V
^
DAI HOC QUOC GIA HA NOI
TRirdNG DAI
HOC KHOA HOC
Tl/
NHlfiN
4e4G*4c4c4e4c4e4(
T£N
D£
TAI
:
NGHlfiN ctru
CAC HOP CHAT CO HOAT TINH SINH HOC TlT
CAC CAY THUdc CUA
VI£T
NAM
MA SO: QT - 06
-19
CHU TRI
f)t
TAX:
TS PHAN MINH GIANG
;.Al
HGC
QUOC GlA
HA
Npi
|
DT
/

CN Nguyen Thi
Tu6
d. Muc tieu va noi dung nghien curu
• Sang lgc
lira
chon mgt so
c^y
thuoc co gia tri su dung cao trong Y hoc
d^n
toe
cua
Vi6t
Nam, giam dinh thuc
vat,
thu hai nguyen
lidu
thuc
vat
dung
cho nghien
cihi.
•
Xtr
ly mSu thuc vat, nghien
cihi
qui
trinh
chiet va phan tach
d^
dieu che

cihi.
e. Cac ket qua dat
dirge
1.
Da tien hanh sang lgc lua chgn cay Nghe xanh {Curcuma aff.
aeruginosa Roxb., Zingiberaceae), mgt cay thuoc mdi
dugc
phat hien,
dang duac trong dai tra, va ducfc
sii diing
trong Y hoc dan tgc de dieu
tri cac benh nhu dau
biing
va tang cudng tieu hoa lam doi tugng nghien
ciiu
cua de tai. Da lien hanh giam dinh thuc vat va thu thap nguyen lieu
thuc vat
phue
vu cho nghien
ciiu.
2.
Da
xay
dung dugc cac qui tnnh chiet va phan tach chgn lgc de
di^u
che
cac
phdn chiS't
c6 hoat tfnh sinh hgc,
//-hexan

qua nghien
cthi
cac hop chdt de bay hoi cua Nghe xanh da xac
dinh dugc tdng so 23 hgp chdt (chii yeu la cac sesquiterpenoid) vdi
P-elemen
- mgt hgp chdt co hoat tfnh
chdng
ung thu la thanh phdn
chfnh cua cac sesquiterpen hydrocarbon, va curzerenon, curcumol,
germacron va curdion la cac thanh phdn chfnh
ciia
cac sesquiterpenoid
chiia oxy, trong so do curzerenon da dugc sit dung lam thuoc lam sang
cho ung thu da va c6 tur cung.
5.
Cac ket qua nghien
ciiu
cac hgp chdt phan
cue
hon da ddn den viec phan
lap
mirdi
bdn hgp
chS't
tinh
kiiiet tii
than re Nghe xanh, curdion (1), axit
palmitic (2), zederon (3),
y0-sitosterol
(4), stigmasterol (5),

Candida albicans va Fusarium oxysporum cf
miic
MIC
tir
25-200
)ag/ml.
Cac phan tach co dinh hudfng khang khudn va khang ndm da ddn den viec
phan lap cac hoat chdt la curdion va zederon.
- Curdion va zederon (cac MIC 25
|ig/ml)
co
th^
dugc phat tridn thanh
cac tac nhan chdng benh thdi re
6
thuc vat gay
bdi
ndm mdc Fusarium
oxysporum.
-
Curdion va zederon da dugc chiing to la cac hgp chdt khang cac vi nam
gay benh cho ngucfi tuong
iing
la Aspergillus niger (MIC 50
|ag/ml)
va
Candida albicans (MIC 25
)ig/ml).
- Curdion (MIC 50
^g/ml),

td la co tac
dung
lie
che'
sir
san
sinh
cua nito oxit (NO).
8. Trong
cac
thur nghiem
v^
hoat tfnh chdng
oxy hoa (dgn gdc tu do) va
gay
dgc te'
bao ddi vdi ba
ddng
te' bao ung thu
ngudi
la
Hep-Gj,
Fl va RD, ca
curdion
va
zederon
diu
khong
(hi
hien

sinh vien
nam
thur
3 da
hoan thanh nghien
ciiu
khoa
hgc
trong khudn
khd
ciia
de tai nay.
f.
Tinh hinh
kinh
phi cua de tai
Tdng kinh
phf
dugc cdp: 15.000.000 ddng
Cac khoan
chi
cila
de tai da
dugc phan
bd nhu sau:
Muc
(%)
109
110
114

