UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA...
Transcript of UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA...
![Page 1: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/1.jpg)
UNIVERSITI PUTRA MALAYSIA
CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND THEIR BIOLOGICAL
ACTIVITIES
CHEW YEAN L I NG
FSAS 1995 7
![Page 2: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/2.jpg)
CHEMI CAL CONST I TUENTS OF GLYCOSHIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND THE I R B I OLOGI CAL
ACTIVITIES
By
CHEW YEAN L I NG
The s is S ubmi t t ed in Fu l f i l me n t o f t he R e q u i r ements f or t h e De gree o f Ma s t e r o f S c i e nce
i n t he Fa c u l ty o f S c i ence a nd Env i r onme n t a l S t ud i e s , Unive r s i t i P e r t a nia n Ma l a ysia
June 19 9 5
![Page 3: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/3.jpg)
ACKNOWLEDGEMENTS
I wou l d like t o ta k e this oppor tun i t y t o e x pr e s s
m y s i nce r e a pprec i a tlon and gra titude t o A s s o c . Prof .
Dr . Ha wa rdi Ra hma ni a nd Ass oc . P r o f . Dr . Sar i a h Meon
for t he inva l ua b l e advice and guida nce thr oughout the
c o u r se o f this pr oje ct, and Asso c . Pr o f . Dr . Mohd.
Aspo l l a h Hj. Suk a r i for his cons t a nt s uppor t .
I wou ld a l s o l ik e t o a c k nowl edge t he coope ra tion
and assistance o f the s ta f f o f the Chemi str y
De pa r tme n t o f UPM : En . Na z r i Ahmad , En . Z a i na l Abld i n
Kas i m , En. Ho hamad Abu S amah, En . Z a i na l Z a ha r i
Z a k a ria , En. Nordin I smail and En . Zainuddin Samad i .
La s t but n o t l e a s t , I wis h to tha n k P r o f . Norio
Aimi o f Chiba Un i ve r s i t y f or the run n i ng o f I H -NMR
and 13C-NHR s pe ctr a , Mr . S. Anth o ny Samy o f the
Bio l ogy De pa r t ment o f UPH a nd En . Ahmad Zainuddin
I br a h i m o f UK M f o r i d e n t i f ying a nd co l l ect i ng the
p lant mate ria l s , and En . K hir Rani a nd the sta f f o f
the Plant P r otecti o n Department o f UPM f or their help
a nd c o o pe r a t i o n d ur ing the bioas say s tudy .
i i
![Page 4: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/4.jpg)
TABLE OF CONTENTS
ACKNOWLEDGEMENTS
L I S T OF TABLES
L I S T OF F I GURES
L I ST OF PLATES
L I ST OF ABBREV I AT I ONS
ABS TRACT
ABSTRAK
PART I EXTRACTI ON AND I S OLAT I ON OF CHEHI CAL COHPONENTS FROM GLYCOSHIS CALCICOLA AND G. RUPESTRIS ( RUTACEAE)
Page
11
v i
vii
x i
x
x i
xiii
I NTRODUCT I ON . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
L I TERATURE REVI EW. . ... . .. . . . . ... . . . ..... 3
OBJECTI VES OF S TUDy . . . . . . . . . . . . . . . . . . . . . 17
RESULTS AND D I S CUS S I ON . . . . . . . . . . . . . . . . . . 18
G l ycosm i s Cal c i co l a B . C . S t one . . . . . . . . . . . . . 1 8 Glycosm i s Calc i cola ( La ngk a v l I s l and ) ... 18 Glycosm i s Calclcola ( Temp ler P a r k ) . . . . . . 3 6
Glycosm i s R u pestr l s R i dley . . . . . . . . . . . . . . . . . 55 Glycosmi s R upes t r i s «S tem B a r k ) . . . . . . . . 5 5 Glycosm i s R upes tri s ( Lea ves ) . . . . . . . . . . . . 83
CONCLUS I ON . . • • . • • . • . • • . • • . • . . . . . . • . . . . . . . . . 8 4
