Tp ch Ha hc, T. 42 (1), Tr.

99 - 104, 2004

Kt qu ban u v nghin cu thnh phn ha hc

cy l ngn (Gelsemium elegans) ca Vit Nam
Phn I - Phn lp v xc nh cu trc cc oxindol ancaloit
n Ta son 21-11-2003
Li Kim Dung, Trnh Th Thy, Trn Vn Sung,
Phm Gia in, Phm Th Ninh
Vin Ha hc, Vin Khoa hc v Cng ngh Vit Nam
Summary
Chemical investigation of the leaves of Gelsemium elegans growing in Vietnam
resulted in the isolation of the oxindol alkaloids: gelsemine, gelsevirine, humantenine and
gelsenicine. Their structures have been determined on the basis of the IR, MS, 1H- and 13CNMR spectroscopy.
I - M u
Theo cc nh thc vt Vit Nam, chi L
ngn n c ta ch c mt lo i (Gelsemium
elegans, Benth.), thuc h M* tin (Loganiaceae). Cy L ngn hay cn c gi l Co
ngn, Thuc rt rut, i tr ng hay on
tr ng tho l loi cy mc leo kh ph bin
min rng ni Vit Nam. Cy L ngn rt c,
i vi chut nht trng liu c LD50 l 200
mg l kh / 1 kg chut. V vy ng i ta khng
hay dng cy n y l m thuc m ch dng t
t hay vi mc ch u c. Ngo i ra,
Trung Quc hay dng r cy n y l m thuc
cha hi hay cha bnh nm tc. Cy L ngn
* c nhiu nh khoa hc trn th gii
nghin cu v th nh phn ha hc. Cc kt
qu nghin cu cho thy cy n y c cha
nhiu oxindol ancaloit c cu trc a dng nh
gelsemin, gelmixin, kuminin, kuminidin....
Gn y, ng i ta cn tm thy oxindol
ancaloit gn vi iridoit t cy L ngn ca Thi
Lan [1, 2]. Cho n nay trong n c cng nh
trn th gii ch a thy c cng b n o v th nh
phn ha hc cy L ngn ca Vit Nam. Tin

h nh nghin cu th nh phn ha hc l cy L
ngn thu hi ti Lng Sn, chng ti * phn lp
v xc nh cu trc ca cc hp cht triterpen l
axit ursonic v cc oxindol ancaloit c cu trc
ha hc khc nhau. B i n y trnh b y kt qu
nghin cu phn lp v xc nh cu trc ha
hc bn hp cht oxindol ancaloit l gelsemin
(1), gelsevirin (2), humantenin (3) v gelsenixin
(4). Cu trc ha hc ca cc cht tch ra c
xc nh bng cc ph ng php ph hng ngoi
(FT-IR), ph khi (EI-MS), ph cng h ng t
ht nhn (1H-, 13C-NMR) v so snh vi cc t i
liu * cng b.
II - Kt qu v& tho lun
Dch chit metanol 85% ca l cy L
ngn sau khi loi dung mi d i p sut gim,
c chit ln l t bng cc dung mi nhexan, etyl axetat v n-butanol. Ct loi dung
mi thu c cc cn dch chit t ng ng. T
cn dch chit n-butanol axit ha bng HCl
lo*ng v kim ha bng NaHCO3 sau chit
li bng CHCl3, thu c ancaloit tng. T cn
ancaloit n y, bng sc k ct th ng, sc k
99

ct nhanh (flash chromatography) v sc k lp

mng iu ch trn silicagel vi cc h dung

mi thch hp * phn lp c cc ancaloit 1,

2, 3 v 4 sch.
17

O
H

16

H
3

14

10
11

13

12

N4

19

Na
R

20

21
18

O
Gelsemin
Gesevirin
17

16

14
3
7

1: R= H
2: R= OCH3

17

15

15
6

O
N
Me
OMe

15

3
20

19

21

Gelsevirin
(2)
3,80 br s
3,41 d (0,7)
2,02 s

Humantenin
(3)
3,62 d (7,4)

12
14ax
14eq

7,38 d (7,4)
6,97 dt (7,6; 7;6;
0,7)
7,16 dt (7,6; 7,6;
0,8)
6,72 d (7,6)
2,81 dd (14,3;2,7)
1,98 br s

7,37 d (7,3)
7,11 dt (7,6; 7,6;
1,0)
7,31 dt (7,6; 7,6;
1,1)
7,01 d (7,7)
2,60 m
1,69 q (6,7)

