Tdi li^u (ham khao
X. Nguyin Minh Khfli, Ph?m Thj N g u y ^ Hdng, Do Thi Phuong (2013), Nghien cixu dOc tinh cdp, tde dyng bdo v^ gan vd tde d^ng gdy d^c t l bdo ung thu ciia xdo tam phdn. Tgp chiDw^c li$u sd 1, 14-20. 2. Vi^n Du^rc Li?u (2004), Cdy thudc vd dpng v^t ldm IhuSc a Vift Nam, Tap I, trang 531. 3. Aggarwal BB, Bhardwaj A, Agganval RS, Seeram NP, Shishodia S, Takada Y (2004), "Role of resveratrol in prevention and therapy of cancer:
preclinical and clinical studies". Anticancer research, 24(5A):2783-2840. 4. Agostinis P, Vantieghem A, Merlevede W, de Witte PA (2002), "Hypericin in cancer treatment; more light on the way", The international Journal of biochemistry and cell biology, 34(3), 221-241. 5. Bhardwaj, A., and Aggarwal, B. B. (2003),
Receptor-mediated choreography of life and death, J. Clin. Immunol. 23, 317-332. 6. Feng L, Zhang LF, Yan T, Jin J, Tao WY (2006), "Studies on active substance of anticancer effect in Polygonum cusptdatum", Journal of Chinese medicinal materials, 29(7), 689-691. 7. Ferenc P, Soldr P, Kleban J, Mikes J, Fedorocko P (2010),
"Down-regulation of Bcl-2 and Akt induced by combination of photoactivated hypericin and genistein in human breast cancer cells". Journal of photochemistry and photobiology. B, Biology, 98(1), 25-34. 8. Ghanim H, Sia CL, Abuaysheh S, Korzeniewski K, Patnaik P et al (2010), "An anti- inflammatory and reactive oxygen species suppressive effects of an extract of Polygonum cuspidatum containing resveratrol", The Journal of Clinical Endocrinology and Metabolism, 95(9), E l - 8 . 9. Han JH, Koh W, Lee HJ, Lee HJ, Lee EO et al (2012),
"Analgesic and anti-inflammatory effects of ethyl acetate fraction of Polygonum cuspidatum in experimental animals". Immunopharmacol Immunoloxicol. 34(2), 191-195. 10. Hsu CY, Chan YP, Chang J (2007),
"Antioxidant activity of extract from Polygonum cuspidatum". Biological research, 40(1), 13-21. 11. Sung B, Parli B, Yadav VR, Aggarwal BB. (2010), "Celastrol, a triterpene, enhances TRAIL-induced apoptosis through the down-regulation of cell survival proteins and up-regulation of death receptors", J Biol Chem. 285(15), 11498-11507. 12. Wiley, S. R., Schooley, K., Smolak, P. J., Din, W. S.. Huang, C. P., Nicholl, J. K., Sutherland, G. R., Smith, T. D., Ranch, C , and Smith, C. A. (1995), "Identification and characterization of a new member of the TNF family that induces apoptosis", Immunity 3(6), 673-682. 13. Zhang Z, Ye T, Cai X, Yang J, Lu W, Hu C, Wang Z, Wang X, Cao P. (2013), "5,7-Dihydroxyflavone Enhances the Apoptosis-Inducing Potential of TRAIL in Human Tumor Cells via Regulation of Apoptosis-Related Proteins", Evid Based Complement Alternat Med. Article ID 434709, 13 pages.
Tap chiDuffc liiu, tap 18, so 4/2013 (Trang 213 - 220)
THANH PHAN HOA HQC CUA QUA INfUC NAC
Phmmg Tki$n Thmmg'*, LeXuan Thuy^, Nguyin Minh Kh&i' Khoa Hoa phdn tich - Tieu chudn, Vien Du(rc li$u
^Trung tdm Kiem nghiem thuoc San La
* E m a i l : p h u o n g t h i e n t h u o n g @ y a h o o . c o m ( N h a n bai ngay 2 9 thang 7 n a m 2 0 1 3 )
T 6 m t a t
Tit djch chi^t methanol qud nQc ndc, dd phdn ldp duge sdu chdt (1-6). Bdng phdn tich p h i (UV, IR, ' H - N M R ,
"C-NMR. MS), ket hgp vdi tlnh chat ly h6a dd xdc djnh cdc chdt lin lugt Id acid ursolic (1), baicalein (2), chrysin (3), oroxylin A (4), luteolin (5), vd apigenin (6). KSt qud cho bilt cac thdnh p h ^ chinh trong qua niic ndc Id cdc flavonon, trong dd chdt c6 hdm lugng ldm nhdt Id chrysin. Ldn ddu tien luteolin duge tim thdy c6 trong cdy niic ndc.
Tir khda: Nuc ndc, thdnh phdn hoa hQC, Acid ursolic, Flavon, Luteolin.
