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UNIVERSITI PUTRA MALAYSIA
THE HYPOGLYCAEMIC AND CHEMICAL STUDIES OF PARKIA SPECIOSA, Hassk
FATHAIYA BTE JAMALUDIN
FSMB 1991 1
THE HYPOGLYCAEMIC AND CHEMICAL STUDIES OF PARKIA SPECIOSA, Has sk
BY
FATHAIYA ETE JN1ALUDIN
The s i s s ubmitted in Ful f i lment o f the Requi rements for the De gree o f Ma s te r of Sc ience in the Faculty
of Food Sc ience and B iotechnol ogy Unive rsitj Pe rtan ian Ma l a y s i a
Jul y , 1991
ACKNOWLEDGEMENTS
I would l ike to take th i s opportunity t o e xpress ny
sinc e re gratitude t o my much r e s p e c t e d supe rv i s o r , D r . Suha i l a
Mohamed and my co-supervi�or, Assoc. Prnf. Dr. Md. Nordin Hj.
La j i s f o r the i r pat.ience, guid ance , encollragement, suggestions
und di scus s ions that t.hey have given me LhroughouL the course
o f th i s re s e arch wo rk .
I am also indebted to Dr. Yazi d Abd. Manaf for his
guidance. d i s c u s s ions and advice on the s tat i s t ical aspec t of
the wor k . My appreciation i s als o dedic a t e d to Mr. Atan Mohd .
Sa r ip , D r . John B . B remner from Unive r s ity of Tasmania and D r .
Lindsay T. Byrne from Unive r s i t y o f We s t e rn Aus t ra l ia for
I
performing the spectra data of the samples. A speci�l thank
a l s o goe s t o Dr . Mawa rdi Rahmani and D r . Asp o l l ah Hj . Suka r i
for the i r help and d i s c u s s ions on the dete rmina t ion o f the
chemical s t ruc ture wo rk; to the Inf o rmat ion Document a ry S y s tem
and Malaysian Med i c a l Institute for p roviding information on
d iabetes mellitus c a s e s .
I am also grateful t o Mr . Z a ina l Abidin Jamin and othe r
staff. from the Depar tmen t of Vet.erilullY Medicine tor t.heir
va luab l e ass istance on the b io a s s ay aspect; to Mr. Zainuddin
S amad i , Mr. Zainal Zahari Zakaria, Mr. Zainal Abidin Kassim,
i i i
Mr . Narz a r i Ahmad f rom the Depar tmen t o f Chemis t ry for the i r
a s s i s t ance and � �ope ra t ion o n the chemic a l re s e a rch ; t o Mis s
Azu r a Ab d . Ja lil, Mi s s Z ak iah Azizi lor lheir help parLicul arly
in hand l ing rat s ; and a l s o t o Mrs . Asma rani Abdu l lah and Mr .
Abd. Hadi I sma il for typing my the s i s .
La s t
thank t o
but not le a s t , 1 would l ike to expre s s my
my husb and Al fakh r i Albabj i , and my three
sp e c ia l
chil dren
l1uhanunad H a s i f , S ak inah and S a rah for the i r l ove , s uppo r t ,
sac r i fice s and unde r s t anding .
Fina l l y , I wish to thank MPKSN and IFS fo r funding the
re s e a rch ; and Un ive r s iti P e r t anian Ma lay s ia for f inanc i a l
s uppo r t through Graduate As s i s t an t ship P rogramme .
