Kết quả ban đầu về thành phần hóa học cao chiết diclometan lá cúc quỳ (tithonia diversifolia (hemsl ) a gray

T. 51(6) 770-773

TAP CHI HOA HQC

THANG 12 NAM 2013

KET QUA BAN DAU VE THANH PHAN HOA HOC CAO CHIET
DICLOMETAN LA CUC QUY
(TITHONIA DIVERSIFOLIA (HEMSL.) A. GRAY)
Tan Thi Thanh Thayl*, Tan Van Hieul, Vu Thi Thu Ha', Trinh Thi Thu?, Trill Van Sung2
Phong Thi nghiem trong diem Cong nghe loc, hoa dau, Vien Hoa hQc Cong nghiep Viet Nam
2 Vien

Hoa hQc, Vien Han lam Khoa hoc va COng nghe Viet Nam
Den Tim soon 15-10-2013

Abstract
Four compounds: myristyl myristate (1), hispidulin (2), eupafolin (3) and tagitinin A (4) were isolated from the
dichlomethane leaf extract of Tithonia diversifolia (Hemsl.) A. Gray. Their structures were elucidated by analysis of IR,
MS, and NMR spectra and comparison with the published data. Compounds 1 and 3 were isolated for the time from
this plant.
Keywords: Tithonia diversifolia, Asteraceae, hispidulin, eupafolin, tagitinin A.
1. DAT VAN DE
Citc quy hay con gqi di quy, sum quy bien diep,
quy dai, huang ducmg dai, huang duang Mexico...
(Tithonia diversifolia) la,rnat loii thuc vat trong ho
Cue (Asteraceae) co ngu8n g'Oc tir Mexico, hien nay
duqc phan be rOng khip a cfic khu vuc can nhiet dai
va nhi'et dai [1]. Clic quS, thuc clang cay bui, cao 12 in, than cif) long, phan thanh nhieu canh, la thuon,
phien co thity, bia co ring. Hoa a du ng9n, tren
cueng dai. Hoa a bia hinh m8i lep, yang ttrai; hoa

gi6a hinh Ong; gift hoa co vay. Cay ra hoa tir thing
12 den thing 2. 0 Viet Nam, cay cue quy hien nay
da duqc to nhien hem, moc pho bin a eat nhieu nai,
dac biet la a vimg Tay Nguyen [2].
Trong y hQc co truyen Chau My, ntrac ep tir re" va
là cay duqc sir dung dieu tri fip-xe [3]; 6 Mexico loai
nay duqc uong de chi-1.a sot ret va dien hinh la chita
cfic khiSi to mfiu va rut ca, bOt tir la sao kho tri vet
tham, vet thuang va Wing da nhiem trong.... 6 Dai
Loan, dich chiet nuac ctia là cay duqc sir dung de tri
dfii thfio &rang (cluing dai nhieu va chung khfit
nhieu) [4] va viec ling dung dm T. diversifolia trong
dieu tri dfii thao dul:mg cling di duqc ding ki bio
118 [5].
Tren the giai, cay cue quy duqc quan tam nghien
elm ye thinh phAn hoa h9c va hoat tinh nhieu nhAt
trong cfic loai thuc vat thuc chi Tithonia [10]. Cfic
hq? chit sesquiterpen lacton vi ditecpen la thinh
phan hda hoc chinh vi co nhieu hoat tinh thit vi cua
'oil nay [10]. Tuy nhien, cho den nay, chtra co cong
trinh nao ye thinh phin h6a hQc cita cay cfic quS, thu

hfii tai Viet Nam duqc cong b6. Bai bar) nay ding be
ket qui !mac du ye vi0c khao sat thanh plan hoa
hQc cua cao chiet diclometan tir la cay cuc quy thu
hai tai Tay Nguyen.
2. THIJC NGHIEM
2.1. Phuong phi') nghien cum
Sic 14r ban mong duce tien hanh tren ban silica
gel Merck 60F254. Sa'c ky cOt sir dung silica gel c6