i l l
![Page 5: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/5.jpg)
EXPERIMENTAL............................... 85 Gene r a l M a t e r i a l s a nd Method s . . . . . . . . . . . . . . 85 Ext r ac t i o n o f Glycosmis Calclcola ( Langk a w i I s l a nd ) . . . . . . . . . . . . . . . . . . . . . . . . . . 8 7
I s o la t i o n o f GC1 (4 4 ) . . . . . . . . .. . . . . . . . . . . 87 I s o l a t i on o f S i t o s t e r o l (48)............ 89 Is o la t i on o f GC6 . . . . . . . . . . . . . . . . . . . . . . . . 9 0
Ext r ac t i on o f Glyco s m i n Ca l c i c o l a ( Te mp l e r Park ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 1
I s o l a t i on o f F l i nd e r s i n e (4 9)........... 9 1 I s o l a t i o n o f De s me t hoxyzanth o phyl l i n e ( 50 ) . . . . . . . . . . . 9 3
Ex t r a c t i on o f Glycosmis Rupestrls . . . . . . . . . . I s o l a t i on o f G l ycoma u r i n ( 2 2 ) . . . . . . . . . . . 9 4 I s o l a t i o n o f 7 -m e t h o xyg l ycoma u r i n (51)............... 9 6 I s o l a t i on o f De s -N-me t hyl a c r o nyc i ne ( 5 3 ) . . . . . . . . . . . . . 9 7 I s o l a t i on o f s t i gmas t - 7-e n o l (54 ).................... 9 8 I s o l a t i o n o f GCIS . . . . . . . . . . . . . . . . . . . . . . . 9 9
PART I I ANTI FUNGAL ASSAYS AGAINST CHAONEPHORA CUCURBITARIUH, COLLETOTRICHUH CAPSICI AND C. GLOEOSPORIOIDES
I NTRODUCT I ON . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
OBJECT I VES OF S TUDy . . . . . . . . . . . . . . . . . . . . . . . . 1 0 6
LITERATURE REVIEW . . . . . . . . . . . . . . . . . . . . . . . . . . 107
MATER I AL S AND METHODS . . . . . . . . . . . . . . . . . . . . . . 1 1 1
P r e pa r a t i on o f P l a n t Extracts . . . . . . . . . . . . . . 1 1 1 Preparation of stock Cultures . . . . . . • . . . . . . . III Preparat i on o f Assay Media................. 1 1 2 Myce lia l Gr owth a nd Spor u l a t i on . . . . . . . . . . . . 1 1 2 S p o r e Ge r m i na t i on . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 4 TLC B i oa s s a y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 5
RESULTS . • . • . • . . . • . . • . . . . . . . . . . • . . . . . . . . . . . . 118
Myce l i a l Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1 8 S p orula t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Spore Ge r m i na t i on . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2 5 TLC B i oa s s a y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 27
D I S CUS S I ON AND CONCLUS I ON . . . . . . . . . . . . . . . . . . 1 3 1
iv
![Page 6: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/6.jpg)
BIBLIOGRAPHY . . • . . • . . . . . . . . • . . . . . . . . . . . . . . . . 133 VI TA . . . . . . . .. .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 3 7
v
![Page 7: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/7.jpg)
L I ST OF TABLES
Tab l e
1 1H-NMR Data (6) o f (44) a nd GC1 . . . . . . . .
2 l3C-NMR Chem i ca l S h i f t Data (ppm )
P a ge
25
o f Gel................................. 32
3 13C-NMR Chem i ca l S h i f t Da t a (ppm ) o f F l i nders i ne w i t h the Observed Va l ues . . .