Tap chtDUffc mu, tdp 18, so 4/2013 213
Summary
Chemical Constituents of the Fruit of Oroxylum indicum
Phytochemical study on the methanol extract from the fruit of the medicinal plant Oroxylum indicum resulted in the isolation of six compounds 1-6. Their sOiictures were identified as ursolic acid (1), baicalein (2), chrysin (3), oroxylin A (4), luteolin (5), and apigenin (6) by analyses of physicochemical and spectroscopic data. This is the first report on the presence of luteolin in the plant O. indicum.
Keywords: Oroxylum indicum. Fruit, Chemical constituent. Ursolic acid. Flavone, Luteolin.
1. Dat van de
Cdy mic nac co ten khoa hpc Id Oroxylum indicum (L.) Vent, (ten d6ng nghia Bignonia indica L.), thuoc hp Niic ndc (Bignoniaceae) [1-3]. Trong y hpc c6 truyin mic ndc cho hai vi thuoc la vo mic ndc (vo thdn va vo re) vd hat mic ndc [1-4].
Cdc vi thulc ndy dupc diing trong mot so bdi thulc chiia kiet ly, tieu chdy, ho, dau Vli, ap-xe vii, myn nhpt, benh sai dau, sot phdt ban, sung viem, viem gan, dau mat do, mpt s6 benh ngodi da nhu eczema, chdm hoa cac not sdn, viem da co dia, ngiia. Cdc nghien ciiu ve thdnh phdn hoa hpc chinh cua vo niic nac vd hat Id cdc flavonoid thupc nhom flavon, chii yeu Id baicalein, chrysin, oroxylin A, apigenin, kaempferol, quercetin, 3,5,7-trihydroxyflavon; 5,7-dihydroxy -3-methoxyflavon, pinostropin vd cac glycosid cua chiing [5-8]. Ngodi ra con co cdc flavonoid nhom flavanon va flavanonol [5], pterocarpan [5], triterpenoid [7], phytosterol, alkaloid vd mpt so chat khdc [5-8].
Nguoi dan van dung qud niic ndc chua gia de lam thiic an vd phong ngira cdc benh ke tren. Tuy nhien, cho den nay vdn chua co nghien ciiu ndo ve thanh phdn hoa hpc vd tac dyng sinh hpc cua qua niic ndc an dupc de chiing minh cho cac tac dyng kl tren. Nhdm gop phan tim hieu gid tri sir dung cua qua mic nac co the sii dyng lam thulc va thuc phdm, chiing toi tiln hanh nghien ciiu phdn lap mpt s6 thdnh phdn hoa hpc trong qua niic nac. Bdi bdo nay trinh bay ket qud phdn lap sdu chat tir qud mic ndc.
2. Nguyen lieu va phinmg phap nghien cihi
2.1. Nguyen lieu
Dupc lifu Id qud mic ndc khodng 3 tudn tuli, dupc thu hdi tai tinh San La ddu nam 2013. Mdu dupc lieu duac TS. Nguyen Quic Huy, Bp mon Thuc vat, Trucmg DH Dupc Hd Npi thdm dinh ten khoa hpc la Oroxylum indicum (L.) Vent., hp Niic ndc (Bignoniaceae).
Mau nghien ciiu hifn dang dupc luu giii tai Khoa Hoa phan tich - Tieu chudn, Vien Dupc lieu. Dupc Ueu dupc nia sach, thai nho, say kh6 truoc khi nghien ciiu.
2.2. Dung moi, hoa chdt
Cac dung moi dung cho chiet xudt vd phdn lap hoat chat: dicloromethan (CH2CI2), ethylacetat (vilt tat Id EtOAc), n-hexan (Hx), methanol (CH3OH), n-buthanol (BuOH)... la cdc hoa chdt cong nghifp dupc mua tir cdc cong ty kinh doanh vdi chdt lupng dam bdo cho thuc nghifm. Ban mong chay sdc ky lop mong silica gel F254 (Merck) va RP18 (Merck).
2.3. Thiet b}, d^ngcyi
Cac loai cpt sdc ky (cpt thiiy tinh), mdy cat quay, may do dp nong chdy Gallenkamp {Sanyo electrothermal digital), mdy do pho tii ngoai (UV) Shimadzu UV-1800, mdy do phi hong ngogi (IR) GX-Perkin Elmer, khoi pho luong phun mil difn tu (EMS-MS) Agilent 1100 LC-MSD Tram, phi cpng huang tir h?t nhdn ('H-NMR, '^C-NMR, DEPT) Bruker AM-500 FT-NMR.
2.4. Phuang phdp nghien cuu
Chiet cac chat tir dupc lifu bdng MeOH
214 Tap chiDitofc lieu, tap 18, so 4/2013
theo phuang phdp dun hii luu each thiiy.
