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3
S T N E T N O C F O E L B A T
N O I T A R A L C E D S I S E H
T i
T N E M E G D E L W O N K C
A ii
S T N E T N O C F O E L B A
T i ii
S E L B A T F O T S I
L v ii
S E R U G I F F O T S I
L v iii
S N O I T A I V E R B B A F O T S I
L x i
K A R T S B
A x ii
T C A R T S B
A x ii i
N O I T C U D O R T N I 1 R E T P A H C
1 .
1 Overview 1
2 .
1 ProblemStatements 2
3 .
1 ScopeO fResearch 3
4 .
1 ResearchObjecitves 3
W E I V E R E R U T A R E T I L 2 R E T P A H C
1 .
2 Diplaziumesculentum(Retz. )Swatrz 4 1
.
2 1. Inrtoduciton 4
1 .
2 2. Theusageo fD .esculentum 5 1
.
2 3. Researcheds tudy on D .esculentum 6
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
4 2
.
2 Evoluitono fex rtacitonmethods 8 3
.
2 Pressu irzedHo tWate rExrtaciton( PHWE) 1 0 1
. 3 .
2 Inrtoduciton 1 0
3 .
2 2. Wate ra sgreens olvent 1 0 3
. 3 .
2 Temperature 1 1
4 . 3 .
2 Pressure 1 3
5 . 3 .
2 Advantage sanddisadvantage so fPHWE 1 4 6
. 3 .
2 Insrtumentaitono fPHWexrtactor 1 5 4
.
2 Freer adical sandulrtavioletr adiaiton 1 8 5
.
2 Anitoxidants 1 9
1 . 5 .
2 Inrtoduciton 19
2 . 5 .
2 Anitoxidan tmechanisms 2 3
6 .
2 Anitoxidan tassays 2 6
1 . 6 .
2 DPPHf reer adicals cavengingacitvtiy 2 6 2
. 6 .
2 Tota lpheno ilccontents( TPC) 2 7 3
. 6 .
2 Tota l lfavonoid scontents( TFC) 2 8 4
. 6 .
2 Theeffect so fTPCandTFConDPPHs cavenging i
v it c
a t y 2 9
7 .
2 Designo fexpe irments( DOE) 30
1 . 7 .
2 Inrtoduciton 30
2 . 7 .
2 Pirnciple so fDOE 31
3 . 7 .
2 Responses urfacemethodology 32
R E T P A H
C 3 MATERIALSANDMETHODS 3 4
3 1. Inrtoduciton 3 4
3 2. Chemical s 3 4
3 .
3 Equipments 3 4
4 .
3 Plan tmateirals 36
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
5 5
.
3 Compairsons tudyo fex rtacitonmethods 3 6 1
. 5 .
3 PHWexrtaciton 3 6
2 . 5 .
3 Autoclaving 3 6
3 . 5 .
3 Boiilng 3 7
4 . 5 .
3 Soaking 37
5 . 5 .
3 Sonicaiton 37
6 . 5 .
3 Soxhle texrtaciton 37
6 .
3 Anitoxidan tassays 3 8
1 . 6 .
3 DPPHs cavengingacitvtiy 3 8
2 . 6 .
3 Tota lpheno ilccontents( TPC) 3 8 3
. 6 .
3 Tota l lfavonoid scontents( TFC) 3 9 7
.
3 Screeningo fpressu irzedho twate rex rtacitoncondiitons 3 9 8
.
3 Opitmizaitonusingdesigno fexpeirment 4 0 9
.
3 DPPHantriadica lefifciency 4 1
0 1 .
3 Separaitono fcrudeD .esculentumPHWexrtact 4 1 1
1 .
3 Acidhydrolysi so fpape rchromatographyf raciton 4 2 2
1 .
3 HPLCanalysi so fhydrolyzedexrtact 4 2
4 R E T P A H
C RESULTSANDDISCUSSIONS 4 3 1
.
4 Compairsons tudyo fex rtacitonmethods 4 3 1
. 1 .
4 DPPHr adicals cavengingacitvtiy 4 3 2
. 1 .
4 Tota lpheno ilccontents 4 6
3 . 1 .
4 Tota l lfavonoid scontents 4 7 2
.
4 Exrtacitonyieldandpressurei nt hePHWex rtactors ystem 4 8 4 3. Screeningo fpressu irzedho twate rex rtacitoncondiitons 5 1 4 1. 3. DPPHs cavengingacitvtiy 5 1 4 .. 2 3 Tota lpheno ilccontents 5 5 4 3. 3. Tota l lfavonoid scontents 5 9 4 4. 3. Correlaitonanalysis 6 3
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
6 4
.
4 Opitmizaitonoft hreer esponse susingr esponses urface y
g o l o d o h t e
m 6 7
1 . 4 .
4 OpitmizaitonusingDesignExpetr 6 7 2
. 4 .
4 Effect so fexrtacitont emperature ,itmeandvolumeo f y
ti v it c a g n i g n e v a c s H P P D n o r e t a
w 7 1
3 . 4 .
4 Effect so fexrtacitont emperature ,itmeandvolumeo f s
t n e t n o c c il o n e h p l a t o t n o r e t a
w 7 6
5 . 4 .
4 Effect so fexrtacitont emperature ,itmeandvolumeo f s
t n e t n o c s d i o n o v a lf l a t o t n o r e t a
w 79
6 . 4 .
4 Opitmizaitonoft hreeparameter swtiht hreer esponses 8 3 5
.
