# Nghien CIPU - Ky thuat
4 Franz von Bruchhausen, Hermann Hager [2010}, Hagers Handbuch der pharmazeutischen Praxis. Springer-verlag, pp. 72.
5. U L, Kadota Y. (2001), Aconitum L., In: Flora of C/ima, Beijing Science Press, St. Louis Misoun Botanical Garden Press, 6, pp. 149-224.
6. Sang Hee Shim, Ju Sun Kim, Sam Sik Kang, Kun Ho Son, KiHwan Bae (2003), /Mkaloidal constituents from Aconitum jaluens, Archn/es of Pharmacal Research. 26, pp. 709-715.
7. Joannes Barnes and Linda A. Anderson (2008), Herbal medicines. Pharmaceutical Press.
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Hop chat polyacetylen va polyphenol phan lap tir re cay den kim {Bidens pilosa L.)
Pham Thanh Ky^, Pham Van Vir^mg'*, Nguyin Thi Bich Thu^ Phan Van Kiem^
^ Truang Dgi hoc Duac Ha Noi, ' Benh Viin Qudn Y17- Qudn Khu 5,
^Vien Duac iieu ^ Vien Hoa sink Bien - Vien Khoa hoc va Cong nghe Vie! Nam 'E-mail: [email protected] Summary
From the roots of Bidens pilosa L. (Asteraceae), two compounds. 7-phenyl-2-heptene-4.6-diyn-1-ol (BP11) and 4-allyl-2,6-dimethoxyphenol-1-0-^-D-glucopyranoside (BP13), were isolated by combination of various chromatography methods Their structures were identified by NMR including ID- and 2D-NMR... The compound BP13 were isolated from the genus Bidens for the first time.
Keywords: Bidens pilosa L , roots, two compounds.
D|t van de
Cdy don kim co ten khoa hpc Bidens pilosa L, thupc hg Cue- Asteraceae, la cay mgc hoang 6 nhieu noi t u mien nui, trung du, dong bdng va nhieu nu'dc tren the gidi nhu: Trung Quoc, Han Quoc, Nhat Bdn, Brazil....vv. Day la loai cdy mi?c quanh ndm, sdc sinh san nhanh va d u g c coi nhu' loai "co dai".
Nghien cdu vh thdnh phan hoa hpc cho t h i y cSy don kim co polyacetylen, polyphenol...
NhOng nhom hop chat nay cua cay dan kim fluvc Chung minh eo tac dung bao ve gan, ha glucose mau vd uc ehe t i bao ung t h u '^'^.
Bk gop phan nghien c d u ve thanh phan hoa hpc r l cdy don kim, bdi bdo nay trinh bay k i t ctuS phan Igp va xac djnh c i u true Id 7-phenyi-2- i»pten-4,6-diyn-1-ol, mpt hgp c h i t thuge nhom polyacetylen vd 4-allyl-2,6-dimethoxyphenol-1- 0"^-D-glucopyranosid, mOt hgp c h i t thuge
^ 6 m polyphenol.
Nguyen lieu nghien CLPU
va phu'O'ng phap
Nguyen lieu
Rd cay don kim dugc thu hai tai Ydn Lac - tinh Vinh Phuc vao thdng 4-5 va thang 9 - 1 1 nam 2010. Mdu nghien cuu da du'gc PGS.TS Vu Xuan Phucng vd ThS. Do The Cudng (Phong Thyc vat - Vipn Sinh thdi va Tai nguyen sinh vdt - Vien Khoa hgc vd Cong nghe Viet Nam) xac dinh ten khoa hpc la Bidens pilosa L., thuoc hg Cue - Asteraceae. M i u tieu ban dugc luu tai Vien Sinh thai vd Tdi nguyen sinh vat.
PhLFong phap nghien c u u
- C h i i t xuat cdc c h i t trong d u g c lieu bdng phucng phap ngam vdi methanol d nhiet dp phong.
- Phan lap cdc chat bang sac ky cpt dung chat h i p phu la silicagel pha thudng c d hgt Id 0,04- 0,063 mm (240 - 430 mesh, Merck) vd pha dao ODS hode YMC (30-50 pm, Fujisilisa Chemical Ltd).
- Sdc ky Idp mong d u g c thye hi^n trdn ban mong trang san silica gel DC-A!ufolien 60 F254 (Merck 1.05715), RPis F254S (Merck). Phat hipn chat bang den t d ngoai d hai budc song 254 vd
TiPCHi DU<?C HQC - 05/21(13 (SO 445 N.\\I 53) 55
# Nghien CLFU - Ky thuat
365 nm, hoac dung thuoc t h u id dung dich H2SO4 10% d u g c phun deu len ban mong, say kho rfii hff ndng tren b i p dien t u den khi hien mau.
