.al. The nitric oxide (NO) scavenging activity and 1-1 Diphenyl,2-Picryl hydrazyl (DPPH) radical scavenging activity was measured using the methods of Sreejayan et.al and Shimada et.al. respectively using spectrophotometer. Vitamin E and normal saline were used as reference standard and control for all four invitro antioxidant measurement assays. The results showed significant antioxidant activity of MEPF in all four in vitro methods used in this study and the IC50,(the half maximal inhibitory concentration, of an inhibitor that is required for 50% inhibition of antioxidant activity) of MEPF was comparable to that of vitamin E, the reference standard compound used in this study. It is concluded that the methanolic extract of leaves of Pandanus fascicularis has significant antioxidant activity.
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
833
ATIOXIDAT ACTIVITY OF METHAOL EXTRACT OF
PADAUS FASCICULARIS Lam.
Sanjeeva
1
, Raveendra Kumar N
1
, Padmalaxmi D
1
, Ananthbabu
1
,
*
Udupa A.L.
2
,
Nkem Ojeh
2
, Subir Gupta
2
, Rathnakar U. P
3
, Vijayalaxmi Patil
3
, Prabhath Kodancha G
3
, Deepa
Benegal
4
,Adarsh Benegal
4
, Sanjana R
Rao
5
, Nisarga S
5
,Shubha H.V
6
.
1
Shanthiram Medical College Nandyal , Andhra Pradesh, India.
2
FMS, UWI, Cavehill, Barbados. West Indies. BB11000
3
Kasturba Medical College, Mangalore-575001, Manipal University, Karnataka, India.
4
Vijayanagar Institute of Medical Sciences Bellary, Karnataka,India
5
Yennapoya Medical College,Mangalore-575018,Karnataka,India.
6
JSS Medical College Mysore,Karnataka,India.
* Corresponding author: Dr.A.L.Udupa,
Faculty of Medical Sciences, University of West Indies, P.B. # 64
Cave Hill Campus, University Drive, Barbados.BB 11000.
Phone:(+1246) 417 4690 (O); Fax: 438 9170
Email: aludupa2002@yahoo.com
Summary
Plants are widely used in traditional and folklore medicine. Use of medicinal plants and their
products are almost doubled over the last decade in developing countries and the present trend of
wide spread interest in alternative therapies is well known. Pandanus fascicularis is used in
traditional systems of medicine to treat varying conditions like rheumatism, fever, headache,
earache and also used as antispasmodic. No evidence of scientific study is available on this plant.
So the present study is aimed at investigating the invitro antioxidant activity of methanolic
extract of Pandanus fascicularis. Present study is aimed at investigating the invitro antioxidant
activity of methanolic extracts of leaves of Pandanus fascicularis(MEPF) by four different in-
vitro methods. The lipid peroxidation was assayed by estimating the thiobarbituric acid reactive
substances (TBARS) in different concentrations of MEPF on normal rat liver homogenates. The
reduced glutathione (GSH) was assayed in liver homogenates of different concentrations of
MEPF using the method of Ellman et.al. The nitric oxide (NO) scavenging activity and 1-1
Diphenyl,2-Picryl hydrazyl (DPPH) radical scavenging activity was measured using the methods
of Sreejayan et.al and Shimada et.al. respectively using spectrophotometer. Vitamin E and
normal saline were used as reference standard and control for all four invitro antioxidant
measurement assays. The results showed significant antioxidant activity of MEPF in all four in
vitro methods used in this study and the IC50,(the half maximal inhibitory concentration,
of an inhibitor that is required for 50% inhibition of antioxidant activity) of MEPF was
comparable to that of vitamin E, the reference standard compound used in this study.
It is concluded that the methanolic extract of leaves of Pandanus fascicularis has significant
antioxidant activity.
Key words: Pandanus fascicularis, antioxidant action, lipidperoxidation, reduced glutathione
nitric oxide scavenging ,diphenyl picryl hydrazyl radical scavenging.
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
834
Introduction
Plants are widely used in various traditional and folklore systems of medicine. Consumption of
medicinal plants and their products has almost doubled over the last decade in developed
countries. At present the trend of wide spread interest in alternative therapies, is well known.
With the recent success of many plant derived drugs such as anticancer agent taxel and it’s
derivatives from Taxus baccata and Taxus brevifolia and antimalarial, artemisinin from the
Chinese wormwood Artemisia annua, the interest is growing.
The development of science and phytochemistry rejuvenated the hopes for remedies in
chronic diseases and this has generated new enthusiasm in the research work to develop herbal
medicine. WHO (World Health Organization) estimated that 80% of the population in
developing countries still relies on plant-based medicine for preliminary healthcare. Now efforts
are being made to develop herbal medicines in research institutes. Although India has the
tradition of alternative therapies like Ayurveda, Siddha and Unani, there are no procedures to test
the safety and efficacy of traditional remedies and to standardize their effective cure. For these
reasons we should increase our efforts in the area of medicinal plant research and should exploit
efficiently for the benefit of humanity.