hat 0,04-0,063 mm.
Pho Ong huang tir hat nhan NMR duqc ghi teen
may Bruker Avance 500 MHz, pill) khoi ESI—MS
duce do tren may Water LC/MS/MS-Xevo TQ. Pho
IR duqc ghi tren may Nicolet 6700 FT-IR.
2.2. MAu thurc vat
MAu thuc vat ducic thu hfii tai tinh Gia Lai vao
thing 4 nam 2012. Tieu ban duac Wu girt. tai Phong
Thi nghiem trong diem Cong nghe 19c hoa dau. Mau
lfi duqc rim sach, phai, sAy kho.
23. Thuc ngh4m
Li cue quy (400 g) di sAy kho, xay nh6 duqc
chiet nhanh (khofing 3 phut) bang diclometan (x
31An) a nhiet d8 phang theo qui trinh trong tai lieu
[13]. LQC dich chiet, loai dung moi dual fip suit
gam. CArt con lai duqc him tan trong Me0H-H20
(7:3, v:v) va chi& phan lop lin Itrqt vii n-hexan va

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Trein Thi Thanh Thiry va Ong sy-

TCHH, T. 51(6), 2013
diclometan. Cit loai dung mei dual ip suit giam thu
dtrqc can cac cao chiet Wang ling.
Cao chiet diclometan (15,7 g) duqc phan tach
bang sic ky cet ten silica gel, dung mei rua giai la
hon hop n-hexan-etyl axetat vii lam etyl axetat
tang din, thu dtrqc 12 phan doan (Fl-F12).

Phan doan Fl (320 mg) duqc rira va ket tinh lai
tong metanol thu duqc chit 1, phan doan F6 duqc
tinh the ten cet Sephadex LH-20 WA dung mei rim
giai metanol thu duqc 35 mg (0,0087 %) chit 2.Chit
3 (40 mg, 0,010 %) thu dtrqc tir phan doan F10 (100
mg) sau khi tinh cite tren cet Sephadex LH-20. Phan
doan F5 duqc tinh the ten OA silica gel vii he dung
re 3:7 thu dtrqc
moi rim giai n-hexan:etyl axetat
hqp chit 4 sach (44 mg, 0,011 %).
- Myristyl myristat (tetradecyl tetradecanoat, 1):
bet matt tring, C28115602, ESI-MS (m/z):
425[M+H]'; ham lacing 0,08% so vii mau kho.
'H NMR (500 MHz, CDC13, Sppm): 4,05 (2H, t,
J= 7 Hz, 0-CH2), 2,29 (2H, t, J = 7,5, CH2), 1,60
(6H, m), 1,28 (40H, brs), 0.88 (6H, t, J= 7 Hz).
'3C NMR (125 MHz, CDC13): 8, 174,0 (-COO),
64,4 (0-CH2), 22,7-34,5 (CH2), 14,1 (2CH3)- Hispidulin (4',5,7-trihydroxy-6-methoxyflavon,
2): tinh the mau yang nhat, C161-11206, ESI-MS (m/z):
301 [M+H]+; ham lucmg 0,009%. IR (KBr) v*max
(cm'): 3423, 1657, 1606.
'H-NMR (500 MHz, CDCI3+CD3OD, Sppm):
7,83 (2H, d, J = 9 Hz, 1-1-2', H-6'), 6,93 (2H, d, J=9
6,55 (1H, s, H-3),
Hz, 11-3', H-5'), 6,57 (1H, s,
3,90 (3H, s, -OCH3).
'3C NMR (125 MHz, CDC13+CD3OD, Sppm):
166,3 (C-2), 103,3 (C-3), 184,1 (C-4), 153,9 (C-5),
132,8 (C-6), 158,8 (C-7), 95,3 (C-8), 154,6 (C-9),
105,7 (C-10), 123,2 (C-1'), 129,3 (C-2' & C-6'),

116,9 (C-3' & C-5'), 162,6 (C-4'), 60,9 (6-OMe);
- Eupafolin (3 ',4',5,7-tetrahydroxy-6-methoxyflavon, 3): bet yang nhat, CI6H1 207, ESI-MS (m/z):
316 [M+H]+; ham luqng 0,01 %. IR (KBr) v*max
(cm"): 3411, 1654, 1602.
'H NMR (500 MHz, CD3OD, Sppm): 7,35 (1H,
s, H-2'), 7,34 (1H, d, J = 8,5 Hz, 11-6'), 6,89 (1H, d, J
= 8,5 Hz, H-5'), 6,51 (111, s, H-8), 6,50 (1H, s,
3,89 (3H, s, 0-Me).
'3C-NMR (125 MHz, CD3OD, Sppm): 166,3 (C2), 103,3 (C-3), 184,1 (C-4), 153,9 (C-5), 132,9 (C6), 159,0 (C-7), 95,3 (C-8), 154,6 (C-9), 105,7 (C10), 123,6 (C-1'), 114,1 (C-2'), 147,0 (C-3'), 153,9
(C-4'), 116,7 (C-5'), 120,3 (C-6'), 60,9 (6-OMe).
- Tagitinin A (3, 10-epoxy-germacrolide-6,7trans-lacton, 4): tinh the khong mau, C19H2807, ESIMS (m/z): 369 [M+H]+; ham lucmg 0,011 %. IR
(cm"): 3495, 2930, 1756, 1663, 1447.
(KBr)

