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Evaluation of antioxidant and phytochemical analysis of Pteris tripartita Sw. A

critically endangered fern from South India

Article · January 2010

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Journal ofFa irylakeBota ni ca l Garde n 9(3): 28 ~ 34 (20 10)

Evaluation of Antioxidant Activity and Phytochemical

Analysis of Pteris tripartita Sw.- a critically endangered
fern from South India
Xavier* Ravi Baskaran and Robert Jeyacbandran
(Oepanment of Plan t Biology aod Plan t Biotechnology.St. Joseph's College, Tiruchirappalli ,Tamil Nadu- 620 002, India)

Abstract: This study is o utlined to probe the i11 ••iJro antioxidant activi ty o f a criticall y endangered fern, P1eri.1· Jriparlila Sw. The plant extracts were screened fo r
their possible antioxidant activity by three comp lementary test systems namely. DPPH free radical scavenging, metal chelati ng assay and phos 1>homolybdcn um
method and the resu lts re lated to the total phenol content. The IC,, values of acetone and methanol extracts in DPPH radica l scavenging assay we re obtained to be
77.04 and 11 3.63 11g/ ml respectively. The acetone extract was analyzed by gas chromatography/ mass s pectrometry (GC/MS) and their bioac ti ve compounds were
identified. llowever, the IC,,, va lues for tbe s tanda rd BHA, BHT, quercetin a nd rutin we re noted to be 25.58, 4 5.56, 18.79 and 24. 7l 1Jglml respectively.
Furthermore. total phenolic co nt~nts , flavon oids. tanni ns a nd vitami n C we re determi ned. Accordingly. the aceto ne extract exhibited highest tota l phenolic
content (67.00:0.28 GAE mg/g). The highest content of total fluvoooids was also observed in ethyl acetate extract (48.24± 1. 28 RE mg/g extract). Owing to having
a ntioxida ti ve components. the extracts exhibited satisfying a ntioxidant ac ti vit ies. Our findings demonstrate that the extracts of P. rriparlita possess s ignificant
antioxidant activi ties and thus it has great pote ntial as a source for na tural health products.
K eywords: i11 vi tro an tiox idaot. DPPH. GC/ MS, natural health products