15

2,30 m

7,45 d (7,3)
7,06 dt (7,6; 7,6;
1,1)
7,29 dt (7,7; 7,7;
1,1)
6,95 d (7,7; 0,5)
2,83 dd (14,5; 2,9)
2,02 ddd (14,5;
5,6; 2,8)
2,31 m

100

19

4: Gelsenixin

Gelsemin
(1)
3,77 s
3,49 s
1,98 br s

11

Bng 1: S liu ph 1H-NMR ca cht 1 - 4 [500 MHz, CDCl3, gi tr

3
5
6ax
6eq
9
10

18

20

O
N
OMe

18

3: Humantenin

16

14

2,28 m

(ppm), J (Hz)]
Gelsenixin
(4)
3,72 dd (4,5; 1,9)
4,41 m
2,65 m
2,37 dd (15,5; 2,0)
7,53 d (7,5)
7,06 dt (7,6; 7,6; 1,0)
7,25 dt (7,6; 7,6; 1,1)
6,87 d (7,6)
2,37 dd (15,5; 2,2)
2,13 ddd (15,0; 5,5;
3,0)
2,71 d (9,0; 7,4)

16
17a
17e
18ax
18eq
19
21exo
21endo
NbMe
NaOMe
N-H

2,45 d (8,3)
3,90 dd (11,1;1,6)
4,07 dd (11,1;1,6)
5,07 dd (11,1;1.0)
4,93 dd (17,8;11,0)
6,25 dd (17,8;
10.9)
2,75 d (10,6)
2,37 d (10,5)
2,25 s

2,45 d (8,4)
3,98 d (11,1;2,1)
4,09 dd (11,1; 2,1)
4,97 dd (17,8; 1,1)
5,14 dd (11,0; 1,2)
6,24 dd (17,8;
11,0)
2,77 d (10,4)
2,35 d (10,3)
2,25 s
3,96 s

2,33 m
4,19 dd (11,0; 1,0)
4,05 dd (11,0; 5,3)
1,65 d (6,8), CH3

2,45 m
4,27 dd (10,0; 3,4)
3,93 dd (10,0; 3,5)
18-CH3 1,28 t (7,4)

5,38 br q (6,8)

2,87 dd (17,0; 9,3)

2,40 dd (17,0; 7,5)

3,41 d (15,0)
3,33 d (15,0)
2,36
3,98

3,94 s

9,25
Bng 2: S liu ph

C
2
3
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
NbMe
NaOMe

13

C-NMR ca cht 1 - 4 [125 MHz, CDCl3, gi tr

Gelsemin
(1)
179,18
69,28
71,91
50,31
54,03
131,67
128,03
121,57
127,89
109,08
140,63
22,67
35,58
37,98
61,28
112,29
138,27
53,77
65,68
40,50

Gelsevirin
(2)
172,85
69,34
72,25
50,88
52,19
128,10
128,19
122,72
127,91
107,05
139,41
23,00
35,81
38,06
61,46
112,99
138,05
54,01
66,07
40,61
63,07

Ph hng ngoi ca cc ancaloit 1 - 4 u c

hp th c tr ng ca nhm cacbonyl 1710 cm-1.
Ph 1H-NMR u c tn hiu ca 4 proton thm
cnh nhau trong khong 6,7 - 7,4 ng vi
H-9, H-10, H-11 v H-12. Ph 1H-NMR ca

Humantenin
(3)
174,30
72,06
61,46
28,16
55,16
129,05
128,02
119,54
125,78
107,21
138,84
25,24
34,43
38,11
66,84
12,74
122,88
136,91
45,52
42,43
63,28

(ppm)]
Gelsenicin
(4)
171,15
74,72
72,12
37,47
55,67
132,03
124,50
123,18
127,88
106,39
137,85
25,48
39,55
42,29
61,86
9,82
26,82
184,41

63,16

cc cht n y vng tr ng cao t ng i phc

tp v c nhiu tn hiu ca cc proton no
(aliphatic proton) b che ph ln nhau, cho thy
y l cc oxindol ancaloit c cha h vng
khng no (aliphatic polycyclic) [2, 4]. Kt lun
101

