Tp ch Khoa hc 2011:19b 56-61

Trng i hc Cn Th

NGHIN CU THNH PHN HA HC CA THN CY

C XC (ACHYRANTHES ASPERA. L) TR VINH
Tn N Lin Hng1, Nguyn Minh Hin2 v Trn nh Lun3

ABSTRACT
From the chloroform and ethyl acetate extracts, we have isolated and identified four
compounds: stigmasterol, the mixture of stigmasterol and spinasterol with the ratio of
1:3, sitosterol 3--O-D-glucopyranoside and quercetin 3-O--D-galactopyranoside.
Their structures were evaluated by spectrum data of MS, 1H, 13C, DEPT NMR and
2D-NMR.
Keywords: Achyranthes aspera. L, Quercetin 3-O--D-galactopyranoside
Title: Study on the chemical composition of Achyranthes aspera L. growing in Tra
Vinh Province

TM TT
T cao chloroform v cao ethyl acetate, chng ti c lp v nhn danh bn hp cht:
stigmasterol, hn hp stigmasterol v spinasterol vi t l 1:3, sitosterol 3-O-- Dglucopyranoside v quercetin 3-O--D-galactopyranoside. Cu trc ca cc cht ny
c ngh cn c vo ph nghim t d liu MS, 1H, 13C, DEPT NMR v ph 2 chiu
NMR.
T kha: Achyranthes aspera. L, Quercetin 3-O--D-galactopyranoside
[1]
1 M U

Cy c xc c tn khoa hc: Achyranthes aspera L., thuc chi Achyranthes, h

Amaranthaceae (Rau gin, Dn). Tn thng thng: c xc, ngu tt nam, nh
khoanh ng (Ti), c nh ln ngu (Thi), h ng.
C xc l loi cy tho, cao gn 1 m, c lng mm nhiu hay t. Thn cng,
phnh ln nhng mu. L mc i, hnh trng hay mi mc, nhn hoc hi c
lng, gc thun, u t hoc nhn, dy 312 cm; cung l di. Cm hoa mc thnh
bng n ngn thn, di 20-30 cm; l bc con hnh gai; hoa mc r xung p st
vo cung cm hoa; di gm 5 phin hnh mi mc nhn, nhng phin pha trong
rt nh; nh 5, nh lp c nhiu tua vin u; bu hnh tr. Qu nang c l bc
cn li, nhn thnh gai d mc vo qun o khi ng phi; v rt mng, dnh vo
ht; ht hnh trng di, dy 1 mm. Ma hoa qu thng 712.
C xc phn b ri rc hu ht cc tnh vng ng bng, trung du ti Vit Nam;
ngoi ra c Lo, Campuchia, Thi Lan, n , Trung Quc v mt s
nc khc.
Trong y hc c truyn Trung Quc, c xc c dng iu tr nhc u, cm
nng, st rt, si niu, vim thn mn tnh. Y hc c truyn n li dng nc