4 DPPHantriadica lefifciency 8 5
6 .
4 Separaitonandi denitifcaitono fcompoundsi ncrudeexrtacts 9 2 1
. 6 .
4 Separaitonbyusingpape rchromatography( PC) 9 2 2
. 6 .
4 DPPHs cavengingactiviite so fpape rchromatogram s s
n o it c a r
f 9 2
3 . 6 .
4 Idenit ifcaitono f lfavonoid scompounds 9 4
5 R E T P A H
C CONCLUSIONSANDFUTUREWORK 9 5
5 1. Conclusion 9 5
5 2. Futurework 9 6
S E C N E R E F E
R 9 7
S N O I T A C I L B U P F O T S I
L 1 10
X I D N E P P
A 1 11
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
7
F O T S I
L T A B L E S
. O
N PAGE
1 .
2 Thes rtucture so fcompoundsi denitifedf romD .esculentum 2 2 3 1. Rangeo fparameter susedf ort heopitmizaitono fanitoxidan tacitvtiy 4 0
1 .
4 ThePHWEcondiitonsf o rhighes tanitoxidan tassays 6 7 2
.
4 ANOVAo fcen rta lcompostiedesignmodel 6 8 3
.
4 TheBox-Behnkendesignmode lwtiht hreer esponses 7 0 4
.
4 Models ummarys taitsitcf o rDPPHs cavengingacitv tiyr esponse 7 1 5
.
4 ANOVA fo rDPPH scavenging acitvtiy response on Box-Behnken n
g i s e
d 7 2
6 .
4 Models ummarys taitsitcf o rTPCr esponse 7 6 7
.
4 ANOVAo fTPCr esponsef o rBox-Behnkenmode ldesign 7 7 8
.
4 Models ummarys taitsitcsf o rTFC 7 9 9
.
4 ANOVAonTFCi nBox-Behnkendesignmodel 8 0 0
1 .
4 Kineitcbehaviou ro fquerceitnandD .esculentum 8 8 1
1 .
4 Theconcenrtaitonneededt or educeDPPHconcenrtaiton by50%and n
o it a c if i s s a l c c it e n i k r i e h
t 8 9
2 1 .
4 Antriadica lef ifciencie so fquerceitnandD .esculentum 9 1
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
8
S E R U G I F F O T S I L
. O
N PAGE
1 .
2 Photo so fD .esculentum 4
2 .
2 Soxhle texrtacitonapparatus 9
3 .
2 Schemaitcdiagramo fpressu irzedho twate rexrtactor 1 6 4
.
2 Theprototypeo fPHWexrtactor 1 7 5
.
2 Thebasicsrtuctureo f lfavonoid 2 0 6
.
2 Thes rtuctureo fDPPHf reer adical s 2 3 7
.
2 Mechanismo fDPPHr adicals cavengingbykaempferol 2 5 1
.
3 Schemaitcdiagramofr esearch lfow 3 5 1
.
4 DPPHs cavengingacitvtiyf romD .esculentumwtihdfiferen t s
d o h t e m n o it c a rt x
e 4 4
2 .
4 Tota lpheno ilccontentsf romD .esculentumwtihdfiferen texrtaciton s
d o h t e
m 4 6
3 .
4 Tota l lfavonoidcontentsf romD .esculentumwtihdfiferen texrtaciton s
d o h t e
m 4 7
4 .
4 PHWexrtact syieldandpressurei nt heexrtacitons ystem 5 0 5
.
4 DPPHr adicals cavengingacitvtiyo fD .esculentum PHWexrtact sa t 5
2
1 oCexrtacitont emperature 5 1
6 .
4 DPPHr adicals cavengingacitvtiyo fD .esculentum PHWexrtact sa t 0
5
1 oCexrtacitont emperature 5 2
7 .
4 DPPHr adicals cavengingacitvtiyo fD .esculentumPHWexrtact sa t 5
7
1 oCexrtacitont emperature 5 3
8 .
4 Tota lpheno ilcconten to fD .esculentum PHWexrtact sa t125oC e
r u t a r e p m e t n o it c a rt x
e 5 5
9 .
4 Tota lpheno ilcconten to fD .esculentum PHWexrtact sa t150oC e
r u t a r e p m e t n o it c a rt x
e 5 6
0 1 .
4 Tota lpheno ilcconten to fD .esculentum PHWexrtact sa t175oC e
r u t a r e p m e t n o it c a rt x
e 5 7
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
9 1
1 .
4 Tota l lfavonoid sconten to fD .esculentum PHWexrtact sa t125oC e
r u t a r e p m e t n o it c a rt x
e 5 9
1 .
4 2 Tota l lfavonoid sconten to fD .esculentum PHWexrtact sa t150oC e
r u t a r e p m e t n o it c a rt x
e 6 0
3 1 .
4 Tota l lfavonoid sconten to fD .esculentum PHWexrtact sa t175oC e
r u t a r e p m e t n o it c a rt x
e 6 1
4 1 .
4 Thecorrelaitono fDPPHs cavengingacitvtiyandt ota lphenolic f
o s t c a rt x e W H P m o r f s t n e t n o
c D .esculentum 6 3
5 1 .
4 Thecorrelaitono fDPPHs cavengingacitvtiyandt ota l lfavonoid s f
o s t c a rt x e W H P m o r f s t n e t n o
c D .esculentum 6 4
6 1 .
4 Thecorrelaitonoft ota lphenoilccontent sandt ota l lfavonoid s W
H P m o r f s t n e t n o
c exrtact so fD .esculentum 6 6
7 1 .
