71

Tp ch Ha hc, T. 42 (1), Tr. 71 - 75, 2004

NGHIN CU THANH PHN HA HC CY KYDIA GLABRESCENS
n Ta son 6-5-2003
Nguyn Quyt Chin, Nguyn Vn Hng, Trn vn Sung
Vin Ha hc, Vin Khoa hc v% Cng ngh Vit Nam
summary
Taraxerol (1), scopoletin (2) and (-)-epicatechin (3) were isolated from Kydia
glabrescens along with -sitosterol and -sitosterol glucoside. The identification of the
isolates were made by spectroscopic techniques. In addition, the
13
C-NMR assignments of
taraxerol were revised, based on 2D NMR data.

i - m u
Cy Kydia glabrescens Masters in Hook l
cy i mc thuc h Bng (Malvaceae) [1].
Trong vic thc hin d n iu tra c bn ti
nguyn thc vt tnh Ha Bnh [2] chng ti 8
thu 9c cc mu ca cy ny mc t nhin ti
huyn Mai Chu, tnh Ha Bnh. Da trn kt
qu sng lc ha hc s b, ng thi 9c bit
cy Vng qu cnh (Kydia calycina) cng chi l
mt cy thuc dn gian ca ta v cng 9c
dng lm thuc Trung Quc v n [3],
chng ti 8 chn cy Kydia glabrescens lm
i t9ng nghin cu su hn. T l ca cy, bn
cnh -sitosterol chng ti 8 phn lp v xc
nh 9c cu trc ha hc ca hp cht
tritecpen taraxerol (1). T v ca cy, bn cnh
-sitosterol glucosit chng ti 8 phn lp v xc
nh 9c cu trc ha hc ca cc hp cht
scopoletin (2) v (-)-epicatechin (3). Cc cht
tch ra 9c xc nh cu trc bng s kt hp
cc ph9ng php quang ph hng ngoi (FTIR),
quang ph t ngoi (UV), ph khi l9ng (MS),
ph cng h9ng t proton v cacbon (
1
H- v
13
C-
NMR) v cc ph cng h9ng t ht nhn hai
chiu COSY, HMQC, HMBC, NOESY. y l
cng trnh nghin cu u tin 9c bit v
thnh phn ha hc ca cy Kydia glabrescens.
II - kt qu v# tho lun
Cao chit metanol ca l v v cy Kydia
glabrescens 9c phn b ln l9t trong cc dung
mi n-hexan, diclometan, etylaxetat v n-butanol.
HO
H
1
3
24
23
27
14
15
30 29
20
O O
2
4 5
6
8
4a
8a
HO
MeO
O
OH
OH
HO
OH
OH 1
1
2
3
5
7 9
10
1'
4'
3'
1 2 3
1 2 3
72
Dung mi 9c ct loi d9i chn khng cho cc
cao chit t9ng ng. T cc cao chit n-hexan
v diclometan ca l 8 phn lp v xc nh
9c cu trc ca taraxerol (1) (0,005% so vi
nguyn liu kh) v -sitosterol. Taraxerol
cng cn 9c pht hin thy trong cao chit
etyl axetat. T cao chit etyl axetat ca v 8
phn lp v xc nh 9c cu trc ca scopoletin
(2), (-)-epicatechin (3) v -sitosterol glucosit.
Taraxerol (1): Ph khi va chm electron
(EIMS) cho pic phn t m/z 426 (32) [M]
+
, ng
vi cng thc phn t C
30
H
50
O. Pic c bn m/z
204 (100) v pic m/z 302 (53) th hin s phn
mnh c tr9ng ca cc hp cht tritecpen thuc
nhm taraxeran c ni i gia C-14 v C-15
[4]. Phn tch cc ph NMR v so snh cc tnh
cht ha l khc cho thy 1 l hp cht 8 bit
taraxerol. So snh ph
13
C-NMR ca 1 vi ph
ca taraxerol ghi trong cng mt dung mi 8
cng b trong t9 liu [5] cho thy cc s liu
khp nhau c bn v gi tr, nh9ng khc nhau
v kt qu gn cc gi tr . l mt vic
th9ng gp phi, do lc ch9a c cc ph9ng
php NMR hai chiu nh9 ngy nay. Phn tch
cc kt ni th hin trn cc ph NMR hai
chiu (COSY, HMQC, HMBC v NOESY) ca
1 cho php chng ti 9a ra kt qu gn gi tr
mi th hin trn bng 1.
Bng 1: Kt qu gn gi tr ph
1
H-NMR v
13
C-NMR ca 1 so snh vi t9 liu [5]
S liu o T9 liu
C

H
(J = Hz)
(CDCl
3
, 300 MHz)