Introduction beginn in g of th e 20 '' century. However, restrict ions o n the

Oxygen is essential fo r the s urviva l of al l on thi s earth. use of th ese c omp o unds a re now be in g impos ed because o f
Durin g th e proces s of oxyge n utiliz ation in norm al the ir c arc ino geni c ity (Mah davi a nd Sal ukhe, 19 95).
physiolog ical and m etabo li c processes approximate ly 5% of C urrentl y, the research o f natura l antiox id an ts like
oxygen gets uni vale ntl y red uced to oxygen deri ved free a lternative sources of sy nthes is antiox id a nts was emerged
radi cal s ( Yu , 1994; Halli well a nd G utte rid gc ( 1989). and the exp lo itat ion of th e va rious seco ndary meta bo li tes o f
However, oxygen -centere d free radi ca ls and o th er reactive the p lant was high lig hted in rece nt years . T hus, the pheno li c
oxygen species ( ROS), wh ich arc continuou sly, produced in co mpounds in pa rti cu lar the flavono ids ha ve dra w n
vivo, res ult in ce ll de ath and ti ss ue damage (Ha lli well and atte nti o n as a potential source ofb ioact ive mol ecul es . Their
G utte ridgc, 1999). Reacti ve oxygen spec ies and ass oc iated fla van nu c leu s struc ture is l inked to t he a nt ioxidant
free radi c als ha ve bee n implic ated in th e etiology of va riou s capaci ty. These substances are able to red uce free radica ls
human di seases in c ludin g inflammation , m e tab o li c like superox ide , peroxyle, a lkoxyle and hydroxy l. The
di sorde rs, ce llular aging and atherosclerosis, hea rt di sease, reduct ion occurs by hyd rogen t ransfer react ion to the
strok e, diabe tes me llitu s, canc e r, mal a ria. rh e umatoid reactive oxygen species (ROS), (Jovanovic eta/.. 1994 ).
arthr it is and HI V/ A TD S (A iho and Le inon e n, 1999; Fern s are used in d i fferent tradit ional med ici nal
Odukoya et a/. , 2005). Therefore, radic al scavenge rs g ive systems of I ndia a nd p lay an imp orta nt ro le in folklo re
promi si ng ind ications of new th erap euti c approaches (Cho med ici ne. S ystem atic s urveys of medic inal use of fern ha ve
e ta/. , 2003). An antioxidant is a compound that inhibits or been scarce ly undert ak en wh ic h a re no t pharmaco logically
delays the oxidation of sub strates even if the compound is eva luated (Ka ndh asamy ei a/. , 2008: Kau sh ik and Dbim ian ,
present in a s ig nificantl y lower co ncentration than is the 1995 ; Di xi t and Bhatt, 1974 : Ma n ic kam and Irudayaraj,
ox idized sub strate. The scave ngin g of reac ti ve ox ygen 1992). Man y spec ies of ferns and fern a llies a re o f eno rm o us
species (ROS) is one of the po ss ib le mec han is ms of action economic utility w ith t he most im portant being the ir
of ant ioxid ant co mpound s. T he res t deal s w ith the medicinal , aesthet ic and food value (Yas uda. 1999).
prevention o f ROS form ation by me tal binding or enzyme Recen tl y. some ferns ha ve been re ported for ant iox idant
inhibition . C hain breaking antioxidants prevent damage by activity such as, three S elagin e/la species (Gayat hri et a/..
in terfering with the free radica l propa gation casca des 2005) , Polypodium leucotomus (Gomba u eta/., 2006) , fi ve
(Malko wsk i, 2008). Synthetic ant iox idant s s uc h as Equisetum sp ecies (M i Jovanov ic et a/., 2007), Abacopteris
buty lated hydroxyto lu ene (BHT), butyla ted hydroxyanisole penangiana (Zhao er al.. 2007 ), six folk medic inal fe rn s
( BHA ) and prop y l ga llate have been us ed s in ce the (Cbang er a/.. 200 7), 3 1 species of fe rns (Ding et a/., 2008).

28
 Evaluation o f Antiox idan t Activi ty a nd Phy tochemical Analysis of Pteris tripartim
Sw.- a critically end an ge red fern fr om South India