Phan doan bdng dung m6i c6ng nghifp Hx, EtOAc, vd BuOH, chay qua sdc ky cpt pha thudn (pha tinh la silica gel), pha ddo (pha tmh Id YMC va RP18) dl tach vd phdn lap cdc chat. Sdc ky lap mong dimg d6 theo doi vet cdc chat tir cdc phan dogn. Kiem tra dp tinh khiet bang SKLM (soi dudi den tit ngoai a buoc song 254 nm vd ha nong a 110°C); do nhiet dp nong chdy va dp quay cue ; kiem tra bdng HPLC. Xdc dinh cdu tnie ciia cdc chdt phdn lap dupc dya tren ket qud phdn tich cdc tinh chat Iy hoa (cam quan, nhiet dp nong chdy, dp quay cyc) vd cdc pho tu ngoai (UV), hong ngoai (IR), phi khoi (MS), pho cpng hudng tit hat nhdn ('H- NMR, '^C-NMR, DEPT) sir dyng chdt chudn npi Id TMS (tetramethyl silan) vd so sanh cdc dii lifu thu dupc tir thuc nghiem voi cac da lifu dd c6ng b6.
2.5. Chiit tach vd phdn lap cdc hgp chdt tif qud nuc ndc
Dupc lieu 1,0 kg qud mic ndc kho dupc cat nho, ngam Ignh bdng 5 lit con 96% trong 3 ngdy (^ 3 ldn). Cac dich chiet c6n dupc gpp lai, cdt thu h6i dung moi thu dupc 80,2 gam cao kho (gpi Id cao con). Cao con ndy lai dupc hoa tan vao nudc (1 lit) cdt thdnh nhu dich rii ldc, chilt phdn dogn ldn lupt voi Hx (0,5 lit X 3 ldn), EtOAc (0,5 lit x 3 ldn), BuOH (0,5 lit X 3 ldn). Cac dich chilt Hx, EtOAc, vd BuOH dupc cdt loai bo dung moi, thu dupc 12,6 gam cao phdn dogn Hx (viet tdt la cao Hx); 32,5 gam cao phdn dogn EtOAc (cao EtOAc), vd 24,7 gam cao phan dogn BuOH (cao BuOH). Cao phan doan EtOAc (30 g) dupc chay qua cot silica gel (12 15 cm), nia gidi bdng hf dung moi CH2Cl2-MeOH theo gradient ning dp 100:1, 50:1, 40:1, 30:1, 20:1, 10:1, 5:1, 4:1, 2:1, 1:1 va 0:1, moi hf 4 lit. Kilm tra thdnh phdn trong djch rira gidi bang TLC, tir dich nia giai dupc phdn thanh 6 phdn doan chinh Id
FEOI.l (4,3 g), FE0I.2 (3,8 g), FE0I.3 (12,5 g), FE0I.4 (4,7 g), FE0I.5 (3,6 g), vd FE0I.6 (6,6 g). Tiep tyc cho phdn doan FEOI.l chgy qua cpt silica gel (4 x 25 cm), nia gidi bdng hf Hx-EtOAc theo gradient 10:1, 9:1,...1:1, 0:1; phdn dich nia gidi boi hf 5:1 din 4:1 dupc tiep tyc cho qua cpt silica gel (2,5 x 20 cm), riia giai vdi he 3:1 thu dupc chat so 1 (27 mg). Phdn doan E0I.3 dupc cho chay qua c6t sephadex LH20 (4 x 25 cm), rira gidi bang dung moi MeOH, thu dupc nhieu phdn doan, trong dd bang TLC xdc dinh dupe phan dogn cd chiia flavonoid (4,2 g) cho cac vet mdu vdng tren sdc ky dd. Sur dyng cpt sdc ky pha ddo YMC (4 x 25 cm), nia gidi bang hf dung mdi MeOH-HzO theo ti If 10:1, 5:1, 3:1, 2:1, 1:1, vd 0:1 thu dupc chdt si 2 (48 mg). Phdn dogn cd chiia cac flavonoid con lai dupc tiep tuc cho chay qua cpt sdc ky pha ddo RP18 (2,5 X 25 cm), riia gidi bdng hf dung moi MeOH-HiO theo ti If 15:1, 10:1, 5:1,2:1, 1:1, vd 0:1 thu dupc chdt sd 3 (112 mg), vd sd 4 (16 mg). Phdn doan E0I.5 cho chay qua cdt sephadex LH20 (4 x 25 cm), rira gidi bang MeOH thu dupc phdn dogn chiia flavonoid (1,3 g). Cdc chat so 5 (7 mg) vd sd 6 (11 mg) thu duoc tir phdn doan ndy sau khi chay cpt RP18, rira gidi bdng hf dung mdi MeOH-H20 theo cdc ti If 10:1, 7:1, 5:1 vd4:l.
Chdt so 1: Chdt sd 1 thu dupc d dgng bpt vd djnh hinh, mdu trdng; nhift dp ndng chdy 258-260 ''C; phi UV (do trong MeOH) X^^x 203 nm; phd IR v^ 3425 (OH), 2864(C-H), 1690 (C=0), 1460, 1039 (C-0).