4 Thei nteracitono fexrtacitont emperatureandvolumeo fwate reffect s n o it c a rt x e s e t u n i m 0 1 t a e s n o p s e r y ti v it c a g n i g n e v a c s H P P D n
o 7 4
8 1 .
4 Thei nteracitono fexrtacitont emperatureandvolumeo fwate reffect s c
s H P P D n
o avengingacitvtiyr esponsea t20minute sexrtaciton 7 4 9
1 .
4 Thei nteracitono fexrtacitont emperatureandvolumeo fwate reffect s n o it c a rt x e s e t u n i m 0 3 t a e s n o p s e r y ti v it c a g n i g n e v a c s H P P D n
o 7 5
0 2 .
4 Thecontou rplo to fDPPHs cavengingacitvtiyr esponsewtih n
o it i d n o c n o it c a rt x e m u m it p o t a n o it c i d e r
p 7 5
1 2 .
4 Thei nteracitono fexrtacitont emperatureandvolumeo fwate reffect s e
m it n o it c a rt x e s e t u n i m 0 2 t a e s n o p s e r C P T n
o 7 8
2 2 .
4 Thei nteracitono fexrtaciton itmeandt emperaturea t50mLwate r e
m u l o
v 7 8
3 2 .
4 Thei nteracitono fexrtaciton itmeandvolumeo fwate reffect son 5
2 1 t a C F
T oC 8 1
4 2 .
4 Thei nteracitono fexrtaciton itmeandvolumeo fwate reffect son 0
5 1 t a C F
T oC 8 1
5 2 .
4 Thei nteracitono fexrtaciton itmeandvolumeo fwate reffect son t
a C F
T 175oC 8 2
6 2 .
4 Thecontou rplotf o ropitmumTFCr esponse 8 2 7
2 .
4 Thedesriablitiyo fopitmizedcondiitonsf o rhighes tanitoxidan t y
ti v it c
a 8 4
8 2 .
4 Kineitc so fquerceitna tdfiferen tconcenrtaitons 8 5
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
0 1 9
2 .
4 Kineitc so fD .esculentuma tdfiferen tconcenrtaitons 8 6 0
3 .
4 Determinaitonoft he itmeneededt or eacht hes teadys tatet oEC50 fo n
it e c r e u
q 9 0
1 3 .
4 Determinaitonoft he itmeneededt or eacht hes teadys tatet oEC50 9 0 2
3 .
4 DPPHs cavengingacitviite so fpape rchromatographys eparaitonf o r e
l u c s e .
D ntumcrudeexrtacts 9 3
3 3 .
4 TheHPLC-UVchromatogram so fcrudeexrtac to fD .esculentum 9 4
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
1 1
S N O I T A I V E R B B A F O T S I L
P
D HP 2 -,2 diphenyl-1-picrylhydrazy l C
P
T Tota lpheno ilccontents C
F
T Tota l lfavonoid scontents E
O
D Designo fexpeirments E
W H P
W H P
r e t a w t o h d e z ir u s s e r
P exrtaciton
r e t a w t o h d e z ir u s s e r P W
A
B Butanol-aceitcacid-water C
P Pape rchromatography L
C
H Hydrochlo ircacid l
C l A 3
M E S
e d ir o l h c ir t m u i n i m u l A
t n e m e r u s a e m r o r r e d r a d n a t S
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
2 1
i r i
C - C i r i A n it o k is d a it f D a r i p a d a E k t r a k D i p l a z i u m e s c u l e n t u m n a ij a K : n a n a k e t r e B s a n a P r i A n a k a r t s e k g n e P n a k a n u g g n e M
n a m u m it p o g n e P
K A R T S B A
n a n a k e tr e b s a n a p r i a n a k a rt s k e g n e
P (PHWE )telah djialankan untuk mengurangkan k
i n a g r o t u r a l e p n a a n u g g n e
p bertoksik untuk mengeksrtak sebaitan bioakit f dar i m
u t n e l u c s e m u i z a l p i
D . Perbandingan antara kaedah-kaedah pengeksrtakan telah n
a k u k a li
d dan keputusan menunjukkan PHWE mempunya ikeupayaan penyingkrian s
a b e b l a k i d a
r , 22 -, diphenyl-1-picrylhydrazyl (DPPH) (70% )dan jumlah kandungan h
c e t a c g µ 3 8 . 8 1 1 ( k il o n e
f i en s tara / mg eksrtak keirng )yang itngg iberbanding dengan p
, n a f a l k o t u a g n e
p endidihan , perendaman dan sonikasi dan pengeksrtakan Soxhlet . i
a y n u p m e m n a m a d n e r e p n a k a rt s k e g n e
P jumlah kandungan lfavanoid keseluruhan i
g g n it r e
t (28.03µgr uitnsetara/ mgeksrtak keirng) .Untukmendapatkan eksrtak PHWE ,
k i a b r e t g n a y a d i s k o it n a i ti v it k a n a g n e
d kaedah permukaan gerak balas dengan reka x
o B k u t n e
b -Behnken t elah dliakukan .Keputusanmenunjukkan keadaanopitmum yang n
a k u lr e p i
d ialahpadasuhu 175oC ,21mini tmasapengeksrtakandengan 50mLai ryang l
e p m a s g 2 a d a p e k n a k h a b m a ti
d D. esculentum yang telah dike irngkan. Keupayaan k
a rt s k e k u t n u H P P D l a k i d a ri t n
a D. esculentummenunjukkannlia iEC50ialah1241.14µg /mL dan masa untuk mencapa ikestablian (TEC50 )ialah 79.83 minti .Maka ,eksrtak
m a l a d t a b m a l i a g a b e s n a k ir o g e t a k i
d penyingkrianradika lbebasDPPH .Flavonoid yang C
L P H i u l a l e m i t s a p l a n e k i d h a l e t k a rt s k e m a l a d i d g n u d n a k r e
t iatiu querceitn dan
. n it e c ir y m
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
3 1
f o s e it r e p o r P e v it a d i x o x it n
A D i p l a iz u m e s c u l e n t u m E x t r a c t B y U s i n g y
d u t S n o it a z i m it p O : r o t c a r t x E r e t a W t o H d e z i r u s s e r P
T C A R T S B A
n o it c a rt x e r e t a w t o h d e z ir u s s e r
P (PHWE )wasi mplementedi nattemptt or educet heuse m
o r f s d n u o p m o c e v it c a o i b g n it c a rt x e n i t n e v l o s c i n a g r o c i x o t f
o Diplaziumesculentum .