Pandanus fascicularis Lam. (Synonyms: Pandanus tectorius, Pandanus odoratissimus Roxb,
Family: Pandanaceae). Vernacular names
1, 2
: Sanskrit- ketaki, Hindi-Kura, Kewda, Ketki,
Gagandhul, Kannada-Tale mara, English-Screw pine) is distributed widely in coastal regions of
Indian subcontinent and Andaman Islands. The plant is a branched palm like shrub, up to 1-3 m
high, rarely erect, stem supported by aerial roots, leaves glaucous-green, 0.9-1.5m, ensiform,
long lanceolate, acuminate with three rows of prickles each on the margins and on midrib
beneath, spinescent. Male flowers in spikes enclosed in large, white fragrant spathes, female
flowers in solitary spadix. Syncarpium yellow or red, drups numerous, each consisting of 5-12
carpels; each carpel 5-12.5 cm long, turbinate and angular20. Propagation is by seeds and
vegetative method. Leaves, flowers, roots, fruits, spadices, bracts are used in leprosy, smallpox,
syphilis, scabies, pain, heat of body, diseases of heart and brain and leucoderma
21
. The tender
floral leaves are used to flavour cream, rice (giving a flavour similar to basmati rice) sherbets,
jellies and sometimes curries
2, 3
. Oil from bracts is used for headache and
rheumatism and considered as stimulant and anti spasmodic
3
. Kewda attar or water prepared by
distillation of spadices is used to flavour sweets, syrups and soft drinks
2
.The flower is pungent,
bitter; improves complexion. The anthers are useful in pruritis, earache, headache, leucoderma,
eruptions, and diseases of the blood. Fruit is useful in relieving “vata”, “kapha” and urinary
discharge and is beneficial in leprosy
3
. A medicinal oil is prepared from the roots is considered
as diuretic, depurative and tonic
3
. Juice obtained from inflorescence is used for rheumatic
arthritis in veterinary medicine
4
. Kewda oil is used in ear ache, head ache, arthritis, debility,
depurative, giddiness, laxative, leprosy, rheumatism, small pox and spasms
5
.
The principle constituent of kewda oil responsible for the characteristic aroma of spadices is b-
phenyl ethyl methyl ether. 2-acetyl –1 pyrroline is a major volatile component in the tender floral
leaves or spathes. Blossoms yield 0.1-0.3% essential oil called kewda oil containing benzyl
benzoate, benzyl salicylate, benzyl acetate, benzyl alcohol, geraniol, linalool, linalyl acetate,
bromostyrene, guaiacol, phenyl ethyl alcohol, and aldehydes. Cirsilineol, n-triacontanol, β-
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
835
sitosterol, β-sitostenone, stigmast-4-ene-3,6dione stigmasterol, campesterol, daucosterol, and
palmitic acid, stearic acid isolated from rhizomes
2,6,7,8
Inflammatory diseases including different types of rheumatic diseases are very common
throughout the world. The greatest disadvantage in the potent synthetic drugs available at present
lies in their side effects, toxicity and reappearance of symptoms after discontinuation. Hence
search for new anti- rheumatic agents that retain the therapeutic efficacy and devoid of adverse
effects are justified.
So Pandanus fascicularis, a plant that was traditionally used for rheumatism with no scientific
proof claimed as yet on its leaf is investigated in this work. Inflammatory state is
associated with free radical formation and cell damage. The present study has been undertaken to
investigate and evaluate methanolic extract of leaves of Pandanus fascicularis for its free radical
scavenging potential. In this study after preliminary phytochemical screening of the extract, its in
vitro antioxidant activity were tested out using lipid peroxidation, reduced glutathione assay,
nitric oxide and DPPH scavenging activity and compared with that of vitamin E the standard
antioxidant compound.
Materials and Methods
Leaves of Pandanus fascicularis used for the investigation were collected from the east coast
road, Chennai-96, in the month of April 2004. The plant was identified and authenticated by
research officer (Pharmacognosy), Central Research Institute (Siddha), Arumbakkam, Chennai-
600106.
Preparation of methanol extract :
The leaves of Pandanus fascicularis were collected, coarsely powdered and was
successively extracted with methanol using Soxhlet extractor. The methanol extract of Pandanus
fascicularis (MEPF) was dried under reduced pressure using a rotary flash evaporator and it was
kept in the refrigeration. The percentage of yield of methanolic extract was 9%.
The methanol extract thus obtained was used for the preliminary phytochemical screening
and pharmacological studies such as for in-vitro antioxidant activity. The liver homogenate (3ml)
of normal rats was used for invitro lipid peroxidation(LPO) and reduced glutathione(GSH)
activity of MEPF after dissolving it in 3ml normal saline in varying concentrations (25g-800
g/ml). Lipid peroxidation (LPO) was assayed by estimating thiobarbituric acid reactive
substances(TABRS) and reduced glutathione (GSH) was assayed in the liver homogenates of
normal rats with different concentrations of methanol extract of Pandanus fascicularis
(MEPF).The experimental protocol was approved by the Institutional Animal Ethics Committee
(IAEC) , Reference No: IAEC-X-3 / CLBMCP / 2004-2005.
PRELIMIARY PHYTOCHEMICAL SCREEIG
9,10
The methanol extract of Pandanus fascicularis leaves was subjected to preliminary
phytochemical screening. This extract was tested for the presence of alkaloids, carbohydrates,
proteins, steroids,sterols,phenols,flavonoids,tannins,gums.mucilage,glycosides,saponins and
terpenes
9,10.
.
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
836
I-VITRO ATI OXIDAT STUDIES:
Lipid peroxidation (LPO)
11
The degree of lipid peroxidation was assayed by estimating the thiobarbituric acid
reactive substances (TBARS). Different concentrations (25-800 µg/ml) of MEPF were added to
the normal rat liver homogenate(3ml). LPO was initiated by adding 100 µl 15 mM ferrous
sulphate solution to 3 ml of liver homogenate. After 30 min 0.5ml of this reaction mixture was
taken in a tube containing 1.5 ml of 10 % w/v trichloroacetic acid (TCA). After 10 min, tubes
were centrifuged and supernatant was separated and mixed with 1.5 ml of 0.67% thiobarbituric
acid (TBA) in 50% acetic acid. The mixture was heated in a boiling water bath at 85 °C for 30
min to complete the reaction. The pink coloured complex formed was measured at 535 nm in a
spectrophotometer. Vitamin-E was used as reference standard. The percentage inhibition of LPO
was calculated by comparing the results of the test with those of controls not treated with the
extracts as per following formula -Percentage inhibition = {(control-test) /(control)} X 100.
Reduced glutathione (GSH) assay
12
:
Liver homogenate(3ml) with different concentrations (25-800 µg/ml) of MEPF was
mixed with 0.5 ml of precipitating buffer (5% w/v TCA in 0.1 mM EDTA). The sample was
centrifuged at 2000 rpm for 10 min and the supernatant was mixed with 2.5 ml of 0.1 M
phosphate buffer (pH 8.0). The colour was developed by adding 100 µl of 0.01% DTNB.