Phd khdi (EMS-MS): m/z = 456 [M]", 455 [M - H]' (KLPT cua C30H10O3 la 254); Phi 'H-NMR (aceton-rffi, 500 MHz): 5,22 (IH, t, J = 3,5 Hz, H-12), 3,15 (IH, m, H-3), 2,23 (IH, d, J = 10,5 Hz, H-18), 1,13, 0,98, 0,96, 0,83, 0,78 (tdt cd diu cdc tin hifu 3H, s, H- 23, 27, 26, 24, vd 25), 0,96 (3H, d, J = 7,5 Hz, H-30), vd 0,89 (3H, d, J= 6,5 Hz, H-29);
Tgp chiDUffc lieu, tap 18, so 4/2013 215
'^C-NMR (aceton-rf«, 125 MHz): 39,5 (H-1), 28,1 (H-2), 78,6 (H-3), 37,8 (H-4), 56,2 (C- 5), 19,1 (C-6), 34,0 (C-7), 40,4 (C-8), 48,5 (C-9), 39,9 (C-10), 24,0 (C-U), 126,3 (C- 12), 139,3 (C-13), 42,9 (C-14), 28,7 (C-15), 25,0 (C-16), 48,3 (C-17), 53,9 (C-18), 39,9 (C-19), 39,9 (C-20), 31,4 (C-21), 37,6 (C- 22), 28,7 (C-23), 15,9 (C-24), 16,3 (C-25), 17,7 (C-26), 23,9 (C-27), 178,5 {C-28), 17,5 (C-29),21,5(C-30).
Chdt s6 2: bot mau vang; nhiet do nong chay 262-265 °C; phd UV (MeOH) X„„ 215, 274, 324 nm; ph5 IR v„,x cm"': 3415 (OH), 3104 - 2949 (C-H), 1660 {C=0), 1616 - 1474 (C=C), 1082 (C-0). Ph6 khdi (EMS- MS): m/z = 269 [M H]' (KLPT cua C15H10O5 la 270); Pha ' H - N M R (500 MHz) va "C-NMR (125 MHz): xem Bang 1.
Chdt so 3: bgt vo dinh hinh; mau vang;
nhiet dp nong chay 258-260 °C; ph6 UV (MeOH) Xmsx 214, 268, 312 nm; ph6 IR Vmax cm-': 3399 (OH), 2939 - 2628 (C-H), 1652 (0=0), 1448, 1170 (C-0). Ph6 khoi (EMS- MS): m/z = 255 [M + H]*, 253 [M - H]"
(KLPT cua CisHioOi la 254); Ph6 ' H - N M R (500 MHz) va '^C-NMR (125 MHz): xem Bang 1.
Chdt so 4: bgt vo dinh hinh; mau vang;
nhiet do nong chay 258-260 "C; ph6 UV (MeOH) "Km 211, 271, 315 nm; pho lRv„ax cm"': 3399 (OH), 2937 (C-H), 1653 (C=0), 1499, 1097 (C-0). Ph8 kh6i (EMS-MS): m/z
= 285 [M -^ H]*, 283 [M - H]" (KLPT cua CisHijOi la 284); Ph6 ' H - N M R (500 MHz) va "C-NMR (125 MHz): xem Bang 1.
Chat so 5: bot vo dinh hinh; mau vang;
nhiet do nong chay 327-329 °C; ph6 UV (MeOH) X„„ 254, 267, 347 nm; pho IR v„„
cm-'; 3413 (OH), 2925 - 2855 (C-H), 1654 (C=0), 1454, 1372, 1265, 1167, 1034 (C-O).
Ph6 khai (EMS-MS): m/z = 287 [M + H]*, 285 [M - H]- (KLPT cua CisHioOs la 286);
Pha 'H-NMR (500 MHz) va '^C-NMR (125
MHz): xem Bang 1.
Chdt s6 6: bgt vo dinh hinh; mau vang;
nhiet dg nong chay 338-340 °C; ph6 UV (MeOH) Xmax 200, 268, 334 nm; ph8 IR v„„
cm': 3398 (OH), 2937 - 2866 (C-H), 1661 (0=0), 1467, 1359, 1176 (C-0). Pho khai (EMS-MS): m/z = 271 [M + H]*, 269 [M - H]- (KLPT cua CuHioOj la 270); Ph6 ' H - NMR (500 MHz) va '^C-NMR (125 MHz):
xem Bang 1.