Khoa Khoa hc T nhin, Trng i hc Cn Th

Cao hc ha hu c K16
3
Cao hc ha hu c K15
2

56

Tp ch Khoa hc 2011:19b 56-61

Trng i hc Cn Th

sc c xc lm thuc li tiu v tr gan. i vi ph n, c xc gip lm gim

ri lon kinh nguyt, au kinh, kinh b, au bng sau sanh.
Trn th gii c mt s cng trnh nghin cu v cy c xc v cng b nhng
hp cht phn cc thuc nhm saponin. nc ta cho n nay ch thy nghin
cu v hot tnh v cng dng ca n m cha thy nghin cu v thnh phn ha
hc ca cy c xc. Do , trong bi bo ny chng ti cng b mt s hp cht
phn cc yu v trung bnh c lp c t nguyn liu c xc thu hi ti
Tr Vinh.
2 NGUYN LIU V PHNG PHP
2.1 Nguyn liu
Mu nguyn liu dng trong nghin cu l phn thn cy c xc c thu hi
x Hng M, huyn Chu Thnh, tnh Tr Vinh vo thng 3/2010. Mu cy
c ThS. Nguyn Th Kim Hu (B mn Sinh hc, Khoa KHTN, Trng HCT)
nh danh khoa hc v mu tiu bn (k hiu CX 1) c lu ti Phng th nghim
Ha hu c, Khoa KHTN, Trng HCT.
Dung mi s dng trong ti l dung mi ng chai xut x Vit Nam
(Chemsol). Silica gel 60 (Merck) dng cho sc k, TLC dng Silica gel F254
(Merck).
2.2 Thit b
Ph khi lng (ESI-MS) c ghi trn my MS 5989 B (Hewlett Pakard). Ph
cng hng t nhn (NMR): 1H-NMR (500 MHz) v 13C-NMR (125 MHz),
COSY, DEPT, HSQC, HMBC c ghi trn my Bruker AM500 FT-NMR ca
Vin Ha hc, Vin Khoa hc v Cng ngh Vit Nam. im nng chy c o
trn my Electrothermal IA 9000 series, dng mao qun khng hiu chnh.
2.3 Chit xut v c lp
Thn cy c xc (10kg) c chit ngm dm vi methanol 98%, ct loi dung
mi di p sut gim thu c cao tng (250 g), tip theo thm 0,5 lt nc ct v
ln lt chit phn b lng-lng cao ny vi petroleum ether (PE), chloroform (C),
ethyl acetate (E) v n-butanol (B) thu c cc cao PE (61,27 g), cao C (60,65 g),
cao E (40,15 g), cao B (33 g). Tin hnh sc k ct cao C, cao E.
T cao C (50 g) tin hnh sc k ct vi h dung mi c tnh phn cc tng dn t
petroleum ether pha t l vi ethyl acetate, x ct vi methanol thu c cc cht
rn mu trng, khi kt tinh li u c dng tinh th trng. Cht (1) (3,8 g) l tinh
th hnh kim, cht (2) l tinh th hnh phin (30 mg) v cht (3) c dng v nh
hnh (20 mg). T cao E (20 g) tin hnh sc k ct vi h dung mi petroleum
ether: ethyl acetate t (9:1) n (0:10) v h dung mi ethyl acetate: methanol t
(99:1) n (30:70), x methanol, thu c 6 phn on chnh c khi lng ln
lt l 3,57; 3,52; 4,56; 3,12; 2,32; 1,13 (g). Tip tc sc k ct phn on s 4
(3,12 g) hai ln trn ct silica gel 60 v 1 ln vi silica gel RP-18, thu c cht
(4) c dng tinh th ht mn mu vng chanh (18 mg).

