Tai lieu tham khio
1. Vfl Vin Chi (2012), Tit diin cay thudc ViftNam, t?p 1, Nxb Y hgc, tr. 1036-1037. 2. Phgm Hoftng Hg (1991), Cdyco Vi^i Nam, Nxb tre. 3. Ashish M., Saket V., Abhinave P. M. (2012), A plant review: Butea monosperma. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 3(1), 700-714. 4. Cherdshewasart W. and Nimsakul N. (2003), Clinical trial of Butea superba, an alternative herbal treatment for erectile dysflmction, Asian Journal ofAndrology, 5,243-246. S.
Indian Institute of Forst Management (2001), Operational strategy for sustainable forestry development with community. 6, Pijush K. D. Amal K. M. (2012), Biodiversity and conservation of dyeyielding plants for justification of its in the local areas of lateritic zone of West Bengal, India, Advances in Bioresearch, 1, 43-53. 7 Sreekumar S., Seeni S., Pushpangadan P.
(1998), Production of 2-hydroxy 4-methoxy benzaldehyde using root cultures of Hemidesmus indicus. Journal of plant Biotechnology, 62 (3), 211-218. 8. Sunatra A. (1931), The remedy paplet of Hwao Khrew tuber of Loung Anusamsutarakromkarphiset, Chiang mat Upatipongsa Press.
Tgp chiDirac li^u, tgp 19, s6 6/2014 (Trang 334 -338)
THANH PHAN HOA HOC CAN CHIET ETHYL ACETAT CAY THOM LOM GAI
Trdn Thanh Hd'' *, Nguyin Vdn Bgi^, Ddi Thf Hdng^, Nguyin Minh Khdi' ' Vien Duac Ueu
^Khoa Hod Hoc- Trudng Dai hoc Khoa hoc Tu nhien- Dgi hoc Quoc gia Hd NQI
•Email: thanhha.tran889(ggmail.com (Nhgn bdi ngay 08 thang 7 ndm 2014)
Tflm tSt
NghiSn ciiu thinh phin hfla hoc cin chiet ethyl acetat ciia ciy thom lom gai Polygonum perfoliatum L. buflc diu di phan l^p dupe b6n h^p chit 3'-0-methyi-3,4-methylenedioxy ellagic acid (I), daucoslerol (2), quercetin (3), quercctin-3-0- glucuronid (4), Ciu triic hoa hgc cua chiing dugrc xic dinh d^ia tren co so cac dCT li^u phfl thyc nghidm MS, NMR vi cic dft lipu phfl NMR di cflng bfl. H(?p chit 1 lin diu tiSn dupe phin lip tir cay niy.
TO" khfla: Thom lom gai. Flavonoid, 3'-0-methyl-3.4-methylenedioxy ellagic acid. Quercetin-3-O-glucuronid.
Summary
Chemical constituents of the ethyl acetat extract of Polygonum peiforlialum L.
Phylochemical study on the ethyl acetate layer of the rhizomes of Polygonum perfoliatum L. resulted in the isolation of four compounds: 3'-0-methyl-3,4-methylenedioxy ellagic acid (1), daucoslerol (2), quercetin (3), and quercetin-3-0- glucuronide (4). Their structures were identified on the basis of physicochemical data and NMR, MS spectral analysis in comparison wilh the published data. The compound (1) was isolated from this plant for the first times.
Keywords: Polygonum perfoliatum L., Flavonoid, 3'-0-metl^l-3.4-methylenedioxy ellagic acid, Quercetin-3-0-glucuromde.