3. Ket qua va Ban luan
Chat so 1 thu duge a dang bgt va dinh hinh man trSng, nhiet do nong chay 257-260
°C. Pha UV ciia chSt so 1 cho thay chi co mgt dinh hap thu tai Xmax 203 imi, chiing to 1 CO lien ket dai trong phan tu. Pha IR cho biet trong phan tii cila 1 co cac nhom OH, C=0, va C-O, tiic la co the co nhom COOH trong phan hi. Tren ph6 '^C-NMR cua 1 c6 tong cgng 30 tin hieu carbon, ket hgp voi pha khoi CO pic phan tii m/z = 456 [M]- phu hgp voi khgi lugng ciia mgt chat co khoang 30 carbon. Ciing tir pha '^C-NMR cho tin hieu mgt carbon thuoc nhom carbonyl co do dich chuyen 5c 178,5 ppm, chiing to 1 co chua nhom COOH. Tii tat ca cac dii lieu thu duge nay cho thay chat sa 1 la mgt triterpnoid co nhom COOH [9]. Tren pha ' H - N M R CO tin hieu cua mpt proton thuge hen ket doi (6H 5,22), phu hgp vai tin hieu co mgt lien ket dai tren pha "C-NMR va DEPT [8c 126,3 (CH) va 139,3 (C)]. TiSp tuc, pha 'H-NMR CO tin hieu cua bay proton thuge nhom methyl (CH3), trong do co hai nhom xuat hien doublet (5H 0,89 va 0,96) va nam nhom xuat hien a dang single (8H 0,78; 0,83; 0,96;
0,98 va 1,13). Nhu vay, chSt 56 1 la mpt triterpnoid thuoc nhom ursan [9]. Ngoai ra, cac tinh chat vat ly chat sa 1 co dang va dinh hinh, nhi^t dg nong chay trong khoing 258- 260 °C cho thay day co thS la acid ursolic [9], mgt chdt co khai lugng phan tii bJng 456. So sanh vcri cac dO lieu da cong b6
216 Tgp chl Duac lieu, lap IS, so 4/2013
trudc ddy (Bdng 1) cho phep khdng dinh chdt si 1 Id ursolic (3-^hydroxy-urs-12-ene-28- oic-acid).
'COOH
Hinh 1. CSng thiic h6a hoc cua chdt s6 1 (acid ursolic)
Chdt so 2 thu dupc cd dang bdt mdu vang, nhiet dp ndng chay Id 262-264 °C. Phi UV cua 2 cho cdc dinh hap thy cyc dgi tgi X^ax 215, 274, 324 nm, chiing td cd li^n kit ddi lien hpp trong phan tii. Phi IR cho bilt trong phan ttr ciia 2 cd cdc nhdm OH (3415 cm"'), 1660 (C=0), vd 1082 (C-0). Tren phd '^C- NMR xudt hifn mudi ba tin hieu carbon cd dp dich chuyin 5c trong khoang 95,1 - 184,2 ppm, gpi y Id cdc carbon cua vong thom vd mpt nhdm carbonyl (C=0), phii hpp vdi phd IR. Phd H-NMR cho cac tin hifu ctia proton tuong duong ndm proton cua vdng thorn tai 8H 7,97 - 7,99 (2H, m) vd 7,56 -7,59 (3H, m) Id cdc proton cua mpt vdng thom bi the mpt
Bang 2. S6 lifu ph6 'H- (500 MHz) va '
vi tri. Dovdng chi thS d mdt vj tri nSn cd tinh ch^t ddi xiing, hai carbon vd hai proton tai cdc vi tri 2' vd 6' doi xiing nen cd cac tin hifu tning nhau tren phd ' H - vd '^C-NMR;
tuong tu nhu vdy Id carbon va proton cua cdc vi tri 3' vd 5'.Tuy tren phd '^C-NMR chi xuat hien mudi ba tin hifu carbon, nhung cd the chat so 2 cd mudi lam carbon trong phdn tii. Ket hpp tdt cd cdc dfl lieu phd UV, IR, '^C- vd ' H - N M R cho flidy chdt sd 2 Id mot flavonoid, phu hpp vdi khdi luong phan tu 270 tim dupc tren khd khli (EMS-MS) cho pic ion m/z ^ 269 [M - H]" tuong iing voi KLPT cua C15H10O5 Id 270. Ngodi ra tren pho ' H - N M R cdn co hai tin hieu proton singlet tai 5H 6,73 va 6,62 ppm cd thi cua vdng thom ho|.c cua lien kit ddi, chiing td c6 mdt hodc hai vi tri ciia vong thom khdng bj the, hodc mdt vi tri cua vdng thom vd vi tri sd 3 khdng bi the. Mpt flavonoid cd mpt vong thom cd bdn proton thi do chi cd thi Id vdng B, va vdng thom cd mdt hodc hai proton Id vdng A. Nhu vay chdt si 2 duoc xdc dinh Id mpt flavonoid cd khli lupng phan td Id 270, cd nhift dp ndng chdy 262- 264 **C, cd vdng B khdng bi thi, mpt ho^c hai vi tri cua vdng A khdng bj the nSn cd the Id 5,6,7-trihydroxyflavon (ten thudng gpi Id baicalein). So sanh vdi cdc dii lifu da dupc cdng bo [10], [11] cho phep khdng dinh chdt sd 2 la baicalein, mpt flavonoid hay gap trong vd, hoa vd hat niic nac [5-8].