57

Tp ch Khoa hc 2011:19b 56-61

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3 KT QU V THO LUN
3.1 Nhn danh cu trc ca cc cht trong cao C
3.1.1 Stigmasterol (cht 1)
Cht (1) l tinh th hnh kim, mu trng, kt tinh trong chloroform (3,8 g). Nhit
nng chy 169 - 170C. Sc k bn mng cho vt Rf = 0,43 khi gii ly bn
mng trong chloroform. Ph 1H-NMR (500, CDCl3), (ppm): 3,52 (1H, m, H-3);
5,35 (1H, d, H-6); 5,16 (1H, d, H-22), 5,04 (1H, d, H-23). Ph 13C-NMR cho thy
c 4 tn hiu 140,79; 121,70; 138,37 v 129,32 ppm ln lt thuc v lin kt i
ti cc v tr C5; C6; C22 v C23. T ph DEPT cho thy hp cht c 29 C trong
c: 6 C dng -CH3, 8 C dng -CH2- v 10 C dng -CH; 3 C t cp (gm 1 >C= v
2 >C<). Khi so snh vi ph ca hp cht stigmasterol trong ti liu tham kho [2],
cc s liu trng khp hon ton.
3.1.2 Hn hp stigmasterol v spinasetrol t l 1:3 (cht 2)
Cht (2) c dng tinh th hnh phin, mu trng, kt tinh trong chloroform. Sc k
bn mng cho vt Rf = 0,39 khi gii ly bn mng trong chloroform.
Ph 1H-NMR (500 MHz, DMSO): ngoi cc tn hiu mi n ca proton methyl
vng t trng cao gip nhn ra khung sterol, cn c cc proton olefin vng
(4,9 - 5,3 ppm), gm 2 loi tn hiu vi cng tch phn theo t l 1:3. C th,
ngoi cc tn hiu c trng ca proton olefin mch h dng HC=CH ti C22 v
C23 ca khung stigmastan nh: 5,14 (1H, d, 9,5 Hz, H-22), 5,11 (1H, d, 8,5 Hz, H23), c tn hiu c trng cho proton ca lin kt i dng >C=CH C5 v C6 l
5,30 (d, 4,5 Hz); v mi 5,01 (m) vi cng tch phn gp 3 ln tn hiu va
nu, l tn hiu ca lin kt i C8 v C7. Ph 13C-NMR (125 MHz, DMSO): cho
thy 58 tn hiu nhng cc tn hiu ca carbon methyl (vng 11,0 ppm - 21,3 ppm),
v cc nhm CH2 v CH rt kh nhn dng trong vng xen ph dy c cc tn
hiu (vng 21,4 - 56,8 ppm). Ph cn c 4 tn hiu ca carbon t cp 35,9 - 44,8
ppm; c 2 tn hiu 70,8 v 71,5 ppm dng carbon hydroxymethin v 8 tn hiu ca
carbon olefin. Ngoi s hin din ca stigmasterol cn c mt mt sterol khc vi
2 lin kt i trong phn t gm HC=C< (117,3 ppm v 139,4 ppm) v HC=CH
trn mch carbon h (138,2 ppm v 129,3 ppm). Cc d liu t ph proton, 13C,
DEPT NMR cho php d on cht (2) l hn hp ca 2 loi sterol: stigmasterol
v spinasterol t l mol ln lt l 1:3.
28
21

19
10

11
9

HO 3 4 5 6

12

18

20

22
23

13 17 16
8
7

29

28
21

26

24
25
27

14 15

Stigmasterol

12

11
19
10

18

20

22

HO 3 4 5 6

26

24
23

13 17 16
9

29

25
27

14 15
8
7

Spinasterol

3.1.3 Sitosterol 3-O--D-glucopyranoside (cht 3)

Cht (3) dng bt mn, mu trng, kt tinh trong methanol. Nhit nng chy
269-270C. Sc k cho 1 vt c Rf = 0,41 khi gii ly bn mng trong h dung mi
ethyl acetate: methanol (9:1).
58

Tp ch Khoa hc 2011:19b 56-61

Trng i hc Cn Th

Ph 1H-NMR (500MHz, DMSO), (ppm): 1,0 (3H, s, H19); 5,32 (1H, m, H6); 3,56
(1H, m H3). Ph 1H-NMR xc nhn s c mt ca mt proton lin kt i 5,32
ppm; nhm proton ca ng vng 3,02 - 4,43 ppm; 6 tn hiu proton ca
methyl c trng ca hp cht sterol trong vng 0,86-1,08 ppm. Trong 13C-NMR
cho thy 35 tn hiu ca nguyn t carbon, trong c 7 tn hiu dng hydroxy
methin (vng t 61,08- 76,89 ppm); c 2 tn hiu 140,17 v 121,82 ppm thuc v
lin kt i ti v tr C5 v C6; c 6 nhm methyl, 12 nhm methylen, 14 nhm
methin, 3 nhm C t cp. Ti 100,77 ppm l carbon anomeric ca ng. T d
liu ph COSY gip nhn din cc tng quan ca proton trong ng, kt hp
vi ph HSQC, HMBC chng ti ngh cu trc ha hc ca cht (3) l sitosterol
3-O--D-glucopyranoside.
S liu ph 1D- NMR ca cc cht (1), (2), (3) c trnh by trong bng (1).
29
28
21
18
12
19
2