l 3C -NMR Chem i ca l S h i f t s (ppm ) o f
48
G l yc oma ur i n ( 2 2 ) ......... . ... . . . ....... 6 3
5 lH-NMR S pectra l Data (6) o f ( 2 2 ) , ( 5 1 ) a nd ( 5 2 ) . . . . . . . . . . . . . . . . . . . . . . . . 70
6 P l a n t Extra c t s I nves t i ga ted f or An t i f u nga l Act i v i t y . ...... . . . ... . .. . ... 1 1 3
7 E f fect o f G. cal c i co l a a nd G. r upes tr i s extra c t s o n Myce l i a l Growth of C. cucurbltarlum, C. capsicl a n d C . gl oeos por i o i des . . . .. . . . .... . . . .. . .. . 1 19
8 E'f fec t o f G. ca l c i co l a a nd G. rupes tr is ext r a c t s o n S pore Germ i na t i o n o f C . cucurbl tarlum, C . caps i c i a nd C . gl o e os pori o i des . . . .. . . . . .. . . . . . .... . 126
9 A n t i f unga l Cons t I t uent s o f E x t r a c t B • • • 129
v i
![Page 8: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/8.jpg)
F i g u r e
1
2
3
4
L I ST OF FI GURES
Schema tic Dia g r am o f Extrac tion and I so l ation P r ocedure o f G . ca lc i cola (Lang k a wl I sland ) . . . . . . . . . . . . . . . . . . . . . .
I R S pe c t r um o f ( 4 4 ) ................... .
IH - NHR S p e c t r u m o f ( 4 4 ) ............... .
Mass S pe c t r um 0 f ( 4 4 ) . . . . . . . . . . . . . .. . . .
5 Mass S pe c t r a l Fragmenta tion P a t t e rns
P a ge
1 9
2 1
2 3
2 6
o f ( 4 4 ) . .. . . . . .... . . . . . . . . .. . . . . . . . . . . . 2 7
6 1 3 - 4 C NMR S pe ct r um o f ( 4} .............. .
7 PHSQC S pe c t r um o f ( 4 4 ) ................ .
8 PHSQC S p e c t r um o f ( 4 4 ) . . .. . . . .... . .... .
9 I R S pe c t r um o f ( 48 ) • • • • • • • • • • • • • • • • • • • •
10 Mass Spe c t r um of ( 48 ) ................. .
1 1 Mass S pe c t r al Fragmenta t i on Pa t t e rns
2 9
30
31
3 4
3 5
o f ( 48 ) ................................ 3 7
1 2 S chematic Dia g r am o f Ex t r ac t i on and I sola t i on P r oced ure o f G. calcicola (Temple r Par k ) . . . . . . . . . . . . . . . . . . . . . . . . . 3 8
1 3
1 4
15
1 6
1 7
1 8
1 9
Mass S pe c t r um o f ( 49 ) .. . . . . . . . . . . . . . . . .
I R S p e c t r um o f ( 49 ) ................... .
1 _ H NMR S p e c tr um o f ( 49) ............... .
NOESY S p e c t r um o f ( 49 ) ................ .
Cou p l ing o f Various P r o tons Obse rved f r om NOESY S p e ct r um o f ( 49 ) . . . • . . . • . . . .
1 3C -NMR S p e c t r um o f ( 49 ) .............. .
I R S pe c t r um o f ( 50 ) . . . . . . . . . . . . . . . . . . . .
v i i
40
4 1
4 3
4 4
45
4 6
4 9
![Page 9: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/9.jpg)
20 IH-NHR Spe c t r u m o f (50) ............... .
2 1 Mas s S pe c t r um o f (50) ................. . 2 2 Mas s Spec t r a l Fragme n ta t i o n P a t t e r n s
51
5 3
o f (50)................................ 5 4
2 3
2 4
2 5
2 6
2 7
2 8
S chema t i c D i a gr a m of Extraction and I s ola t i o n P r ocedure o f G. r upes t r is S t em Ba r k ,a nd Leaves . . . . . . . . . . . . . . . . . . .
Ma s s Spec t r um of ( 2 2 ) . . . . . . . . . . . . . • . . . .
I R Spec t r um o f ( 2 2 ) ................... .
1�-NMR S p e c t r um o f ( 2 2 ) ............... . 1 3C-NMR S pe c tr um o f ( 2 2 ) . . . . . . . . .. . . . . .
Ma s s S p e c t r um o f (51) ................. .
