Tgp chi Khoa hgc 2012:21a 114-118 ^ ™ ^ ^ ^ ^ ' ^<^^rhg
KET QUA NGHIEN ClTU THANH PHAN HOA HOC CUA THAN CAY CO XU^dC {ACHYRANTHES ASPERA, L)
Ton Nit Lien Huang", Nguyin Minh Hiin' vd Trdn Dinh Ludn^
ABSTRACT
Continuing to the chemical study on Achyranthes aspera L. (Vietnamese: Nguu tdt, family Amaranthaceae), after the previous paper described the result study on materials of Tra Vinh province, we now focus on the raw materials collected in Vinh Long province. From the different polarity extracts, we have isolated and identified four compounds:
dodecanoic acid stigmasterol, a-D-glucopyranosyl-(I-^2)-p-D-fi^tofuranoslde. and quercetin Their structures were evaluated by spectrum data of MS, H. C. DEPT NMR and2D-NMR.
Keywords: Achyranthes aspera. L, Stigmasterol, a-D-Glucopyranosyl-(1^2)'p-t>- fructofuranoside, Quercetin
Title: Study on the chemical composition of Achyranthes aspera L.
TOM TAT
Trong qud trinh nghien cuu thanh phdn hoa hoc cho cdy Co xudc, (Nguu tdt nam, Achyranthes aspera L , hg Rauden, Amaranthaceae). tiip theo bai bdo dd cong bo trudc ddy khi khdo sdt cdy Co xudc thu hdi d Trd Vinh. chung loi tiin hdnh co lap chdt tren nguyen lieu dugc thu hai a Vinh Long. Tie cdc cao co dg phdn cue khdc nhau, chung toi dd CO lap vd nhdn danh them bdn hgp chdf dodecanoic acid, stigmasterol. a-D~
glucopyranosyl-(I~*2j-P-D-fructofuranoside vd quercetin. Cdu true cua cdc chdt ndy duac di nghi cdn cu vdo cdc dit lieu phd MS, 'H. " C , DEPT-NMR vd phd 2 chiiu NMR.
Tu khda: Achyranthes aspera. L, co xudc, sterol, dodecanoid acid, a-D-glucopyranosyl- (l—*2)-fi-D-fructofuranoside, quercetin
1 MO DAU
Ciy Cd xudc cd ten khoa hpc: Achyranthes aspera L., thudc chi Achyranthes., ho Amaranthaceae (Rau gien. Din). Ten thdng thudng: Cd xudc, Nguu t i t nam, Nha khoanh ngii (Tai), Cd nhi lin ngu (Thii), Ha ngvi.
Cd xudc la logi ciy thao. cao gin 1 m, co long mem, than ciing, phinh len d nhiing miu. Li mgc ddi, hinh triing, nhan hoac hoi cd Idng, gdc thudn, diu tCi, diy 3—12 cm; cudng la dai. Cum hoa mpc thinh bdng dan d ngpn than, dai 20-30 cm, hoa mpc ru xudng ap sit vio cudng cum hoa. Q u i nang cd la bac cdn Igi tgo hinh gai nhpn de mic vao sd vii khi chgm phai; vd qua rit mong, dinh vao hgt; hat hinh tning dii, day 1 mm. Mua hoa qui thing 7-12. Hinh ding cd xudc rit gin gidng vdi ciy nguu tit, cung l i loai dupe thao rit hiiu dung d nhieu nude Trung Qudc, An do. Nepal,. .-
' Khoa Khoa hpc Iv nhien. Truong Dgi hpc CSn Tho - Cao hpc h6ahihicaK16. Truong D?ii hpc C4n Tho
^ Cao hpc hoahOucoKlS. Truong Dgi hpc C4n Tho
Tgp chi Klioa hgc 2012:21a 114-118 Truong Dgi hgc Cdn Tha
C i y cd xudc d i dupe sir dpng Uong y hpc cd truyen nude ta tir lau, vi nhiing cdng dung huu hieu trong dieu trj dau khdp, nhuc diu, sot ret, viem than, sdi nieu va tri gan. Ddi vdi phu nu, cd xudc giiip lam giim rdi logn kinh nguyet, dau do kinh b l , dau bung sau sanh. Ciy cd xudc vira la phuong thuoc vira la mdt nguyen liSu tdt trong nhieu bai thudc hien dang cd ten Uong danh muc thudc dieu tri cua Benh vien Dgi hpc Y H i Ngi, Vien dupe lieu Trung uong.
Tidp theo ket qua nghien cim ve thinh phin hda hpc ciia cay cd xudc thu hai d T r i Vinh m i chiing tdi da cdng bd trudc diy, (Tdn NO Lien Huong et al., 2011), hi?n nay nghien curu dugc thuc hien tren nguyen lieu thu hogch d VTnh Long. Do do, trong bai bao n i y chiing tdi cdng bd them mdt sd hgp chit phin cue trung binh va kha vira cd lap duoc.