6'

5'

17

13

24
23

26
25
27

16

2'

OH

10

14

15

3
1'

3'

20

OH
HO 4'
HO

11

22

5
4

Sitosterol 3-O-D-glucopyranoside
3.1.4 Quercetin 3-O--D-galactopyranoside (cht 4)
T ph 1H-NMR (500 MHz, CD3OD-d4) nhn thy c 12 tn hiu proton. Trong
vng 6,0-8,0 ppm cho thy c 5 proton ca vng thm gm 2 proton v tr meta
vi nhau (J=2,0 Hz) thuc vng A, vng B vi 3proton trong 1H v tr meta
v para vi 2H cn li, Jmeta=2,5Hz; Jorto= 8,5Hz. ph 1H v COSY xc nh cc
tn hiu proton ti (3,0-4,0 ppm) l proton ca phn ng. Da vo HSQC xc
nh c tn hiu H 5,19 ppm l proton annomer (d, J=7,5 Hz), ng vi C
105,1 ppm.
T ph 13C-NMR v ph DEPT-NMR: Hp cht (4) gm 21 C, trong c 1
carbon carbonyl 179 ppm; 1 carbon anomer ti 105 ppm. Trong vng (60-70 ppm)
c 5 tn hiu gm 4 carbon hydroxymethin v 1 nhm OCH2- (62,0 ppm) nn kh
nng y l ng glucose hoc galactose dng pyranoside vi nhm CH2 cui
mch. Cn c vo COSY, cho thy hng s ghp cp ca cc proton H4 , H3 , H5
u nh (J = 3,5 Hz), nn phn ng c ngh l galactose. Da vo HSQC,
HMBC xc nh c tng quan ca proton anomer ti H 5,19 ppm l tn hiu
doublet (J=7,5 Hz), vi carbon ti C 132 ppm (ng vi C3), nn d on ng
dng -pyranoside lin kt glycosid vi khung flavonol ti C3. Da vo cc d liu
ph 1D v 2D-NMR ca hp cht (4), chng ti ngh cng thc ca hp cht l
Quercetin 3-O--D-galactopyranoside. Cc s liu ph ca cht ny hon ton
khp vi ti liu tham kho [3].

59

Tp ch Khoa hc 2011:19b 56-61

Trng i hc Cn Th

OH

HO

OH

2'

1'

2
6
5

OH
OH

O
O

OH
O

HO
OH

Quercetin 3-O-D-galactopyranoside
Bng 1: Ph NMR ca cc cht (1), (2), (3) so vi ti liu tham kho
TT

Cht (1)
1

H (Hz)

1
2
3
3,52 m
4
5
6 5,35 d (5,0)
7
8
9
10
11
12
13
14
15
16
17
18
1,01 s
19
0,89 s
20
21
22 5,16 d (7,5)

13

37,2
31,6
71,7
42,3
140,7
121,6
31,9
31,9
50,1
36,5
21,0
39,7
42,2
56,8
24,3
28,9
55,9
12,0
19,4
40,5
21,23
138,3