8/ec lznum orientale Linn (Lai e/ at., 20 I 0), Marsilea quadrifolia p e rfo rme d a t solvent bo il ing tempera ture. Afte r t he
(Ripa el a/., 2009), Cheilanthes anceps Sw. (Chowdhary er a!., extrac ti o n, the so lve nt was rem oved from the solute mi xture
2010), Pteris ensiformis Burm (We i et a!. , 2007) and Preris by reduce d pressu re w ith ro tary evaporator and s to red at
multijlda (Shyur et a/., 2005; Wang et at., 2009). 4 °C fo r f urther use.
Th e ph yot c he mi ca l po tential o f pte rid ophytes is
re lati ve ly une xplo red mo re. a ltho ug h pterid o phytes possess DPPH radical scavenging activity
g reat eco no mic po te ntia l due to so m e th erapeuti c va lues
F ree radi ca l scavenging ac t iv ity of PT ext racts was
(Asakawa, 1982 ; As akawa, 1995 ; C he n et a/., 200 5;
de termined us ing the DPPH method (B lios, 1958) . The
Dhim a n, 1998; Gogoi, 2002; Redd y et a/., 200 I ; Sing h et
a nti oxid a nt capac it ies o f the samp les us ing DPP H as sa y
a/., 200L Sing h et a l., 2008; Vas udeva , 1999) . C hemica l
we re compa red w it h tb ose of BH A. BHT, rut in and que rcetin
invest iga ti o n of more tha n 30 spec ies of the P teridac e ae has
and the blank.
been re po rte d (C as till o et a/., 1998). Ph ytoc hemi cal
C belating capacity
investigatio ns o n the P teris genu s led to iso lation of va ri o us
The c he lat ing of ferr o us ions by vario us extrac ts of
pheno lic comp ounds (Jes uda ss et al., 2 000) . fl avo nol
Ptripartita was estima ted by th e m ethod of Din is er a/.,
g lycos ides (Salatino and Prado , 199 8; Tana ka eta /. , 1978),
( 1994 ). Briefl y the extracts (250 ll L) were ad ded to a
ka uran es (Woe rd e n b a g era/., 1 99 6 ) and
so luti o n of 2 mmol / L FeC I2 (0 .05 mL) . Th e reaction was
ptero s insesquite rpenes (Sa ito et al., 1990; Mu rakami eta/. ,
initiated by the a ddi t io n of 5 mmol / L fe rroz ine (0.2 mL) a nd
198 0). Acc ording t o C ha ndra et al.. (2 008) Pteris trip artita
the mixture was s haken v igorous ly and left sta nding a t room
Sw. ( PT) is a c ritica ll y endangered fern from So uth Indi a
te mpe rature fo r I 0 mi n . Absorba nce of the so lution was t he n
and it s fr o nd s be in g taken inte rnall y during c hildbirth in
m eas ure d spectroph otometrica lly at 562 nm. Th e chelat ing
Bo uga in ve lle . Thu s. in this study, in vitro ass ays were
a cti vity of th e ex tracts wa s eva lua ted usi ng EDTA as
per form e d to evalua te the a n tiox ida nt ac ti viti es o f crude
st a nd ar d. The resu lts we r e expresse d as m g EDTA
ex tracts and phy toc he mi ca ls o f P.tripartita were ide ntified
equi va lent/ g extrac t.
us ing GC/ M S technique.

Materials and Methods Phosphomolybdenum assay

The a ntioxida nt acti vity of PT sa mples wa s evaluated
Chemicals
by the ph osph omolybdenum me th od ( Prie t o et a/., 1999) . An
Fe rri c c hlorid e, l , 1-d i phe n y 1-2 -pi c ry l-h y dr azyl
aliquot o f I 00 11 L o f samp le so lution (in 1 mM d imethy l
( DPPH), fer rozin e, ammo nium mo ly bdate. vitamin C,
sul foxid e, DM SO) was comb ined wi th I m L of reagent
Buty la ted hydroxyani so le (BHA), Buty lated hydroxyto luene
soluti o n (0 .6 M sul phuric acid , 28 m M so dium phosphate,
( BHT), querc etin, mtin, sodiu m ph osphate, sodi um ni trite,
a nd 4 m M a mmoniu m mo lybdate) in a via l. The vials were
sodium h ydroxide and AlC l , were purc hased from Himed ia,
cappe d a nd incubated in a wa ter ba th a t 95"C for 90 min.
Merc k or S ig ma. All other reagents used were of ana lytical
Aft er the sa mp les had coo led to room tempera ture, th e
g rade.
ab sorban ce of the mi xture was measured a t 695 nm against a
b lank. Th e res ults re ported (ascorb ic acid equi valen t
Preparation ofplant extracts anti o xida nt ac tiv ity) a re mea n va lues expressed as mg of
The aeri al part of Pteris tripa rlita were co llec ted fro m ascorbic ac id e quiva le nts/g ex tract.
A lakar hill s of Tamil Nadu and authentification of th e Dete rmin ation o f total phenolics a nd tannin s
species w as made by Dr.A .Be nnia min , Bota ni ca l S urvey of Th e to tal pheno l ic co nte nt was determ ined by Fo lin-
In d ia, Arunac ha l prades h. Its voucher s pec ime n has been c ioc alte u meth od. Us in g th e same extract th e tannin s were
depos ited in th e Dep artme nt o f Pl ant Bi o logy and Plant estimated a ft e r treatm e nt w ith polyv in yl po lypyrro lidone
Bi otec hnol ogy o f S t.Joscp h's co lle ge , Tiru cbira pa lli. The
(P V PP). The am o unt of to tal phe no li cs an d tanninS were
sha de dri e d ae rial p art o f Ptripartita ( PT) we re g ro und to a
ex pressed as galli c ac id eq uiva le nts (GA E) in mg p er g
fin e powder and sox hle t extracted sepa rate ly w ith hexane,
ex tract (S iddhu raju a11d Becker, 2003) .
ace to ne, et hy l acetate, c hlo roform, e th a no l, me th ano l and
Estim ation of total flavonoids
water. Th e sox hle t procedure co ns isted of g roun d PT
The flavono id con tents of PT ex trac ts were quant ified
sampl es (I 0 g ) pl aced in s ide a thimble loaded into th e
a nd it acts a s maj o r ant iox id a nts in pl an ts red uc ing
soxhle t extractor. The to tal extrac ting time wa s 6 h, a nd th e
ox idati ve s tress. Tb e content was e stima ted as per the
total amo unt of so lve nt wa s 6 0 ml m a inta ined co ntinu ous ly
method described by Zhi shen er a/., ( 19 99). Initiall y 500j.l-L
r e flu xing over th e sa mple. Th e so lve nt assays we re
of all the plant extrac ts we re take n io diffe re nt te st tubes. To