C
(CDCl
3
, 100 MHz)
HMBC

C
(CDCl
3
, 25 MHz)
1
1,3 m
1,1 m
37,77 t 38,1
2 1,6 m 27,18 t 27,3
3 3,19 dd (10,3; 4,9) 79,08 d C-23, 24 79,2
4 38,78 s 39,1
5 0,8 m 55,57 d 55,7
6 1,6 m 18,82 t 19,0
7
2,03 m
1,3 m
41,36 t C-5, 6, 8 35,3
8 39,0 s 38,9
9 1,45 m 49,31 d 48,9
10 37,59 s 37,9
11 1,45 m 17,52 t 17,7
12 1,60 m 33,73 t 35,9
13 38,03 s 37,9
14 158,12 s 158,1
15 5,53 dd 116,89 d C-8, 13, 16, 17 117,0
16
1,90 m
1,70 m
37,77 t C-14, 15, 17, 18 36,9
17 35,8 s 38,1
18 0,95 m 48,8 d 49,4
19 1,60 m 36,7 t 41,4
20 28,81 s 29,0
73
21 1,30 m 33,12 t 33,9
22 1,40 m 35,15 t 33,2
23 0,97 s 28,01 q C-3, 5, 24 28,1
24 0,80 s 15,43 q C-3, 5, 23 15,6
25 0,92 s 15,46 q C-1, 5, 9, 10 15,6
26 1,09 s 25,92 q C-7, 8, 9, 14 30,1
27 0,91 s 21,33 q C-12, 13, 14, 18 26,0
28 0,82 s 29,85 q C-16, 17, 18, 22 30,1
29 0,91 s 33,36 q C-19, 20, 21, 30 33,5
30 0,95 s 29,93 q C-19, 20, 21, 29 21,5
Scopoletin (2): thu 9c t cao etylaxetat
ca v cy. Trong khi phn lp bng sc k ct
trn silicagel, 2 9c ra gii ra cng vi mt cht
khc v ng kt tinh vi n d9i dng tinh th
hnh kim trong n-hexan/axeton. Phn tch bng
sc k bn mng ch thy mt vt Rf = 0,54
(CH
2
Cl
2
/ MeOH 95 : 5) pht hunh quang xanh
d9i UV
354
. Tinh ch tip bng sc k lng cao p
iu ch trn ct ng9c pha Grom-Suphir 110
C18 vi h dung mi MeCN / H
2
O (6 : 4) cho 2
sch. Ph khi phn gii cao ion ha bi electron,
thc d9ng (HRESIMS) cho pic m/z 193,0555
[M+H]
+
, suy ra cng thc phn t C
10
H
8
O
4
. Phn
tch cc ph NMR v so snh cc tnh cht ha l
khc cho thy 2 l hp cht 8 bit scopoletin
thuc nhm cc hp cht cumarin. Kt qu phn
tch cc ph NMR ca 2 ph hp vi cc s liu
8 9c cng b [6].
(-)-Epicatechin (3): thu 9c t cao etyl
axetat ca v cy. Ph khi phn gii cao ion
ha bi electron, thc m (HRESIMS) cho pic
m/z 289,073 [M-H]
-
, suy ra cng thc phn t
C
15
H
14
O
6
. Hp cht 3 cho cc ph UV v IR c
tr9ng cho cc hp cht flavan. Phn tch cc ph
NMR v so snh cc tnh cht ha l khc cho
thy 3 l hp cht 8 bit (-)-epicatechin. Kt
qu phn tch cc ph NMR ph hp vi cc s
liu 8 9c cng b [7]. Khc vi (+)-catechin,
dng (2R-trans), c gc quay cc d9ng,
(-)-epicatechin (3), dng (2R-cis) c []
D
- 50,4
0
(c 0,25; EtOH) (t9 liu [8] -68
0
, trong EtOH).
Theo t9 liu [8], taraxerol c hot tnh khng
vi sinh vt, chng vim v chng khi u;
scopoletin c hot tnh chng co tht c trn;
(-)-epicatechin c hot tnh chng vim ty.
iii - thc nghim
im nng chy 9c o trn my Boetius.
Gc quay cc o trn my t ng Perkin-Elmer
model 241 polarimeter. Ph UV ghi trn my
UV 1201. Ph hng ngoi ghi d9i dng vin
nn KBr trn my FTIR Impact-410. Ph khi
EIMS ghi trn my HP 5989B MS. Ph khi
phn gii cao ion ha bi in t thc d9ng v
m (HRESIMS) 9c ghi trn my Q-ToF-2.
Ph NMR 9c ghi trn my Brucker Avance
DPX-300 MHz vi TMS l cht chun ni.
Sc k bn mng 9c tin hnh trn bn
mng trng sn silicagel Merck 60 F
254
, hin
hnh bng thuc th CeSO
4
. Sc k ct 9c tin
hnh vi silicagel (230-400 mesh, Whatman).
Sc k lng cao p 9c thc hin trn h thit
b gm bm cao p Waters 600E v PDA