4,0
100.0
KHOA QUAN
Lt
(Ky va ghi ro ho ten)
CHU TRI
Dt
TAI
(Ky va ghi ro ho ten)
Phd Chu nhiem Khoa Hoa hoc
PGS.
TS Trdn Thi
Nhir
Mai
il.
'iim
TS Phan Minh Giang
TRlTCfNG
DAI HOC KHOA HOC
Tl/NHlfiN
PHOHI^UTRUONU
PGSJS.^Aw^iu^am.
Outline Report
a. Project title
Study on biologically active compounds from Vietnamese medicinal
plants
Code
No.: QT-06-19
b.
Head of the project

2.
A selective extraction and fractionation procedure was established for the
preparation of the biologically active n-hexan (3.14% yield on the basis of
the dry material), ethyl acetate (0.6%) and
/i-butanol
(0.14%) extracts.
3.
A suitable chromatographic isolation method was developed for the
systematic chemical and biological study of the
/z-hexan
extract of the
rhizomes of C. aff. aeruginosa. By using this method the main
constituents curdion and zederon were isolated in the yields of 0.1% and
0.05%,
respectively, of the dry material. This method will be suitable for
the development of large-scale isolation of the biologically active curdion
and zederon, as well as pharmaceutical preperations containing different
groups of sesquiterpenoids.
4.
The study of the volatile compounds of C. aff. aeruginosa determined in
total 23 compounds (mainly sesquiterpenoids), of which
P-elemene
- an
anticancer compound - was the main constituent of the sesquiterpene
hydrocarbons, and curzerenone, curcumol, germacrone, and curdione
were the main components of the oxygenated sesquiterpenoids, of which
curzerenone was used as a clinical drug for cervix and skin carcinomas.
5.
The study of the more polar compounds led to the isolation of fourteen
pure compounds from the rhizomes of C. aff. aeruginosa, curdione (1),

xylem
of plants caused by Fusarium oxysporum.
- Curdione and zederone were demonstrated to be active against the
human pathogenic fungi Aspergillus niger (MIC of 50
^g/ml)
and
Candida albicans (MIC of 25
|ig/ml),
respectively.
- Curdione (MIC of 50
^g/ml),
ethyl acetate (MIC of 200
^ig/ml),
and
n-butanol (MIC of 50 fig/ml) extracts showed inhibitory effect against
Staphylococcus aureus. They can be selected for the study toward the
discovery of active compounds against methicillin-resistant
Staphylococcus aureus (MRSA) and vancomycin-resistant (VRSA)
activities.
7.
The hepatoprotective compounds were isolated and identified in the
rhizomes of C. aff. aeruginosa such as furanodiene, germacrone,
curdione,
13-hydroxygermacrone,
and aerugidiol. Furanodiene and
13-hydroxygermacrone
were demonstrated to inhibit the nitric oxide (NO)
production.
8. In the antioxidant (radical scavenging activity) and cytotoxic activity
against the human cancer cell lines

5
2.5 Phan tach phdn chiet n-hexan va phan lap cac sesquiterpenoid 6
2.6 Phuong phap
thii
hoat
tinh
sinh hoc 6
2.6.1
TTiir
hoat tfnh khang khudn va khang ndm in vitro 6
2.6.2 Thijf hoat tfnh chdng oxy hoa in vitro 6
2.6.3
Hiir
hoat tfnh gay ddc
te'
bao in vitro 6
2.7 Hdng sd vat ly va du kien phd cua cac hgp chdt dugc phan lap 7
2.8 Ket qua va Thao luan
11
2.8.1 Phan lap va xac dinh cdu tnic cac sesquiterpenoid
tir
Nghe xanh
11
(C.
aff.
aeruginosa,
Zingiberaceae)
2.8.2 Danh gia cac hoat tfnh sinh hoc cua Nghe xanh 22
(C.
aff. aeruginosa, Zingiberaceae)

Gas chromatography
GC-MS: Gas chromatography - Mass spectroscopy
UV: Ultraviolet spectroscopy
IR: Infrared spectroscopy
NMR: Nuclear magnetic resonance
ID
NMR: One dimenssional nuclear magnetic resonance
2D NMR: Two dimenssional nuclear magnetic resonance
^H-NMR:
Proton nuclear magnetic resonance
^^C-NMR:
Carbon 13 nuclear magnetic resonance
DEPT: Distortionless enhancement by polarization transfer
NOE:
Nuclear Overhauser effect
NOED: Nuclear Overhauser effect difference spectroscopy
NOESY: Nuclear Overhauser effect spectroscopy
'H-^H
COSY: Proton - proton correlation spectroscopy
HMQC:
Heteronuclear multiple quantum correlation
HSQC:
Heteronuclear single quantum correlation
HMBC:
Heteronuclear multiple bond correlation
CD:
Circular dichroism
1
Ldl MCJ
DAU