x e n o s ir a p m o
C rtaciton method swere done and t he resutl sshows t hat t he PHWE had r
e h g i
h 22 -, diphenyl-1-picrylhydrazy l(DPPH) free radica lscavenging acitvtiy (70% ) o t d e r a p m o c ) s e l p m a s y r d g m /t n e l a v i u q e n i h c e t a c g µ 3 8 . 8 1 1 ( s t n e t n o c c il o n e h p l a t o t d n
aautoclaving ,boiilng ,soaking,sonicaitngandSoxhle textraciton .Soakingexrtaciton had . )t c a rt x e y r d g m /t n e l a v i u q e n it u r g µ 3 0 . 8 2 ( s t n e t n o c s d i o n o v a lf l a t o t t s e h g i h e h
t By
x o B g n i y l p p
a -Behnken design o f response surface methodology , the op itmized o
it n a t s e b e h t r o f n o it i d n o
c xidan tacitvtiyo fPHWEwa sa t175oC ,21minute sexrtaciton d
n u o r g d e ir d f o g 2 o t d e d d a e m u l o v r e t a w L m 0 5 d n a e m
it D .esculentum .TheDPPH
e h t e r e h w d e z y l a n a s a w y c n e i c if f e l a c i d a ri t n
a D .esculentumexrtact shadEC50valueo f L
m / g µ 4 1 . 1 4 2
1 and itme to reach steady state (TEC50 )wa s79.83 minutes .Thus ,the , C L P H h g u o r h T . s l a c i d a r e e r f H P P D e h t g n i g n e v a c s n i w o l s s a d e z ir o g e t a c s a w t c a rt x
ehei denitifed lfavonoidsi nt hecrudeex rtact swerequerceitnandmy irceitn. t
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
4 1
1 R E T P A H C
N O I T C U D O R T N I
2 .
1 Overview
o t d e s u e r a h c i h w s t c a rt x e r o s l a ir e t a m t n a l p y n a s a d e n if e d e r a s e n i c i d e m l a b r e H
l a c o l s e s s o p d lr o w e h t t u o h g u o r h t s e r u tl u c l l A . h tl a e h s
’ n a m e v o r p m i o t r o s s e n ll i t a e rt
0 0 0 2 n a h t e r o m e r a e r e h T . ) 0 0 0 2 , r e t s o F
&
e ti h W ( s e n i c i d e m s e v it a n r e tl a s a s t n a l p
( h a m it a F p i c a k s a h c u s e u l a v l a n i c i d e m n i h c ir y l h g i h a i s y a l a M n i s t n a l
p Labisia
a il i m u
p ) ,pegaga (Centella asiaitca) ,misa ikucing (Orthosiphon stamineus )and ma s (
k e t o
c Ficu sdetloidea)( Bernama ,2009) .
e h t e r a s l a c o l g n o m a e g a s u s l a b r e h f o s n o s a e r e h
T avaliablitiy and lowe rcos t
s i e c r u o s l a r u t a n f o n o i s s e r p m i e h t s u l p , p u o r g e m o c n i r e w o l e h t y b e l b a d r o f f a h c i h w
e v it c a e h t h g u o h t n e v E . s e n i c i d e m n r e d o m e z i s e h t n y s y ll a c i m e h c o t d e r a p m o c e f a s
0 6 n a h t e r o m d e t u b ir t n o c e v a h s t n a l p m o r f d e t c a rt x e s d n u o p m o
c -70% in the modern
e h t y d u t s o t t u o d e ir r a c e b d l u o h s h c r a e s e r r e h tr u f ; s t n e m p o l e v e d s e n i c i d e m
s a b A ( y d e m e r s a s t c e f f e e d i s d n a y ti c i x o t e h t ,s s e n e v it c e f f
e -Husin ,2001 .)
m u i z a l p i
D esculentum i samong t housand so fPteirdophyte sspecie swheret hei r e
r a s d n o r
f eaten and beileves t o have an itmalaira leffects , rteatj aundice ,consitpaiton , ,
e h c a r a
e to rteatf ever ,measle sand dermaitits( Baa se tal,. 1990 ;Kala ,2005 ;Roostiae t .l
a ,2008) .Loca lcommun tiyi nMalaysiacollectst heediblef ernst os elli nt hemarke to r n a c d n a s d l e if t e w , s k n a b r e v ir d e d a h s y ll a it r a p t a w o r g y ll a m r o n y e h T . e m o h t a k o o c
g e w o N ( s e ti s r e ir d n o d n u o f e b o s l
a e ta.l ,2003) .