detect ca Waters 996, ct ng9c pha Grom-
Suphir 110 C18, 12 m, 300 40 mm.
L v v ca cy Kydia glabrescens 9c
thu hi vo thng 6 nm 1999 ti x8 P C,
huyn Mai Chu, tnh Ha Bnh. Tn cy do TS
V Xun Ph9ng xc nh; tiu bn (s 2308)
9c l9u gi ti Vin Sinh thi v Ti nguyn
sinh vt, Vin Khoa hc v Cng ngh Vit Nam.
1. X l v chit tch cht t l
L v v cy 9c phi kh trong bng rm
nhit phng v nghin thnh bt. Bt l
(2,4 kg) 9c chit vi metanol. Dch chit sau
khi c trong chn khng loi phn ln metanol
9c phn b ln l9t trong n-hexan, etyl axetat
74
v diclometan. C kh cc dung dch cho cc
cao t9ng ng: hexan (18,9 g), etyl axetat (14 g)
v diclometan (1 g).
Cao hexan (18,9 g) 9c tch s b trn ct
silicagel vi cc h dung mi l hexan, hexan/
diclometan (1 : 2) v (1 : 4) thnh 3 phn on
gp. Phn on 2 (2 g) kt tinh trong diclo-
metan/metanol cho 1 (17 mg). Phn cn li
9c tch tinh trn ct silicagel, ra gii bng
h dung mi gm n-hexan v l9ng tng dn
diclometan v etyl axetat, thu 9c thm 1 (50 mg)
v -sitosterol (100 mg).
Cao diclometan (1 g) 9c tch trn ct
silicagel, ra gii bng h dung mi gm hexan
v l9ng tng dn etyl axetat cho 1 (62 mg).
Cc phn on sau (62 mg) cha cht chnh l
-sitosterol. Tng s 1 thu 9c qua cc ln
chit tch l 129 mg (0,005% so vi nguyn
liu kh). Kim tra trn bn mng (n-hexan/
diclometan 1 : 4) cho thy cht 1 cng c trong
cao etyl axetat.
2. X l v chit tch cht t v
Bt v (3,6 kg) 9c chit vi metanol.
Dch chit sau khi c trong chn khng loi
phn ln metanol 9c phn b ln l9t trong
n-hexan, etyl axetat v butanol. C kh cc
dung dch cho cc cao t9ng ng: hexan (8,1 g),
etyl axetat (7,8 g). Dch chit butanol qua kim
tra s b cho thy ch cha cc cht mu
polyphenol nn khng 9c nghin cu tip.
Cao etyl axetat (7,8 g) 9c tch s b trn
ct silicagel vi h dung mi gm diclometan
v cc l9ng metanol tng dn (ti 20%) thnh
15 phn on gp. Phn on 2 (220 mg) kt
tinh trong etyl axetat cho -sitosterol (39 mg).
Phn on 4 (100 mg) ch cho mt vt trn bn
mng (Rf = 0,54, CH
2
Cl
2
/ MeOH 95 : 5) v kt
tinh t nhin trong dung mi chy ct d9i
dng cc tinh th hnh kim (13 mg). Kim tra
trn HPLC cho thy y l mt hn hp gm 2
cht c ph UV khc nhau. Hn hp ny 9c
tch bng sc k lng cao p iu ch trn ct
ng9c pha Grom-Suphir 110 C18, 12 m, 300
40 mm, MeCN / H
2
O (60 : 40), 20 ml/min cho 2
(3,1 mg) v mt cht sch khc c thi gian l9u
cao hn (4,5 mg). Cu trc ca cht ny ang
9c nghin cu tip.
Phn on 10 (200 mg) kt tinh trong
metanol nng cho -sitosterol glucosit (51 mg).
Phn on 12 (426 mg) 9c kt tinh trong etyl
axetat nng cho cht 3 (63 mg).
Taraxerol (1): Tinh th hnh kim mu trng,
nc 281 - 282
o
C; TLC: Rf = 0,53 (n-hexan/
CH
2
Cl
2
1 : 4); []
25
D
+ 1,4
o
(c 1,2; CHCl
3
); IR
(KBr)
max
3489 (OH), 2939 (CH), 1474 (CH
2
),
1386, 1037, 1003 cm
-1
; EIMS (70 eV) m/z 426
[M]
+
(32), 411 (16), 302 (53), 287 (47), 204
(100), 189 (31);
1
H-NMR (CDCl
3
; 300 MHz) v
13
C-NMR (CDCl
3
; 75 MHz): xem bng 1.
Scopoletin (2): Tinh th hnh kim mu
trng (trong metanol). nc 203 - 204
o
C; TLC: Rf
= 0,54 (CH
2
Cl
2
/MeOH 95 : 5); UV (EtOH)
max