Absorbance was noted at 412 nm using UV spectrophotometer. The percentage reduction was
calculated by comparing with control. Vitamin-E was used as reference standard.
itric oxide (O) scavenging activity
13
:
Nitric oxide scavenging activity was measured by using spectrophotometer. Sodium
nitroprusside (5 mM) in phosphate buffer saline was mixed with different concentrations of
MEPF (25-800 g/ml) dissolved in normal saline and incubated at 25°C for 30 min. A control
without test compound but with equivalent amount of sodium nitroprusside was taken. After 30
min 1.5 ml of the incubation solution was removed and diluted with 1.5 ml of Griess reagent (1%
sulphanilamide, 2% phosphoric acid, and 0.1% napthyl ethylene diamine dihydrochloride). The
absorbance of the chromophore formed during diazotization of the nitrite with sulphanilamide
and subsequent coupling with napthyl ethylene diamine dihydrochloric acid was measured at 546
nm. Vitamin-E was used as reference standard.
1-1 Diphenyl, 2-picryl hydrazyl (DPPH) radical scavenging activity
14
:
DPPH scavenging activity was measured by spectrophotometric method. 0.1 mM
solution of DPPH was prepared in ethanol and 1 ml of this solution was added to 3 ml of
MEPF in normal saline at different concentrations (25-800 g/ml). Equal amount of normal
saline was added to the control. The mixture was shaken well and incubated at room
temperature for 30 min. The absorbance was read at 517 nm using a spectrophotometer.
Vitamin-E was used as reference standard. All the assays were read at a particular nm using
spectrophotometer, UV –1601 Shimadzu
Statistical tests: In in-vitro experimental methods the percentage of inhibition and the IC50
value or the half maximal inhibitory concentration, representing the concentration of an
extract that is required to inhibit 50% of oxidant activity were calculated and compared with
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
837
that of the standard antioxidant compound vitamin E and the control(normal saline) used in
this experiment. Results were presented as mean ± SEM and the correlation coefficient denoted
as “r”, indicating the potency of the test compound as compared to the standard. The
antioxidant activity is considered as significant if the “r” value is >0.5. The p’ value is not
mentioned here (though it is p<0.001) since here it indicates the lesser activity than the
standard compound used.
Results
Preliminary phytochemical screening:
The results of preliminary phytochemical screening of the methanol extract of Pandanus
fascicularis leaves showed the presence of alkaloids, carbohydrates, phenols, steroids, sterols,
proteins and glycosides.
In vitro antioxidant methods:
Lipid peroxidation:
MEPF inhibited the ferrous sulphate induced lipid peroxidation in a concentration
dependent manner. The IC
50
value of MEPF was found to be 669.23 µg/ml(r= 0.61). The IC
50
value of vit-E was 411.3 µg/ml (Table - 1). The regression coeifficient “r” is the ratio between
vitamin E and MEPF indicating the potency of MEPF as compared to vitamin E.
Reduced glutathione assay.
MEPF inhibited the oxidation of reduced glutathione in a dose dependent manner. The
IC
50
values of MEPF was found to be 697.1µg/ml (r= 0.59). The IC
50
value of vit-E was 414.25
µg/ml (Table-1).
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
838
Table 1: Effect of MEPF on Lipid peroxidation (LPO) and oxidation of GSH in rat
Liver homogenate (Mean ± SEM, n=6)
Table 2: Effect of MEPF on nitric oxide scavenging activity and free radical scavenging
activity by DPPH reduction action (Mean ± SEM, n=6)
Percent (%)Inhibition Percent (%)Inhibition
Nitric oxide scavenging activity Free radical scavenging activity
by DPPH reduction
Drug (MEPF/Vit. E)
Concentration (µg/ml)
MEPF Vit. E MEPF Vit. E
25 5.3 ± 0.04 24.20 ± 0.16 8.27 ± 0.04
26.30 ± 0.09
50 12.3 ± 0.08
39.46 ± 0.02 17.55 ± 0.05
37.66 ± 0.02
100 20.4 ± 0.06 50.82 ± 0.01 29.35 ± 0.07 47.30 ± 0.03
Percent (%) inhibition Percent (%) inhibition
Lipidperoxidation(LPO) in
liver homogenate
Oxydation of GSH in liver
homogenate
Drug (MEPF/Vit. E)
concentration (µg/ml)
MEPF Vit. E MEPF Vit. E
25 2.67 ± 0.04
28.31 ± 0.05 4.25 ± 0.14 25.66 ± 0.16
50 10.47 ± 0.02
40.84 ±0.02 12.12 ±0.06 41.86 ± 0.01
100 22.56 ± 0.02
57.54 ±0.02 25.91 ±0.05 55.36 ± 0.08
200 35.45 ± 0.02
67.95 ±0.02 35.91 ±0.57 68.40 ± 0.12
400 44.49 ± 0.01
81.42 ±0.10 43.71 ±0.15 75.75 ± 0.08
800 59.77 ± 0.01 97.25 ±0.03 57.38 ±0.24 96.56 ± 0.12
IC
50
669.23± 0.04 411.3± 0.14 697.1 ±0.11 414.25 ±0.15
r=Vit.E/MEPF 0.61 0.59
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
839
200 31.8 ±0.06 67.82 ± 0.01 43.03 ±0.04
60.60 ± 0.10
400 43.2 ± 0.08
81.05 ± 0.01 52.49 ± 0.50
79.19 ± 0.08
800 56.1 ± 0.02
96.18 ± 0.01 66.98 ± 0.03
98.77 ± 0.03
IC
50
713.01± 0.07
415.89 ±0.02 597.19± 0.50 404.98 ± 0.18
r=Vit.E/MEPF 0.58 0.68
Discussion
Indigenous drug system can be a source of a variety of new drugs, which can provide
relief to pain, fever and inflammation but their claimed reputation has to be verified on a
scientific basis. The present investigation revealed that the extract of Pandanus fascicularis
leaves (MEPF) has a significant antioxidant activity in all the four invitro models used in this
study.