vuc hien nay tap
trung vao viec phat hien cac hgp chdt
"ddn
dUcmg'
cd hoat tfnh sinh hpc cao tir
nguon thien nhien va tdi uu hoa cac hgp chdt
''ddn
ducrtig'
tir thien nhien su
dung cac phuong phap
chuye'n
hoa hoa hgc. Mot sd nghien
ciiu
gdn day
ciia
chiing tdi da phat hien dugc cac
enf-kauran
diterpenoid tir cay Khd
sam
cho
la
{Croton tonkinensis Gagnep., Euphorbiaceae) va chalcon cardamomin tir cay
Rieng rirng {Alpinia conchigera
Griff.,
Zingiberaceae)
th^
hien cac tac dung
chdng viem manh thdng qua viec
lic
che yeu td phien ma

lam thudc va thuc pham. Y hoc dan tdc Viet Nam thudng sir dung
cac loai Nghe {Curcuma) nhu Nghe vang {Curcuma longa L.), Nghe den
[Curcuma zedoaria (Berg.) Roscoe], Nghe
re
vang {Curcuma
xanthorrhiza
Roxb.),
Nghe ten ddng {Curcuma aeruginosa Roxb.) va Nghe udng {Curcuma
aromatica Salisb.) trong dieu tri cac benh ung thu, viem loet, xO gan va an udng
khdng tieu. Trbng mot chuong
Uinh
tie'p tuc nghien
ciiu
ve cac hgp chat cd hoat
tfnh sinh hgc i\s cac cay thudc dan tdc Viet Nam, Nghe xanh {Curcuma aff.
aeruginosa Roxb.), mdt loai Nghe mdi dugc phat hien, da dugc
lira
chon
lam
ddi
tugng nghien cuu ve hoa hgc va hoat tfnh sinh hoc.
Nghe xanh (C. aff. aeruginosa Roxb.) la loai cay thao cao tir 1-2 m. Vc
mat thuc vat hgc cay Nghe xanh tuong tu nhu cay Nghe ten dong (C.
aeruginosa Roxb.) [8]. Tuy nhien, didm khac biet giua Nghe xanh va Nghe
ten ddng la
Idt
cdt ciia than rl Nghe xanh thdm hon cua Nghe ten ddng, hoi
nga sang mau xanh muc Ciru Long. Hien nay cay dang dugc trdng d Ha Ndi,
Vmh Phiic va Hai Phdng di thu than
re

ciiu
cua de tai la
cay
Nghe xanh {Curcuma aff. aeruginosa
Roxb.,
Zingiberaceae) dugc
lira
chgn
uen
co sd tac dung chira benh Uong Y hpc
dan tdc ciia Viet Nam. Cac loai Nghe {Curcuma) thupc hp Zingiberaceae thudng
dugc
sir
dung mdt
each
an toan trong thuc pham va Y hgc dan tpc do dd cac hgp
chdt cd hoat tfnh sinh hpc dugc phat hien tir cac loai nay se cd
trie'n
vpng phat
tridn
thanh cac tac nhan dieu
Ui
cac loai benh khac nhau.
Phuong phap nghien ciru he thdng ve hoa hpc dugc bdt dau bdng chiet chpn
lpc cac ldp chdt chfnh
sir
dung cac dung mdi hiiu co theo thu' tu dp phan
cue
tang ddn. Mpt phuong phap phan tach cac phdn chiet dugc phat trien tren co sd
cac khao sat sdc ky; phuong phap phan tach nay cho phep gpp cac hgp chdt

NMR)
va hai chidu (2D NMR) vdi cac ky thuat khac nhau. Cdu tnic lap thd mong ddi
va tuyet ddi ciia cac hgp chdt nay dugc xac dinh bdng phan tfch cac phuong
phap phd nuclear Overhauser effect mdt chieu (NOED) hoac hai chieu
(NOESY) va phd circular dichroism (CD).
Cac phdn chiet dugc thir nghiem cac hoat tfnh sinh hgc nhdm dinh vi cac
hgp chdt cd hoat tfnh sinh hpc cdn dugc xac dinh. Cac hgp chdt tinh khiet dugc
phan lap dugc ihix nghiem mgt sd hoat tfnh sinh hgc (hoat tfnh khang khuan va
khang ndm, chdng oxy hoa va gay ddc te' bao) trong mpt mdi tuong quan vdi
hoat tfnh cua cac phdn chie't va tac dung dugc ly hgc dan tpc cua
cay
thudc
dugc nghien ciru.
Phuong phap nghien
cixu
nay dugc tdm tdt trong So dd
1.
So do 1. Qui trmh phan lap cac hgp chat co hoat tinh sinh hgc tir
Nghe xanh (Curcuma aff.
aeruginosa^
Zingiberaceae)
Curcuma aff. aeruginosa Roxb.
(Zingibearceae)
I
Cac phdn chiet
c
phan tich sac ky
hoat
tinh
sinh hoc