(log

) 227 (4,14); 255 (3,63); 259 (3,61, vai);

298 (3,66); 345 (4,05) nm; IR (KBr)
max
3285
(OH), 1703 (C=O), 1561 (aromat), 1506, 1291,
1143, 858 cm
-1
; HRESIMS m/z 193,0555
[M+H]
+
(tnh cho C
10
H
9
O
4
: 193,0501);
1
H-NMR
(CDCl
3
; 300 MHz) 7,85 (1H, d, J= 9,5 Hz, H-
4); 7,19 (1H, s, H-5); 6,79 (1H, s, H-8); 6,17
(1H, d, J = 9,5 Hz, H-3); 3,90 (3H, s, 6-OMe);
13
C-NMR (CDCl
3
; 75 MHz) 161,2 (s, C-2);
151,8 (s, C-6); 151,1 (s, C-8a); 145,9 (s, C-7);
144,6 (d, C-4); 113,3 (d, C-5); 112,0 (s, C-4a);
109,9 (d, C-3); 103,7 (d, C-8); 56,69 (q, 6-OMe).
(-)-Epicatechin (3): Tinh th hnh kim mu
trng (trong EtOAc nng), nc 245
o
C; TLC: Rf
= 0,12 (CHCL
3
/MeOH 9 : 1); []
25
D
- 50,4
0
(c
0,25; EtOH); UV (EtOH)
max
(log

) 231
(4,296); 279 (3,717) nm; IR (KBr)
max
3455
(OH); 1628 (aromat); 1521 (aromat); 1259,
1144, 1091; 795 cm
-1
; HRESIMS m/z 289,0733
[M-H]
-
(tnh cho C
15
H
13
O
6
: 289,0712);
1
H-NMR
(CDCl
3
; 300 MHz) 6,97 (1H, s rng, H-2);
6,80 (1H, dd, J = 8,1; 1,3 Hz, H-6); 6,75 (1H, d,
J = 8,5 Hz, H-5); 5,94 (1H, d, J = 2,0 Hz, H-8);
5,92 (1H, d, J = 2,0 Hz, H-6); 4,81 (1H, cm
pic trng nhau, H-2); 4,17 (1H, s rng, H-3);
2,86 (1H, dd, J = 17,0; 2,4 Hz, H-4b); 2,73 (1H,
dd, J = 17,0; 4,4 Hz, H-4a);
13
C-NMR (CDCl
3
;
75 MHz) 158,01 (s, C-9); 157,68 (s, C-5);
157,39 (s, C-7); 145,95 (s, C-3); 145,79 (s, C-4);
138,31 (s, C-1); 119,4 (d, C-6); 115,9 (d, C-5);
115,3 (d, C-2); 100,13 (s, C-10); 96,4 (d, C-8);
75
95,9 (d, C-6); 79,8 (d, C-2); 67,5 (d, C-3);
29,2 (t, C-4).
Cng trnh n%y Ac ho%n th%nh vi s h
tr t%i chnh ca D n cp nh% nAc 1999 -
2000 iu tra c bn t%i nguyn thc vt tnh
Ha Bnh.
t#i liu tham kho
1. Phm Hong H, Cy c Vit Nam (An
illustrated flora of Vietnam), Quyn I., Tr.
5300. NXB Tr, H Ni (1999).
2. Trn Vn Sung. Bo co kt qu thc hin
D n cp nh n9c 1999 - 2000 iu tra
c bn ti nguyn thc vt tnh Ha Bnh.
3. V Vn Chi. T in Cy thuc Vit Nam,
Tr. 1334. NXB Y hc (1996).
4. K. Shiojima, Y. Arai, K. Masuda, Y. Takase,
T. Ageta, H. Ageta. Chem. Pharm. Bull.
Vol. 40, Tr. 1683 - 1690 (1992)
5. N. Sakurai, Y. Yaguchi, T. Inoue. Phyto-
chemistry, Vol. 26, P. 217 - 219 (1987).
6. Aldrich Library of 13C & 1H FT NMR
Spectra, 2, 1314 (1992).
7. B. E. Breitmaier, W. Voelter, Carbon-13
NMR Spectroscopy. VCH Publishers, New
York, P. 455 (1987).
8. Dictionary of Natural Products on CD-ROM,
Version 9 : 1, Chapman & Hall/CRC (2000).