Recent studies suggest that inflammation and tissue damage are due to the liberation of
free radicals
15
. The free radicals have been implicated in the pathophysiology of various clinical
disorders including inflammation, acute hypertension and cancer etc.
16
. Normally endogenous
intracellular antioxidants protect the tissue from injury by free radicals
17
.Therefore development
of antioxidant drug could be beneficial as adjunct to anti-inflammatory therapy. Phytochemical
screening revealed the presence of phenols, which could be responsible for its anti-inflammatory
and antioxidant activity. Phenols are very important plant constituents because of their
scavenging ability due to their hydroxyl groups. The phenolic compounds may contribute
directly to antioxidant action.
The MEPF showed significant protection against ferrous sulphate induced LPO which
could cause by absence of ferryl perferyl complex. It is generally assumed that ability of the
plant phenolic compounds to chelate iron in LPO system is very important for their antioxidant
property. Therefore an attempt was made to determine the role of iron chelation, since the
inhibition of ferrous sulphate induced LPO could also be due to chelation of iron. So it can be
concluded from the present study, the extract offers protection against ferrous sulphate induced
LPO by either metal chelation or absence of ferryl perferyl complex, which is essential for
inhibition of LPO
18
. GSH is a non enzymic mode of defense against the free radicals. Thiols
especially cystein and glutathione are important in leukocyte functioning
19
.
The study on nitric oxide scavenging demonsrates that the methanol extract of Pandanus
facicularis is a potent scavenger of nitric oxide. NO generated from sodium nitroprusside reacts
with oxygen to form nitrite ions which can be estimated by the use of Griess reagent. Scavengers
of NO compete with oxygen leading to reduced production of NO
13.
The free radical scavenging activity of the plant extract MEPF was evaluated based on
the ability to quench the synthetic DPPH. Because of the odd electrons DPPH shows a strong
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
840
absorption band at 517 nm in visible spectrum. As this electron becomes paired off in presence
of free radical scavenger, the absorption vanishes and the resulting decolourisation is
stoichiometric with respect to the number of electrons taken up
20
. The bleaching of DPPH
absorption is representative of the capacity of the test compounds to scavenge free radicals
independently. The results revealed that the test compound /extract is an electron donor and
could react with free radicals to convert them to more stable products and terminate the radical
chain reaction. In the present study it can be concluded that the MEPF has significant antioxidant
activity. Since the antioxidants have been demonstrated to be useful in inflammatory disorders
21
the claimed beneficial effects of Pandanus fascicularis in traditional medicine in various
rheumatic disorders could be due to its antioxidant activity. This enables us for further research
to find out the active principle responsible for the antioxidant action and its isolation to be used
in rheumatic disorders.
References
1. Rastogi PR, Mehrotra BN, Sinha S, Srivastava M, Bhushan B. Compendium of Indian
Medicinal Plants. 1
st
ed. Lucknow; CSIR: Publications and Information Directorate., 1989; 4
: 533-534.
2. Prajapati ND, Purohit SS, Sharmak A, Kumar T. 1
st
ed. A handbook of medicinal plants.
Jodhpur; Agrobios., 2003: 378-379.
3. Kirtikar KR, Basu BD, Blatter E. 2
nd
ed. Indian Medicinal Plants. New Delhi; Indian Book
Center ., 1991; 4 : 2591-2593.
4. Charterjee A, Pakrashi SC. The treatise of Indian Medicinal Plants. 2
nd
ed. New Delhi;
National Institute of Science Communication., 2001: 6 ; 9-10.
5. Raina VK, Kumar A, Srivatsava SK, Shyamsunder KVN, Kahol K. Essential oil composition
of Kewda( Pandanus odorotissimus) from India.internet, Journalwebsite.
6. Ambasta SP, Ramachandran K, Saxena SN.The Useful Plants of India. New Delhi;
Publications and Information Directorate, CSIR., 1994 ; 2: 423-425.
7. Chopra RN, Nayar SL, Chopra IC. Glossary of Indian Medicinal Plants. 1
st
ed. New Delhi;
CSIR., 1996: 184-185.
8. Basu BDM. Indian medicinal plants. 2
nd
ed. New Delhi; periodical experts book
agency.,1991: plata no 991.
9. Kokate CK. Text book of Practical Pharmacognosy. 4
th
ed. Delhi: Vallabh prakashan; 1977:
107-121.
10. Harbone JB. Phytochemical methods. 1
st
ed. London; Chepman and hall., 1973: 60-66.
11. Okhawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric
acid reaction. Anal Biochem 1979; 95: 351-354.
12. Ellman GI. Tissue sulfhydryl groups. Arch Biochem Biophys 1959; 82: 70-77.
13. Sreejayan N, Rao MNA. Nitric oxide scavenging by Curcuminoids. J Pharm Pharmacol
1997; 49 :105.
14. shimada K, Fujikawa K, Yahara K, Nakamuray T. Antioxidative properties of xanthin on
auto oxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 1992; 40: 945-
946
Pharmacologyonline 1: 833-841 (2011) Sanjeeva et al.
841
15. Elaine M, Conner, Mathew, Grisham. Inflammation, free radicals and antioxidants. utrition
1996; 12(4): 274-277.
16. Hemnani T, Parihar MS. Reactive oxygen species and oxidative DNA damage. Indian J
Physiol Pharmacol 1998;42: 440.
17. Shenoy R, Shirwaikar A. anti-inflammatory and free radical scavenging studies of Hyptis
suaveolens(Labiateae). Indian Drugs 2002;39(11): 574-577.
18. Govindarajan R, kumar MV, Rawat AKS, Mehrotra S. Free radical scavenging potential of
Picrorhiza kurrooa Royle ex Benth. Indian J Exp Biol 2003; 41: 875-879.
19. Yamini B, Tripathi, Sharma M. Composition of anti oxidant activity of alcoholic extract of
Rubia Cordifolia with Rubiadin. Indian J Biochem Biophys 1998; 35: 313-316.