Phd cdng hudng tir proton
(*H-NMR)
va cacbon 13
(^^C-NMR)
dugc ghi
tren cac may Bruker
AV
500 spectrometer va JEOL
JNM-a
400 NMR
spectrometer.
Sdc ky ldp mdng (TLC) dugc thuc hien tren ban mdng Uang sdn Merck
Alufolien DC 60
F254.
Cac vet chdt dugc phat hien bang
each
phun thudc thir
10%
H2SO4
trong 50% EtOH hoac
vanilin/H2S04
dac 1% sau dd ho ndng d
200°C,
va bang den tir ngoai d budc sdng
X
254 nm. Chdt hdp phu
dimg
cho sdc
ky cdt la silica
gel

dugc thu thap d
Phii
TTiuy,
Gia Lam, Ha Ndi.
20 kg than rl nghe xanh tuoi dugc rira sach, thai lat mdng, phoi khd trong
bdng dam, sdy d 45-
50°C,
rdi xay thanh bdt min, thu dugc 5 kg bdt khd.
2.4 Dieu che cdc phdn chiet
tit
thdn ri Nghe xanh
(C.
aff. aeruginosa)
Bdt than
re
Nghe xanh khd (5 kg) dugc ngam chiet trong metanol 6 Idn
6
nhiet do phdng, mdi ldn trong 3 ngay. Sau khi lgc bd ba, dich chiet metanol
dugc cdt loai dung mdi dudi ap sudt giam xudng cdn 1/10 the tfch, roi pha
loang vdi nudc cdt. Dich
metanol-nudc
dugc chiet chgn lpc vdi n-hexan,
EtOAc va n-butanol, cac dich chie't dugc lam khan bdng
Na2S04,
sau dd dugc
cdt loai dung mdi cho cac phdn chiet tuong
iing
la phdn chie't n-hexan (157 g,
hieu sudt 3,14% so vdi lugng nguyen lieu khd). etyl axetat (30 g, 0,6%) va
n-butanol

g),
bang CC tren silica gel vdi he dung mdi n-hexan-EtOAc (13:1 va 9:1) va kcl
tinh lai trong n-hexan cho 1,65 g zederon (3) va 40 mg hdn hgp cua
/?-sitosterol
(4) va stigmasterol (5). Nhdm phan doan 5 (3,1 g) dugc phan tach bdng sdc ky
cdt CC tren ODS
(MeOH-H20,
3:2
va
7:3) sau dd dugc tinh che bdng HPLC
dieu che' tren cdt ODS
(MeOH-H20,
7:3) dd cho
13-hydroxygermacron
(6)
(53,9 mg) va aeruginolacton (7) (21,2 mg). Nhdm phan doan 6 (4,0 g) dugc
phan tach bdng CC tren ODS
(MeOH-H20,
3:2 va 7:3) va
ti^p
tuc bdng HPLC
dieu che'
uen
cot ODS
(MeOH-H20,
3:2) dd cho guaianediol (8) (18,2 mg),
isoprocurcumenol (9) (42,3 mg) va procurcumenol (10) (95,7 mg). Nhdm phan
doan 7 (1,7 g) va 8 (1,7 g) dugc phan tach bdng CC tren ODS
(MeOH-H.O,
3:2

diphenyl-1-picrylhydrazyl (DPPH) trong dung dich etanol bao hoa [10].
2.6.3
Thic tdc
dung gdy
dgc
tebdo in vitro
Tac
diing gay
ddc te' bao dugc
thii
theo phuong phap dang dugc thuc hicn tai
Vien nghien
cihi
ung thu Qudc gia Hoa Ky (National Cancer Institute, NCI)
[11] vdi ba ddng te' bao ung thu ngudi la
Hep-G2
(ung thu gan),
R
(ung thu
mang tir cung) va RD (ung thu mang tim).
2.7 Hang so
vdt
ly vd
du:
kien
phd
cua cdc hop
chdt
duac phdn lap
Curdion (1):