2 9 Ma s s S p e c t r a l Fr a gme n ta t i on P a t t erns
5 6
58 5 9
6 0
6 2
6 5
o f (51) . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . . . 6 6
3 0 I R S pe c t r um of (51).................... 6 8
3 1 1H -NHR S pe c t r um o f (5 1). . . . . .. . . .. . . . .. 6 9
3 2 NOESY s pe c t r um o f ( 5 1 ) . . . . . . . . . . . . . . . . . 7 2
3 3 P r o t o n Cou p l i ng P a t t e r n Obs e r ved i n NOESY S pe c t r um o f (51)................. 7 3
3 4 Ma s s S p e c t r um o f (53).................. 7 5
3 5 I R S p e c t r um o f (53).................... 7 6
3 1 -6 H NMR S pe ct r um o f ( 5 3 ) . . . . . . . . . . . . . . . . 7 7
37 Mass Spectral Fragme n tat i o n Pat ter n s o f ( 53) . . . . . . . . . . . . . . ....... . ... . . ..... 7 8
3 8 I R S p e c t r um o f (5 4) . . . . .. . . . . . . . . . . . . . . 8 1
3 9 Mas s S p e c t r um o f (54 ).................. 8 2
4 0 H i s t o g r a m show i ng e f f e c t o f G. calclcola a nd G. r upest ris e xt r a c t s o n myc e l i al gr owth o f C. c u c u rb i tar i um, C. capslcl and C. gloeosporioides . . . . . . 1 2 0
4 1 H i s t o g r a m sho w i n g e f f e c t o f G . calcicola a nd G. r u�es tris e x t r a ct s on s po r ula t i on o f C . ca ps l c l and C. gloeosporlolde� . . . . . . . . . . ... . . . . . . . . 124
v i i i
![Page 10: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/10.jpg)
LIST OF PLATES
P la te Page
1 Ch i l l i A i s a hea l thy f r u i t . Ch i l l i B sh owi n g s ympt oms o f a n thr a c n o s e . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 0 5
2 E f f e c t o f CHCl 3 e x t r a c t B o n myce l i a l g r owth o f Chaonephora cucurbi tar i um . . . . 1 2 2
3 Ef f e c t o f CHCl 3 e x t ract B o n myce l i a l g r owth o f Col l e tot r i chum caps i c i . . . . . . . 1 2 2
4 E f f e c t o f CHC 1 3 e x t rac t B o n myce l i a l g r owth o f Col l e totr i chum gl oeos por l o l des . . . . . . . . . . . . . . . • . . . . . . . . 1 2 3
5 E f f e c t o f CHCl 3 e x t rac t F o n s p o r e g e r m i na t i on o f Col l e t ot r i ch um caps i c i . . 128
6 CHC 1 3 e x t rac t B , GC7 (F l i nd e r s i ne (49» and Gce (de s me th oxyzanthophy l l i ne (50) w i th s ome i mpur i t i e s ) on TLC p late show i ng zones of i nh i b i t i on a f t e r s p r a y i ng w i th Asper g i l l u s fl a vus . . . . . . . 1 30
i x
![Page 11: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/11.jpg)
br CHC13 CDC13 d d d d t EtOH FAB I R m m Me MeOH m.p. MS NMR PLC ppm s TLC w
LIST OF ABBREVIATIONS
br oad c h l o r o f or m d e ut e r a ted c h l or o f o r m d o ub l e t d o u b l e t o f d o u b l e t d o u b l e t o f t r i pl e t e t hano l Fa s t At om i c B ombar dment I n f r a Red med i um ( i n IR yata ) mult i ple t ( i n H -NMR d a ta ) me thyl me t hano l melting point Ma s s S p e c t r u m Nuc l e a r Magne t i c R e s onance P r e pa ra t i ve Th i n La ye r Chr oma t ogra phy pa r t per m i l l i on strong Thin Layer Chromatography weak
x
![Page 12: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/12.jpg)
Abs t ra c t o f t h e t he s i s p r e s ented t o t h e Sena t e o f Un i v e r s l t i Pe r tani an Ma l a ys i a i n f u l f i l ment o f
r e q u i r e ments f o r t he d e g r e e o f Ma s ter o f S c i ence .