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
5 1
s e n i c i d e m e v it a n r e tl a f o n o it a r a p e r p e h t r o f s l a ir e t a m t n a l p f o n o it c a rt x E
v it c e l e s w o l , d e t c a rt o r p e r e w y l s u o i v e r
p tiy and low exrtaciton yield ,and also requrie e s e h t e m o c r e v o s a h s s e c o r p n o it c a rt x e r e w e N . s t n e v l o s c i n a g r o c i x o t f o s t n u o m a e g u h
n o it c a rt x e r e t a w l a c it ir c b u s d n a n o it c a rt x e d i u lf l a c it ir c r e p u s s a h c u s s k c a b w a r d
o r e r r e H
( e ta.l ,2006) .Atlernaitve green solvent i n replacing organic solvent ssuch a s y l h g i h e r a s d n u o p m o c l a r u t a n g n it c a rt x e n i d e s u y l n o m m o c , r e t a w d n a e d i x o i d n o b r a c
n o n h ti w r e t a W . s y a d a w o n e l b a r u o v a
f -toxictiyand non-lfammablecharacteirsitc smake m
r e t n i s t c a rt x e t n a l p e h t e l d n a h o t y s a e t
i so fprocessing ,analyzing and produciton . , o p s e r C
&
s o lr a C ( h c r a e s e r t n e s e r p e h t n i t n e v l o s s a n e s o h c n e e b s a h r e t a w , e r o f e r e h T
. ) 0 8 9 1 , s n i k n e J
&
k n i y e o n S ; 5 0 0 2
2 .
1 Problems tatements
1 . 2 .
1 Localcommuniitesi n poo rcountire suseD .esculentum to rteatleprosy ,askin y
b d e s u a c e s a e s i
d Mycobacterium leprae (Sarke r& Hossain ,2009) .Previou s o n d n a s l a m i n a n o s t n a l p e h t g n i d e e f f o m o t p m y s y ti c i x o t s w o h s d e h c r a e s e r
i h s n a v m o S ( s t c a rt x e t n a l p e h t f o y ti v it c a l a i b o r c i m it n
a e lta. ,2006 ;Kankamo le t
l
a. ,2006 ;Nanasomba t&Teckchuen ,2009) .Doest heanitoxidan tacitvtiyoft he
? e s a e s i d n i k s e h t g n it a e rt n i d i a s t c a rt x e
2 . 2 .
1 Theorganicsolvent saret oxic ,expensiveandno tenvrionmentf irendlyi nt erm s o
r e r r e H ( l a s o p s i d e t s a w d n a n o it c a rt x e f
o e . lta . 2006) . So , wha t are the
? s t n e v l o s e v it a n r e tl a
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
6 1 3
.
1 Scopeo fresearch
e h
T research focuse son the ex rtaciton o fD .esculentum acitve compound s d n a s d o h t e m n o it c a rt x e r e t a w s u o ir a V . y ti v it c a l a c i d a ri t n a H P P D n i e t u b ir t n o c h c i h w
o c r o f e d a m e r e w n o it c a rt x e t e l h x o
S mpairson .Pressu irzed ho twate rexrtaciton swere e r e w s t c a rt x e l l A . e d a m s a w n o it a z i m it p o d n a s r e t e m a r a p f o e g n a r a n i t u o d e ir r a c
l a t o t , y ti v it c a g n i g n e v a c s H P P D f o e s ir p m o c h c i h w s i s y l a n a t n a d i x o it n a o t d e t c e j b u s
o c s d i o n o v a lf l a t o t d n a s t n e t n o c c il o n e h
p ntents .The op itmized exrtac twa stested on e r e w t c a rt x e e d u r c e h t n i s d n u o p m o c e v it c a o i b e h t d n a y c n e i c if f e l a c i d a ri t n a H P P D
. d e if it n e d i d n a d e if ir u p
4 .
1 Researchobjecitves
: w o l e b d e z i m e ti e r a s e v it c e j b o e h T
.i To determine the bes t an itoxidan t acitvtiy o f D . esculentum from vairou s .
s d o h t e m n o it c a rt x e .i
i To screen the anitoxidan tacitvtiy o fpressu irzed ho twate rexrtaciton from .D m
u t n e l u c s
e a tvairou scondiiton. .i
ii To opitmized the condiiton o f pressu irzed ho t wate r exrtaciton from D . m
u t n e l u c s
e .
. v
i Tos tudytheantriadica lefifciencyo fopitmizedD .esculentumexrtact . .
v Toi dentfiyt he lfavonoid sgroupi nt heopitmizedD .esculentumexrtact .s
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
7 1
2 R E T P A H C
W E I V E R E R U T A R E T I L
1 .
2 Dipla izumesculentum(Retz. )Swartz
1 . 1 .
2 Introduciton
m u t n e l u c s e .
D (Retz. )Swatrz (Fig .2.1 )also known a sAthy irum esculentum s a n w o n k ( s n r e f e l b a t e g e v s i h T . e a e c a ir y h t A f o y li m a f e h t n i s i l e p o C ) . z t e R
( pucuk
u k a
p in Malaysia )grow swlidly a tmois to rwe tarea ssuch a sparitally shaded irve r i
o l p i d e h T . d n u o r g d a e t s e m o h r o s k n a
b d plan toccur sthroughou tAsia and Oceania ) ij i F d n a n a p a J , h s e d a l g n a B , a e n i u G w e N a u p a P , d n a li a h T , s e n i h p il i h P , a i s e n o d n I (
g e w o N
( e tal,.2003 ;Sarke r&Hossain ,2009 ;Takamiyae ta.l ,1999 .)
f o o t o h P : 1 . 2 e r u g i
F D .esculentum
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
8 1 .