- Pho cgng hudng t u nhan (NMR): ^H-NMR (500 MHz) va "C-NMR (125 MHz) do tren may Bruker AM500 FT - NMR Spectrometer, d Vien Hod hgc, Vien Khoa hgc va Cong nghe Viet Nam.
- C h i i t xuat vd phan lap cac hgp c h i t Re cay don kim sau thu hai d u g c rua sach, thai nho, phoi khfi va s i y d 60°C sau do nghien nho thu d u g c 3,0 kg bpt tho. Bgt nay dug'c ngam c h i i t 3 lan, t i n ddu 48 g i d , 2 ldn sau 24 gid, moi ldn dung 10 lit methanol. Gpp cae djch c h i i t , c i t thu hfii dung mfii d u d i ap s u i t giam thu dug'c 158,0 gam dich co dam dac. Djch nay d u g c phan tan vdo 2 lit n u d c c i t , t i i n hanh chiet phdn bfi ldn l u g t vdi clorofomi (3 x 1,5 lit), ethylacetat (5 x 1,5 lit), n-butanol (5 x 1,5 lit).
Cac dich chiet clorofomi, ethylacetat, n-butanol d u a c c i t thu hoi dung mfii d u d i dp s u i t giam ddn khi kiet dung mfii thu d u a c cac can theo t h u t y : c i n Pi (55,0 g), can P2 (25,0 g), cdn Pa (28,0 g) va Idp n u d c con lai.
Ldp n u d c du'gc phan lap tren sac ky egt trao doi ion (Dianion HP-20), logi bo d u d n g vd muoi Bang 1: D u lieu phd NMR cua hap chat BP11
vo ea bdng n u d c c i t sau do rua giai vdi he dung mfii M e O H / H 2 0 (50/50, 100/0). Sau khi eat thu hoi dung moi d u d i ap s u i t giam thu d u a c 2 phan doan t u a n g ung la PSA (15,8 g) vd P3B (10,5 g). Can P3B (10,5g) tiep tuc duge phan tdch tren egt silicagel pha t h u d n g vdi he dung mfii rua giai eloroform/methanol co do phdn c y c tang dan (20 : 1 , 10 : 1 , 5 :1) sau do rua giai l i n l u g l bdng acid formic va methanol dup'c 5 phdn dogn t u a n g u n g . t u P13A d i n P13E. Hgp e h i t BP13 (14 mg) dug'c phan lap t d phan dogn P13B s u dung sdc ky cpt silicagel pha t h u d n g va cot silicagel pha dao vdi he dung moi m-a giai thich hgp.
Mat khac, tien hdnh phan tach can P I (55 g) tren egt silicagel pha t h u d n g vdi he dung moi co dg phdn c y c tang dan hexan/aceton (50 : 1 , 25 : 1 , 10 : 1 , 1 .1) d u g c cac phan doan t u a n g ung. T u can cua phan dogn (10:1) tiep tuc phan lap tren cpt silicagel pha dao, n>a giai vdi hd dung moi thich hgp d u a c hap c h i t BP11 (30 mg).
K e t q u a v a t h a o l u a n
Hgp c h i t BP11 thu dugc la chat ran khfing mau.
Ket qua do pho NMR dugc trinh bdy d bang 1:
Vl tri C 1 2 3 4 5 6 7 8 9 10 11 12 13
5 c (ppm) 56,3 145,5 110,0 80,0 77,4 74,3 77,3 121.2 132,3 128,3 128,4 128,3 132,3
dc ^''(F>pm) CDCIs 62,6 145,4 108,8 81,28 79,79 73,8 74,5 121,7 132,4 128,4 129,1 128,4 132,4
Su"(ppm,J,Hz) 4,25 dd (5,0,2.0) 6,4511(16,0; 5,0) 5,9011(16,0:2,0)
7.49 dd (8,0; 2,0) 7,30 ddd (8,0; 8,0; 2,0) 7,33 ddd (8,0; 8,0;2,0) 7,30 ddd (8,0; 8,0; 2.0)
7.49 dd (8,0; 2.0) 5*c 7-phenyl-2-heptene-4,6-diyn-1-ol trong chemdraw,
Pho ^H-NMR x u i t hien cdc tin hieu trong vLing 5 7,30^-7,49 (5H), trong do eo 2 tin hieu co c u d n g do manh t u a n g dng vdi 2 cap proton doi xdng, dieu nay khang djnh s y co mat cua 1 vong benzen the mono. Ngoai ra con co s y
V o trong CDCh, 125 MHz. ^500 MHz.