CHEMI CAL CONSTI TUENTS OF GLYCOSH I S CALCICOLA AND G. RUPESTRI S (RUTACEAE) AND THE I R B I OLOGI CAL
ACTI V I T I E S
Cha i r man : Fa c u l t y :
By
CHEW YEAN L I NG
June 1 9 9 5
A s s o c i a te P r o f e s s o r Dr . Ma va r d i Ra hmani S c i ence and Env i r o nment a l Stud i e s
I nves t i ga t i on o f the l e a f and t v i g s e x trac t s o f
Gl ycosm i s ca l c i col a ( R u taceae ) f r om t vo d i f f e r ent
l oca t i ons has l ed to t he i s o la t i on o f f i ve compound s .
The s t r u c t u r e s o f t h e s e compound s we r e e l uc i d ated by means o f s pec t r o s c o p i c methods ( I R , uv, I H-NMR , 1 3 C_
NMR , MS ) and by compa r i s on w i t h p r e v i o u s s t ud i e s.
G. ca l c i co l a c o l l e c t ed f r om Langk a v i I s l and
y i e l d ed �e t hy l ge r ambu l l i n I s i t o s t e r o l and anoth e r
unid�ntl f led c ompo und . From ano t her s e pa r a te
x i
![Page 13: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/13.jpg)
collection of the same plant from Templer Park, a new
alkalo id (desmethoxyzanthophyll i ne) and f 1 i nders i ne
have also been isolated. A similar study on the bark
of another species, G. rupestris, collected from
Langkawi Island afforded
m e t h o xyg lyc o m a u r i n) ,
a new alkaloid (7-
g l yc o m a u r i n , d e s-N-
methylacronycine and stigmast-7-enol, while the leaves
of the plant gave an unidentified compound.
The antifungal activities of these plant extracts
were evaluated using the poison food and spore
germination techniques. The plant extracts exhibited
varying degree of fungitoxic response on the growth of
the fungal pathogens of chilli (Capsicum annuum L.).
The chloroform extract of G. calcicola collected from
Templer Park has been shown to be most effective in
inhibiting the mycelial growth, sporulation and spore
germination of Chaonephora cucurbi tarium,
Col1etotrichum capsici and C. gloeosporioides.
The antifungal compounds in this extract were
identified using TLC bioassay technique against
Aspergillus flavus. The results indicated that the
activity of the extract is due to the presence of its
major components: f lin d e r s i n e a n d
desmethoxyzanthophylline.
xii
![Page 14: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/14.jpg)
Abs t rak t e s i s yang d ikemukaka n kepada S e na t Un i ve r s i t i P e r ta n i a n Ma l a ys i a ba g i memenuh i ke pe r l ua n
I j a za h Ha s te r Sa i ns .
KANDUNGAN KI HI A DARI GLYCOSHIS CALCICOLA DAN G. RUPESTRIS (RUTACBAE) DAN AKT I VI TI BI OLOGI NYA
O l e h
CHEW YEAN L I NG
J u n 1995
P e nge r u s i : P r o f e s s o r Had ya Dr . Ha wa r d i Rahma n i Fa k u l t i S a i ns dan Penga j i a n A l a m S e k i ta r
Kaj i a n k e a t a s e k s t r a k d a u n d a n r a n t i ng
Gl ycosmis ca l cico l a ( R utaceae ) t e l a h mengha s i lka n
l i ma k ompa un . S t r uktur s e ba t i a n t e r s e but te l a h dapa t
d i t e n t u k a n de ngan menggunaka n ka edah s pe k t r osko p i dan
pe r ba nd i ngannya d e ngan k a j i a n -kaj i a n ya ng t e l a h
d i l a k u k a n t e r d a hu l u .
G. calcjcola ya ng d l kump u l ka n dar i P u l a u
Langka wi te lah membe r l ka n me t i l g e r ambu l i n
s i t o s t e r o l d a n s a t u s e ba t l a n ya ng be l um d lke n a l
pa s t i . Dar i pengumpu l a n ber a s i ngan s pe s i e s ya ng sama
x i i i
![Page 15: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/15.jpg)
di Templer Park, satu alkaloid baru
(desmetoksizantofilina) dan flindersina telah berjaya
dipencilkan . Kajian yang serupa ke atas kulit batang
spesis yang lain, G. rupestris, yang dikumpulkan dari
Pulau Langkawi telah member ikan satu alkaloid baru ( 7 -
metoksiglikomaurin), glikomaurin, des-N
metilakronisin dan stigmast- 7-enol, manakala daunnya
pula member ikan satu sebatian yang belum dikenal
pasti .