1. Dat van de miln dich vd chdng con triing [3, 5-7,10].
Chi Polygonum hg Rau ram (Polygonaceae) eo Cay thdm 16m gai (Polygonum perfoliatum khoang 300 loai, phan bo chii yeu d phia bdc ban L.) hay con ggi la cay rau ma ngg, phan bo d cdu, ti-ong dd d Viet Nam co khoang 25 loai. Bhutan, Trung Qudc, An Dg, Indonesia, Nhat Nhieu lodi dugc sii dung trong y hgc dan gian vdi Bdn, Han Qudc, Nepal, Philippin, Nga va Vi?t cong dung khang viem, luu thong mau, chiia ly. Nam [1, 2]. Ci nude ta cay mgc d cac tinh miln lgi tilu [I, 2]. Theo cac nghien ciiu da edng bo, mii vd trung du, ddi khi thdy d cae viing ding thdnh phdn hda hgc eua cac loai thugc chi nay bao bdng, nhdt id phia bdc. Thdm ldm gai cd vj chua, gom anthraquinon, coumarin, flavonoid, lignan, tinh mat, cd tac dung thanh nhigt, trir thap, tieu polyphenol, sesquiterpen, triterpen vdi nhieu tac dgc, khoi ngiia; dugc diing trong dan gian trj cam dyng dugc ly dang chii y nhu chong khdi u, chong sot, viem rugt, tieu chay, kilt ly, myn nhgt, Id oxy hda, chdng viem, chdng HIV, chdng suy gidm loet, chdc ddu, rdn cdn, trj khoi u, phti nl, viem
334 Tgp chi Duac lieu, tap 19, so 6/2014
khdp dgng thip va nhiem trung [1, 2]. Thanh phan hda hgc ciia thdm 1dm gai bao gdm cac hgp chat flavonoid (kaempferol, quercetin, rutin, avlcularin, quercetm-3-a-^-D-glucuronic mefliyl, quercetin-3-O-^-D-glucuronid butyl, kaempferol- 3-0-rutinosid), triterpen (acid ursolic, betulin), cac phytosterol, acid beo, acid phenolic (protocatechuic, gallic, ellagic, 3,3-di-O-methyl ellagic, ferulic, vanillic, clorogenic.) [3, 6, 7], Trong so cac hgp chdt dugc phan Igp, nhilu hgp chat cd tac dung sinh hgc dang chii y nhu: Hgp ch^t 8-oxo-pinoresinol cd kha ndng gay dgc doi vdi te bao ung thu bieu md tuyin vii (Bcap-37), ung thu rudt kit (RKO), ung thu bieu md t l bao gan (SMMC-7721), ung thu tuyin tiln liet (PC3) va nhdn te bao bach cau (K562) [10]; quercetin- 3-f?-^-D-glucuronid chirng minh tdc dung chdng Igi virus ciim A [6], la tdc nhdn mdi hd trg cho vi?c dieu tri b?nh Alzheimer [5].
Trong chuong trinh nghien cihi thanh phan hda hgc va sang Igc hogt tinh sinh hgc mgt so lodi cdy thugc chi Polygonum nham gdp phan tgo CO sd khoa hgc cho vi?c su dung eac loai cay nay lam thuoc mgt each hi?u qua va an todn, dap irng nhu cau chiia b?nh cua nhan dan chiing tdi da thu dugc nhung kit qud ban dau trong viec phan lap cdc hgp chat tir phan dogn chiet ethyl acetat ciia than rl cdy thom lom gai.
2. Nguyen lieu v^ phuong phap nghien ciru 2.1. Nguyen li^u
Mdu nghien cuu Id than rl cua cay thom lom gai {Polygonum perfoliatum L.) thu tgi Ba Vi- Hd Ngi vdo thang 07/2013, dugc sdy khd d nhiet dg 50°C roi xay thdnh bgt. Mau do cdc nha khoa hgc Khoa Tai nguyen dugc lidu - Vien Dugc li?u gidm djnh ten khoa hgc. Tieu ban mau so DL- 300713 dugc luu tgi Khoa Hda thuc vdt- Vi?n Dugc li?u.
2.2. Phmmgphdp nghien cuu 2.2.1. Phucmg phap phan Igp cac hgp chdt:
sac ky Idp mdng (TLC): Sdc ki ldp mong dugc tiiyc hi?n tren ban mdng trdng sdn DC- Alufolien 60 ?2Si (Merck 1,05715). Phat hi?n
chat bang den tii ngoai budc sdng 254 nm hogc dimg thuoc thu hien mdu Id dung dich vanilin/
H2SO4 1%, H2SO4 10% phun diu len bdn mdng, sau do say kho roi ha ndng tren bep dien tir d n C C den khi hien. Hodc su dyng thudc thir Id FeCyethanol 5%
Sac ki cgt (CC): Sac ky cgt dugc tien hdnh vdi chat hap phu la silica gel pha thudng (cd hgt 63- 200 fun vd 40-63 fim, Merck).