n y c chng minh thm qua ph 13C-NMR

v ph DEPT: pha tr ng thp c tn hiu ca
nhm cacbonyl ( 170-180, C-2), 4 nhm CH
ca nhn thm ( 106-132, C-9, C-10, C-11 v
C-12). Pha tr ng cao hn l tn hiu ca
nhm OCH2 ( 61,3-66,8, C-17), Nb-CH2 (
65,7-66,1, C-21) v cacbon vng no ( 23,055,0). Ph khi va chm electron (EI-MS) ca
cc cht 1 - 4 u c pc ion phn t.
Ph khi va chm electron (EI-MS) ca
cht 1 c pic ion phn t m/z 322 [M]+ (40)
(C20H22N2O2) v mt s pic quan trng khc
m/z 279 [M-43]+ (40), 251 [M-71]+ (20), 108
[M-214]+ (100). Pic m/z 120 [M-202]+ do to
th nh ion 1-metyl-3-vinylpyridinium. Ph 1HNMR ca cht 1 c tn hiu ca 4 proton thm
cnh nhau 7,38 - 6,72 v tn hiu ca nhm
cacbonyl C179,16 (C-2) ph 13C-NMR chng
minh cho s c mt ca vng oxindol. Pha
tr ng cao hn l tn hiu ca nhm
metylenoxy ( 61,28, C-17) v nhm CH2-N (
65,68, C-21). Dublet kp ca vinyl proton
6,25 (H-19) v cp dublet 5,07 (Ha-18),
4,93 (Hb-18) cho thy phn t c nhm =CH2
cui mch. iu n y c khng nh thm qua
tn hiu ca nhm =CH2 ( C 112,29) trong ph
13
C-NMR v ph DEPT. Hai ph n y cng cho
bit phn t c 20 nguyn t cacbon trong
c: 1 nhm CH3 ( 40,50, Nb-CH3), 4 nhm
CH2, 10 nhm CH v 5 cacbon bc bn (Cq).
Qua phn tch ph v so snh vi s liu *
cng b trong t i liu [2], * xc nh c cu
trc ca cht 1 l gelsemin. Gelsemin l mt
ancaloit rt c, c bit i vi tim v t ng
i ph bin trong chi L ngn (Gelsemium).
Ph khi EI-MS ca cht 2 c s phn mnh
ph hp vi cht 1, nh ng c hn mt nhm
metoxy (OCH3), c th hin qua pic ion phn
t m/z 352 [M]+ (40) v mt s pic khc m/z
321 [M-31]+ (90), 309 [M-43]+ (35), 291 [M-3130]+ (16), 278 [M-74]+, 108 [M-31-214]+(100).
Pic m/z 323 [M-C2H5]+ (14), 292 [M-C2H5OCH3]+(14), 239 [M-82-31]+, 176 [M-176]+ to
th nh do c s chuyn dng trans [2]. Pic m/z
321 [M-OCH3]+ (90), 291 [M-OCH3-NO]+ (14),
gi cho thy nhm metoxy gn vi nit ca
nhn indol. Kt lun n y c khng nh thm
102

qua ph NMR. So snh ph 1H- v 13C-NMR

ca cht 2 vi cht 1, ta thy hai cht ch c s
thay i nhn indol: ( H 6,72, c 107,05, C12) v c thm mt nhm metoxy ( H 3,96, c
63,07). Cc tn hiu ca h vng monoterpen
u ph hp vi ph ca gelsemin. Qua phn
tch ph v so snh vi s liu * cng b trong
t i liu [2, 3], * xc nh c cu trc ca
cht 2 l gelsevirin hay cn c tn l Nametoxygelsemin.
Ph EI-MS ca cht 3 c pic ion phn t
m/z 354 (60) v mt s pic quan trng khc
m/z 339 [M-15]+ (8), 323 [M-31]+ (100), 178
[M-176]+ (10), 122 [M-232]+ (70). Ph 1H- v
13
C-NMR ca cht 3 c nhn oxindol ging vi
cht 1 v 2, nh ng li c dublet ca nhm
metyl cnh ni i 1,65 (d, J = 6,80 Hz, H318), quartet ca vinyl proton 5,38 (q, J = 6,8
Hz, H-19) v singulet ca nhm N-CH 3
2,36. Ph 13C-NMR ca cht 3, c tn hiu
ca nhm CH2-N chuyn v pha tr ng cao
hn ( 45,52, C-21) v nhm metylenoxy
chuyn v pha tr ng thp hn so vi cht 1, 2
( 66,84, C-17), cho thy phn t cht 3 c s
chuyn v ca ni i mch nhnh (C18-C20).
Kt lun n y c khng nh qua tn hiu ca
nhm =CH ( 122,88, C-19) v Cq cnh ni i
( 136,91, C-20). Ph 13C-NMR v ph DEPT
cng cho bit cht 3 c 21 nguyn t cacbon,
trong c 3 nhm CH3, 4 nhm CH2, 9 nhm
CH v 5 cacbon bc 4 (Cq). Qua phn tch ph
v so snh vi s liu * cng b trong t i liu
[2, 4], * xc nh c cu trc ca cht 3 l
humantenin. Cht n y cng * c tm thy t
cy L ngn ca Thi Lan [2].
Ph EI-MS ca cht 4 c pic ion phn t
m/z 326 (60) v mt s pic khc m/z 321
[M-31]+ (90), 309 [M-17]+ (5), 295 [M-31]+
(100), 150 [M-176]+ (90), 122 [M-176-28]+
(100). Ph 1H- v 13C-NMR ca cht 4 c nhn
oxindol ph hp vi ph ca cht 2 v 3, nh ng
khng c tn hiu ca vinyl proton (H-19) m
thay v o l tn hiu ca nhm etyl c thy
r qua mt triplet ( 1,28, t, H3-18) v hai
dublet ca dublet 2,87 v
2,40 (Ha-19;
Hb-19). Ph 13C-NMR v DEPT cho thy phn