cgng h u d n g cua cdc proton cua mgt n i l doi trans tgi 6 6,45 ( I H , tt, J = 16,0, 5,0 Hz, H-2), 5 , 9 0 ( 1 H . tt, J = 16,0, 2,0 Hz, H-3) vd hai proton con Igi vdi 5 4,25 (2H, dd, J ~ 5,0, 2,0 Hz) tuang dng vdi mgt nh6m oximethilen.
56 TAP CHi DUgfC HQC - 05/2013 (SO 445 NAM 53)
» Nghien CLPU - Ky thuat
Phdn tieh p h i "C-NMR va DEPT eho t h i y co 13 tin hieu cgng hudng eua carix)n, trong do co 2 tin hieu eua CH tgi 6 145,4 vd 108,8 dac Inj-ng eho n6i doi frans ngodi vong. Hai tin hieu cua CH vdi cu&ng do manh tai S 132,4 vd 128,4 xdc nhan cho 2 cap carbon doi xung cua vong benzen tgi C-9, C-13 va C-10, C-12. Mgt tin hieu cgng hudng cua CH tai 5129,1 tuang ung vdi C-11, tin hieu con Igi cGa 1 cartx)n bac bon 5 121,7 Id eua C-8 eua vong benzen co g i n nhom the. S y co mat cua 4 carbon bgc 4 vdi do djch chuyen hoa hoc 5 73,8 -^81,28 3gc trung cho 2 lien k i t ba trong phdn t u BP11 vd 1 nhdm -CH2- tgi 5 62,6 va ket hap vdi cac gid trj proton dgc tacng cho viing tham noi tren cho phep l(hang djnh trong phan t d cua BP11 co mpt vong Bang 2: D u liiu pho NMR cua hgp chit BP13
thom lien k i t vdi 1 day carbon co 2 lien ket 3, mpt lien ket doi dgng trans va mgt nhdm oximethilen d ddu mgch. T u cdc d u kien pho neu tren, hgp chat BP11 dugc nhdn dang la 7-phenyl-2-hepten-4,6- diyn-1-ol vdi cfing thuc phan t u la C13H10O.
Hinh 1: Cau true phin twcua hgp chat BP11 Hgp c h i t BP13 d u a c t i i n hanh do pho NMR de phan tich xac dinh c i u true. K i t qua do p h i NMR d u g c trinh b a y d bang 2:
WtriC 1 2,6 3,5 4 a P r V 2' 3' 4' 5' 6 2.6-OCH3
•&
136.5 153,8 107,5 134,9 40,4 137,8 115.9 105,3 76,0 78,3 71.7 78,2 62,8 56.7
**'
138.39 154,17 107,53 134,71 41.34 138.65 116.16 105,58 75,74 77,83 71,35 78,32 62.61 56,99
SH"' "^ mutt. (J in Hz)
6.54 (2H, s)
3.36(2H, d, 6,5) 5,98 (IH.m) 5.10(1H, dd, 12,0, 1,5) 5.08(1H. dd, 10.0; 1,5)
4,e4(1H. d. 8,0) 3,48 (1H,m) 3,43(1H, d, 2,5) 3,42(1H, d, 2,5) 3,22 (IH.m) 3.80(1H, dd, 12,0,2,0) 3,68(1H, dd, 12,0; 5,0)
3.84 (6H, s)
HMBC (H -f C)
1.2,4.5
3, 5, p, Y a
a, 3 1 r, 3'
4 2.6 'Scdotrong pyridin-d5 t^i 88,55 MHz, 22,5 MHz
a trong CD30D, b 125 MHz, c 500 MHz Tren pho proton cua hgp chat BP13 t h i y tuat hien tin hieu cua 1 vong tham the 4 vi tri mpt each doi xdng tgi 5H 6,54 (2H, s, H-3; H-5);
<^c tin hipu proton dac trung cho 1 phdn t u flirt'ng glucose trong khoang 5H 3,22 - 3,84 (6H;
H-2'; H-3'; H ^ ' ; H-5' vd 2 x H-6') va tin hipu 1 praton anome tgi 5H 4,84 ( I H , d, J = 8,0 Hz) ; mOt n i i dfii ngodi mgch the mpt vi tri trong
T*PCHi DlTQC HQC - 05/2013 (SO 445 NAM 53)