2.2.2. Phuong phdp xdc dinh cdu tnic hda hoc cac hgp chat:
Diem ndng chay do tren mdy Boetius.
Pho khdi lugng phun mil di?n tii (ESI-MS) dugc ghi tren he thing LC/MSD Trap Agilen Series 1100.
Ph6 cgng hudng tir hat nhan 'H-NMR, '^C- NMR, USQC, HMBC duac ghi ti^n may Bruker Avance-500 MHz, chuan ngi TMS (tetrametyl silan).
2.3. Chiet xudt vaphdn lap
1,5 kg bgt dugc li?u dugc ngam vdi cdn 90% d nhi?t dQ phdng (3 lan, moi Ian 4 ngay). Cdc djch chiet dugc ggp lai va cat loai cdn nude dudi ap suat giam thu dugc can chiet tong (290 g). Cgn chiet nay dugc phdn bd lai lan lugt trong cac dung mdi cd dg phan cue tdng dan: n-hexan, ethyl acetat, «-buthanol; cat loai het dung mdi dudi ap suat giam thu duge cdc phan cdn chiet tuong img
«-hexan (17,0 g), ethyl acetat (100 g), «-buthanol (47,3 g). Cat ki?t dich nude cdn Igi dudi ap suat giam d 80°C cho phan chiet nude (105 g).
Phan tdch can ethyl acetat (70 g) bang sac ky cgt silica gel cd hat 63-200 )im vdi he dung mdi rira gidi gradient ethyl acetat/methanot 100:1, 20:1, 15:1, 10:1, 4:1 va 2:1 (v/v) thu dugc 9 nhdm phan dogn ki hieu tir I- IX.
Phan doan 1 (1,50 g) dugc tinh chl tren silica gel ed hgt 40-63 pm, riia giai vdi he dung mdi dichlomethan/ methanol (20:1) cho cdc chat 1 (8 mg), 2 (20 mg) va 3 (8 mg).
Phan doan 11 (0,52 g) dugc tinh chl tren silica gel cd hgt 40-63 |4m, rira giai vdi h? dung mdi gradient diclomethan/methanol (10:1) cho chat 4 (30 mg).
Tgp chi Duac lieu, tdp 19, sd 6/2014 335
Bing l.D&li|uph3 IH, 13C-NMR cua chflt 1 v i 4 C h i t l
C 1 2 J 3 3a 4 5 6 6a
r
2' 3' 4' 5' 6' 6'a 3'-0CH,
8c"* (ppm) 1127 131,0 138.3 104,3 1500 103,8 116,0 158,3 110,9 141,6 1402 150.0 112,1 116,0 157,6 60,9
" do trong DMSC
•SOOK
5H"(ppm)
6,39 (2H, s)
-
7,535 (IH, s)
- .
7,533 (1H,S)
4,05 (s)
-d.. 'nSMHz.
IHz
Chit 4 C
2 3 4 5 6 7 8 9 10
r
2' 3' 4' 5' 6' 1"
2"
3"
4"
5"
6"
Sc**(ppm) 158,5 135,8 179,4 162,9 99,9 166,1 94,7 159,2 105,7 122,7 116.2 145.9 149.8 118,1 122,8 104,4 75,6 77,6 73,4 78.1 176.3
SB"(ppm)
- - -
6 , 2 1 ( l H , i i , J - 2 , 0 H z ) 6,40(1H, d , / - 2 , 0 H 2 )
-
7,98(lH,(i,.'=2,0Hz)
-
6.88(IH,d,J=8,5Hz) 7,51(lH,d<i,,/-2,5;8,5Hz)
5,33(1H, il,J^7.5Hz) 3,58 (m) 3,53 (m) 3,53 (m) 3,64 (m)
'dotrong CDiOD-d^ '')25MHz.
'500MHz
3. Ket qua va ban luSn
Chat 1: chat ran mdu trang, hi$n mdu tim hong vdi thudc thu vanilin/H^SOj 1%, nhi^t do ndng chdy 296-298''C.