'H NMR (CDC13, 500 MHz, Sppm), 6,27 (1H,
d, J = 3,5 Hz, H-13a), 5,58 (1H, ddd, J = 3,0, 5,5,
8,5 Hz, H-8a), 5,54 (1H, d, J= 3,5 Hz, H-13b), 4.57
(1H, ddd, J= 2, 6,5, 10,0 Hz, 11-613), 4,25 (111, dd, J
= 7,5, 9,5 Hz, H-la), 4,08 (1H, ddd, J = 3,0, 6,5,
10,0 Hz, H-7a), 2,44 (1H, m, H-2A), 2,44 (1H, m,
H-2'), 2,08 (1H, m, H-4P), 2,08 (1H, m, H-2b), 2,08
(1H, m, H-5b), 2,08 (111, m, H-5a), 1,96 (11-1, dd, J=
6,0, 14,0 Hz, H-9b), 1,82 (1H, m, 9,0, 13,0, H-9a),
1,44 (3H, s, H-14), 1,11 (31-1, d, J = 6,5 Hz, H-15),
1,08 (311, d, J = 7,0 Hz, II-3), 1,06 (3H, d, J = 7,0
Hz, H-4').
'3C NMR (CDC13, 125 MHz, Sppm): 176,34
(C-1'), 169,42 (C-12), 137,07 (C-11), 121,57 (C-13),
105,73 (C-3), 81,77 (C-6), 81,46 (C-10), 78,26
(C-1), 70,39 (C-8), 47,84 (C-7), 47,04 (C-2), 44,34

(C-4), 37,79 (C-5), 34,62 (C-9), 34,03 (C-2'), 24,99
(C-14), 19,18 (C-15), 18,72 (C-3'), 18,35 (C-4').

OH 0
2: R = H
3: R = OH

4

Ifinh 1: Cau truc ciia chAt 1-4
3. KET QUA VA THAO LUAN
Ket hcrp thong tin tir phe 'H-, '3C-NMR va pic
ion phan tir tai m/z 425 [M+H]+ ten phe ESI-MS
dm 1 la C28115602. Tren
suy ra cling thirc phan
pito 'H-NMR cua chat 1 xuat hien tin hreu triplet tai
8 0,88 (J= 7 Hz) viii tY le tich phan là 6H ttrcmg (mg
vii cac proton dm 2 nhom metyl du mach. Hai
nhOrn metyl co ding de djch chuyenho,c cho
thay hqp chat 1 co ciu triic mach Wang doi can
ming. Tin II iOu triplet dm 2 proton a 8 4,05 (J =

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TCHH, T. 51(6), 2013

Ket qua ban ddu ve thanh phdn...