iv
TABLE OF CONTENTS Page
ACKNOWLEDGEMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii
LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . viii
LIST OF FIGURES . . . . . . . . . " . . . . . . . . . . . . . . . . . . . . . . x
LIST OF SCHEMES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii
LIST OF ABBREVIATIONS • • • • • • • • • t • • • • • • , • • • • • • • • • • • • • • • • xiv
ABSTRACT
AB STRAK
CHAPT ER
1
2
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xvi
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . , . . . xviii
INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
LITERATURE REVIEW 6
General Backg round ahout Pa rkia specio s a 6
Cyc lic Polysul fide s in Parkia s pec io s a . . . . . . . 7
Cyclic Polys ul fide s from Other P l,nts . . . . . • . . 10
P recursors of Cyclic Poly sul fides in S eeds of Parkis specioss . . . .. . . . . . . . . . . . . ... . . .. . . . 11
Anticancer Agent from Pa rkia specio s a . . . . . . . . 1 3
The Story about Diabetes . . . . . . . . .. . . . . . . . . . . . 1 3
Factors Causing Abnormally High Glucose Levels 1 7
Problems Existing in Diabetics .... ...... 21
Therapeutical Principl e in the Management of Diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Dietary and Exercise Therapy . . . . . . .. . ... 23
Insulin Therapy . . . . . . . . . . . . . . . . . . . . . . . . . 23
Sulfonylurea Agents . . . . . . . . . . . . . . . . . . . . . 26
v
3
Page
B iguanide De r ivat ive s . . . . . . . . . . . . . . . . . • . 3 2
Other New Oral Hypoglyc aemic Compounds Used in the T r e a tment o f Diabetic Animals 36
MATERIAL S AND METHOD . . . . . . . . . . . . . . . . . . . . . . . . . 37
Plant Material .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. 39
Phytochp�ical Screening Te s t ................. 40
Extrac t ions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . • . . 40
Isolat ion and Pur ific a t ion of the Biologically Act ive CHC l � Ext rac t ......................... 41
Chroma tographic Frac t ionat ion of CHCl� Ext r act of Empty Pods . . . . . . . . . . . . . . . . . . . . 4 2
Chromatographic Frac tionat ion of the CHC 1 3 Ext ract o f S e e d s . . . . . . . . . . . . . . . . . . . . . . . . . 4 6
Hypoglycaemi c Act ivi ty S tudies . . . . . . . . . . . . . . . 4 7
Sc reening Te s t o f a l l Hypoglycaemic Act iv ity
the Extract s for
B l ood Gluco s e Dete rmina tibn - Gluc o se Oxida s e Me thod
E f fe ct o f CHC l 3 Ext ract s on Norma l and
4 7
4 9
D iabe t ic Rat s . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 0
Dete rmination of the Activity of the Fractions CHC 13 Extract o f Empty Pods 51
Determinat ion of the Act iv ity of the Frac t ions of CHC l 3 Ext ract o f Seeds . . . . . . 5 2
Blood Gluc o s e Dete rmination - Reflo lux II M Method . . . . . . . . . . . . . . • . . . . . . . . . . 52
De te rmina tion of Hypoglycaemic Act ivity o f S - 4 ( A ) , P - 7 ( A ) , P-9(3AB) and P-9 ( 4 B ) in Alloxan-Diabe t ic Rats . . . . . ..... ...... ... 53
Compari s on S t udy be tween the E f f e ct s o f Gl ibencl amide ( Ant idiabe t ic D rug ) and S - 4 ( A ) as well as P- 7 ( A ) on the B l ood Gluco s e L eve l s in A l l oxan D iabe t i c Rat s . . 5 3
v i
4
5
Page
De tarmination of Do s e -Re sponse R e l a t ionship o f Fre sh S e e d s and Empty Pods Mat e r i a l . . . . 54
RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . . . . . . 55
Phytochemica l S c reening Te s t . . . . . . . . • . . . • . . . . . . 55
C l inic a l Technique s for A s s e s sing Hypoglyc aemic Ac t iv i t i e s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Hypoglycaemic Ac t ivi t i e s o f Ch l o roform Extrac t s 56
E f fe c t s o f Ch loroform Extrac t s on N o rma l and Diabe t ic Rat s 59
De te rminat ion o f the Ac t iv ity o f the Fract ions . . 64
Hypoglyc aemic Ac t ivity of S-4(A) and P - 7 ( A ) in A l loxan-Diab e t ic Rat s . . . . . . . . . . . . . . . . . . . . . . . . . . 6 7
C omparison Study Be twe en the E f f e c t s o f G l ibenc l amide (Antidiab e t ic Drug ) and S-4(A) as we l l a s P - 7 ( A ) o n the B lood Gluc o s e Leve l s in A l loxan-Diab e t i c Rat s . . . . . . . . . . . . . . . . . . . . . . . . . . 69
D e t e rminat ion of D o s e - Re spon s e Re l a t ion sh ip o f Fre sh S e e d s and Empty Pods . . . . . . . . . . . . . . . . . . . . . 7 1
S t ructure E luc idat ion o f S - 4(A ) . . . . . . . . . . . . . . . . 7 8
S t ructure Eluc idat ion o f P-9(4B) . . . . . . . . . . . . . . . 90
S t ruc ture Eluc idat ion o f P - 7 ( A ) . . . . . . . . . . . . . . . . 92
S t ruc ture E l uc idat ion o f P-9(3AB) , . . . . . . . . . . . . . 96
General D i s c u s s ion ............ ................. 98
CONCLUS ION
BIBLIOGRAPHY
APPENDICES
BIOGRAPHICAL
. . . . . . . . . . " " . . .. . . . . . . . . . . . . . . . . . . . .