Akti vi ti antikulat ekstrak tumbuhan tersebut
telah dikaji dengan menggunakan teknik 'poison food'
dan pereambahan spora. Ekstrak tumbuhan yang dikaji
menunjukkan aktiviti antikulat yang berbeza terhadap
pertumbuhan kulat patogen eili (Capsicum annuum L.).
Ekstrak klorofom G. calcicola yang dikumpulkan dari
Templer Park didapati paling berkesan mereneat
pertumbuhan misellum, pensporalan dan percambahan
spora Chaon ephora cucurbitarium, Colletotrichum
capsici dan C. gloeosporioides.
Sebatian antikulat dalam ekstrak ini telah dapat
dikenal pasti dengan menggunakan kaedah biocerakinan
TLC terhadap Aspergi 11 us f 1avus. Keputusannya
menunjukkan bahawa aktiviti ekstrak tersebut adalah
disebabkan oleh kehadiran komponen utamanya:
flindersina dan desmetoksizantofilina.
xiv
![Page 16: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/16.jpg)
PART I
EXTRACT I ON AND I SOLAT I ON OF
CHEMI CAL COMPONENTS FR OM
GLYCOSH IS CALCICOLA
AND
G. RUPESTRIS
(RUTACEAE)
1
![Page 17: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/17.jpg)
2
INTRODUCTION
Glycosm i s i s a genus o f f o r t y s pe c i e s o f sma l l
t r e e s and shr ubs o f the f am i l y Ru taceae , f ound mos t l y
i n S o uth-Ea s t As i a and S o uth o f Ch i na . F o u r t e e n
s pe c i e s a r e r e po r t e d t o occur i n Ma l ays i a ( Wh i tm or e ,
1 9 8 3 ) . G. calc i cola B.C . S t one and G. rupes t r i s
R i dl e y a r e shr ubs wh i ch g r ow o n l i me stone r oc k s and
l i me s t one h i l l s r e s pe c t i ve l y . G . calc i cola can be
f o und i n Lang k a w i , P e r a k , S e l angor and K e l antan
whe reas G . r upes tr i s occurs in P e r l i s , Kedah , P u l a u
Adang and Langkaw i .
Membe r s o f the genus a r e a r oma t i c and a r e used
med i c i na l l y , ch i e f l y the r o o t s ( B ur k i l l , 1 9 6 6 ). G .
penta phylla Co r r . i s used f o r the t r e a tment o f f ever
in India, vhile G. cochlnchlnensis Plr r e found i n the
Dutch I nd i e s
de c oc t i on o f
i s used t o t r e a t s w o l l en s p l een . A
the l e a ve s and r o o t s o f the l a t t e r ,
a l s o , i s used a s a s t i mu l ant t o d i ge s t i on .
![Page 18: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/18.jpg)
3
L I TERATURE REV I EW
A numbe r o f stud i es o n t he c hem i c a l c o nst i t u e n t s
o f Gl ycos m i s spe c i es have b e e n r e por t ed . T h e spec i es
wh i ch have b e e n i nvest i ga t ed i n d e t a i l i nc l ude G.
pen taphy l l a, G . ma ur l t i ana, G . b l 1ocu l a r l s, G.
arborea, G. cyanocarpa a nd G. ch l or osperma . These
i nvest i ga t i o ns r e ve a l ed t h e presence o f va r i o us
c ompounds be l o ng i ng t o t he a l ka l o i d , c o umar i n a nd
a m i d e gro u ps .
One o f t h e p l a n t s t h a t had been e xhaust i ve l y
i nvest i ga te d was G . pen taphy l l a a nd i t f u r n i shes a
numbe r o f a c r i d on e , ca r ba z o l e a nd qu i n o l one bases .