CH3),
0,98 (3H, d,
7
= 7,0 Hz,
I5-CH3),
1,58
(IH,
dddd,
7
= 4,1 Hz, 3,2 Hz, 3,1 Hz,
3,0 Hz, Ha-3), 1,66 (3H, s,
I4-CH3),
1,88
(IH,
dqq, J
=
2,1 Hz, 6,6 Hz, 6,7 Hz, H-
11),
2,07-2,16 (3H, m, 2H-2, Hp-3), 2,34
(IH,
dddd, J = 7,6 Hz, 7,1 Hz, 4,1 Hz,
2,2 Hz, Ha-4), 2,39
(IH,
dd, J = 14,0 Hz, 2,2 Hz, Hp-6), 2,69
(IH,
dd,
/
= 8,4 Hz,
8,0
Piz,

9:1).
IR (KBr):
v„,,
3500-2500, 1703 cm'.
EI-MS:
m/z 256
{C,^U,^0„
W).
Zederon (3):
Tinh
the'
hinh kim, mau trang, dnc
150-152°C,
[a]"+265,8°
(r 1,0, CHCI3).
Rf
= 0,77 (TLC, silica gel,
/i-hexan-EtOAc,
4:1).
IR (KBr):
v„3,
1698, 1457, 1275, 1056, 921 cm'.
EI-MS:
m/z 246
(Ci^Hi^Oj,
M^).
'H-NMR
(CDCI3):
5
1,29

16,4 Hz, Hb-9), 3,81
(IH,
s, H-5), 5,48
(IH,
m, H-l), 7,08
(IH,
brs, H-12).
"C-NMR
(CDCI3):
6 10,2 (q, C-13), 15,1 (q, C-15), 15,7 (q, C-14), 24,6 (t, C-2),
37,9 (t, C-3), 41,8 (t, C-9), 63,9 (s, C-4), 66,5 (d, C-5), 122,1 (s, C-10), 131,0 (s, C-
7),
123,2 (s, C-11), 131,1 (d, C-l), 138, d (C-12), 157,0 (s, C-8), 192,1 (s, C-6).
H6n hop cua p-sitosterol va stigmasterol (4/5):
Tinh
th^
hinh kim, mau tring,
di^m
nong chay
134-136°C.
Rf
= 0,43 (TLC, silica gel,
«-hexan-etyl
axetat, 9:1).
IR
(KBr):
v^^
3434,
1648, 1380, 1050, 1022,
958 cm'.

7 =
7,3
Hz,
27-CH3),
0,84
(3H,
t,
7
=
7,6
Hz,
29-CH3),
3,51
(IH,
tt,
7 =
11,7
Hz,
5,1
Hz, H-3),
5,35
(IH,
br
d,
7 =
5,1
Hz,
H-6),
1,0
(3H,

d,
7
=
6,5
Hz,
2I-CH3),
1,0
(3H, d,
7 = 6,5
Hz,
I9-CH3),
3,51
(IH,
tt,
7
= 11,7
Hz,
5,1
Hz, H-3),
5,35
(IH,
br
d,
7
=
5,1
Hz,
H-6),
5,0
(IH,

23,1
(t, C-28),
24,3
(t,
C-
15),
26,1
(t, C-23),
28,2
(t, C-16),
29,1 (d,
C-25),
31,4
(t, C-7),
31,7
(t,
C-2),
31,9
(d, C-8),
33,4
(t,
C-22),
39,8
(t,
C-12),
36,2
(d, C-20),
36,5
(s, C-10),
37,3

I5-CH3),
1,58 (3H, s,
I4-CH3),
1,76 (3H, s.
I2-CH3),
2,92
(IH,
d, 7 = 10,8 Hz, Ha-9), 3,81
(IH,
d, 7 = 10,8 Hz, Hb-9),
4,12(1H, d,7=
12,4Hz,
Ha-13),
4,23
(IH,
d, 7= 12,4 Hz,
Hb-13),
4,62
(IH,
br d, 7 = 6,8 Hz, H-l), 4,93
(IH,
br d, 7 =
11,2
Hz, H-5).
"C-NMR
(CDCI3):
8 16,1 (q, C-15), 17,2 (q, C-14), 18,3 (q, C-12), 24,6 (t, C-2),
29,1 (t, C-6), 38,6 (t, C-3), 56,0 (t, C-9), 63,2 (t, C-13), 125,6 (d, C-5), 126,9 (s, C-
4),
132,0 (s, C-7), 133,6 (d, C-l), 136,1 (s, C-10), 140,2 (s, C-11), 207,8 (s, C-8).