SKETCH . . • , . • . . . . " . . , . • • . . . • • . . • . .
v i i
105
108
121
129
Table
1
2
3
4
5
6
7
8
9
1 0
1 1
12
1 3
14
15
1 6
LIST OF TABLES
Number of Cases and Deaths Attributed to Diabetes Mel l itus in Malaysia ( 1 9 8 4 - 1 98 8 )
Antimicrobial Effects of Compounds [1) and
[ 3 ] . . . . . . . . . . . . . • . . . . . . . . . . . . . . . . . . . . . . .
Clossificnlion of Diabel!'o .............. .
Indicat ions for Insul in . . . . . . . . . . . . . . . . . .
Oral Hypoglyc aemic Sulfony lurea Agen t s . . .
Adverse Effects of Su1fonylureas • . . . . • • •
Ora l Hyp.oglycaemic Biguanide Der ivat ives •
Indications for Treatment of Diabetes with B iguanide s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Treatment with Biguanides is Cont ra-indi c a ted in ............................ .
S ome o f New Comme rc ia l ly Oral Hypoglyc aemic Compound s Used Exp e r imenta l ly in the Treatment of D iabetic Animal • . . . . . . . .
Ana ly s i s o f Va riance on G luc o s e ( Seeds Ext ract-s) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Analys i s o f Va rianc e on Gluc o s e ( Pods Extrac t s ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . , . . .
Analysis of Va riance on Glucose ( Seeds Extrac t s on Norma l Ra ts S tudies ) . . . . . . . . .
Analysis of Variance on Glucose (Pods Ext rac t s on Normal Rats S tudie s ) . . . . . . . . .
Analysis of Variance on Glucose ( Seed Frac t ions S tudi e s ) . . . . . . . . . . . . . . . . . . . . . . .
Ana lysis of Va riance on Gl ucose (Pods Fract ions S tudie s ) . . . . . . . . . . . . . . . . . . . . . .. .
v i i i
Page
2
9
16
2 4
27
32
33
34
3 5
36
1 2 3
123
1 2 4
124
1 2 5
1 2 5
Table
1 7 Analy s i s o f Va�iance o n Gluc o s e ( Pu r e Compounds S tud i e s ) . . . . . . . . . . . . . . . . . . . . . . .
18 Analy s i s of Var iance on Gluc o s e ( G l iben-c lamide S tud ies ) . . . . . . . . . . . . . . . . . . . . . . . . .
1 9 Analy s i s o f Va rianc e o n Gluc o s e ( S e e d s
Page
126
12 6
Do s es S tud ie s ) . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
20
21
22
Analy s i s o f Va r ianc e on Gluc o s e (Pods Do s es S tudies ) . . . . . . . . . . . . . . . . . . . . . . . . . . .
Carbon Chemic al Shifts �-Sitosterol and S t igma s t e ro l
( ppm) o f
Ana ly s i s o f t u r e S tudies )
Va riance On Gluc o s e (Admix-
ix
127
83
128
LIST OF FIGURES
Figure
1 Cy c l ic Poly su l f ides I s o l at e d from Parkia
2
3
specio s a See d s . . . . . . . . . . . . . . . . . . . . . . . . . . .
Lent inic Acid and Epi lent inic Ac id . . . . . . .
Oxida t ion Produc t s Containing sul fide s and 12-Membered Containing a-Sul fur Atoms
Cyclic Po lyHe terocyc l ic
in Chondria cal ifornica ................ . ............. .