D esculentumusuallygrowsi nal argebunchwtihup irghts temandcangrowup r
e h t a e f e h T . h g i h r e t e m e n o o
t -ilkef rond sarecu lred andcanbedividedt ot woort hree w
a s e r a f a e l e h T . s e m
it -ilkeshapeont heedgeandsharp att he itp whichi sabou teigh t d
n a g n o l r e t e m it n e
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2 t u o b a n i y ti d i m u h h g i h t a s e r o p s m o r f n w o r
g o C (French ,1986) .D .esculentum
:) e s a b a t a D t n a l P , A D S U ( w o ll o f s a s i e e rt y n e g o l y h p
m o d g n i
K Plantae-Plants m
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o i s i v i
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2 . 1 .
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e h t ll a h g u o r h t s n r e f e l b i d e s i h t f o s t o o h s g n u o y e h t t c e ll o c s e it i n u m m o c l a c o L
d e ir f e r e w s d n o r f g n u o y e h T . t e k r a m e h t n i d l o s e b o t r o n o it p m u s n o c e m o h r o f r a e y
h g u o h t h a b a S , e g n a R r e k c o r C t a e v il o h w e l p o e p g n o m a s e i v o h c n a d n a c il r a g h ti w
s a d e k o o
c masakl emakinPeninsula rMalaysia( Nowege ta.l ,2003 .)I nBangladesh,t he o s l a s i t I . s t n e i d e r g n i y r r u c s a r o s e h s if , s p m ir h s h ti w d e k o o c e r a s d n o r f g n u o y
s a s i e g a s u r e h t o n A . s e s a e s i d l a n i m o d b a r o n o it a p it s n o c t a e rt o t e v it a x a l a s a d e m u s n o c
i k s r o f y d e m e
r n disease ssuch a sleprosy ,a contagiou sbacte ira disease (Sarke r&
.) 9 0 0 2 , n i a s s o H
g n ir u d g n i d d e b l a m i n a s a d e s u e b o t d e r u o v a f e r o m s i s d n o r f d e ir d , r e v e w o H
a r s i M ( a i d n I n i e n o z e v r e s e R e r e h p s o i B i v e D a d n a N t a s r e g a ll i v e h t y b s r e t n i
w e ta.l ,
e h T . ) 8 0 0
2 rhizome spatr sareplacedi n t hegrain storest oavoid f romi nsect sandpest s
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
9 1
, a y a l a m i H f o e l p o e p e h t y b s e l b a t e g e v s a n e t a e e r a s e ir t n u o c t s o m n i s a s d n o r f e h t d n a
i t e r p U ( a i d n
I e tal,.2009).
3 . 1 .
2 Researcheds tudyonD .esculentum
o h c r a e s e r o n s i e r e h
T nseverelyt oxiccompound exrtacted f rom D .esculentum e r e w ) e d i s o c y l g c i n e g o n i c r a c ( e d i s o li u q a t p f o s n o it a rt n e c n o c e h T . e t a d s i h t o t d e h s il b u p
w o l d n a h g i h e h t n o d e s a b a i d n I n i d e t a g it s e v n
i -atltiudearea .Mos tsample sshowed no n
i e d i s o li u q a t
p D .esculentumwtihanexcepitonf o rasamplef romhigh-atltiudeareao f li
s r a
H -Ganort iwhichcontained19mg/kgvegetables(Somvansh ie tal. ,2006) . g
n i d e e f , y ll a c i n o r
I D .esculentum causest oxictiy symptoms t o rabb ti sand died e
n o it n e m s A . g n i d e e f f o s y a d 1 1 r e tf
a dbySethe ta.l,( 2000 )“Theeffect swerer educed e v it a r e n e g e d h ti w s r e t e m a r a p l a c i g o l o t a m e a h n i s n o it a n r e tl a , s t h g i e w n a g r o d n a y d o b
s e r b if l a i d r a c o y m d e t p u r s i d d n a s n a g r o l a r e c s i v d n a n i a r b n i s e g a h r r o m e a h d n a s e g n a h c
d e e f , s d n a h r e h t o e h t n O .
” tr a e h n
i ing frozen D .esculentum on rat scaused mlid l a c i g o l o h t a p o c i n il c f o e p y t e t a r e d o m h ti w y ti l a t a f d e g a r u o c n e t u b s t c e f f e c i g o l o h t a p
.) 4 0 0 2 , i h s n a v m o S
&
r a w g n a G ( s g i p a e n i u g n i s t c e f f e
f o s t n e t n o c e r u t s i o m e h
T D .esculentumi shigh upt o90-93%.I n100g o fedible f
o n o it r o
p D .esculentum contained 88k Jenergy ,3.6g o fprotein ,4.4mg o f rion and .)
6 8 9 1 , h c n e r F ( C n i m a ti V f o g m 5
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c 0g )
β n i h g i h t u
b -carotene( 74.04ppm) ,off reshvegetables .