20. Blois. Antioxidant determinations by the use of stable free radicals. ature 1958; 26: 1199-
1202.
21. Francis Cheng, Christopher P. Zhao, Andris Amolins, Malgorzata Galazka,Leon Deneski. A
hypothesis for the in vivo antioxidant action of salicylic acid. Biometals, vol9(3): 285-
290,2007.
... The therapeutic effect of natural substance is directly related to their chemical constituent contents. The pandanus oils have been used traditionaly for earache, headhace, arthritis, debility, giddines, laxative, rheumatism, small pox, and spams (Kumar et al., 2011). Its leaves is rich with essensial oils contained, such as ether (37.7%), terpene-4-ol (18.6%), α-terpineol (8.3%), 2-phenylethyl alcohol (7.5%), benzyl benzoate (11%), viridine (8.8%), and germacrene (8.3%) together with small amount of benzyl salicylate, benzyl acetate, and benzyl alcohol. ...
... Furthermore, P. tectorius leaves are used in treatments for cold/flu, hepatitis, dysuria, asthma, boils, and cancer (Meileur et al., 1997). It was rich by phenolic compounds such as alkaloids, flavonoids, saponins (Kumar et al., 2011), phenols (vanilin, benzoic acid methyl ester, and a new benzopuran derivative), terpenes (phytosteroid mixture), eudesmin, kobusin, desmin and hydrofuran (Jong and Chau, 1998). While, physcion, daucosterol, β-sitosterol, camphosterol, palmitic acid and steric acid in rhizomes have been reported by Venkatesh et al. (2012), and its root extracts was rich antioxidant compound from lignans, bezofuran derivate, α-terpineol, β-carotene, β-sitosterol, vitamin C, tangeritine, germacrene-B, and vanidine (Adkar and Bhaskar, 2014). ...
... They have reported that the active chemicals constituents from this plant were phenolics, flavonoids, alkaloids, glycosides, carbohydrate, steroids and triterpenoid. In addition, Kumar et al. (2011) reported the presence of alkaloids, carbohydrates, proteins, steroids, sterols, phenols, tannins, terpenes, flavonoids, gums and mucilage, saponins, and glycosides in the P. tectorius leaves. ...
Article
Full-text available
Pandanus tectorius fruits have been used as major food in Micronesia, but not fully exploited in South East Asia. In Malaysia and Indonesia, P. tectorius fruit is wasted and still not utilized either as a source of food as well as for research. The aims of the present study were determined the phytochemical content, antibacterial and antioxidant activities, total phenolic content (TPC) and cytotoxicity properties of cores and keys parts of P. tectorius fruits extracts against some normal (RAW and L-6) and cancer cell lines (HeLa, HepG2 and MCF-7). Samples were collected from Setiu Wetland, Terengganu, Malaysia. Extracts were obtained by successive extraction using hexane, ethyl acetate and methanol. The antibacterial activity was performed by disc diffusion method against gram positive (B. subtilis and S. aureus) and gram negative (E. coli, and P.aeruginosa) bacteria. Actives extract was continued for antibacterial kinetic study. The TPC and antioxidant were analyzed by Folin-Ciocalteu and DPPH free radical scavenging assay, respectively. Qualitative phytochemical analysis result confirmed the presence of phenolics, flavonoids, terpenoid, steroids, saponins and glycosides as chemicals constituents in P. tectorius fruits extract. It was supported by high TPC content in this extracts. The ethyl acetate extract from cores part (PEC) showed the highest antioxidant capacity (IC50= 0.8±0.20 mg/mL), while ethyl acetate extract from keys part (PEK) was displayed highest antibacterial activity with inhibition zone of 10-15mm, but less strong inhibition as compared to the investigated commercial antibiotics (amphicilin, penicillin, gentamycin, and tetramycin). Antibacterial kinetic study was further proven that PEK has good antibacterial activity. Moreover, cytotoxicity study revealed that all of extracts did not have any cytotoxic activity against all selected cell lines (IC50 value > 30 μg/mL). Result founded that both keys and core parts of P. tectorius fruits were found to be rich by phenolics content and potent as antioxidant and antibacterial agents. Since it is not toxic against selected cell lines, the mechanism study on antiinflammatory, antidiabetic, antiatherosclerosis could be carried out.
... Oil Plants are widely used in the various traditional system of medicine like Ayurveda, Siddha, and Unani for their analgesic, and antiinflammatory, antipyretic activity. 77 50 Piper betel Linn, Piperaceae, Vettalai ...
... In all these animal models both extracts have shown significant anti-inflammatory activity. 77 After initial phytochemical screening of methanolic (MEPF) and aqueous extracts (AEPF) of the plant, the extracts were screened for possible acute antiinflammatory properties in vivo models. Acute anti-inflammatory properties of both the extracts were tested in rodent models by carrageenan induced paw edema, albumin induced plantar edema, acetic acid induced vascular permeability and castor induced diarrhoea. ...
Article
Full-text available
Rheumatoid arthritis is an immune mediated inflammatory disease (IMID). Rheumatoid arthritis occurs when our immune system attacks the tissues near joints, this is due to release of certain chemicals and enzymes that begin to eat away the cartilage and bones. Rheumatoid arthritis affects all the joints in the body, some forms of arthritis can also affect the body’s internal organs. The symptoms of Rheumatoid arthritis include inflammation, pain, swelling and stiffness of the joints, it can also lead to deformity and disability of the joint in severe cases. There are several causes for Rheumatoid arthritis these causes are unknown but some include genetic factor, family history, age, environmental factors, hormones, smoking etc. This paper gives an overview of the medicinal plants with phytoconstituents which can be used in the treatment of RA and different treatments available for Rheumatoid arthritis. These medicinal plants can be extracted and phytoconstituents can be isolated, pharmacological studies can be carried out and the plants can be further explored for future studies. Even modern drugs used for the better improvement of the symptoms, offer only temporary relief and produce severe side effects, so researchers rely on natural remedies, for treatment of various diseases, with efficacy and safety, and with fewer side effects.