'C-NMR (125 MHz) cua cSc ch^t s6 2-6
Vjtri
1 3 4 5 6 7
Chits62' 6«(ppm);
= Hz)
6.91;s (ppm) 162,9 104,5 iez,i 153,6 131.8 147,0
Ch^ls63»
CH(ppni);
do boi (J = Hz)
6,79; s
6,28; d (2,0) 6c (ppm)
164,7 106,2 183.1 163,4 99,9 165,1
Chdt 564-^
6H (ppm); d$
boi (J = Hz)
6,67; s 6c (ppm) 158,6 105,4 184,0 153,7 132,7 165,5
Chits6
6K (ppm); dp bpi (J = Hz)
6,54; s
6,21; bfs
&"
6C (ppm) 166.5 104.0 184,0 163,4 100,3 166,2
Chdts&e' 6H(ppm];d$
b$i(J = Hz)
6,59; 3
6,21; brs 6c (ppm) 166.4 104.0 184,1 159,6 100,3 166,3
Tgp chl Duac lieu, tap IS, so 412013 217
6 9 10
r
1 3' 4' 5- 6- OCH.
tMK 70H 60H
6,63;!
8,04 7,58 7,58 7,58 8,04 m m m m m
12,66; s 10,57; s 8,81; s
94,0 149,8 104.3 129,3 126,2 129,1 131.8 129,1 126,2
6,57; d (2,0)
8.06, dd (8,0,1.5}
7,60; m 7.60; m 7,60; m 8.06, dd (6,0; 1,5)
12.89; s 9,78; bts
94,9 158,9 105,6 132,3 127,2 130,0 132,7 130,0 127.2
6,57;!
7.92; bra (8,0) 7,55; m 7.55; m 7,55; m 7,92; br (8,0)
3,91; s 95,3 154,5 105.9 132.2 127,2 130,0 132,8 130,0 127.2 60,9
6,44; br!
7,38; bra
6,90; brd (8,5) 7,39; d (8,5)
955 159,6 105,5 123,8 114.3 147.2 151,1 116,9 120,4
6,46; bis
7,85; d (8,0) 6,93; d (8,0)
6,93; d (8,0) 7.85; (1(8.0)
95J 163,4 105,5 123,4 129,6 117,2 162,9 117,2 129,6
' Do trong DMSO-rf^; ** Do trong aceton-t/a (CD3COCD3); " Do trong CD30D-CDC13; ** Do trong CD3OD.
C ^ kj hi0u; brs (broad singlet), d (doublet), dd (double of doublet), m (multiplet), s (singlet).
H3C0-
OH 0 OH 0 OH 0
4 5 6 Hinh 2. Cong thiic h6a hgc ctia cdc chdt s6 2 ~ 6
Chat so 3 thu duge cung la dang bgt mau vang, nhi?t dg nong chay 283-285 °C, them nua, cac ph6 UV, IR, ' H - va "C-NMR cua 3 tuong tu voi cac pho cua chdt s6 2 (xem phy Ivc 3), cho thiy 3 cung la mgt flavonoid va c6 ciu tjo tucmg tu vcri 2. DiSm khac nhau 14 tren pho ' H - N M R cua 3 co them mgt proton thuoc v6ng thom, tong cgng co tSt ca tam proton ciia vong therm hay lien kSt doi. Hon niia, CO hai tin hieu proton cua vong therm xuit hi?n a dang doublet 8H 6,57 va 6,28 (dSu
CO J = 2,0 Hz), trong khi mgt tin hi?u single tai 5H 6,79 van la cua proton cua vj tri s6 3.
Dieu nay chting to vong A ciia 3 co hai proton tai cac vi tri s6 6 va 8 [10]. Nhu vay so vcri chat so 2 thi chSt s6 3 khong co tihom OH t^ vj tri so 6, do do 3 duge xae dinh 14 chrysm (5,7-dihydroxyflavon). Ph6 EMS-MS ciia 3 cho pic m/z = 255 [M + H]* va 253 [M - H]', phii hgp vcri khdi luong phan tu 254 cua chrysin (C15H10O4). Vay chit s6 3 duge xae dinh 14 chrysin (5,7-dihydroxyflavon),
218 Tgp chl Duac lieu, tap IS, so 412013
mdt thdnh phdn hda hpc chinh cua nuc nac [5-8].
Chat sd 4 thu dugfc cung la dang bdt mdu vdng, nhiet dp ndng chdy 283-285 °C. Cdc phd UV, IR, ' H - vd '^C-NMR cua 4 tuong tu vdi cac phd cua chat sd 2, cho thdy 4 cung la mpt flavonoid vd cd cdu tao tuorng ty vdi 2.