29
 Evalua tion of Antioxidant Ac tivity and Phytochemical Analysis of Pteris triparlita
Sw.- a criticall y endangered fern from South India

each ex trac ts 2 m l of disti li ed water was added. Then I 50~~ L

of NaNO, was added to al l the te st tubes fol lowed by Results and Discussion
incuba ti on at room tempe rature fo r 6 minu tes . A fte r DPPH radical scaven ging activity
incuba tion I 50 ~L of AIC I, ( I 0%) was added to al l the test DPPH assay has been extens ive ly used fo r screening
tubes including th e blank. A ll the tes t tubes were incubated a ntiox idant ac tivity beca use it can accomm odate many
for 6 minute s at room temp erature. T hen 2 ml ofNaOH was samples in a short period and is sens it ive enough to detect
added to all the test tubes wh ich we re made up to 5 ml using act ive ingredients at low concentrations (Sa nchez-Moreno,
d istilled water. The co ntents in all the test tubes we re 2002). DPPH· is a stable nitrogen-cen tered free radica l the
vo rtexed we ll and they we re allowed to stand for I 5 minutes colo r of w hic h c hanges from v io let to ye ll ow upon redu ction
at room temperature. T he pink colour deve loped due to the by e ither the process of hydrogen- or e lec tro n- donat ion.
presence of flavo no ids was read spectrophotometrically at Substances wh ich a re able to perform th is react ion can be
5 10 nm. The amou nt of flavo no ids was ca lc ul ated in rutin cons idered as antiox idants and therefore radica l scave ngers
equ iva lents. (Dehpour e t a/.. 2009) . Var ious extracts of P. tripartita
Estimation of Vitamin 'C' s howed a wide variat ion of IC ,. ranging from 77.04 to
Total vitamin C/asco rb ic acid content was est ima ted in 133 .68i!g/ ml (Table l; Fig. I). Th e highes t an tiox idan t
o rder to determine the antiox idant prope rty of the plant act ivity was detected in acetone extrac t (77.04 ~tg/ ml). The
under study as ascorb ic acid is a natural a ntiox idant. Its IC, 0 va lues of other extracts were followed by methanol
content of PT extracts was determ ined by the me th od
( 113.63i!g/ ml ), water ( ll6. 2711g/ml), e thyl acetate
formulated by Yen and Chen, ( 1995 ). I 0 mg of sa mple was
(1 1 6.82 ~t g / ml), ethano l ( 117 . 37 J.tg/ m l), ch loroform
di sso lved in I 0 m l of l "1.1 meta ph osphoric ac id. Samp le is
(129.87J.tg/ m l) a nd he xa ne extrac t ( 133.6811g / ml ).
then filtered through Wba tman No . I filter paper and from
respective ly. The IC,11 value of standard com po unds were of
this 1 m l of su pernatant was taken and 9 ml dichloro
quercetin (18.79 11g/ m l), ru tin (24. 7 1 J.tg/ ml) , BHA (25.58
indophenols (Dip ) dye solution was added. Formation of
J.tgl m l) and BHT (45.56 11g/ m l). T he pheno li c compounds
p ink co lor afte r adding of Dip dye so lution shows pre sence
present i n t he ex t r act may contri bu te directly to
of v itamin C. Th e co lor thus formed was read
aotioxidative act ion (Dub e t at.. 1999).
spectropho tomet ri ca ll y at 5 15 nm. Amount of vitamin C was
ca lculated in ascorbic acid equivalent (AAE). Water was Fig. l . DPPH radical sc.lVengingactivity
used as b lank.
GC-MS analysis 160
N
co
The aceto ne ex tract of P.tripartita was analyzed by 140 .,;