Phd ' H - N M R cua 1 chl xuat hien 2 tin hieu singlet tai SH 7,535 (IH, s, H-5') va 7,533 (IH, s, H-5) dugc xdc dinh la hai proton cua vdng benzene.
Tin hieu singlet tai S^ 6,38 (2H, s, H-3a) cho thay sy cd mat cua 2 proton methyl en edioxy. Ngodi ra con cd nhdm methoxy methyl dugc xac dinh tgi 5 H 4 , 0 5 ( 3 H , S ) .
Phd ''C-NMR va DEPT cua hgp chat 1 cho tiidy CO 15 C trong do 12 tin hieu thugc hai vong benzen dugc xac djnh tai <5c II2,7(C-1), 131,0(C-2), 138.3 (C-3), 150,0 (C-4), 103,8 (C-5), 116,0 (C-6), 110,9 (C-r), 141,6 (C-2'), 1402 (C-3'), 1500 (C-4'), 112,1 (C-5'), 116,0 (C-6'), trong do co 10 carbon bgc 4 va 2 carbon methyl. Ngodi ra, hai carbon este ngi vdng xudt hi?n tgi Sc 157,6 (C-6'a), 158,3 (C- 6a), rat ddc tnmg cho cap este ndi vdng trong cac
ddn xuat cua ellagic acid [11]. Phd C-NMR vd DEPT cdn cho thay su xuat hi?n cua nhdm CH2 lien kit vdi 2 nguyen tu O tgi <5c 104,3 (C-3a) va nhdm methoxy tai ^c 60,9 (s, 3-OCH3).
Dya vao dir Ii?u phd tren ding thdi so sanh nhiet do ndng chay va so sanh vdi cac dii kien phd da dugc cdng bo trudc dd [4, 11], cau true cua chat 1 duge xdc dinh la 3'-0-methyl-3,4- methylenedioxy ellagic acid.
Chat 2; Chat rdn mau trdng, nhiet do ndng chay 283-286''C. Hgp chat ndy cho gid tii Rf = 0,42 (TLC, silica gel, dichlomethan: methanol 9:1, v/v), vd Rf= 0,33 (TLC, RP-18 silica gel, chlorofom;
methanol 1 :10, v/v), vet chat hi?n mau tim v6i H2SO410%, f. Tir nhihig dir ki?n hda ly h«n chung toi gid thiet 2 la mgt steroid co ten la daucosteroL De khang dinh chac chan dieu nay ehung tdi da cho 2 chgy sac ki Idp mdng vdi chat chuan lii daucoslerol thi cho cimg mdt gia trj Rf Tif do chiing toi nit ra kit lugn hgp chat 2 la daucosteroi.
336 Tgp chi Duac lieu, tdp 19, so 6/2014
Chat 3: Tinh the hinh kim mau vang, cd nhi?t dg ndng chay 313-314X. Hgp chat nay cho gia tii Rf = 0,73 (TLC, silica gel, dichlometiian:
aceton 7:3, v/v), vd Rf = 0,30 (TLC, RP-18 silica gel, metfianol - H2O 1:1), v^t chat hi?n mau den vdi thuoc thu FeCls/ethanol 5%. Dua vao cac d|ic trung hda 1^ tren chung tdi dy doan hgp chdt nay la mgt flavon cd ten ggi la quercetin. Dk khdng djnh chdc chan dilu nay chiing toi d3 cho 3 chay sac ki ldp mdng vdi chdt chudn la quercetin thi cho Cling mdt gid tri Rf. Tir dd chung toi nit ra kit lugn hgp chit 3 la quercetin.
Chat 4: tinh the hinh kim mau vang, nhiet dg ndng chay 212-2 H-C.
Tren pho khoi lugng ESI-MS cho pic ion phan td m/z 479 [M+H]^ tuong iing vdi khdi lugng phan tii M = 478 dvC. Phan tich phd ' H - NMR, '^C-NMR vd DEPT cua chdt 4 cho cdc tin hifu d ^ tnmg ciia mdt flavonol glucosid, Dieu nay dugc khang dinh bdi cdt tin hieu proton tren pho H-NMR vdi tin hi?u cua 5 proton trong 2 vdng tham d •5H 7,98 (IH, d, J = 2.0 Hz, H-2'), 7,51 (IH, d d , 7 = 2 . 5 ; 8,5 Hz, H-6'), 6,88 (IH, d.