1,80
(3H, s,
I3-CH3),
1,89
(IH,
m, H-3), 2,07
(IH,
m, Ha-6), 2,24
(IH,
m, H-3), 2,26
(IH,
m, Hb-2), 2,31
(IH,
m, Ha-9), 3,05
(IH,
m, Hb-9), 3,08
(IH,
m, Hb-6),
4,24
(IH,
brd,
7= 9,5 Hz, H-5), 4,77
(IH,
br s, H-l).
"C-NMR
(CDCI3):
8 8,6 (q, C-13), 16,8 (q, C-15), 17,3 (q, C-14), 25,3 (t, C-6)
25,6 (t, C-2), 38,4
(t,
C-3), 51,3 (t, C-9), 109,8 (d, C-8), 123,4 (d, C-5), 127,4 (s, C-

I4-CH3),
5,52
(IH,
br s, H-6).
"C-NMR
(CDCI3):
8 21,7 (q, C-14), 21,94 (q, C-13), 21,95 (q, C-12), 22,1 (t, C-
2),
23,1 (q, C-15), 25,6 (t, C-8), 37,8 (d, C-11), 41,0 (t, C-3), 43,2 (t, C-9), 50,9
(d, C-5), 51,3 (d, C-l), 75,8 (s, C-10), 80,8 (s, C-4), 121,9 (d, C-6), 150,1 (s, C-7).
Isoprocurcumenol (9):
Bot vo dinh hinh,
[a]"-62,5°
(c 1,82,
CHCl,).
UV (MeOH):
X^,^
nm (log
e)
248 (3,75).
CD (MeOH): Ae (nm): -1,15 (224), +0,94 (265), -0,51 (308)
(c
5,30x10"'
M).
'H-NMR
(CDCI3):
8 1,16 (3H, s,
I5-CH3),
1,36
(IH,

(t, C-14), 135,4 (s, C-7), 142,4 (s, C-10), 144,8 (s,
C-11),
204,2 (s, C-8).
Procurcumenol (10):
Bot vo dinh hinh,
[a]"+60,9° (r
0,86,
CHCI3).
UV (MeOH):
X„,^
nm (log e) 258 (4,30).
CD (MeOH): Ae (nm): -58,47 (214), -62,25 (237), +44,39 (278), +28,39 (334)
(c
7,69x10''M).
'H-NMR
(CDCI3):
8 1,15 (3H, s,
I5-CH3),
1,67 (3H, s, 12-CH3), 1,68 (3H,
s,
I3-CH3),
1,80 (3H, d, 7 = 1,2 Hz,
I4-CH3),
2,09
(IH,
dd, 7
=
16,0 Hz, 12,4
H/„
Hb-6),

8 1,32 (3H, s,
I5-CH3),
1,70
(IH,
ddd, 7 = 13,6 Hz,
11,6
Hz,
6,4 Hz, Ha-2), 1,80 (3H, s,
U-CHj),
1,80
(IH,
m, Ha-6), 1,91 (3H, s,
I2-CH3),
1,91
(IH,
m, Ha-3), 1,93 (3H, s,
I3-CH3),
1,94
(IH,
m, H-5), 2,04
(IH,
dd, 7
=
11,6
Hz, 4,8 Hz, Hb-3), 2,13
(IH,
m, Hb-2), 2,56
(IH,
br d, 7 = 14,0 Hz, Hb-6),
4,18

1707, 1651, 857 cm'.
'H-NMR
(CDCI3):
8 0,92 (3H, s,
I4-CH3),
1,38 (3H, s,
I5-CH3),
1,52
(IH,
m,
Ha-3),
1,69 (3H, m, 2H-2, Hb-3), 2,10 (3H, s,
I3-CH3),
2,47
(IH,
s, H-5), 2,62
(IH,
d, 7 = 16,8 Hz, Ha-9), 2,99
(IH,
d, 7 = 16,8 Hz, Hb-9), 3,57
(IH,
br s, H-
1),
7,0
(lH,s, H-12).
"C-NMR
(CDCI3):
8 8,8 (q, C-13), 14,4 (q, C-14), 24,3 (q, C-15), 27,8
(I,
C-2),

Hz, H-l), 7,05
(IH,
br s, H-12).
'^C-NMR
(CDCI3):
8 8,7 (q,
C-13),
12,7 (q, C-14), 26,7 (t, C-2),
31,2
(q,
C-15),
38,3 (t, C-3), 39,1 (t, C-9), 45,6 (s, C-10), 59,8 (d, C-5), 69,8 (s, C-4), 78,4 (d, C-
1),
118,7
(s, C-7), 120,4 (s,
C-11),
139,5 (d, C-12), 165,3 (s, C-8), 197,9 (s, C-6).
Aeruginon (14):
Bot vo dinh hinh,
[«]"
+173°
(c 0,76,
CHCI3).
'H-NMR
(CDCI3):
8 0,82 (3H, d, 7 = 6,4 Hz,
I2-CH3),
0,88 (3H, d, 7
=
6,4 Hz,