The r o ot ba r k has be e n r e po r t ed t o e la b o r a t e t hese
a l k a l o i ds . I n 1 9 6 6 , Gov i ndacha r i e t . a l i so l a t e d a
known compound, sk i mm i a n i ne ( 1 ) , C1 4 H1 304 N , m.p .
1 7 60C a nd t hr e e new a c r i d on e a l k a l oids: n or a c r o nyc i ne
( 2 ) , C1 9 H1 7 03N , m . p . 200oC; des-N-me thyl a c r onyc i ne
( 3 ) , C1 9 H1 7 03 N, m . p . 2 68 - 2 7 0oC a nd des-N
me t hylnoracr o nyc i ne ( 4 ) , C1 SH1 5 03 N , m . p . 2 4 6 °C . I n
1969 Cha k ra bo r t y r e ported t he iso l a t i o n o f
g l yc o z o l i ne ( 5) , a
c a r ba zo le d e r i va t i ve .
![Page 19: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/19.jpg)
Me.O
OMe. o OH
OMe
(1) (2)
o OR Me.O
(3) R = Me ( 5 ) R = H ( 4 ) R = H ( 6 ) R = OMe
I n con t i nuat ion o f their s t udies on
4
Me Me.
the
carba zol e a l k a lo i d s of G . pen taphyl l a, Cha kra bort y e t
a l . ( 1 9 74 ) re ported the s t ruc t ura l s t ud i e s o f
g l ycozo l i d i ne (6) , m.p . 1 6 0- 1 6 2 0C from the root bark
of th i s p l a n t . Subse que n t l y , Das and Chowdhury
( 1 9 78 ) d i scovere d a nove l
g l ycoso lone
q u i no l one a l k a l o i d ,
Further
![Page 20: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/20.jpg)
5
s t udy o n the r o ot bark of th i s s pec i es by Chowdhury et
a l . ( 1 9 8 7 ) a f f o rded c a r ba z o le (8), C1 ZH1 9 N , m.p. 2 4 5 0C a nd 3 -methyca r ba z o le (9) , C1 3 H1 1N , m . p . 2 0 7 oC .
Th i s i s the f i rst repo r t o f t he iso la t i on o f c a r ba z o le
fr om a p l a nt source .
OMe.
( 7)
(8) R = H
( 9 ) R = Me
R
An a n t i ba cter i a l c a r ba z o l e a lka l o i d ,
g l yc o z o l id o l (10) , C1 4 H1 3 N02 ' m.p . 2 4 0oC was
d i sc o vered f r om the r o o t o f G. pen ta phylla by
Bha t tacha ryya et al . ( 1 9 8 5 ) . The c ompo und has been
f o und to be a c t i ve a ga i nst some Gra m-pos i t i ve a nd
![Page 21: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/21.jpg)
6
Gram-negative bacteria. In the same year, he and
Chowdhury reported t he struct ure o f a n o t her carba z o l e
a lka o l i d , g l yc o z o l ida l ( II ) , m. p . l S50C . In 1 9 9 2 ,
Jash et a l . i so l a ted two car baz o l e
g l ycoz o l i c i ne (12) ,
source .
I H
(10) Rl = OMe; R2 = Me; R3 = OH
( II ) Rl = R 3 = OMei R 2 = CHO
(12) Rl = H; R 2 = Me; R 3
= OMe
(13) Rl = OMe; R2
= CHOi R 3 = H
a lkal o i ds ,
and
![Page 22: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/22.jpg)
7
An i nve s t i gat i on o f the l eave s of G . pentapbylla
by Bhattachar yya a nd Chowd hur y ( 1 9 85 ) l e d t o the
i solat i on of a qu i n o l on e a l ka l o i d , g l ycol one ( 1 4 ), C 1 6 H 1 9NO) , m . p . IISoC . Fur t he r chem i ca l study on t he
l e ave s o f t h i s s pe c i e s by K a ma r u z zaman e t a l e ( 1 9 8 9 )
a f forded a c a r ba zole alk alo i d , mupami me (15),
C 1 9H 19N0 2 ' m .p. 15 2oC . Th i s i s t he f i r s t C I 8
a l k a l o id r e po r ted' from t h e genus Gl ycosm i s.