t ca cht 4 c 19 nguyn t cacbon, trong

c 2 nhm CH3, 4 nhm CH2, 8 nhm CH v 5
cacbon bc 4 (Cq). Ph 13C-NMR ca cht 4
pha tr ng thp, ngo i tn hiu ca nhm
cacbonyl C 171,45 (C-2) cn c tn hiu ca
nhm -C=N C 184,41 (C-20); pha tr ng cao
c tn hiu ca nhm metyl 9,82 (C-18),
metylen 26,82 (C-19) v tn hiu ca nhm
metoxy 63,16 (Na-OCH3). Qua phn tch
ph v so snh vi cht 2 v 3, * xc nh
c cu trc ca cht 4 l gelsenixin. Dn xut
19-oxo-gelsenixin cng c tm thy t cy
L ngn ca Thi Lan [2]. Ng i ta * xc nh
c cu hnh tuyt i ca gelsenixin bng
nhiu x tia X (X-ray) v ph nh sc trn
(CD).
Mt iu l th l , da v o kt qu nghin
cu cu trc ha hc cc monoterpen indol
ancaloit c phn lp t chi L ngn, cc nh
khoa hc Nht Bn v Thi Lan * a ra gi
thuyt qu trnh sinh tng hp cc Gelsemium
ancaloit. Cu trc ca cc ancaloit n y * c
a ra mt cch c lp da trn c s pht
sinh sinh hc v kt hp vi ph khi v ph
cng h ng t ht nhn phn gii cao mt chiu
v hai chiu. Tt c cc ph ng php thoi bin
thng th ng nhm xc nh cu trc ca
Gelsemium ancaloit u khng th nh cng.
III - Thc Nghim
1. Ha cht v thit b
Ph cng h ng t ht nhn o trn my
Bruker Avance 500 MHz ti Vin Ha hc:
dch chuyn ho hc ( ) c tnh theo ppm,
tetramethylsilan (TMS) cho 1H-NMR hoc tn
hiu dung mi CDCl3 cho 13C-NMR c dng
l m ni chun, hng s t ng tc (J) tnh bng
Hz. Ph khi EI-MS ghi trn my Hewlett
Packard 5989B MS Engine vi nng l ng ion
ho 70 eV ti Vin Ha hc. im chy o trn
my BOTIUS (Heiztisch Mikroskop) ti Vin
Ho Hc. Sc k lp mng phn tch
(SKLMPT) c tin h nh trn bn mng
nhm silicagel Merck 60F245 trng sn, d y
0,2 mm. Silicagel c ht 0,040 - 0,063 mm c
dng cho sc k ct.