tiit
Nghe xanh (C. aff.
aeruginosa, Zingiberaceae)
Trong mot thdng bao trudc chung tdi da thdng bao ve ket qua xac dinh
mudi tam sesquiterpenoid
dS
bay hoi
tir than
re Nghe xanh (C. aff. aeruginosa)
sir
dung cac ky thuat GC va GC-MS, trong sd dd bay sesquiterpenoid da duoc
phan
lap
bang
each
ket hop ky thuat chung
cS^t
Kugelrohr va phan tach
sac
ky
cot tren silica gel [4]. Trong de tai nay mot phuong phap chiet va phan tach
mdi da duoc phat
tri^n
d^
nghien
ciru
cac hop
chS't
cd hoat tfnh sinh hoc
tir

lang
dfo.
Bang phuong phap chiet chon loc nay cac
phSn
chiet
/i-hexan
(hieu sua't 3,14% so vdi luong nguyen lieu khd ban dau), etyl axetat (0,6%)
va
Az-butanol
(0,14%) da duoc
di^u
che.
Phan chiet
/i-hexan
duoc phan tach bang cac phuong phap
sac
ky cot
gradient tren silica gel,
Ircn
ODS va HPLC dieu che tren cot ODS
d^
cho
mudi bon
hgfp
chat tinh khiet, curdion (1), axit palmitic (2), zederon (3),
/^-sitosterol
(4), stigmasterol (5),
13-hydroxygermacrone
(6),
aeruginolacton (7), guaianediol (8), isoprocurcumenol (9), procurcumenol

afT.
aeruginosa Roxb. (Zingibearceae)
ihAnrekh6(2kg)
i
n-hexan
Dg&m chie't
bing MeOH,
ohi^t
dd
ph^og
Ph^n chi^'l
MeOH (CA)
phAo b6
giira
?I20
va dung mdi
bihi
ca
EtOAc
n-BuOH
f
CAI
157
g
CA2
30
g
CA3
7g
Hi

4g
CA26-27
1.7
g
CA28-31
l>7g
5 g, CC, silica
gel,
/i-hexan-HiOAc(25:l)
lOg.CC,
sUicage!,
/j-hexau-KiC)Ac
(1
M.9:l)
1,2
3,4,5
6,7
8,9,10 11,12 13,14
dieu kien sic ky:
CC,
ODS gel.
MeOH-HjO
(3:2, 7:3} va
m.
prep. HPLC, ODS gel,
MeOH-HiO
(7:
i)
b.
peep.

sesquiterpenoid
chifa
oxy vdi furanodien (7,0%), curzerenon (12,8%), curcumol (5,6%),
germacron (11,2%) va curdion (6,9%) la cac thanh phan chinh. Trong nhom
phan doan 2 ham lugng cac sesquiterpenoid chira oxy da tang len nhieu lan so
voi trong nhom phan doan 1 (72,7% so vdi 43,5%), vci curzerenon (23,6%),
12
germacron (10,1%), curdion (29,4%) va curcumol (7,0%) la cac thanh phan
chfnh, trong khi do cac sesquiterpen hydrocarbon
chi
chiem 2,4%. Ngoai ra ca
hai nhom nay con chiia cac metyl este
ciia
cac axit beo o luong nho (tuong
iing
la3%val%).
Bang 1: Cac thanh phdn de bay hoi cua cac nhom phan doan 1 va 2
STT
1
2
3
4
5
6
7
8
9
10
11
12

phan doan
1"
Ham
luong
7,1
2,2
0,9
1,6
4,0
1,7
2,7
3,7
7,0
0,6
1,6
4,4
12,8
5,6
11,2
6,9
1,2
1,4
0,4
Hgp cha't
P-Elemen
P-Cayophyllen
y-EIemen
Aromadendren
a-Humulen
y-MuuroIen

13
14
Nhom
Th5i
gian liru
(phut)
11,92
12,18
14,08
14,82
15,32
16,08
17,31
18,59
18,87
20,13
20,93
23,67
25,21
26,49
phan doan
2*"
Ham
luong
0,1
0,3
0,6
0,1
0,3
2,4

GC-MS:
45'C-150"C,
lO'C/phiit,
dDcng
1
phiit,
150'C-250'C,
6'C/phut,
diing
15
phut.
*"*
% TIC duoc
do
irfin c6l
mao
qu^
HP-5.
13
Phdn lap vd xdc dinh cdu
triic
cdc sesquiterpenoid
Curdion (1):
Curdion (1) dugc phan lap sac ky vdi hieu suat 0,1 % so vdi lugng than
it
kho
tir
nhdm phan doan 2 chiia
chii
yeu la curzerenon (GC-MS: 23,6%) va curdion