2 NGUYEN L l £ U VA PHU'ONG PHAP 2.1 Nguyen lieu
M i u nguyen lif u dimg Uong r^hien ciru la phan thin ciy cd xudc dugc thu hii v i o thing 3/2010 tgi tinh Vinh Long.
2.2 Thief bi
Phd khdi lupng (ESI-MS) dupe ghi Udn miy MS 5989 B (Hewlett Pakard). Phd cpng hudng tir hgt nhin (NMR): ' H - N M R (500 MHz) v i '^C-NMR (125 MHz).
COSY, DEPT, HSQC, HMBC dugc ghi Uen may Bruker AM500 FT-NMR ciia Vien Hda hpc, Vien Khoa hgc v i Cdng nghd Viet Nam.
2.3 Chiet xuat va c6 lap
Bpt khd tu than ciy cd xudc (2,1 kg) dupe chiet vdi ethanol, cat loai' dung mdi dudi ap suit giim thu dugc cao tdng (204 g). Vdi mdi 100 g cao tdng cho them 0,5 lit nude cit va lin lupt chilt phin bd Idng-Idng cao nay vdi cic dung mdi petroleum ether (PE), chloroform (C), ethyl acetate (E) v i w-bulanol (B), cd quay dudi dung mdi ciia dich chiet thu dupe cic cao PE (41 g), cao C (19,2 g), cao E (6,2 g), cao B (21 g). Tien hanh sic ky cgt cao PE, cao C, E.
Tir cao PE (4Ig) tiln hanh sic ky cdt vdi he dung mdi cd tinh phin circ ting dan tir petroleum ether pha ty le vdi ethyl acetate, x i cgt vdi methanol, thu dupe 13 phan doan P. I — P.13. Tilp tuc sic ky vii lin Uen 2 phan doan P.3 v i P . I O thu dugc hai chit, khi kit tinh lai d i u cd tinh thi Uing. Chit (1) tinh thi hinh kim (4,20 g), chit (2) dgng bgt min (70 mg).
Tir cao C (19 g) tiln hanh sic ky cdt vdi he dung mdi PE:E ting din dp phin cue tir (9:1) din (0:10) v i he dung mdi E:M tii (99:1) d i n (30:70), sau dd xa cdt vdi methanol thu dupe 12 phin dogn, C1-C12. Tilp tuc sic ky cgt phin dogn CIO (3,12 g)hai lin Uen cpt silica gel, thu dupe chit (3) tinh thi miu ving (15 mg).
Tiln hanh sic ky cpt cao E (2 g) vdi he dung mdi PE:E ting din dd phan cue [tir (40:60) din (0:100)] va he dung mdi E:M tang din dp phin cue tii (90:10) den (30:70), sau dd xa cdt bing methanol, thu dugc 7 phin dogn: E1-E7. Tiep tuc sic ky cgt ddi vdi phan dogn E.4 (4,12 g) hai lin tren cdt silica gel, thu dupe chat (4) (25 mg).
Tcn> chi Khoa hoc 2012 21a 114-llS TnangBai hoc do Tha
3 K E T QUA VA THAO L U ^
3.1 Nhan danh caQ t n i c cua cac chat trong cao PE 5.1.1 stigmasterol (chAt 1)
Chit (1) la tinh thi hinh kim, mau trSng, ItSt tinh trong chloroform (4,2 g). Nhiet do nong chay 169-170"C. Sjc ky ban mong cho vit % = 0,43 khi giai ly bang CHCI3. Khi so sanh ph6 cua hcyp chat vod stigmasterol trong tai lieu tham khao (J.
Goad va T.H-Akihisa), cac so li?u trung khap hoan toan.
3.1.2 Dodecanoic acid (chdt 2)
Chit (2) la tinh thi dang bot, mau trSng, kit tinh trong E (70 mg). SSc ky ban mong cho vit R^ = 0,31 khi giai ly ban mong trong hexane.
Ph6 ' H-NMR (500 MHz. CDClj, 6 ppm) cho tong cpng 3 loji proton cua - C H j , - CH2- va -COOH. Tir 0.85 den 2,36 ppm la proton gSn vai carbon bao hoa. Pho
"C-NMR (125 MHz, CDCI3, 8 ppm) kit hap voi ph6 DEPT-NMR cho thjy chit (2) CO 12 carbon. Trong do co 1 nhom carbonyl -COOH (5 = 179,7 ppm), 10 nh6m - C H 2 - (S = 22.7—34,0). 1 nhom -CH3 (5 = 14,1). Vay chit (2) la dodecanoic acid.