... The overall Pandanus genus contains about 600 species distributed mainly in subtropical and tropical regions; there are around 30 to 40 species of Pandanus in India (5). P. odoratissimus is said to be a restore health, strength, or well-being, promoting a feeling of well-being in tropical climate (6). ...
... Pharmacological studies revealed its anxiolytic (Bhatt and Bhatt 2016), anticoagulant (Omodamiro and Ikekamma 2016), antihyperlipidemic (Zhang et al. 2013a), and diuretic (Tan et al. 2014) properties. The leaves are also used in traditional medicine in the treatment of heartache, earache, arthritis, debility, giddiness, laxative, rheumatism, smallpox and spasms (Sanjeeva et al. 2011), while the root decoction is used to treat hemorrhoid (Wen et al. 2011). The fruits, male flowers and aerial roots are also employed to combat digestive and respiratory disorders (Omodamiro and Ikekamma 2016). ...
Article
The Pandanus tectorius methanolic (PTM) and P. tectorius aqueous (PTA) extracts were investigated for their potential in vivo anti-inflammatory activity using carrageenan-induced rat paw edema. Parameters including paw thickness measurement, histopathological, and immunohistochemical cyclooxygenase (COX)-2 expression analyses were measured and analyzed. PTA at 500 mg/kg significantly reduced inflammation after induction of carrageenan (Carr), with mean paw thickness change of 0.110 ± 0.024 mm at sixth hour post-induction and histopathological mean inflammatory grade of 1.80 ± 0.20. The reduced immunohistochemical COX-2 expression using PTA at 500 mg/kg was determined with mean final 3,3′-diaminobenzidine (DAB) intensity of 63.70 ± 2.08. The profiling of metabolites by liquid chromatography–mass spectrometry (LC–MS) revealed presence of ethyl caffeate and dihydroconiferyl alcohol as putative secondary metabolites of PTA which were the major peaks and have been reported to possess anti-inflammatory activities. This research has provided scientific insights of utilizing P. tectorius as a potential anti-inflammatory agent containing secondary metabolites which may be pharmacologically relevant.
... The overall Pandanus genus contains about 600 species distributed mainly in subtropical and tropical regions; there are around 30 to 40 species of Pandanus in India (5). P. odoratissimus is said to be a restore health, strength, or well-being, promoting a feeling of well-being in tropical climate (6). ...
... The essential oil obtained from inflorescences of P. odoratissimus (kewda) contains the following chemicals 7.5% 2-phenylethyl alcohol, 18.6% terpene-4-ol, 8.3% germacrene-B, 8.3% α-terpineol, 8.8% viridine, 37.7% ether, and 11% benzyl benzoate along with lesser amount of benzyl acetate, benzyl salicylate, and benzyl alcohol on gas chromatography. The phytochemical screening of aqueous and methanolic extracts of Pandanus have shown the presence of proteins, lignans, mucilage, coumestrol, alkaloids, isoflavones, glycosides, phenolic compounds, carbohydrates, tannins, steroids, flavonoids, sterols, saponins, terpenes, gums, vitamins, and amino acids [10,11]. Mainly due to the increase in resistance of dermatophytes to the azoles and other antifungal drugs and due to the side effects of long-term usage of currently available antifungals, new antifungal agents are to be explored. ...
Article
Objective: The present study aimed to evaluate the antifungal activity of Pandanus odoratissimus oil against dermatophytic fungi, and it was compared against the two commonly used antifungal agent’s fluconazole and griseofulvin.Methods: A total of seven strains of dermatophytes were tested for antifungal activity using oil extracted from the flower of P. odoratissimus by using agar-well diffusion method and the zone of inhibition was compared with antifungal agent’s fluconazole and griseofulvin. Minimum inhibitory concentration (MIC) was determined using the tube-dilution method.Results and Conclusion: The zone of inhibition varied from 16.32 to 19.76 mm for fluconazole, 12.12–18.16 mm for griseofulvin, and 2.5–9.59 mm and 7.63–12.88 mm for 2.5 mg/ml and 5 mg/ml of P. odoratissimus extract, respectively. Epidermophyton floccosum and Trichophyton violaceum showed the lowest MIC value of 0.15 mg/ml. The results of our study have shown that the extract from P. odoratissimus can work significantly better against fungal diseases caused by dermatophytes. It was also found that it acts as a perfect alternative to the currently available antifungals such as fluconazole and griseofulvin.
... The different fraction of Pandanus conoideus extracts have antioxidant activity with IC 50 5, 25 to 53, 47 µg/mLthat has a positive correlation with the total phenols (r 2 = 0.645) and flavonoids (r 2 = 0.709) [13]. Pandanus fascicularis also has significant antioxidant activity [14]. Pandanus odoratissimum root extract have antioxidant activity and positive correlation with the content of phenol and flavonoid [15], as well as aquadest and methanol extracts of Pandanus foetidus that has antioxidant activity too [16]. ...
Conference Paper
Full-text available
Breast cancer is one of cancer types that can lead to human death. Beside that, Indonesia has a lot of potential anticancer herbs, among them is from genera Pandanus. Anticancer potency can be determined by cytotoxic activity on the cancer cells. A species of genera Pandanus that can be found in Indonesia is Pandanus tectorius. Pandanus tectorius is abundant in the area of Gunungkidul, D.I. Yogyakarta but its anticancer potency is not explored yet. So, its important to explore anticancer potency that possible on Pandanus tectorius. This research aimed to study the cytotoxic and apoptosis induction activity of the leaf and fruit extracts of Pandanus tectorius on breast cancer cells (T47D). The plant materials were sampled in Gunungkidul, D.I. Yogyakarta. Extraction was done by maseration method using ethanol, chloroform, and aquadest independently. The extracts were subjected to Thin Layer Chromatography for identifying compound groups; cytotoxic activity was assessed by the MTT assay in triplicate. Statistical analysis was conducted by ANOVA, LSD test with Tukey HSD and probit analysis to determine the IC 50. Extracts with the lowest IC 50 value were subjected to double staining method (Ethidium-Bromide Acrydine-Orange) to differentiate necrotic from apoptotic death pathway of cell death. The results showed that IC 50 values from low to high were aquadest leaf extract (61.28 µg/mL), ethanolic leaf extract (105.09 µg/mL), chloroform leaf extract (222.76 µg/mL), and ethanolic fruit extract (773.69 µg/mL), while the IC50 values of chloroform fruit extract and aquadest fruit extract could not be determined. Aquadest and ethanolic leaf extracts showed a low apoptosis induction ability, suggesting that the cell death was through non-apoptosis pathway tautophagy pathway. It might be concluded that aquadest and ethanolic leaf extracts has potency on cytotoxic activity towards breast cancer cell in T47D cell culture.