4 Homologue o f 1 , 2, 4-Tri thio lane in A l l ium
5
6
7
8
9
1 0
cepa . . . . .. . . . . . . .. . . . . . . . . .... . " . . . . . .. . .. . .. . .. .
Compound s with 2-Su l fur Cont aining Amino Ac ids and 1-Sul fur Conta ining Imino Ac id in Se eds of Parkia spe c i o s a .. . . . . . . . . . . . . . . .
Oral Glucose Toleranc e Te s ts ......... . . .. .
Inje c t ion Sites for Insulin . . . . . . . . . . • . . . .
The Ana ly t i c a l TLC of Crude Seeds Extrac t I in 10 0 % CHC l3 • • • • • • • • • • • • • • • • • • • • • • • • • • • • •
Effect o f Dif ferent Chemical Solven t s Extract o f f. spec io s a on Blood Gluc o s e Leve l s i n All oxan-Diab e t ic Ra t s . D a t a are me ans ± SE (n .. 4 ) . . . .. . . . . . .. . . . .. . . . . . . .
E f fe c t o f Chloro form Extrac t s o f P . spe c io s a o n Norma l and Diabe t ic Rat s . Data are Me ans ± SE (n-4) . .. . . . . . . . . . . . ....... .
1 1 Determination o f Seven Se eds Frac t ions for Hypoglycaemic Activity. Data a r e Means ±
Page
8
10
12
1 2
14
15
25
41
6 0
6 7.
S E ( n - 4 ) . . .. . . ...... . . . . . . . . ............ 65
12 Dete rmination of S ix Empty Pods Frac t ions for Hypog lyca emic Ac t ivity . Da ta are Means
13
:I::: SE en • 4 ) . ... .. . . .. .. . .. . .. .. ........ .. . . ........ .. .... .. 6 6
Hypoglyc aemic Ac t ivity o f P-9 ( 3AB) and P- ( 4B) . Data
S-4 ( A) , P-7 ( A) , a r e Me an s ± S E
( n = 4 ) .. .... , .. . . . . .................. . , . .. . . ...... . ,.... . .. . .. 6 8
x
Figure
14
15
16
Comparison Study Between the Effects of G1ibenclamide (Antidiabetic Drug) and S-4 ( A ) a s we l l as P - 7 ( A ) on the Blood Gluco s e Leve l s in Al l oxan-Diab e t i c Ra t s . Data a re Me ans ± SE ( n = 4 ) . .. . . . . . . . . . ... .. . . . . . . .
Do s e-Responsp Relationship of Fresh nnd G round Sppch on Blood G 1 \l( osp Lev? Is of
Al loxan- Diabpti( Rats. Datd arp Moans 1 SE (n "" 4 ) . . . . . . • . • • . . . . . . . . . . . . . . . . . . . . . . . . .
Do s e -Re s pon s e Re l a t ion ship o f F r e s h and Ground Emp ty Pods on B l ood Gl uc o s e Leve l s of Al loxan-Diabe tic Rat s . Data are Means ± SE (n = 4 ) • . • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •
1 7 Percent Lowering of Blood Glucose a t Each Dose Point of Fresh and Ground Seeds. Data
Page
70
73
7 4
are Means ± SE (n = 4 ) . . . . .. . . . . . . . . . . . . . . 76
18
19
20
21
Percent Lowering of Blood Glucose at Each Dose Point o f Fresh and Ground Empty Pods. Da ta are Means ± SE ( n = 4 ) . . . . . . . . . . . . . . .
Repre s entatives S te ro ids
The Ca rbon S ke l e t on s and Numb e r ing S y s tem o f P e rh ydrocy c l opentenophenanthrene Ring S y s tem . . . ... . . . . . . ... . . . . . . .. . ....... . . . . .
The Chemic a l S t ruc ture s and Numb e r ing System of S - 4 (A) .. . .......... . . . . . . . . .. . . .