Uuc
ciia 1. Ca'u
hinh
lap thd tuong doi cua curdion da
dirge
chiing tdi xac dinh bang
cac phan tfch cac ph6 NOED va NOESY. Ca'u
hinh
lap the tuyet ddi
ciia
curdion
dugc phan lap ttr C.
wenyujin
da dugc xac dinh bang
ph6 nhi6u
xa lia X
[121.
Sesquiterpenoid nay da dugc chirng to la cd hoat tfnh manh bao ve gan
1131.
Axit palmitic (2):
Axit palmitic (2) dugc phan lap
tir
nhdm phan doan 2 dudi dang tinh
the
hinh kim, khdng mau, dnc
52-54''C.
Tren ph6 IR xuat hicn dai hap thu
ciia
nhdm
hydroxy cua axit carboxylic (dai rong

vong epoxit (1275, 1056 va 921
cm"*).
Cdng thiic phan tu ciia 3 dugc xac dinh la
14
Ci6H,803
bang phan tfch phd EI-MS (m/z 246,
M^).
Cac phan tfch phd 'H-,
"C-
NMR,
*H-*H
COSY va HMQC da cho cac
dfl
kien phd
phii
hgp vdi cau triic cua
zederon dugc phan lap tif C. zedoaria [14].
Ca^u
hinh tuong ddi cua zederon dugc
chiing tdi xac dinh bang phd NOESY. Ca'u triic lap
\hi
tuyet ddi cua hgp chat
nay da dugc xac dinh bang phd nhieu xa tia X [14].
P-Sitosteroi (4) va stigmasterol (5):
P-Sitosterol (4) va stigmasterol (5)
dirge
phan lap tir nhdm phan doan 4 dudi
dang mot hdn hgp kel linh hinh kim, mau trang, dnc
134-136*'C.
Phd

te, p-sitosterol dugc diing dd didu Ui chiing
phinh
to tuyen tidn
li6t.
Cac steroid nay cung cd tac dung ddi vdi noc ran [16], lam giam
miic
cholesterol
va
lie
che
sU
phat tridn
ciia
ung thu ruot ket
[17].
13-Hydroxygermacron
(6):
Hgp cha't 6 dugc phan lap dudi dang bdt vd djnh hinh tir cac phan tach sac ky
nhidu
1^
nhdm phan doan 5, sir dung cac ky thuat sac ky vdi pha dao. Cac phan
tfch phd
*H-
va
*^C-NMR
da cho ihiy 6 la mot
dSn xuSft
cua germacron vdi
15
nhdm hydroxy gan vao vj

hydroxygermacron (6) cdn dugc phat hien la cd hoat tfnh bao ve gan trong cac
thii nghidm
in vitro va in vivo nhu da dugc phat hien ddi vdi curdion (2) |20|.
Tuy nhien trong khi curdion dugc cho la thd hien tac dung qua co che bao ve lc
bao gan chdng lai ddc tfnh cua
D-GalN
(D-galactosamin),
thi co che tac dung
cua
13-hydroxygermacron
cd thd la bang
each tie
che
sir
san sinh
TNF-a
(tumor
necrosis
factor-a).
Aeruginolacton (7):
Hgp chat thien nhien mdi aeruginolacton (7) dugc phan lap dudi dang hot
25
vd dinh hinh vdi do quay
cue am
([a]^
-8,1°) tir nhdm phan doan 5 bang cac ky
thuat sac ky vdi pha dao. Cdng thiic phan tu cua 7 la
C,5H2o03
da dugc xac dinh
bang phd khdi lugng phan giai cao HR-FABMS ion

la 16,8
(q),
17,3 (q) va 8,6 (q)], mot nhdm xeton
[5^
171,9 (s)], mdt cacbon
xctal
|8(-
109,8 (s)], va cac cacbon cua ndi ddi
[6c
123,4 (d), 127,4 (s), 133,0
(s),
131,7
(d),
134,2 (s) va 162,4 (s), va bdn nhdm metylen
[8^
25,3 (t), 25,6 (t), 38,4
(t),
51,3
(t)].
Tren co sd cac phan tfch NMR ca'u
Uiic ciia
7 dugc gia thiet la mot
germacran lacton.
Su cd mat cua vdng y-lacton gdn vdi vdng mudi cacbon d C-7 va C-8 dugc
xac djnh bang cac tuong tac HMBC (Hinh 1) giua H-13
(^^
1,80) va C-7
|5,.
162,4 (s)],
C-U