... The different fraction of Pandanus conoideus extracts have antioxidant activity with IC 50 5, 25 to 53, 47 µg/mLthat has a positive correlation with the total phenols (r 2 = 0.645) and flavonoids (r 2 = 0.709) [13]. Pandanus fascicularis also has significant antioxidant activity [14]. Pandanus odoratissimum root extract have antioxidant activity and positive correlation with the content of phenol and flavonoid [15], as well as aquadest and methanol extracts of Pandanus foetidus that has antioxidant activity too [16]. ...
Conference Paper
Full-text available
Breast cancer is one of cancer types that can lead to human death. Beside that, Indonesia has a lot of potential anticancer herbs, among them is from genera Pandanus. Anticancer potency can be determined by cytotoxic activity on the cancer cells. A species of genera Pandanus that can be found in Indonesia is Pandanus tectorius. Pandanus tectorius is abundant in the area of Gunungkidul, D.I. Yogyakarta but its anticancer potency is not explored yet. So, its important to explore anticancer potency that possible on Pandanus tectorius. This research aimed to study the cytotoxic and apoptosis induction activity of the leaf and fruit extracts of Pandanus tectorius on breast cancer cells (T47D). The plant materials were sampled in Gunungkidul, D.I. Yogyakarta. Extraction was done by maseration method using ethanol, chloroform, and aquadest independently. The extracts were subjected to Thin Layer Chromatography for identifying compound groups; cytotoxic activity was assessed by the MTT assay in triplicate. Statistical analysis was conducted by ANOVA, LSD test with Tukey HSD and probit analysis to determine the IC 50. Extracts with the lowest IC 50 value were subjected to double staining method (Ethidium-Bromide Acrydine-Orange) to differentiate necrotic from apoptotic death pathway of cell death. The results showed that IC 50 values from low to high were aquadest leaf extract (61.28 µg/mL), ethanolic leaf extract (105.09 µg/mL), chloroform leaf extract (222.76 µg/mL), and ethanolic fruit extract (773.69 µg/mL), while the IC50 values of chloroform fruit extract and aquadest fruit extract could not be determined. Aquadest and ethanolic leaf extracts showed a low apoptosis induction ability, suggesting that the cell death was through non-apoptosis pathway tautophagy pathway. It might be concluded that aquadest and ethanolic leaf extracts has potency on cytotoxic activity towards breast cancer cell in T47D cell culture.
... and the writhing model of mouse and compared with the analgesic action of codeine and aspirin. Pandanus aqueous extract revealed significant analgesic activity by both central ( < 0.001) and peripheral ( < 0.001) mechanisms in this study, which is comparable to that of codeine and aspirin, and this favors the use of Pandanus aqueous extract in rheumatism and rheumatoid arthritis in traditional medicine [23]. ...
Article
Full-text available
Pandanus odoratissimus Linn. (family: Pandanaceae) is traditionally recommended by the Indian Ayurvedic medicines for treatment of headache, rheumatism, spasm, cold/flu, epilepsy, wounds, boils, scabies, leucoderma, ulcers, colic, hepatitis, smallpox, leprosy, syphilis, and cancer and as a cardiotonic, antioxidant, dysuric, and aphrodisiac. It contains phytochemicals, namely, lignans and isoflavones, coumestrol, alkaloids, steroids, carbohydrates, phenolic compounds, glycosides, proteins, amino acids as well as vitamins and nutrients, and so forth. It is having immense importance in nutrition. A 100 g edible Pandanus pericarp is mainly comprised of water and carbohydrates (80 and 17 g, resp.) and protein (1.3 mg), fat (0.7 mg), and fiber (3.5 g). Pandanus fruits paste provides 321 kilocalories, protein (2.2 g), calcium (134 mg), phosphorus (108 mg), iron (5.7 mg), thiamin (0.04 mg), vitamin C (5 mg), and beta-carotene (19 to 19,000𝜇g) (a carotenoid that is a precursor to vitamin A). Pandanus fruit is an important source of vitamins C, B1, B2, B3, and so forth, usually prepared as a Pandanus floured drink. Traditional claims were scientifically evaluated by the various authors and the phytochemical profile of plant parts was well established. The methods for analytical estimations were developed. However, there is paucity of systematic compilation of scientifically important information about this plant. In the present review we have systematically reviewed and compiled information of pharmacognostic, ethnopharmacology, phytochemistry, pharmacology, nutritional aspects, and analytical methods. This review will enrich knowledge leading the way into the discovery of new therapeutic agents with improved and intriguing pharmacological properties.