22 The Differences of the C-Chemical Shifts of
7 7
7 9
80
82
the Sterols . . . . . . . . . . . . . . ... . . . . . . . . . . . . . . 84
23 Hypoglycaemic Activity of Authentic �sitosterol. Stigmasterol and Mixture of Those Two ( 66 mg : 34 mg) Compared to the Is olated S-4(A). Data are Means ± SE (n - 4 ) • • . • . . • • • • • • • • . . • • • . . • . • • • • • • • • • • • •
24 Thp Chemical St ructu re of Stigmast-4-pn-3-onp
xi
8 7
95
F igure
25
2 6
2 7
2 8
The Effect o f the A l c oholic Ext ract o f H. s a l i c ornic� on Blood Gluc o s e Leve l o f Norma l and A l loxan-Diabe t ic M i c e . . . . . . . . . .
Effect o f S ingle Oral Adm ini s t ration o f C S-0 4 5 on Pla sma Gluc o s e in Diabe tic Mice. B lood Samples Obtained in Nonfasting State. Data Bre Me ans ± SE ( n - 4). 0, Cont rol; . , C5-0 4 5 . "'p < 0.05 v s . c ontrol . . . . . . .
Effect o f C S -045 on Plasma Gluc o s e Nonf ast ing S tate i n Normal Rats. D rug was Ora l ly Admin i s t e re d a t Doses o f 100 ( . ) and 1000 (�) mg / kg; 0, Contro l . Data a re Means ± 5E ( n .. 6 ) • • • • • • • • • • • • • • • • • • • • • • • •
Dose-Re s pons e Curve of C S-045 in Diabe t ic Mic e . Maximum Pe rcent Decre a s e C a lculated for Pla sma Gluc o s e Leve l at Each Do s e Point . Blood S amp l e s Obta ined in Nonfa sting State. Data aru Means ± SE with Numbe r of Anima l s in Parenthe s e s . . . . . . . . . .
x ii
Page
9 9
103
103
104
Scheme
1
2
3
4
LIST OF SCHEMES
The Extractions of P. speciosa . • . . . • . . . • .
The Screening Studies of A l l the of �. speciosa on Rats for Their
Extracts Hypogly-
caemic Activity . . . . . . . . . . . . . , . . . . . . . . . . . .
The Effect of CHCl� Extracts on Normal and Diabe t ic Rats . . " . , . . . . . . . . . . . . . . . . . . . . , .
Determination of Dose-Response Relation-ship of Fresh Seeds and Pods Materials . . .
xiii
Page
57
58
61
72
IR
1H_NMR
CDCL3
TMS
s
d
t
q
m
br-s
dd
BBD
HPLC
UV
MS
TLC
std. dev.
pet - ethe r
CHC13
EtOA"
CH.lC12
LIST OF ABBREVIATIONS
Infrared Spectra
Proton Nuclear Magnetic Re s onance Spectra
Deuterated Chloroform
Tetramethylsilance
singlet
doublet
triplet
quartet
multiplet
b road s ingle t
dOt"blet-doublet
Carbon- 13 Nuclear Magnetic Re s onance Spectra
Broad Band Decoup l e d
High Performance Liquid Chromatography
Ultra-violet Absorption
Mass Spectra
Thin Layer Chromatography
Standard deviation
Petroleum ethe r
Chloroform
Ethyl act!tate
Dichloromethane
xiv
NH4CHC1;, Ammonical chloroform
MeOH Methanol
S Seeds
p Empty pods
T Test
G S t andard
Fract Fraction
Gliben Glibenc.lamide
Sitos Sitosterol
Stigma Stigmasterol
xv
Abstract of the thesis s ubmitted to the Universiti Pertanian Malaysia as fulfilment of the for the degre � of Master of Science.
Senate of rsquirement
Chairman
Faculty
THE HYPOGLYCAEMIC AND CHEMICAL STUDIES OF PARKIA SPECIOSA, Hassk
B y
FATHAIYA BTE JAMAL UDIN
July , 1 9 9 1
Dr. Suhaila Mohamed
Food Science and B iotechno logy
The admin i s t rat ion of the chloroform ext rac t s o f Pa rk ia
spe c io s a ( pe t a i papan) to a l l oxan- treated d iabe t ic rats
p roduc ed a s igni f icant depre s s ion in b l ood gluc o s e leve l s .