GC-MS (GC-Thermo ul tra and M S -DSQ II, Ita ly) equipped e 120

with TR- 5 (The rm o) column. Th e carrier gas was helium. ~ 100

~ 80 "' • Senesl
The te mperature program was set as fo ll ows: 85 "C hold for 3 a..
0
0
60 "'.,.;
min , raised at 6"C/ min to I 9o•c and 8 · c / min to 250 ·c, a nd
J
---..--..--Jjjj
40
bold for i 0 min. T he injector a nd detector temperatures we re
20
se t at 250 and 200 "C, respecti ve ly. The in terface 0
temperature was set at 230"C. Mass spectra were taken at 70
eV. Th e mass range was sca nned from 50 to 600 amu.
Plant extracts
I d en tifica tion of components
PTH= hexane extract: PTC= chloroform extract: PTA= ace tone extract:
The spectrum of the unknown compo nent was
PTEA= ethyl acetate extract: PTE= e-thanol extract: PT M= methanol extract;
compared wi th the s pectrum of the known components PTW= water extract. BHA ( Butyl3tcd h yd rox yanisolc). B HT (l3utylatcd
stored in N IST a nd Wiley 9 libra ri es. The name and hydrox y tol ucnc), querce- tin and ru t in were used as standards.
mo lecu lar weight of the components of th e test material s Chelating capacity
were ascertained. Bi va lent transi tion me tal ions play an important ro le a s
Statistical ana lysis catalysts of oxidative processes, leading to the format ion o f
A ll exper imen ts were carried out in tripl icate and their hydroxyl radi ca ls and hyd ro peroxide d ecomposition
results are expressed as mean ± SE (n = 3) . The statisti cal reactions v ia Fenton c hemi stry (Halliwell, 1997). ln th e
analyses were perfor m ed by one way ANOVA w ith S PSS prese nce of chelating agents , the complex format ion is
13.0 so ft wa re a nd re lationships were cons idered to be disrupted wi th the resu lt that the red co lour of th e co mp lex
stati stically s ign ificant w hen P<O.OS. is dec reased. Measureme nt of colour reductio n, the refore,

30
 Evaluation of Antioxidant Activity and Phytochemical Analysis of Pteris tripartita
2oto~m3-4WJ Sw.- a critically endangered fern from South lndia

allows the es timation of the chclat ing activity of the and phosphomolybdenum as say. re spectively ( Loo et a/ ..
coexisti ng chelato r. The tran siti on metal ion. Fe2+ possess 2008).
the ability to move sing le electrons by virtue of wh ic h it can Table: Antioxidant effect s of P.1ripar1ita extracts in DP PH radical
scavenging activity, chelating capacity and phosphomolybdenum a ssays
allow the formation a nd propagation of many radical
reaction s, eve n starting with relatively non-reactive Pl:tnl CXITaC IS
OI'PH assay Metal cb elating act ivity Phns phomol_ybl.en um assay
(I'IJ/1111) (mg EDTA/g cxtra<t) ( n:tg AAE/g utrtll1)
radicals (About- Enein et al., 2003). The c helating effects
o n ferrou s io ns by various so lvent extracts of aerial parts of Hexane 133.68 13.8(bj). 42 l t7.93±1~.47