Article
Pandanus odorifer (Forssk.) Kuntze commonly known as Kewda is an economically important essential oil bearing plant belonging to the family Pandanaceae. Kewda flower distillation industry in Ganjam district (Odisha) accounts for nearly 90 % of the production of commercially important Kewda perfumes in the country and 50 % of that of the world. Economically, it is an important natural bioresource for the perfumery industry due to the exquisite fragrance it possesses. Besides the extensive use of Kewda male flowers for perfume production, other parts of the plant are also used in fibre, food, pharmaceuticals and handcraft industries. Hence, the plant contributes to be the livelihood of the local people of Ganjam district by providing an alternative source of income for the poor coastal villagers. Reports on the molecular characterization of Kewda germplasm using molecular markers are scanty. Molecular marker based assessment of genetic diversity in Kewda is very important for germplasm characterization, conservation and future improvement of the taxon. Hence, in the present study, 43 highly polymorphic molecular markers i.e. 13 ISSRs (Inter Simple Sequence Repeats) and 30 SSRs (Simple Sequence Repeats) were screened on 84 Kewda accessions collected from different zones of Ganjam district, Odisha. Both ISSR and SSR markers showed moderate level of polymorphism i.e. 68.63 % and 74.36 % respectively. Dendrogram was constructed by UPGMA (Unweighted Pair Group Method with Arithmetic Mean) cluster analysis for both the ISSR and SSR markers and the result was confirmed by Principal coordinate analysis (PCoA). Analysis of molecular variance (AMOVA) and population genetic study (POPGENE) was also done. SSR system was found to be more effective than ISSR system in evaluating the genetic diversity in Kewda accessions. The information obtained from genetic diversity analysis could be used for future breeding programmes for Kewda improvement to meet its ever increasing demand in the perfumery industry.
Article
The ethanolic extract of H. suaveolens was tested to study the effects on the inflammatory reaction, using the technique of Carageenan induced paw edema in albino rats. The extract showed significant anti inflammatory activity comparable to the reference standard Ibuprofen. Antioxidant investigations of the ethanol extract along with its fraction using nitric oxide induced free radical assay methods showed good free radical scavenging activity thereby supporting its anti-inflammatory properties.
Article
The chemical composition of the essential oil obtained by hydrodistillation of staminate inflorescences of kewda (Pandanus odoratissimus L.) has been compared with the kewda oil from the local market. Both the oils were subjected to high-resolution GC and GC–MS, which resulted in the identification and quantification of 85 and 59 components, amounting to 98.7% and 98.6% of the total oils, respectively. The major components of the hydrodistilled kewda oil were 2-phenyl ethyl methyl ether (37.7%), terpinen-4-ol (18.6%), α-terpineol (8.3%) and 2-phenyl ethyl alcohol (7.5%), whereas the major constituents of kewda oil from the local market were 2-phenyl ethyl alcohol (33.2%), 2-phenyl ethyl methyl ether (16.1%), benzyl benzoate (11.0%), viridine (8.8%) and germacrene B (8.3%). Copyright © 2004 John Wiley & Sons, Ltd.
Article
METHODS for measuring antioxidants and appraising antioxidant activity appear to be of two general types. If the chemical nature of the antioxidant is known, one may strive for a test specific for the compound or group of interest; for example, the nitroprusside test for sulphydryl groups. Alternatively one may observe the inhibition of some natural oxidative process such as the β-oxidation of fats, as a function of the added antioxidant.
Article
The autoxidation of soybean oil in a cyclodextrin emulsion system was studied in the presence of an emulsion stabilizer consisting of polysaccharides such as xanthan, tragacanth gum, and methylcellulose. Xanthan strongly inhibited the peroxidation of soybean oil containing tocopherols but showed no antioxidant activity on soybean oil without tocopherols in the emulsion. Xanthan did not have hydrogen-donating ability but expressed Fe2+-binding activity. The Fe2+-binding activity corresponded to the pyruvate content of xanthan. Depyruvated xanthan did not inhibit effectively the autoxidation of soybean oil. The Fe2+-chelating structure of xanthan is discussed.
Article
This paper presents a new hypothesis for the physiological antioxidant action of salicylate. Current theories have focused on the radical scavenging nature of salicylate. This explanation may have limitations because it is unlikely that salicylate reaches the necessary concentrations to effectively prevent damage to cell components. We propose that salicylic acid decreases the flux of hydroxyl radicals through chelation, which causes a redox deactivation mechanism of iron Fenton reaction centers. This is due to voltammetric results which indicate that the iron-salicylate complex does not have the thermodynamic driving force to act as an effective Fenton reagent necessary for the production of damaging oxygen-containing radicals. Furthermore, despite the more facile thermodynamics associated with Fenton-type processes at acidic pH values, the complex maintains Fenton inactivity due to a pH-sensitive redox potential shift that follows asE Fe[Sal] = 0.793 - (0.059 pH). This is important since inflammation sites are acidic relative to healthy tissue. This redox potential shift is unique to salicylates when compared with other common iron chelation agents such as EDTA. Further evidence for the lack of Fenton-type reactivity of the iron-salicylate complex is offered in the form of oxidation studies of calf thymus (CT) DNA by hydrogen peroxide. Salicylate prevents the iron-catalyzed oxidation of CT-DNA strands as indicated by the detection of the constituent bases by HPLC. However, salicylates were not able to prevent the copper-catalyzed oxidation of CT-DNA. These results are predicted by the cyclic voltammetry of copper-salicylate, which confirms that it is an effective Fenton-type catalyst, further adding to the proof that salicylate acts by redox deactivation of iron, not by hydroxyl radical scavenging. Finally, the iron-salicylate e.m.f. suggests that it may also act as a superoxide dismutase, which indicates another possible important antioxidant feature.
Article
The reaction of lipid peroxides in animal tissues with thiobarbituric acid was dependent on pH of the reaction mixture as was the case for linoleic acid hydroperoxide. The optimum pH was found to be 3.5. Taking this fact into consideration, a standard procedure for the assay of lipid peroxide level in animal tissues by their reaction with thiobarbituric acid was developed as follows. Ten percent ( tissue homogenate was mixed with sodium dodecyl sulfate, acetate buffer (pH 3.5), and aqueous solution of thiobarbituric acid. After heating at 95°C for 60 min, the red pigment produced was extracted with n-butanol-pyridine mixture and estimated by the absorbance at 532nm. As an external standard, tetramethoxy-propane was used, and lipid peroxide level was expressed in terms of nmol malondialdehyde. Using this method, the liped peroxide level in the liver of rats suffering from carbon tetrachloride intoxication was investigated. The results were in good agreement with previously reported data obtained by measuring diene content.