Phd 'H-NMR ciia 4 gidng cua 3 la cd nam tin hieu proton cua vdng B khdng bj th^ (8H 7,92; 2H, brd va 7,55; 3H, m), mdt proton vong thom (5H 6,57; s) vd mdt proton cua vi tri sd 3 (5H 6,67; s), tuy nhien, diem khdc Id c6 them mpt tin hi?u proton thudc nhdm methoxy (OCH3) gdn vac vdng therm (5H 6,57). Tren phd '^C-NMR cung cd tin hieu cua nhdm methoxy tai 6c 60,9. Dieu ndy chiing td chdt sd 4 cd mpt nhdm methoxy (OCHs) the vdo vdng A tai vi tri sd 6 nen cdng thiic phan tii la C16H12O5 va cd KLPT la 284, phii hpfp vdi phd EMS-MS cho cac pic m/z = 285 [M + H]"" va 283 [M - H]' va cdc cac dii lieu phd da duac cdng bd [11].
Vdy chat sd 4 dupc xdc djnh la 5,7- dihydroxy - 6 - methoxyflavon, ten thdng thudng Id oroxylin A, Id chdt thudng gap trong niic nac [5], [6], [11].
Chat sd 5 thu dupc cung cd dang bpt mau vang, nhift dp nong chay 327-329 "C, ket hpp vdi cdc pho UV, IR, 'H- vd '^C-NMR cua 5 cho bik day cflng la mpt flavonoid.
Phd ' H-NMR cua 5 khac vdi phd ciia cac chdt 2-4 la cd tin hieu proton ABX ciia vdng thom tai 6H 7,38 (IH, brs); 6,90 (IH, brd, J=
8,5 Hz), va 7,39 (IH, d, y = 8,5 Hz). Dac dilm tren phd ' H - N M R cua 5 gidng vdi phd cua chat s6 3 la van cd cac tin hi?u ciia cdc proton tai vj tri sd 3, 6 va 8, ldn lupt tai SH 6,54'(IH, s), 6,21 (IH, brs), vd 6,44 (IH, brs). Nhu vdy cd thS thay nhdm ABX thupc vong B, tiic la d cac vj tri sd 3 vd sd 4 cua vdng B dd du(7C t h i Tren phd '^C-NMR cua 5 chi cd tin hieu cua mudi ldm carbon nen cdc nhdm the vao vdng B sg Id hydroxyl
(OH), suy ra chdt sd 5 la 3',4',5,7- tetrahydroxyflavon, cd cdng thiic phan tu la C15H10O6 vd cd KLPT id 286. Di6u ndy phu hpp vdi phd EMS-MS cua 5 cho cac pic m/z
= 287 [M + H]* va 285 [M - H]-, ddng thdi cdc cac du: liSu phd ciia chdt nay cflng phii hpp vdi cdc sd lieu dd duac cdng bo [12], [13]. Do dd 5 duac xae dinh la 3',4',5,7- tetrahydroxyflavon, cdn gpi Id luteolin. Mac du luteolin la mpt flavonoid nhdm flavon rat hay gap trong thuc vat [12], [13], nhung ddy Id ldn ddu tien chdt ndy dupc phan lap tu' cdy nuc nac.
Dang bpt mau vang, nhiet dp ndng chay 338-340 "C, kk hpp vdi cdc phd UV, IR, 'H- vd '^C-NMR cua 6 cho bi6t day cflng la mdt flavonoid. So sanh pho 'H-NMR ciia 6 vdi 5 thay cung cd tin hieu cua cdc proton tai vi tri sd 3, 6 vd 8, ldn lu^ tai 5H 6,59 (IH, s), 6,21 (IH, brs), vd 6,46 (IH, brs); nhung cd diem khac tren phd 'H-NMR cua 6 la cd tin hieu proton ABAB cua vdng thom tai 5H 7,85 (2H, d, J = 8,0 Hz) vd 6,93 (2H, d,J= 8,0 Hz). Nhu v|iy cd the thay nhdm ABAB thupc vdng B, tuc la dupc thS d vj tri sd 4'. Dieu nay phii hpp vdi phd '^C-NMR cua 6 xudt hi?n mudi ba tin hi?u carbon do sir triing dp dich chuyen 5c cda nhdm ABAB (C2' triing vdi C6', C3' trimg vdi C5') do tinh chat ddi xiing. Cflng do cd mudi ba tin hi|u carbon nen nhdm the vdo vi tri so 4' cua vdng B cung duac xdc djnh la hydroxyl (OH), suy ra chat sd 6 la 4',5,7-trihydroxyflavon, cd cdng thiic phdn tu Id CisHjoOs vd co KLPT la 270, phii hap vdi pho EMS-MS cho cac pic m/z = 287 [M + H]"" va 285 [M - H]'. Cdc cac dfl- li^u phd cua 6 cung phu hop vdi cdc sd lipu da dupc cdng bd [12], [14]. Vdy chdt sd 6 dupc xae dinh la 4',5,7-trihydroxyflavon, cdn gpi Id apigenin, mdt flavon dd dupc phan lap tir vd nuc ndc [5], [7], [8].