Oth e r s olvent extrac t s f a i l e d to induc e any re duc t ion in the
b lood gluc o s e l eve l s o f the d iabe t ic anima l s . In norma l rat s
s e rum g luco s e l eve l s we re not changed a f t e r the admin i s t rat ion
of chloroform extracat s ( 4 0 0 mg / kg body we ight) c omp a rab le to
tho s e admin i s tered with s a l ine s o lut ion .
The de t e rminat ion o f do s e - re spon s e re l a t ionship of fre sh
and ground s e e d s and emp ty pods showed tha t the minimum
effect ive do s e of raw ma t e r i l s nec e s s a ry to produce a
significant (p < 0.0001) decrease in serum glucose concentra-
tion wa s 25 mg / kg body we ight .
xvi i
Extrac t ion and isolation work on the act ive f rac t ions o f
the mothe r l iquor� r e s ulted in the is olation o f pure c ompounds:
5-4(A) f rom pe tal seeds , c ons i s t e d o f a mixture o f p-slto s te rol
(6 6 % ) and s t igma s te rol (34%) ; P-7(A ) f rom empty pods was
propose d to be s t igma st-4-en-3-one; P-9 (4B) f rom pe t a i pods wa s
E' luc idated as a s t igma sterol while P-9 ( 3AB ) was a c ompound from
tho pods which could not. 1)(' ldont.ifiud. Tht) S-'t{A) nlld r-7(A)
5smple s we re found to have strong hypoglyc aemic activity . The
occ urrence of p-s i t o s t e rol, s t igmas t e rol and s t igmas t-4-en-3-
one we re reported for the f i r s t t ime in Pa rkia spe c io s a. The s e
naturally occurring c ompounds we re sugge s te d f o r new potent ial
o rally hypoglycaemic agent s .
A c omparative s tudy on the hypogly c aemic e f f e c t o f
gl ibenc lamide (antidiabetic d rug ) , with 5-4(A) and P-7(A )
showe d gl ibenc lamide to have 1 1 1% hypoglycaemic ac tiv ity a t 5
rng /kg body we ight . A 2 : 1 mixture of �-s ito stero l and
s t igma ste rol displaye d 83% hypoglyc aemic ac t ivity while
s tigmast-4-en-3-one had 84% e f f e c t at a dose o f 100 mg/kg.
Howeve r, p-s ito s t e rol o r s t igmaste rol individually was found t o
pos s e s s n o hypoglycaemic prope rty when te s te d i n i s o lation.
Th is finding suggests 8-4(A) produced a Ilynergiatic effect to
cau s e hypoglyc aemic ac tivity. The behav ioural syne rg ism has
not been repo rted before in work related to the s e s te ro l s .
xv i i
Ab s trak te s i s Pe rtanian Ma l ay s ia Mas te r S a in s .
y ang d ik emukakan kepada Senat Unive r s i ti s ebaga i memenuh i kepe r luan unt uk I j a z�h
Penge rus i
Fakulti
KAJIAN HIPOGLISAEMIK DAN KIMIA TERHADAP PARKIA SPECIOSA, Ha s sk
O l e h
FATHAIYA JAMAL UDIN
Jula i , 1 9 9 1
Dr . Suhaila Mohamed
Sa ins Makanan dan Bi oteknologi .
Pentadbi ran ek s t rak k l o r ofom dari P a rkia spe c i o sa (peta i
papan ) me lalui mulut k epada t ikus yang dij adikan kenc ing mani s
dengan d i racuni a l loxan te l a h menunjukk an penurunan kandungan
g luko s dalam s e rum darah . L a in - la in eks trak pe la rut d i dapati
tidak memb e r i apa - ap a k e s an te r hadap tikus kenc ing man is . Bagi
t ikus no rma l didapa t i k andungan gluk o s s e rum tidak b e rubah
s e lepas pentadb i ran eks trak k l o ro fom (400 mg / kg b e rat badan )
s e rupa dengan tikus yang d i b e r i s a l ine .