J= 8,5 Hz, H-SO, SH 6,40 (IH, d, J = 2,0 Hz, H- 8), 6,21 (IH, d, y = 2,0 Hz, H-6). Trdn pho "C- NMR cung cho cac tin hieu 5 nhdm CH nhan thom d <5c 99,9 (C-6), 94,7 (C-8), 116,2 (C-2'), 118,1 {C-5') va 122,8 {C-6'), 4 carbon cd dg chuyin djch dc 166.1, 162.9, 149,8, 145,9 ppm dgc tnmg cho cac carbon C-7, C-5, C-4', C-3' cd lien ket tryc tilp vdi oxy. Cac tin hieu tren cho phep khang dinh phan aglycon trong hgp chat 4 la quercetin [8].
Tin hieu ciia mgt gdc acid glucuronic the hi?n rd d 5n 5,33 (IH, d, J = 7,5 Hz, H-1") vd 3,51- 3,64 (H-2", H-3", H-4", H-5"), <5c 104.4 (C-I"), 78,1 (C-5"), 75.6 (C-2"), 77,6 (C-3"), 73.4 (C- 4"), 176,3 (C-6"). Acid nay dugc khdng djnh tgo lien ket 0-glycosid vcri aglycon quercetin d vi tri C-3 (^c 135.8) thdng qua dii kien pho HMBC.
Tir cdc du' lieu pho tren c6 the du doan hgp chat 4 cd phan aglycon vdi cdu true tuang ty quercetin va phan glycon la gdc acid glucuronic
Tilp tuc so sanh vdi cac dir ki?n phi da dugc cong bo trudc do [8], cau tnic eua chat 4 duge xac dinh laquercetin-3-O-^-D-glucuronid.
Hinh I. Cic hgp chit dugc phan lip tir cin ethyl acetat cay thdm Iflm gai
4. Ket luan than rl eay thom lom gai bao gdm 3'-0-methyl- Bdng cdc phuang phap sde ky kit hgp vdi 3,4-methylenedioxy ellagic acid (I), daucosteroi cdc phuang phdp phd, da phan lap vd nhgn dgng (2), quercetin (3), quercetin glucoronid (4). Hgp ciu tnic cua 4 chat chii yeu bdng cac phuang chat 1 lan dau tien duge thong bdo phan Igp tir phdp pho (MS, NMR) tir c5n ethyl acetat ciia lodi nay.
Tgp chi Duac Hiu, tdp 19, s6'612014 337
Tki lif u tham khao
I. Phsm Hoang Hp (1999), Cdy co Vi^t Nam. Nha xuit ban Tre, Hi Ngi. 2. Dfl Tit Lgi (2006), NhOng cay thudc vd vi thudc Vi?t Nam (tdi bdn Idn thu I), Nhi xuit ban Khoa hpc ky thuat Hi Nfli. 3. Atta-ur-Rahman (Ed.; (2000). Bioactive nahirai products derived from Polygonum species of plants: their structures and mechanisms of action. Studies in Natural Products Chemistry, 22, 607-642. 4. Aua-Ur-Rahman, Ngounou F. N., Iqbal Choudhary M., Shahid M., Talat M., Nur-E-Aiam M., Seema Z., Lontsi D., Ayafor J. F. Sondengam B. L. (2001), New antioxidant and antimicrobial ellagic acid derivatives trom Pteleopsis hylodendron, Planta Medica, 67, 335- 339, 5. Ho L., Ferruzzi M. G., Janle E. M„ Wang J., Gong B., Chen T v., Lobo J , Cooper B., Wu Q. L., Talcott S. T., Pereival S. S„ Simon J. E., Pasinetti G. M. (2013), Identification of brain- targeted bioactive dietary quercetin-3-0-glucuronide as a novel intervention for Alzheimer's disease. The FASEB Journal 27(2), 769-81 6. Narasimhulu, G., Rao P.V. and Reddy K.S. (2014), The genus Po/ygoram (Polygonaceae): An ethnopharmacological and phylochemical perspectives-review, International Journal of Pharmacy and Pharmaceutical Sciences , 6(2), 21-45, 7. Wang K.-J., Zang Y.-J., Yang C.-R,R. (2006), Recent advance on the chemistry and bioactivity of gtvwis Polygonum, Natural Product Research and Development, (18), 151- 164. 8. Agrawal P. K., (1989), Carbon-13 NMR of flavonoids, Elsevier Science Publishers B. V., 152-153. 9. Elisabetta C. Pier L, C , Guido F., Barbara R, Roberto C.