5. Ket luan
Trong cong trinh ndy, nhdm nghidn cihi
Tgp chi Duoc lieu, tdp 18, so 4/2013 219
dd phdn lap dupfc sau chdt tir qud mic ndc . qua mic ndc Id cdc flavon, chdt cd ham luong Bdng viec phdn tich phd (UV, IR, ' H - N M R , ldn nhdt la chrysin. Lan dau tien luteolin '^C-NMR, MS), kk hpp vdi cdc tinh chdt ly duprc tim thdy cd trong cay mic ndc.
hda da xdc dinh cac chat ldn lupt Id acid L&i cdm on: Cdc ldc gid chdn thdnh Bg Khoa ursolic (1), baicalein (2), chrysin (3), hgc vd Cong ngM da tdi trg kinh phi cho nghien oroxylin A (4), luteolin (5), vd apigenin (6). cmi ndy thong qua Nhl?m v?/ Nghf dinh thu hgp Kit qud cho bilt cdc thdnh phdn chinh trong tac vdi Nhgt Bdn (sd 37/2010/HD-NBT).
T^i li^u tham khao
1. Vi|n Dupc li?u (2004), Cdy thudc va dong vQt ldm thudc o Vi?t Nam. NXB Khoa hgc vi KJ thuSt, t|[p 2,480- 484. 2. D6 xAt hq\ (2001), Nhihig cdy thudc va v/ thudc Vi^t Nam. NXB Y hgc, HA Ngi, 726-728. 3. VO ySn Chi (2012), Tu- diin cdy thudc Viet Nam. NXB Y hgc, tap 2, 377-378. 4. Bg Y t6 (2009), Dirge diin Viet Nam IV.
NXB Y hgc, tr. 856. 5. Harminder, Singh V., Chaudhary A.K. (2011), "A review on the taxonomy, ethnobotany, chemistry and phaimacology of Oroxylum indicum Vent", Indian Journal of Pharmaceutical Sciences 73(5), 483-490. 6. Babu et al. (2010), "Gastroprotective flavonoid constituents from Oroxylum indicum Vent", BiQorganic& Medicinal Chemistry Letters 20, 117-120. 7. Yan R. et al. (2011), "Antioxidant flavonoids from the seed oiOroxylum indicum", Fitoterapia 82(6), 841-848. 8. Wei X. et al. (2012), "Chemical constituents of seeds of Oroxylum indicum", Zhongguo Zhang Yao Za Zhi 38(2), 204-207. 9. Thuong P.T. et al. (2005),
"Inhibitory effect on TNF-a-induced lL-8 production in the HT29 cell of constituents from the leaf and stem of Weigela subsessilis. Archives of Pharmacol Research 2S(10), 1135-1141. 10. Chen L. J. et al. (2003), "Isolation and identification of four flavonoid constituents from the seed of Oroxylum indicum by high-speed counter- current chromatography". Journal of Chromalograply A 988(1), 95-105. 11. Yadav A.K. et al. (2012),
"Simultaneous determination of flavonoids in Oroxylum indicum by RP-HPLC", Medicinal Chemistry Research, 22(5): 2222-2227 12. Ha T.J. et al. (2012), "Isolation and identification of phenolic compounds from the seeds of Perilla frutescens (L.) and their inhibitory activities against a-glucosidase and aldose reductase" Food Chemistry. 135(3), 1397-1403. 13. Andersen O.M., Markham K.R. ed. (2006), Flavonoids: Chemistry.
Biochemistry, and Applications. Taylor & Francis, p. 52. 14. Markham K.R., Temai B (1976), "'^C NMR of flavonoid-Il. Flavonoids other then flavones and flavonol aglycones". Tetrahedron 32, 2607-2612.
Tap chiDircfc lieu, tap 18, so 4/2013 (Trang 220 - 224)
TRITERPENOID PHAN LAP TlT CAO C H I E T w-HEXAN C U A C A Y I M U A B A
Trhn Thanh Hd' * Nguyin Minh Khoi', Phgm Thiiy Duan^, Nguyin Van B^i^, Nguyin Thf Hd', Bill Mpng Hoa^
Vien Duac lieu
^Truang dai hgc Khoa hgc Tu nhien - Dgi hgc Qudc gia Hd Ngi
*Email: [email protected] (Nh^n bai ngay 18 thang 6 nam 2013)
T6m tSt
Tir cao chi^t «-hexan ciia ph^n tren mat ddt cay mua bi da phan lap duge ^-sitosterol, daucosterol, hai triterpen acid ursolic va acid betulinic. Ciu tnie h6a hgc ciia chiing duge xdc djnh dya tren co sd cdc dfl ki?n ph6 thyc nghifm MS, NMR vd cdc da ki?n ph5 NMR d5 cong b6. Day Id 1^ dku tiSn thanh h6a hgc cOa cdy mua bh thu hdi cr Vi?t Nam dirge cfing b6.
Titklida: Cdy mua bd, Melastoma candidum D.Don.. Acid ursolic. Acid betulinic.
220 Tap chiDuffc lieu, tdp 18, so 4/2013