Penentuan sukatan bij i p e ta i mentah dan kul it p e t a i menta h
juga te lah dijalankan. Do s min imum dari bahan mentah untuk
menghAHilkan pen lrunan glukos serum yang be rprti (p < 0.0001)
ialah 25 mg/kg berat badan .
xix
Pengek s t r akan dan pemis ahan t e rhadap p e c ahan y ang akt if
t e l ah mengha s i lk,n komponen tulen : S - 4 ( A ) , t e rkandung s eb a t ian
� - s ito s te ro l (6 6%) dan s tigma s te ro l ( 34 % ) daripada b i ji p e t a i
dan P - 7 ( A ) d ikenali s ebagai s t i gma s t - 4 - en- 3 - one , P - 9 ( 4B ) adalah
s t igma s te rol dan akhi r s eka l i P - 9 ( 3AB ) daripada kul it p e t a i
tidak d ikena l i akan k imia dan nama s t ruk turny a .
4 (A ) dan P- 7 ( A ) t e lah menunj ukkan k e s an
Komponen S
hipogl i s a emik
be rbanding dengan kawa l an . Penemuan � - s it o s te ro l , s t igma s t e r o l
dan s t igma s t - 4 - en- 3 - one di dalam P a rk ia spe c io s a adalah pertama
kali di rekodkan . Komponen-komponen te rb itan semulajadi ini
dic adangkan s ebaga i ubat kenc ing man i s yang baru .
S e t e ru snya pe rbandingan kajian d i antara g l ib enc l amide
( ubat kenc ing mani s ) dengan S - 4 ( A ) dan juga P - 7 ( A) dij a lankan .
Keputusan menunj ukkan g l ibenc l amide t e l ah menurunkan kandungan
glukos dalam s e rum darah sebany ak 1 1 1 % pada n i l a i da s 5 mg / kg
berat badan . Seba t i an �- s i tos t e r o 1 dan s t igma s t e r o l (2 1)
menunjuk -kan 8 3 % k e s an h ipo g l i s aemik sementara s t igma s t - 4 - en- 3 -
one mempunya i 8 4 % ke s an hipo g l i s aemik pada n i l a i d o s 100 mg/kg
be rat badan . T e tap i , � - s ito s t e ro l a t au s t igma s te roL s ahaja
t idak memberi apa- apa k e s an hipo g l i s aemik . Penemuan ini
men cadangkan S - 4 (A) mempunyai kesan s ine rgi s t ik untuk
menyebabkan penurunnn glukoR dalam sprllm darah. Tingknh laku
s inergi sme ini a da lah pe rtama kali direkodkan d i dalam ka jian
yang berkai tan dengan komponen s te ro l ini .
xix
CHAPTER 1
INTRODUCTION
The t e rm natura l p r oduc t i s referred t o thos e organ i c
compounds o f natural origin t h a t are unique to one o rgan ism o r
common to a small number o f c l o s e l y re lated o rganisms ( Mann ,
1977 ) .
P r imit ive man found quite a numbe r o f p lant extrac t s
effec t ive a s medic ine s for the re l ie f o f p a in o r a l l eviation o f
the s ymptoms o f d i s e a s e , a s p o i s on s f o r the war fare and hun t ing
uses as effective drugs fo r euthanasia and as na rcotic s ,
hal luc inogen s o r s t imu lant s t o r e l ieve the t e dium or al l eviate
the fat igue and hunger. Today, such uses are st il l be ing
practiced widely especially in remote areas but the acceptance
of herbal me dic ine are continuously spreading into urban commu
nitie s .
Between 1815 and 1860 (Mann, 1977), many of the s e ac t ive
p rinc ip l e s we re i s o lated inc luding mo rphine , s trychn ine ,
quinine , c a f fe ine , nic o t ine , c o de ine , c amphor and c o c a ine . I t
i s wo rth no t ing that during 1 9 5 0's s t ruc ture e luc ida t ion and
natural product reseorch have become much mo re fac i le with the
advent o f nuc lear magnetic resonance, mass spectrome try , and x
Ray crystallography.
1
2
Diabetes was first recognised almost 4 ,000 years ago.
This disease has a tremendous impact on the health sy scem of
individual s in the world. The e stimated number of patients who
suffered from diabetes mellitus in Malay sia between 1984 to
L988 according to the Information Documentary Sy stem' s are
pre sented in Table 1. The incidence of diabete s i s increasing
every year . However, deaths in Malay s ia appeared to be
decreasing by 1% per year. According to the Malay s ian Medical
Institute , the total annual e conomic budget for diabetes was
e stim�ted at approximately 1/4 million r inggits in 1 988 .