(2003), Flavonoids analysis of four Vicia species of Narbonensis complex in two different vegetative phases, Caryologia, 56 (3), 365-371. 10. Wang K. W„ Zhu J, R., and Shen L. Q(2013), A new lignan with anti-tumour activity from Polygonum perfoliatum L., Natural Products Research, 27(6), 568-73. 11. Li X -C„ Hala N. E., Charles D, H., Alice M. C. (1999), NMR assignments of ellagic acid derivatives, Magnetic Resonance in Chemistry, 37(11), 856 - 859.
Tgp chl Duac Heu, tap 19, sd 6/2014 (Trang 338 - 342)
CAC HOfP CHAT DI VONG TlT CAY LAC TIEN
Nguyin Tan Phdt'*, Phiing Van Trung', Biii Trpng Dat", Phan Nhgt Minh', Nguyin Minh Scm\
Le Thi Viet Hod', Nguyin Nggc Hgnh'
' Vi^n Cdng ngh§ Hod hQC - Vi?n Hdn ldm Khoa hoc & Cdng ngh? Vi^t Nam
^Truong Dgi hoc Su Pham TP. HCM
*E-mail: [email protected] hogc ntphat(gict.vast.vn (Nhan bai ngay 01 thdng 8 ndm 2014)
Tflm tit
Tir djch chiet methanol ciia phan tren mil dit ciy lac tien (Passiflora foetida L.) dugc thu hai a tinh An Giang, 3 hgp chil dj vflng acid indole-3-acetic (I). acid pyroglutamic (2), adenosin (3) va 1 polyol myo-inositol (4) d i dugc phan lip. Cau Iriic ciia cic hgp chit dugc xac djnh boi pho cgng hucmg tir hat nhan (ID va2D-NMR) v i s o sinh v6i dii li?u phfl dugc c6ng bfl trong tii li$u tham khio.
Tir khfla: Cdy Igc tien. Passijlora foetida. Acid indole-3-acetic. Acid pyroglutamic. Adenosine. Myo-inositoL Summary
Heterocyclic Compounds from Passijlora foetida L.
Three heterocyclic compounds including indole-3-acetic acid (I), pyroglutamic acid (2). adenosine (3), and one polyol m^'o-inositol (4) were isolated from the methanol extract of the aerial part of Passijlora foetida L,. Their structures were elucidated by (ID and 2D-NMR) and comparison with the published data.
Keywords: Passijlora foetida, lndole-3-acetic acid. Pyroglutamic acid. Adenosine. Myo-inositol.
1. Dat van de triing, khdng khudn, khang ndm, giam dau [7], Theo y hgc cd truyen, cay Igc tien cd vj nggt, [8], [9], [10]... Cac tac gia Huynh Ldi, Tran hoi dang, tinh mat, cd tdc dung thanh nhiet, giai Hiing da phan lap dugc 2 hgp chat: vitexin, 2"- dgc, Igi thiiy, diing lam thudc an than, ehiia mat xylosylvitexin tu than Id lge tien; da xay dyng ngu, suy nhugc than kinh, mat gan, tim hoi hop, phuang phap djnh lugng vitexin trong cay Igc mat ngu [1], [2]. Ngoai ra, lac tien co tae dung tien bdng HPLC [3], [4]. Trong cdc cdng bo chong tang sinh te bdo ung thu, diet ky sinh trudc, chiing tdi da phan lap dugc 9 hgp chat tir
338 Tgp chi Duac lieu, tap 19, so 6/2014