P.tripartita are s hown in Table I. Mo reover. all the extracts Ollorofonn 129.87 t3.31±0. tl 25.86%4.1 1

exh ibited the c helating abil ity to metal io ns. Howeve r, the Act.>tone 77.04 t1.78±0.24 22.56±3.34

maximum amount of chelating capaci ty was obta ined from Ethyl ace tate 116.82 t4.14±0.tl 26.50±7.02
ethanol extracts ( 15. 66 mg/ g). Geckil eta/., (2005) proved Ethanol 11 7.37 t5.66<0.06 36.56±1.72
that the ethanol ex tract is an exce ll ent metal che lator. Me thlll'lll 113.63 t4.50±c0.1 2 24.60±4.44
Further, th e samples had metal che lating ac ti v ity in the
Wotc:r 116.27 9.22.<7.03 21.26~5.0 1
o rder ofPTE>P TM> PTE A> PTH >PTC> PTA> PTW w ith the
BIIA 25.58
rates of 15 .66, 14.50, 14 .1 4. 13.80, 13.31, 11. 78 and 9.22
BHT 45.56
mg/g compared to that of EDTA (Table 1; Fig. 2). Chelatin g
Qut.."'J'CCtin 18.79
agents are effect ive as seco ndary antioxidants because they
Rutin 24.71
reduce the redox potential thereby sta bilizing the oxid ized
Values a re expr essed a s mean±SE (n ~ 3 ).
fo rm of the metal ion (Gulci n eta/. , 2007).

Fig. 2. i\letal chdating capaci~-

Recovery percent and phytochemical
18
contents
v
16 j 15.66 14.5 The yield percentage of p la nt extracts were tabulated
~
14
..

tl'lll 11
12 in table I. Plant extracts ha ve been known since ant iqu ity to
.....
"" 10
possess notab le bi o log ical activ ity, in cl uding antiox idant ,
~ 8 •sones1
fil Ei antibacte ria l and an tifunga l propert ies. There is a growing
""
E 4
interest in tbe use of natural products in the human fo od and
2
0 animal feed industries as co ns umer resistance to synthetic
PTI' PTC PTA PTEA PTE PTM PlW add iti ves in creases (Ito et a/., 1985 and 1986). Ph enol ics,
Plant extract s
flavonoids , tannin s, vita m in s, terpenoids. essen tial o il s and
extracts from the plant are natural ly occurring antiox idants;
Phosphomo lybdcnum assay
there by, tbe commercial de velopmen t o f plants as new
The quantitative antioxidant capacity o f the extracts of
so urce s of antiox idants to enhance health and food
PT were meas ured spectrophotom etri cally through t he
preservation is of present plea (Berge r, 2007; Micha lak ,
phos phomolybdenum me thod, wh ich i s based o n th e
2006: Fritz et al. , 2003; Prakash et a l., 2007; J ayapraka sha
reduction of M o(Y I) to Mo (V) by the antioxidant
et al. , 2002). The amount of total phenolics, total
compounds and the formation of g reen phosphate/ Mo(Y)
flavonoids, tannins and vitamin 'C' in P.tripartita extrac ts
complex w ith the maximal absorption at 695 nm . The
Fig. 3.Total phenolic content in different extracts of P.lripartila Sw.
t ran sfers occur at different redox potentials in the t wo
assays an d also depe nd on th e s tructure of the antioxidant 80.0

70.0 67
(Lee eta/. , 1998). Th is method had bee n used s ucc ess fully 62.43
60.0 57.39
to determine th e antiox idant activity of plant extracts v

(Konyal iog lu el a/., 20 05) and also detects ant ioxidants ~

.. 50.0
36.77
s uch as asco rb ic acid, some ph eno li cs, a-tocop hero l, and .....
"" 40.0 33.27
...""