Bang 1: PhdNMRcua cbat (1) so v4i1taili|u tham khao (J. Goad vaT. H. Akihisa) 5 ppm T T
1 2 3 4 5 6 7 8 9 10 11 12 13 14
C h a t ( l ) ' H ^J. Hz)
3,52 m
5,35 {5.0)
"C 37,2 31,6 71,8 42,3 140,7 121,7 31,9 31,8 50,2 36,5 21,1 39,8 42,3 56,9
Stigmasterol
"c
37,2 31,6 71,8 42,2 140,7 121.7 31,8 32,1 50,2 36,5 21,1 39,8 42,3 56,8
T T 15 16 17 18 19 20 21 22 23 2 4 25 26 2 7 28 29
Chat ' H (J, Hz)
1.01 J 0.89 J
5.16 ( * 7 , j ; 5.04 (rf. 7,5)
\.53 brd 0.79 d 0.81 d 0.80 r
^
" C 24,3 2 8 , 8 54,4 12,0 19,3 40,5 21,2 138,3 129,3 51,2 31,9 19,0 21,1 25,4 12,2
Stismasterol
"c
24,3 28,9 56,1 12,0 19,4 40,5 21,2 138,3 129,3 51,2 31,8 19,0 21,1 25,4 12,2 3.2 Nhan danh cau true ciia cac c h i t trong cao C va cao E
3.2.1 Quercetin (chat 3)
Chit (3) la tinh thi dang bot mau vang, tan trong methanol. SJc ky ban mong cho vet % = 0,3 khi giai ly ban mong trong E:M = 85:15.
Khoi pho CO peak ion gia phan tu m/z = 303 [M+H]* phu hop voi cong thUc phan tu CiiHisO, (khoi lu(?ng phan tu la 302 u). Ngoai ra, manh ion co m/z = 285 la ung vai ion du^c hinh thanh tir ion gia phan tu mat di mpt phan tu HiO.
Tap chi Khoa hgc 2012:21a 114-118 Truong Dgi hoc Cdn Tha
Tir phd ' H-NMR (500 MHz, DMSO) nhan thiy cd 12 tin hieu proton. Trong vimg tir trudng: (6,0-8,0) ppm cd 5 proton cua vdng tham gdm 2 proton d vi tri meta vdi nhau {J= 2,5 Hz) thudc vdng A, va 3 proton khac thupc vdng B, Uong do 1 H d vi fri meta \kpara vdi 2 H cdn lai, (J;„«„ = 2,0 Hz; J^rio = 8,5Hz).
Trong phd '^C-NMR v i DEPT-NMR: chit (3) gdm 15 C, Uong dd cd I carbon carbonyl 175,8 ppm. Vimg 5ppm (135,7 - 163,8) gdm 7 carbon bic 4 k l ndi ddi va mang nhdm t h i oxy tuong iing vdi cac vj tri 2, 3, 5, 7, 9, 3', 4'. Tgi Sppm: 103,0 v i 121,9 la 2 carbon bic 4 ke ndi ddi >C= ung vdi C,o, C,-. Vung Sppm (93,3-119,9) l i vung tin hieu ciia 5 carbon olefin dang =CH- tuong irng vdi Ce, Cg, Cx, Cy, Cf,-.
Dua v i o cac dO lieu phd, chiing tdi d l nghi cdng thiic cua hop chit (3) la Quercetin. Cac sd lieu phd ciia chit nay hoin toin khdp vdi tii li6u tham khao (John Goad).
Quercetin a-D-Glucopyranosyl-( 1 —*2)-^-D- fiuctofiiranoside
3.2.2 a-D-Glucopyranosyl-(I—*2)-^-D-fructqfuranoside (chdt 4)
Chat (4) cd tinh the dgng hgt hinh khdi, khdng miu, tan tdt trong methanol, kit qui sic ky ban mong cho vet R;-= 0,55 khi giai ly ban mong trong h? etyl acetate pha Uong methanol v i nude E:M:N = 4:1:1. Chit (4) cd nhiet dd ndng chiy:
184-185°C.
Trong phd ' H - N M R (500 MHz, CD3OD) cho thiy hpp chit (4) co 14 H, trong dd tin faidu d vimg (3,0-4,0) ppm l i nhung proton cua phin dudng, tin hieu tgi 5,41 ppm la proton anomer. Phin dudng cd 13 H vay Uong phin tir chit (4) cd thi gdm 2 gdc dudng. Phd '^C-NMR (125 MHz, MeOD) va DEPT-NMR cd 12 tin hieu carbon, trong dd cd 8 mui duong cua 8 carbon > C H - ; 3 mui i m ciia 3 carbon -CH2OH va 1 carbon tii c i p >C<.