7Hz) dac trung cho tin hieu cua nhom oxymetylen

(0-CH2) ma ben canh nhom CH2 nay là mot nhom
CH2 khac. Tung tv nhis very tin hieu triplet a 8 2,29
(.1 = 7 Hz) ding dac trtmg cho nhom CH2 et gift men
cacbon bac 4 va met nhom CH2. Tren pho con xuat
hien cam tin hieu cua 46 proton a di) dich chuyin
trong vung 8 1,25-1,62 ppm. PhO DC-NMR bao gom
tin hieu cila nhom cacbonyl este (COO) a 8 174
ppm, 1 tin hieu CH2-0 a 8 64,4 ppm, tin hieu CH3 a
8 14,1 ppm va cum tin hieu cua cac CH2 trong vung
22,7-34,4 ppm. Ket hop cac phuang phfip phO va so
sfinh veri tai lieu tham khao [6], cong thirc ciu tao
chit 1 dugc xac dinh la tetradecyl tetradecanoat
(myristyl myristat).
Chat 2 din tai dual dang bOt mau yang nhat. PhO
IR dm hqp chit nay cho thiy dai hap thu rang
3423 cm"' gqi y sir c6 mat ctia cac nhom hydroxyl.
Dai hap thv a 1657 cm"' dac trtmg cho nh6m
carbonyl clang ot,ti-chtra b5o hoa. Ph6 ESI-MS a the
de) ion am cho pic ion phan tir [M-H]" tai m/z 299
trong khi tren ph8 ESI-MS the di) ion dtrcmg xuAt
hien cac pic [M+Na]' va [M+H]+ tai cac gifi tri m/z
ttrcmg Ung la 323 va 301.
cac dir lieu ph6, cong
thirc phan dm chat 2 dugc dir doan là C161-11206.
Tuy nhien tren ph6 "C-NMR (lac trung cho ciu trim
cua hqp chit khung flavon chi xuat hien 14 tin hieu
Ong twang. Dieu nay cho thay hop chat 2 c6 2 cap
CH twang dtrcrng tai 8 129,3 (C-2'/C-6') va 116,9
ppm (C-3'/C-5'), goi St rang nhan thorn & von B co
1 nh6m the a C-4'. Ngoai ra tren pito con xuat hien

hai tin hieu co di) dich chuyen h6a hoc & 8103,3 va
95,3 ppm cila 2 nhom CH nhan thorn khfic. Tin hieu
Ong Wang cua nhom C=0 dac trung cua nhom
cacbonyl xuAt hien tai 8 184,1 ppm. Tin hieu a 8
60,9 ppm dugc gin cho nhom metoxy. Tren ph8 'HNMR ton tai 2 tin hieu dm 2 cap proton nhan thorn
duplet c6 ttrong tac ortho (J = 9,0 Hz) tai 8 7,83 va
6,93 ppm, ttrcmg ung vai tin hieu cong htrong cua
H-2'/H-6' va H-3'/H-5'. Du lieu nay hoan toan phu
hcrp vbi dir lieu phO '3C-NMR. Tin hieu don co
cuang de) tich phan là 3H a 8 3,90 dugc xac dinh la
nhom metoxy. Tir dir lieu phO pan tich a ten, ket
hqp vbi so lieu phi') der cong bo [7], cong thirc ciu
tao cua hqp chit 2 dugc xac dinh la 4',5,7trihydroxy-6:methoxyflavon (hispidulin). Hispidulin
dugc biet den là hop chat co hoat tinh chong ung
thu, chOng virus ding chit St 18, 10]. Chit nay dugc
tach ra tir cay cue quSt dm An DO va mot vai loci
thirc vat khac [9, 10].
Chat 3 thu dugc dual dang bOt mau yang. Twang
tir vbi hap chat 2, phi!) IR dm hop chat nay ding cho
thay cac dai hap thu & 3411 cm' va 1654 cm-' dac
trtrng cho sir c6 mat twang ung cua cac nhom

hydroxyl va carbonyl dang a,/3-chtra bao hoer. PhO
ESI-MS a che de) ghi ion am va &rang lAn Itigt cho
cac pic [M-H]" tai m/z 315, [M+Na]+ va [M+H]+ tai
cac gia tri m/z twang 1rng la 339 va 317. Tir dir lieu
pho khOi, ket hqp vai ph6 NMR, cong thirc phan tir
dm chit 3 dugc xac dinh la C16111207 Ara c6 nhieu
han mot nguyen tir oxi so veri chit 2. Tren pho "CNMR xuAt hien 16 tin hieu cong hue:mg dac trung
cho cAu trim dm hop chit khung flavon bao g6m 1