J I
E
30.0 •scri"'l
carotenoi ds ( Prieto et a! .. 1999). Th e a ntioxidant capacity ..,
..:
20.0
of PT extracts was found to decrease in the order hexane>ethanol>ethyl

-I
10.5
10.0 3.0
acetate >chloroform>methanol>acetone>water. How eve r, the
0.0
hexa ne ext ra cts show ed hi g hest phosphomolybdenum
PHI PTC PTA PTEA PTE PTM PTW
reduction ( 117.93 g AAE/ g) among var iou s extracts (Tab le
Plant extracts
1) . Hydrogen a nd elect ron transfer from ant iox idant
a nalytes to DPPH· and Mo (VI) co mplex occ ur in the DPPH are shown in Tab le 2. The highest tota l phenolic content was

31
 Evaluation of A nt ioxidant Ac tivity and Phytochemical Analysis of Prcris rripartira
Sw.- a critically endangered fern from South India

obse rved in acetone ex tract (67.00±0 .28 GAE mg/g) The development o f a rapid a nd sens iti ve technique is
followed by methanol extract (62.43± 5.42 GAE mg/g), important fo r th e ana lys is of natural orga ni c mater ia ls.
ethanol extract (57.39± 2.8 1 GAE mg/g) , water extract w here th e ava ilable sa mple size is u sually very sma ll (Watts
(3 6. 77±0 .43 GAE mg/g) , ethy l ace tate extract (33 .2 7±0.60 and Rene de Ia Rie, 2002) . Applicat ion of modern
GAE mg/g) , chl orofor m ext ra c t ( I 0.50±0.17 GAE mg/ g) c hromatograph ic tec hni que s, for exa mple HPTLC , LC and
a nd hexa ne extrac t (3.0 1±0.1 7 GAE mg/g) , respectively GC has enab led more precise a na lysis of pharmaceu ticals o f
(F ig.3). The hi ghest va lue of pheno li c co nte nt indi ca tes that natu ra l origin (Maci ejew icz et at., 200 I ). The GC -M S study
the plant has high a ntiox idant acti v ity. of P.tripartita acetone extract h as s h ow n s i x
Among var ious extracts of PT, the ta nnin contents of phytochemica ls w hich contrib utes to the antioxida nt
acetone and methanol extracts were found to be 63.05 and activity of the plant (F ig.4; Tab le 3 ) . Six ph ytocompone nts
58.40 GAE mg/ g extract resp ec ti vely. The f lavonoid wh ic h present in the aeria l part of PT were hexadecan e
co mpou nd s co nta in hy droxy l fun c ti o na l groups, are (7 .24%), pentadeca nol ( 4. 97%), octadecanoic acid , methy l
responsible for a ntiox id ant effect in the p la nts ( Das and ester(8.08%), a- Caryophyllene(4 . 11%),5-Pheny l-2-
Pereira. 1990; Younes, 198 1). The flavonoid content of PT phenyl methyl a mino-4H -i midazo l[ I ,2c ]pyrimidin-7-one
ex tracts calcu la ted as ruti n eq ui va lent. Flavonoid s act as (3.68%) and 1,3 Bis(4-tert- Butylphenyl)2-phenylpropa n- 2-
a nti ox idan t agents by direct free radica l scave ng in g, ol (5.43%), respective ly. Th e majo r component o f acetone
transition metal che latio n a nd maintena nce o f endogenous extract of Ptripa rtita was oc tadecanoic acid, methyl ester
antioxidants su c h as th e g lu tathione a nd s uperoxi de (8.08%).
dismutase systems (Tourino et a /. , 2005). The high es t Fig . 4. Identificatio n of phytocompo unds in acet one extract of P.tripartila
fl avo noids content of PT was observed in ethyl acetate Sw. us ing GC-MS
extrac t (48.24 RE mg/ g) and c hloroform extract po ssesses
39 .02 RE mg/g ex tract. Vitamin C (ascorbic ac id) is called
an a ntiox id ant because, by donating its e lec tron s, it Jtl j
prevents other compound s from being oxidized. However, f() i
~
10 ~
by the very na ture o f thi s reaction, vitam in C itse lf is
:l
ox idized in the process. Antioxid ant effects of vitamin C w .:
=