23 5,04 d (7,5) 129,2

24
25
26
27
28
29
1
2
3
4
5
6

60

1,53 brd
0,79 s
0,81 s
0,80 t

51,2
31,9
19,0
21,0
25,4
12,3

Stigma SpinaCht (2)

sterol[2] sterol[2]
13
13
1
13
C
C
H (Hz)
C
37,2
37,1
37,1
31,6
31,5
31,4
71,8
71,0 3,60 m
70,8
42,2
39,0
40,2
40,3
140,7
140,8
29,6 5,30 d (4,5) 121,4
121,7
31,8
117,4 5,01 m
117,3
32,1
139,3
139,4
50,2
49,4
50,1
36,5
34,2
36,5
21,1
21,5
21,1
39,8
39,5
39,6
42,3
43,3
43,3
56,8
55,1
56,7
24,3
23,0
23,0
28,9
28,4
29,6
56,1
55,9
56,0
12,0
12,0 0,90 s
18,7
19,4
13,0 0,55 s
19,3
40,5
40,7
40,7
21,2
21,0
20,4
138,3
138,1 5,11 d (8,5) 138,0
138,2
129,3
129,4 5,14 d (9,5) 129,2
129,3
51,2
51,2
51,1
31,8
31,8
31,8
19,0
21,3 0,79 s
18,8
21,1
19,0 0,85 s
19,7
25,4
25,3
25,3
12,2
12,2 0,80 t
18,7

, ppm (J = Hz)
Cht (3)
1

H (Hz)

13

C
37,0
29,3
3,56 m 76,3
39,5
140,1
5,32 d (5,0) 121,8
31,6
31,6
50,0
36,4
20,8
38,4
42,1
56,5
24,0
27,9
55,8
11,6
19,1
35,9
18,9
33,7
25,9

4,43 (7,5 )
3,52 d (5,5 )
3,4 dd (4,0; 3,0)
3,64 d (5,5)
3,73 d (3,0 )
3,66 m
3,54 m

45,7
28,9
18,6
19,7
22,8
11,7
100,7
76,8
73,4
70,0
76,7
61,0

Tp ch Khoa hc 2011:19b 56-61

Trng i hc Cn Th

Nh vy t thn cy c xc thu gom ti Tr Vinh, chng ti c lp v xc nh

cu trc ca 4 thnh phn c hm lng cao trong cy l: 3 hp cht gm
stigmasterol,
sitosterol
3-O--D-glucopyranoside,
quercetin
3-O--Dgalactopyranoside v hn hp stigmasterol v spinasterol vi t l 1:3. Cc kt qu
ny l ng gp mi v thnh phn ha hc ca cy c xc ti Vit Nam.
Cc nghin cu trn nhng phn on cn li ang c tip tc thc hin.
Cc cht sterol v sterol glycosid ny c tc dng tt trong chuyn ha ng
huyt, cn quercetin glycosid c vai tr khng oxy ha rt tt v c ch pht trin
t bo ung th [3, 4].
TI LIU THAM KHO
[1] Huy Bch, ng Quang Chung, Bi Xun Chng, Nguyn Thun Dong, Trung
m, Phm Vn Hin, V Ngc L, Phm Duy Mai, Phm Kim Mn, on Th Nhu,
Nguyn Tp, Trn Ton, Vin dc liu, 2004, Cy thuc v ng vt lm thuc Vit
Nam, Tp II, Nh xut bn khoa hc v k thut H Ni, Trang 575/II.
[2] John Goad and Tosh Hiro Akihisa, Analysis of Steroid.
[3] Malgorzata Materska, 2008, Quercetin and its derivatives: chemical structure and
bioactivity a review, Pol. J. Food Nutr. Sci.
[4] P.J.D. Bouic, S. Etsebeth, R.W. Liebenberg, C.F. Albrecht, K. pegel, P.P. Vajaarsveld,
1996, Beta-sitosterol and beta-sitosterol glucoside stimulate human peripheral blood
lymphocyte proliferation: Implications for their use as an immunomodulatory vitamin
combination, Int. J. Immunopharmac., Vol.18, No.12, pp. 693-700
[5] Ripu M. Kunwar, Y. Uprety, C. Burlakoti, C.L.Chowdhary and R.W., 2009, Bussmann
Indigenous use and ethnopharmacology of medicinal plants in far-west Nepal.,
Ethnobotany Research & applications, Vol 7, pp. 005-028.

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