S a r k a r a nd Cha k ra bor t y ( 1 9 7 7 ) u nd e r t ook t h e
e xam i na t i 'on o f t h e f l owe r h e a d s o f G . pen taphyl l a
f r om wh i ch t h e y r e ported the i so l a t i on a nd s t r uc t u r e
o f g l ycophym i ne (16), C15 H1 2N 2 0 , m . p . 25 4 °C a nd a n
am i d e , g l ycom i d e ( 17), C1 4 H1 3N O , m . p . 1 2 0°C . I n 1 9 7 9 ,
they i s o l a ted a new qu i na zo l i ne a l k a lo i d ,
glycophym o l i ne ( 18 ) , C1 6 Hl 4N20 , m.p. l 650C .
Wor k on t he s e e d s o f G. pen taphyl l a by
Muk he r j e e et a l e ( 1 9 8 3 ) had y i e ld ed a car ba zo l e
d e rivative, glycozolinine ( 1 9) , m .p. 2 3l-2 3 2°C . From phys i ca l and che mic a l e v i d e nc e , i t s s t r uc t u r e wa s
i d e n t i f i e d a s 6 -h yd roxy - 3-me t hylc a r ba zole .
![Page 23: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/23.jpg)
8
Me-O
o
Me.O Me.O
(14 ) (15)
o
(16)
(17)
![Page 24: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/24.jpg)
9
o Me.. HO
�N -o� N�CH2 _
(16) (19 )
A chem i ca l i nv e s t i ga t i on o f t he r o o t s o f
G. maur i t i ana by Ras t og i e t a l . (1980) r e s u l ted 1n
t h e i s o l a t i on o f two a lka l o i d s :
I-hyd roxy-3-me t h oxy-2- ( 3-me t h y l bu t - 2-enyl ) -N-me thyl a c r i d a n -
9-one (20), m . p . 1 34 - 1 3 6o C a nd 4 , 8 -d i me t h oxy- 3- ( 3-
me t hy l but- 2 - e ny l ) -N -me thyl-2 -qu i n o l o n e ( 21 ) . The
s te m bark of G. ma u r i t i ana c o l l e c ted in G i r i ta l e ,
n o r t h c e nt r a l o f S r i La nka was s h own t o c o n t a i n
c a r ba zo l e a lka l o i d s : g l ycomaur i n ( 22 ) , m . p . 1 9 5 - 1 9 6oC
a nd g l ycoma ur r o l ( 23 ) , m . p . 1 4 9 - 1 50o C by Kumar e t a l .
(1989).
o OH
(20)
Me.O
( 21 )
![Page 25: UNIVERSITI PUTRA MALAYSIA CHEMICAL ...psasir.upm.edu.my/8599/1/FSAS_1995_7_A.pdfUNIVERSITI PUTRA MALAYSIA CHEMICAL CONSTITUENTS OF GLYCOSMIS CALCICOLA AND G. RUPESTRIS (RUTACEAE) AND](https://reader031.fdocuments.net/reader031/viewer/2022021817/5a9e6a347f8b9a62178b52da/html5/thumbnails/25.jpg)
10
I H
(22 )
(23)
A new 5 - hydroacr i d one wa s i s o l a t e d by Bowen e t
a l . ( 1 9 7 8 ) f r om t he l eaves o f G . bilocularis. It was
cha r a c te r i zed as 1 , 5 -d i hyd r oxy-2 , 3 -d i me t h oxy-10-
me thyl -9-a c r i d o ne (24 ), m . p . 2 0 6 - 20 7 oC . I n add i t i o n ,
t hey a l s o i s o l a t ed f ive k n own c ompounds: a r b o r i ne
(25) , m . p . 1 5 4 - 1 5 6 oC; a r bor i n i ne ( 2 6 ) , m . p . 1 7 4 -
1 7 6 0Ci s k i mm i a n i ne (1) , k ok us a g i nine ( 2 7 ) , m . p . 1 70 -
1 7 2 °C a nd g l yce r i ne (28), m . p . 1 4 5- 1 4 6 oC .