2. X l mu thc vt v ph!ng php chit

Mu cy L ngn c thu hi v o thng 8
nm 2000 ti ng Vn, Lng Sn. Tn cy do
TS. Ng Vn Tri xc nh, tiu bn s 2199
c l u ti Vin D c liu, H Ni. 600 g L
ngn kh c ngm chit vi dung dch
MeOH / H2O (85 : 15). Dch chit c loi bt
dung mi d i p sut gim, sau chit ln
l t vi n-hexan, EtOAc, v n-butanol. Loi
dung mi thu c 3,5 g, 12 g v 15,4 g cn
dch chit t ng ng. Ha tan cn dch chit nBuOH bng dung dch axit HCl (5%), lc b
phn khng tan, dch axit c kim ha bng
NaHCO3 v chit li bng CHCl3 (4 ln). Ct
loi CHCl3 d i p sut gim thu c 2,1 g
ancaloit tng (h m l ng 0,035% so vi l
kh). 2,1 g cn ancaloit c phn tch bng
SKC trn silicagel, dung mi gii hp:
CHCl3/MeOH (98 : 2
80 : 20) thu c 43
phn on (100 ml / phn on). Cc phn on
c sc k lp mng gn nhau c gp li
th nh 7 phn chnh (F-1 - F-7). Cc phn on
c tinh ch tip bng sc k ct nhanh trn
silicagel, hoc sc k lp mng iu ch vi
cc h dung mi thch hp thu c cc
ancaloit 1-4 sch.
Gelsemin (1): Cht du, nc. 1780C, c tch
ra t phn on F-4 bng sc k ct nhanh trn
silicagel, dung mi CHCl3 / MeOH (95 : 5). H m
l ng: 0,013% (so vi nguyn liu kh). Rf =
0,52, dung mi CHCl3 / MeOH (90 : 10). Ph
hng ngoi ( *KBr cm-1): 3249,3; 2921,7 (N-H);
1709,6 (C=O); 1618,4; 1473,1; 1231,0; 1097,1;
909,1; 735. Ph khi EI-MS (70 eV) m/z (%):
322 [M]+ (40), 279 (40), 251 (20), 146 (15), 134
(16), 120 (15), 108 (100), 93 (8), 77 (10). Ph 1H- v
13
C-NMR: Bng 1 v 2.
Gelsevirin (2): Cht bt m u trng, nc. 2450C
c tch ra t phn on F-3, bng sc k lp
mng iu ch, h m l ng 0,0013%. Rf = 0,40,
CHCl3 / MeOH (95 : 5). Ph hng ngoi ( *KBr
cm-1): 2930,0; 2247,9; 1732,9 (C=O); 1625,9;
1472,1; 1331,7; 1084,3; 917,1; 756,6. Ph khi
EI-MS (70 eV) m/z (%): 352 [M]+(40), 321
(90), 309 (25), 291 (12), 278 (12), 250 (10), 108
(100). Ph 1H- v 13C-NMR: Bng 1 v 2.
103

Humantenin (3): Cht v nh hnh mu nu,

thu c t phn on F-2; Rf = 0,50, CHCl3 /
MeOH (95 : 5), h m l ng 0,0028%. Ph hng
ngoi ( *KBr cm-1): 2929,2; 1719,6 (C=O);
1614,6; 1461,5; 1326,5; 1116,4; 912,4; 747,3.
Ph khi EI-MS (70 eV) m/z (%): 354 [M]+
(100), 339 [M-15]+ (8), 323 [M-31]+ (100), 308
(6), 178 (12), 122 (80). Ph 1H v 13C-NMR:
Bng 1 v 2.
Gelsenixin (4): Cht du, c tch ra t phn
on F-1 bng sc k ct nhanh trn silicagel,
dung mi CHCl3 / MeOH (98 : 2), h m l ng
0,005%. Rf = 0,43, CHCl3 / MeOH (95 : 5). Ph
hng ngoi ( *KBr cm-1); 2929,0; 2245,0; 1726,4
(C=O); 1620,2; 1323,6; 1111,2; 914,5; 733,3.
Ph khi EI-MS (70 eV) m/z (%): 306 [M]+
(100) (C20H22N2O), 291 [M-15]+ (7), 277 (7),
263 (18), 223 (25), 120 (20), 108 (16), 70 (45).
Ph 1H v 13C-NMR: Bng 1 v 2.

104

Chng ti xin chn th=nh cm n TS Ng

Vn Tri (Vin DCc liu, H= Ni) v vic thu
hi v= xc nh mu thc vt.
T&i liu tham kho
1. Tt Li (1991). Nhng Cy thuc v V
thuc Vit Nam. Nh xut bn Khoa hc v
K thut, H Ni, Tr. 366.
2. Dhavadee Ponlux, Sumphan Wongseripipanata,
Sanan Subhadhirasakul, Hiromitsu Takayyama, Masaki Yokota, Koreharu Ogata
(1988). Tetrahedron, Vol. 44, P. 5075 - 5094.
3. Schun Yeh, Geoffrey A. Cordell (1986).
Journal of Natural Products, Vol. 49, P. 483 487.
4. Long-Ze Lin, Schun Yeh, Geoffrey A.
Cordell, Chou-Zhou Ni, and Jon Clardy
(1991). Phytochemistry, Vol. 30, P. 679 - 683.