, m a n t e i V n i s e l b a t e g e v d li w n o h c r a e s e r e h t y b d e tr o p p u
S D .esculentum
y f a e l n e e r g e c n i s w o l s i h c i h w e l p m a s f o g 0 0 1 / e t a l o f g µ 7 2 y l n o d e n i a t n o c s t c a rt x e
o f h g i h n i a t n o c o t d e v e il e b e r a s e l b a t e g e
v latecontents .Vegetable sarenatura lsource s
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
0 2
e t a l o f f o g n i k c a l d n a m a n t e i V s a h c u s y rt n u o c r o o p n i s r e g a ll i v e h t o t s t n e ir t u n f o
r e c n a c d n a r a l u c s a v o i d r a c f o k s ir e h t e s a e r c n i d l u o c y r a t e i d n a m u h n i n o it a rt n e c n o c
e l g O ( s e s a e s i
d e t a.l ,2001) . Study in Southern Thaliand ha s found ou t tha t D . m
u t n e l u c s
e contain sheat-stableandheat-lablieanttihiamin .Therei sonly rtaceamoun t h
s e r f n i d e n i a t n o c e n i m a i h t f
o D .esculentum(Taungbodh tiham ,1995) . e
h
T D .esculentum exrtact shave no anitmicrobia lacitviite swhere t hereare no l
o m a k n a K ( d e t s e t s n i a rt s l a ir e t c a b l a r e v e s f o n o it i b i h n i h t w o r
g e ta.l ,2006 ;Nanasomba t
n o s g n i d n if e v it a g e n e h t e ti p s e D . ) 9 0 0 2 , n e u h c k c e T
& D .esculentum ,tii ss tli lbeenuse d
a rt y b ) 9 0 0 2 , n i a s s o H
&
r e k r a S ( s e s a e s i d t a e rt o t y d e m e r s
a iitona l medicine
e h t t a e rt t n a l p e h t s e o d w o h n o t u o d n if o t s u o e g a t n a v d a e b d l u o w t i ,s u h T . s r e n o it it c a r p
.s e s a e s i d
4 . 1 .
2 Fern sa smedicines
. d lr o w e h t t u o h g u o r h t s e i c e p s s n r e f r o s e t y h p o d ir e t P f o s d n a s u o h t e r a e r e h T
s e i d e m e r l a n o it i d a rt s a d e s u e r a e m o
S to rtea tvairou sdiseases .Lygodium japonicum s a h s s e n ll i s u o ir a v g n it a e rt n i s e n i c i d e m e s e n i h C l a n o it i d a rt d e u l a v f o e n o , ).
b n u h T (
d n a s l a c i d a r e e r f H P P D , n o it a d i x o r e p e m o s o p il t s n i a g a s e it i v it c a t n a d i x o it n a g n o rt s
i L ( g n i g n e v a c s e d i x o r e p n e g o r d y
h e tal,. 2006) .Fou rAdiantum fern sspecie sshowed t
s n i a g a s e it i v it c a l a i b o r c i m it n a t n e t o
p Escherichia coil , Trichophyton rubrum and s
u e r r e t s u ll i g r e p s
A ha ssuppo tr t he use of t hese fernsi n rtadiitona l rteatment samong h
g n i S ( a i d n I n i s l a c o
l e tal,.2008 .)
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
1 2 2
.
2 Evolu itono fextracitonmethods
r e h t o n a o t n i e c n a t s b u s e n o m o r f s t n e n o p m o c g n i v o m e r f o s s e c o r p a s i n o it c a rt x E
. ) 9 9 9 1 , n o s a M ( s a g r o t n e v l o s g n i s
u D .esculentumweref reshlyused ,boliedwtihwate r n
a e n o t e c a , e n a x e h ( s t n e v l o s c i n a g r o h ti w d e k a o s r
o d alcohols ) p iro r to analyse s
n a w a r I
( e tal,. 2006 ;Taungbodhtiham ,1995 ;Rahma te tal,. 2003 ;Somvansh ie tal,.
l o m a k n a K ; 6 0 0
2 e tal,. 2006 ;Nanasomba t& Teckchuen ,2009) .There i sno recen t e
t a d s i h t o t p u t n a l p s i h t n o d e il p p a n e e b s a h d o h t e m n o it c a rt x
e .
t c a rt x e e t a i d e m m i d n a t s a f d e s u y lt s o m s r e n o it it c a r p l a n i c i d e m l a b r e H
d e s u a c g n it a e H . r e t a w h ti w d e li o b e r a s l a ir e t a m t n a l p e h t e r e h w s i n o i s u f n I . n o it a r a p e r p
e n a r b m e m e h t f o t u o e s u f fi d o t s t n e u ti t s n o c s t n a l p g n i w o ll a d n a p x e o t ll a w l l e c t n a l p e h t
.l l e
c Maceraiton i samethod o fexrtaciton which soflty break sup t heplan tmateira lby m o o r t a n o it a ti g a h ti w ) s k e e w ( e m it g n o l a r o f r e t a w s a h c u s d i u q il a n i g n i k a o s
. ) 6 0 0 2 , n a m a a R ; 0 0 0 2 , r e h c e e B
&
n e k r e M ; 9 9 9 1 , n o s a M ( e r u t a r e p m e t
a rt x e t e l h x o S l a n o it n e v n o
C citon i sa standard and well-estabilshed exrtaciton n o it c a rt x e r e t a l e h t o t n o s ir a p m o c a s a o g a s e d a c e d e c n i s d e s u n e e b s a h t a h t e u q i n h c e t
e h t n i t n e v l o s d e t a e h e h t e r e h w s s e c o r p n o it a ll it s i d n o s i e l p i c n ir p c i s a b e h T . s d o h t e m
t d r a w p u w o lf k s a lf s
’t n e v l o
s o t hecondenser( Fig .2.2) .Thecondensedsolventi s iflled e h t o t n i k c a b s d a o l n u d n a l e v e l m u m i x a m s e h c a e r t i l it n u r o t c a rt x e t e l h x o S e h t n i
l li w s e t u l o s e h t e li h w l li t s i d e r l li w t n e v l o s e h T . s t c a rt x e e h t g n i y r r a c k s a lf t n e v l o s
k s a lf t n e v l o s e h t n i n i a m e
r . tIi scheap and simpleex rtaciton method sbu trequriel ong .)