Dua vao phd HSQC, xic dinh duoc carbon anomer 5c 93,7 ppm iing vdi proton anomer 6H 5,41 ppm (IH, d, J=3,5 Hz). Can cii vao phd COSY, tuong tic ciia proton niy vdi proton ciia 4 nhom hydroxymethin, I nlidm -CH2OH va dO lieu J ndi tren, nen xac dinh mdt gdc dudng d dang or-glucopyranose. Theo phd HMBC, proton anomer cd tuang quan vdi carbon tu cip tai 5c 105,4 ppm, C-2'. Mgch cdn Igi cd 2 nhdm —CH2OH ma mdt nhdm cd proton giy tuong quan vdi carbon tir cip (5H 3,63 ppm cd tin hieu giao vdi 6c 105,4 ppm), chiing td dudng cdn lai la dudng y^fiTictofiiranoside.
Phd khdi lupng (FT-MS) khdng thiy xuit hien peak ion phin tir nhung cd cac peak: m/z = 274 (100 % ) , ion m/z = 149 tuong iing vdi [CsHpOsf hay [CgHiaOe- CH2OH]* va peak cd m/z = 130 tuong img vdi [C5H6O4 ] ' hay [CgHioOs -
Tapchi Khoa hfc 2012:21a 114-118 Trucitg Dsi hx Cio The
CHsOH]". Vay suy ra cong thuc phan tu chat (4) la C n H i i O n voi it nhit 2 nhom - CH;OH trong phan rtt, va peak co m/z = 274 tuong ling vai [M - CH2OH - 2 HzO]'. K i t hop nhung du kien tren, chit (4) duoc djnh danh la a-D- GlucopyranosyH 1 —»2)-^-D-iructoturanoside
Bang2: P b S N M R c i a c h a t O ) Sppm Bang 3: Ph6 NMR cna c h i t (4) 6 ppm
2 3 4 5 6 6.18 (i^. 2.0_/
7 6.40W.-2,0y 8 9 10 1' 2' 7,69 (d. 7.5j 3' 4'
5' 6,89 Idd: 7.5:2) 6' 7,54 (d: 7 5/
4 KET LUAN 156.1 135,7 175.8 160.7 98.2 163.8 93,3 147.7 103.0 121.9 115.6 145.0 146,8 II 5.0 119.9
156.2 133,5 177,5 161.1 98.6 164.1 93,6 156.2 103.9 121.6 116.2 144.8 148.4 II5.2 121,1
1 2 3 4 5 6 2' r
3' 4' 5' 6'
5.41{rf:3,5) 3,45 (fltf; 4,0; 10,0)
3,75 m: 3.84 m 3.63 m ; 3.74 tn
4.11 ( J ; 8.0) 3,74 m, 3.86 m
93.7 73.3 74.7 71.4 74.4 62,3 64.1 105.4 79.4 75.8 83,8 63.4
2'. 3 3 2 5.6 4 4 2'
-
r3', 6' 4'. 6' 5'
Nhu vay chiing tdi da cd tap va de nghi c i u tnic ciia 4 hpp chit la stigmasterol, dodecanoid acid, a-D-gIucopyranosyl-(I—>2)-^-D-fiTJCtofiiranoside va quercetin.
Cic ket qui niy li ddng gdp mdi ve thanh phin hda hoc cua cay cd xudc viing Vinh Long. Quercetin cd vai Ud khang oxy hda rit tdt v i lic c h i phat trien te bao ung thu, (theo Kai Jin Wang et al., 2007).
Cic nghien ciiu Uen nhiing phin dogn cdn Igi dang dupe tiep tuc thuc hi^n.
TAI LIEU THAM KHAO
D 5 Huy Bich, D3ng Quang Chung, Bui Xuan Chuong, Nguyen Thuan Dong, Do Tmng Dam, Pham v a n Hien. Vu Ngpc Lo, Pham Duy Mai, Pham Kim Man, Doan Thi Nbu, Nguyen Tap, Tran Toan. Vien duoc lieu. Cay ihuoc vd dong vgt lam thudc a Viet Nam, Nha xuit ban khoa hpc va ky lhu|t Ha Noi, 2004, T|?) II.
Ton N u Lien Huong. Nguyen Minh Hien, Tiin Dinh Luan, Nghiin cuu thdnhphdn hda hgc cua than cdy Co xudc (A chyrcmthes aspera L.) thu a Trd Vinh, Tap chi Khoa hpc Truong DHCT, 2011, so 19b. 56-61
John Goad and Tosh Hiro Akihisa, Analysis of Steroid. Springer Publisher, 1* edition, 1997 Kai Jin Wang. Chong Ren Yang and Ying Jun Zhang, Phenolic antioxydants from Chinese toon (fresh young lea\es and shoots ofToona sinensis). Food Chemistry, 2007, 101, pp.