Table 1
Number of Cases and Deaths Attributed to Diabetes Mellitus in Malaysia
(1984 - 1988)
Year No. of cases Deaths
1 98 4 10125 2 6 4
198 5 1 4 7 6 7 2 61
198 6 17 301 2 6 1
198 7 1 7 901 2 4 5
1988 1 78 50 2 2 7
Source: Information Documentary System.
3
In 1976, the Nat iona l Commi s s ion on Diabe t e s repo r t e d that
5% of the populat ion of United S t a t e s ( app roximate l y 10 mil lion
peop l e ) have diabe t e s . The inc idenc e is inc r e a s ing b y 6 % p e r
ye a r . The i r t o t a l annua l e c onomic budget o f d iabe t e s wa s
e s t imated at approxima t e ly 8 b i l l ion d o l l a r s in 1979 ( Davidson,
1981) .
In Ma lay s ia , almost 680 0 spec i e s o f s e e d p l an t s and 6 0 0
spec i e s o f s e e d le s s p l an t s have been recorded ( Hal t um , 1954 and
Keng , 1970). In l oaa l t radit ional me dical p ra c t i c e s , many o f
the s e p lant s have been u s e d f o r remedies of var ious i l lne s s
inc luding for diabete s . In fac t , it wa s repo rted that at l e a s t
15% of the s e p l an t s had been c la ime d b y va riou s people to have
medic inal u s e s ( L a t i f � a l . , 1984 ). Neve rthe le s s , this a re a
o f r e s e a rch is s t i l l l ack ing i n Ma l ay s ia . For ins tance , Kumi s
kuc ing ( O r tho s iphon s t amineu s ) , Aka r putawa l i ( T ino spo ra
c r ispa ) , Lempedu tanah ( Curanga �) and J e r ing ( P ithe c e l l o
b ium je r inga ) which has been t radit iona l ly u s e d t o t re a t
catarrh o f the b l adde r , hype rtens ion and d iabe t e s have s t i l l
not been t e s ted bio logica l l y and inve s t igated chemi c a l l y .
On the othe r hand , e venthough the re are many substances
that have been iso l a t ed and cha rac terized , the investigation of
their potential biological effects a re still not ca rried out
(Kj ae r , 1977).
4
Therefore, the implication of doing this kind of research
is to opt imize the cult iva t i on and gene t ic s elec t ion o f the
food and plants . Addit iona l l y , new oral h ypoglyc aemic p roduc t s
can b e synthe s i zed comme rcially f rom the he rbal plant s . In the
yea r 1988 , about 74, 2 30.5 x 103 amoun t of d rug s have been
synthe s ized by the private s e ctor in the coun t r y .
The re h a s been chemical inve s t i gat ion o n s e e d s but n o t on
empty pods of Park ia spe cio s a ( Haask ) ( Gmelin e t a l . , 1 9 8 0 ) .
The re are not much studies carried out regarding their medic i-
nal chemistry . This research at tempts t o investigate the
a l l e ge d hypoglycaemic a ct ivi t y o f Parkia spec iosa .
The object ive s and s c ope of the r e s earch a re :
i ) to inve s t igate the hypogly caemic activity of P .
spe c i o s a o n no rma l and a l l oxan diabe t ic ra t s .
i i ) to i s olate the ac t ive comp ounds from the active
f rac t ion s .
i i i ) to elucidate the structures of the isolated a ct i ve
c ompounds .
5
The pha rmacologic a l inve s t igat ions t o s tudy the e f fect o f
P . spe c io s a e .t rac t s on o the r s y s tems i n r a t s s uch a s neuro
pha rmac ological s tudie s , c a rdiova s cular s tudie s , mo rpho logic a l
and b iochemical change s , e f f e c t o f ext r ac t s o n smoo th and
ske l e tal mus c l e s e t c . we re not done . Neve r the le s s the s e field
of s tudi e s are o f int e re s t f o r future inve s t igat ions .