6 0 0 2 , r e ll e W
&
g n a W ( t n e v l o s f o t n u o m a h g i h s u l p e m it n o it c a rt x e
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2 2
s u t a r a p p a n o it c a rt x e t e l h x o S : 2 . 2 e r u g i F
n o it a c i n o s a rt l
U -assisted exrtaciton ha sbeen appiled in solven texrtaciton fo r e
tt e
b ryield o fexrtacts .Thep irnciple ilesi son t hesoundwavet hati nducecavtiaitona l e h t , s ll a w l l e c e h t r a e n e s p a ll o c s e l b b u b e h t n e h W . h t a b r e t a w n i s e l b b u b g n i m r o f
e d i s t u o m e h t t r o p s n a rt d n a s d n u o p m o c e h t g n i v l o s s i d s ll a w e h t o t n i n e v ir d s i t n e v l o s
s a M
( on ,1999) .
g n it n e m e l p m i ) E F S ( n o it c a rt x e d i u lf l a c it ir c r e p u s s i d o h t e m n o it c a rt x e t n e c e R
s i e d i x o i d n o b r a C . s t n i o p l a c it ir c o t p u e r u s s e r p d n a e r u t a r e p m e t h g i h t a t n e v l o s
1 3 ( e r u t a r e p m e t l a c it ir c w o l , y ti c i x o t w o l o t e u d d e z il it u y l s u o m r o n
e oC )and pressure
, ) E L P ( n o it c a rt x e d i u q il d e z ir u s s e r p e h t s i E F S n a h t e l p i c n ir p r e l p m i S . ) r a b 2 7 (
l a c it ir c d n a g n il i o b f o e g n a r e h t t a ) l o n a h t e . g . e ( s t n e v l o s c i n a g r o g n it n e m e l p m i
) E W H P ( n o it c a rt x e r e t a w t o h d e z ir u s s e r p , E L P f o t a h t o t t p e c n o c r a li m i S . e r u t a r e p m e t
r g a s
i eenatlernaitvemethodduet of easiblitiyusedo fwate rass olvent( Turner ,2006 .)
© T hi s it em is p ro te ct ed by o rig in al co py rig ht
3 2 3
.
2 Pressurizedho twaterextraciton( PHWE)
1 . 3 .
2 Introduciton
, r e t a w l a c it ir c b u s s a n w o n k o s l a n o it c a rt x e ) W H P ( r e t a w t o h d e z ir u s s e r P
s e r p r o n o it c a rt x e r e t a w d e t a e h r e p u
s su irzedl owpolartiywaterr eferst ot hewate ratt he 0
0 1 s i h c i h w e r u t a r e p m e t l a c it ir c d n a g n il i o b e h t n e e w t e b e r u t a r e p m e
t oC and 374oC .
f o e s u e h t n i s r e h c r a e s e r f o t s e r e t n i t a e r g s g n ir b t n e v l o s s a r e t a w f o n o it a t n e m e l p m i e h T
e if l a r e v e s n i n o it c a rt x e W H
P ld so fappilcaitons( Smtih ,2002) .
e t s a w d il o s l a t n e m n o ri v n e d n a s li o s n o d e il p p a n e e b e v a h s n o it c a rt x e W H P
, ) s H A P ( s n o b r a c o r d y h c it a m o r a y l o p f o n o it c a rt x e e h t e l p m a x e r o F . t n e m t a e rt
o f e u d i s e r s e d i c it s e p d n a s n a r u h p o z n e b , ) s B C P ( s l y n e h p i b d e t a n ir o l h c y l o
p r the
f o t s a v a n e e b s a h e r e h t , y lt n e c e R . e t s a w l a p i c i n u m d n a s li o s l a ir t s u d n i f o t n e m t a e rt
t n a l p m o r f s d n u o p m o c l a r u t a n f o n o it c a rt x e n o E W H P f o n o it a c il p p a n i t s e r e t n i
o r e r r e H ( E W H P f o n o it a c il p p a n o e n o d n e e b d a h s w e i v e r w e f A .s e c ir t a
m e tal,.2006 ;
Pronyk&Mazza ,2009 ;Smtih ,2002 .)
2 . 3 .
2 Watera sgreens olvent
n o n , s s e l e t s a t t s o m l a , s s e lr u o d o s i r e t a
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&
k n i y e o n S ( t n e v l o s n o it c a rt x e y l d n e ir f l a t n e m n o ri v n e
e r e h t d n a e l b a li a v a y li s a
e forecould reducet hecost sfor t heproduciton o fcommercia l e t s a w d n a l a s o p s i d ’ s t n e v l o s c i n a g r o e h t r o f t s o c e h t o s l a d n a s d n u o p m o c l a r u t a n
n e e r g e v it a n r e tl a n a s i r e t a w , s ti f e n e b e s e h t o t e u D .) 5 0 0 2 , o p s e r C
&
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