j =~
have be en demonstrated in ma ny experiments i11 vitro
(Padayatt y e/ al. . 2003). Th e content of ascorbi c acid in PT
extracts were found to be 236.93 mg/ g (PTH), 229.00 mg/g l
(PTC), 218.96 mg/g (PTM), 2 14.86 mg/ g (PTEA), 201.03
mg/ g (PTA), 200.0 0 mg/g (PT E) and 195.13 mg/g (PTW), 10~

respective ly (Ta bl e 2). However, the highest amount of 0 ~

0
1 1i I
lji ' 1 I I ,b I I I tL I I ,b I I 12!; I I 1111 > > > 'll > >

v itamin C was observed in hexa ne ex tra ct (236.93 mg/g). Tme tll'tnt

Vitamin C a lso inhibi ts the conversion of nitrites; chem icals T able 3 . Phytocompounds ident ified in acetone extr ac t o f P.tripartita by GC-MS.
found in foods and processed meats, into nitrosom ines, Conclusion
dan gerous cancer ca using compound th at ca n lead to cance r Relt> ntion Mol ecular Peak
No. Na me of the compounds
tim c (min) weight area 0/ 0
of tb e stomach , bl add er and co lon (John Wort and Smart,
HexadccatlC 4.71 254 7.24
2004).
2 Pcntadecanol 17.68 226 4.97
Table 2 Estimation ofsolvent ex tract recove ry, tota l pbenols.llavo noids,
tannins and v itamin ' C' in P.t ripartila aerial part extracts 3 Octadecuooic ac id. metbyl ester 20.52 295 8.08

Phytochemical analysis using GC-MS 4 a Caryophylknc 21.08 204 4.11

'l'o hll phenols tlunmuids 'l'llllnln VI tamin ' <.:"'

5-Phenyl-l-phcnyl methyl 31.76 31 6 3.68
Plant e~tnets Ex.tr::u :t Yield~ (:AE n'!V~ ltf:m~/R GAF. m~~ AI\ F. II~/~
ani no41 1-imidazol[ I ,2c]pyrimidin-7 -one
'Y. e:~trart C'i:trliiCf extr a&: I l''Si r:u·t
II 1,3 32.57 400 5.43
llexanc l.l\2 3 01+0.1 7 lfi.92 H .26 (1.9iJJl14 216.9~-ld IR Bis(4-tcrt-Buiylphcnyl)2-pbenylpropan-2 -o I

ChiNuflmll 4.93 10.50•0.17 39.02.%4.2) 5.91 ill.45 ~l9.0fkt::l3.20

Ace~u~ n t8 67.o<hJJ.2S 17.48_±.1).59 hJ.05-iO...S2 :!01 .03:iA.36

As far as we know, t hi s is the firs t study conce rning th e
Erhylat.-et.:.nc 493 )3.27~.60 41(24± 1.2S 29.52±0.67 2t4.RIU2.56 fr ee radica l scavenging capac ity of P.tr ipartita Sw. In
Ethanul t7.76 57.3~.01 15.2fh.0.63 5).\J8:±-2.f<9 200.00<629 conclusion , s ix compounds were identified in acetone
62.4 l..S.42 t6.21Wl.4o 5~ .41M.I.l1 21~.90:!4.02
Mcth"n11 ll.SY extract a nd possessed good free radi cal s ca venging
W<Jtt'r 22.71 16.77>0.41 II.XO±O. II 32.QI.!.0.61 195.13..H.43
act iviti es. These effec ts can be re lated mai nly presen ce of
Each va lue in the t•ble except extrac t yields was exp ressed as m~an±SE
(n= 3 ). % "' percentage; GAE = g allic acid eq u ivalent; RE= rutin eq uiv a len t ; their ph e nolic and flavonoids con tents. Th ese resu lts are
AAE= ascorbic acid eq•tivaleot.

32
 Eva luat ion of An tioxidan t Ac t ivity and l'hytochcmical Ana lys is of Pteris tripartita
20 I O.i:fO!.n3 - 4WJ Sw.- a c r iticall y endangered fe rn fro m South India

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