tin hieu C=0 (8 184,1 ppm), 9 tin hieu C bac 4 va 5
tin hien CH nhan tham (95,3-123,7 ppm) va 1 tin
hieu cua nhom OCH3 (8 60,9 ppm). Ket hqp dir lieu
NMR vai dir lieu MS cho phep nhan dinh so vai 2,
hop chit , 3 c6 cAu trite twang tir nhtmg co them mot
nhomthe .0H. Tren ph Al H-NMR, 2 proton co de
dich chuyen 6,51 va 6,50 ppm twang ter tren pho hop
chit 2 dugc gin cho H-8 va H-3. Tin hien dm 2
proton nhan thorn c6 ttrcmg tic ortho (d, J = 8,5 Hz)
tai 8 7,34 va 6,89 ppm, twang ung vai tin hieu cong
htrang dm H-6' va H-5'. Tren ph8 proton khong
xuAt hiOn twang tac meta vi the tin hieu dan cua
proton tai 87,35 (Iliac gin cho H-2'. Tin hieu clan
vai cueing de) tich phan la 3H a 8 3,90 la dm nhom
methoxy. Ket hqp dn. lieu va so sfinh phi') veri chit 2,
cAu true dm chit 3 dtrqc xac dinh la 3%4%5,7tefrahydroxy-6-methoxyflavon (eupafolin). Hqp chit
nay cling dugc cong bo co hoat tinh gay dac to bao
tren clang to bao ung thisbieu m8 KB [7].
Him chat 4 c6 cong thirc phan tier C19112807, dtrqc
dua ra dva vao viec ket hap cac ph8 ESI-MS, 'HNMR va '3C-NMR. PhO IR dm hqp chit nay (lac
trung cho khung sesquiterpen lacton chtra bao Ma
yeti cac tin hieu a so song 3495, 1756, 1663, 1598,
1447 cm'. Tin hieu a vung 1756 cm'' thuang dtrqc
gap 6. nhom carbonyl dm cac hqp chit y-lacton
[11]. Tren ph6 'H-NMR xuAt hien tin hieu dm 4
nhom metyl co dO dich chuyen Ma hoc et 1,44-1,08
ppm. Cap tin hieu duplet a di) dich chuyen hoa hoc
6,27 va 5,54 ppm veil J = 3,5 Hz la dau hieu nhan
dang cho nhom exocylic metylen cua khung
sesquiterpen lacton [11]. Ph6 '3C-NMR cho thiy str

c6 mat dm 19 tin hieu cacbon, bao Om: 4xCH3, 4 x
trong d6 CO 1 nhom cacbonyl.
CH2, 6xCH,
Phan tich phi)
pho va so sfinh veri tai lieu tham khao cho
phep xac dinh cAu trim hap chit 4 la 3,10-epoxygermacrolide-6,7-trans-lacton (tagitinin A) [9].
TaFitinin A dugc ding 1)6 là hqp chit co hoat tinh
chong tieu dtrang thong qua con &rang hoat hoa thy
the tang sinh peroxisome [12].
4. KET LUAN
Tim dich chiet diclometan cua la cay cue qujt
(Tithonia diversifolia) TA' phan lap dugc 4 hqp chat

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Tran Thi Thanh Thtiy va Ong sir

TCHH, T. 51(6), 2013

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10.Dictionary of Natural Products, version 18:1,
Copyright©1982-2009 Chapman & Hall/CRC 2009.
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Sciences, 10(3), 183-188 (2007).
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diversifolia act as peroxisome proliferator-activated
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Chaves, P. G. B. D. Nascimento, J. E. Lichston, M.
G. Constantino, E. M. Varanda, F. B. Da Costa.
Constituents of glandular trichomes of Tithonia
diversifolia: Relationships to herbivoty and
antifeedant activity, Phytochemistry, 69, 2052-2060
(2008).

1-4. Bang cac phuang phfip ph6 IR, MS, NMR va so
cac dir lieu pho da ding 1)6, cAu triic dm
sfinh
cac hop chat nay &roc xfic dinh la: myristyl myristat
(1),, hispidulin .(2), eupafolin (3) va tagitinin A (4).
Chat 1 va 3 Ian du duce tim thay trong loai cue
quSt.
Liri cam cm: Car lac gia xin than thanh cam on BO

Cong thwang cid tai try kinh phi cho cac nghien ceru
thong qua dd tai, ma so 043-2013/HD- DTCNHD.

TAI Lieu THAM KHAO
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Cay có Viet Nam, tap III, trang
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4. T. Miura, K. Nosaka, H. Ishii, T. Ishida.
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diversifolia, in KK-Ay diabetic mice, Biol. Pharm.
Bull. 28, 2152 (2005).
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mellitus, processes for the preparation of same, and
usage of same, US Pat. 5,773,004 (Fukuoka 1998,
814-01).

Lien he: Trill Thi Thanh Thu),
Phong Thi nghi'e'm trong ditm Cong nghe loc, boa dau
S6 2 Pham NO Lao, Ha N6i
Email:

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