khoang 5H 5,08 - 5,98 (3H, H-p; 2xH-y) vd mpt nhom a-methilen so vdi noi doi tgi vj tri cgng hu'dng 6H 3,36 (2H, d, J = 6,5 Hz). D i i u nay hodn toan phii hgp vdi cac k i t qua phdn tieh pho carbon va DEPT vdi 2 nhom methoxy tgi vi tri cgng hudng 6c 56.99 (2-OCH3; 6-OCH3); cdc tin hieu oximethin va oximethilen dgc trung cho phan t d dudng glucose d u g c xac djnh bdi cdc
57
• Nghien CLFU - Ky thuat
tin hidu carbon cfing h u d n g tgi Sc 105,58 (C-1'), 78,32 (C-5'), 77,83 (C-3'), 75,74 (C'-2) vd 62,61 (C-6'); cac tin hidu cCia vong tham t h i 4 lan v d i 4 tin hieu carbon bde 4 Sc 154,17 (C-2; C-6), 138,39 (C-1), 134,71 (C-4) vd 2 nhom methin tgi Sc 107,53 (C-3; C-5); 1 noi dfii ngodi mach the 1 l i n vdi 1 nhom metiien, 1 nhom methin d u g c xac djnh lan l u g t bdi cac tin hidu carbon cgng h u d n g tgi Sc 116,16 (C-y), 138,65 (C-p) va mgt nhom methylen canh nSi dfii xdc dinh bdi tin hidu carbon cgng h u d n g tai vi tri Sc 41,34 (C-a).
O H
Hinh 2: Cau true phan twcua hgp chit BP13
HO HO
Hinh 3: Cac twang tie HMBC chinh eua hgp chat BP13
Cdc tin hieu proton d u g e gan vdi cac tin hipu carbon t u a n g dng d y a tren viec phdn tfch pho HSQC (bang 2). B e xac djnh vj trf cac lien k i t , tien hanh do p h i HMBC. Ket qua phdn tich pho HMBC cho t h i y proton cOa carbon C-a (5c 41,34/ 8H 3,36 (2H, d, J = 6,5 Hz)) c6 t u a n g tac
vdi cdc tin hipu cart)on tai Sc 138,65(C-p), 116,16 (C- y) vd 107,53 (C-3, C-5); proton tgi vf trf carbon s6 1 ' cua phan t u d u d n g (105,58 (C- 1')/ 5H 4,84 ( I H , d, J = 8.0 Hz) t u a n g tdc vdi C- 1 tgi vj tri cgng h u d n g 138,39 (hinh 3). Ket hgp so sanh v d i s6 lieu carbon eua hgp chat 4-allyl~
2,6-dimethoxyphenol-glucoside^^^ cho phep chung ta xdc djnh d u g c c i u true phan t u cOa BP13 Id 4-allyl-2,6-dimethoxypltenol-1-0-^-0- glucopyranosld v d i cong t h u c phan t d Ci7H240e t u a n g d u a n g vdi so khoi 356.
K i t luan
T u dich chiet methanol re cdy dan kim da phdn lap d u g c 1 hgp chat thuge nhom polyacetylen la 7-phenyl-2-hepten-4,6-diyn-1-ol (BP11) v d 1 hgp c h i t polyphenol Id 4-allyl-2,6-dimethoxyphenol-1- 0-j3-D-giucopyranosld (BP13) Cau tnJc hoa hgc cua eac hgp chat nay d u g c xac dinh bang phuang phap pho cgng hudng t d hat nhan (NMR) vd doi chieu vdi tai lipu tham khao. Trong do, hgp c h i t BP11 da d u g c phdn l^p t u phan tren mgt dat cay dan kim ^^, hgp chat BP13 lan dau tien dugc phan lap t d chl Bidens.
Tai lieu tham khao 1. Miyase T., Kuroyanagi M., Noro T , Ueno A., Fukushima S., (1985), "Studies on Sesquiterpenes fn^m Macroclinidium tntobum Makino 11", Chem. Phan Bull.. Vol 33(10), pp: 4445-4450.
2. Ubillas R. P., Mendez 0. D., Jolad S. D., Luo J., King S. R., Carlson T. J., Fort D. M. (2000),
"Antihyperglycemic acetylenic glucosides from Bidens pilosa", Planta Med- 66: 82-83
3. Wang R, Wu QX, Shi YP (2010), "Pdyacetylens and flavonoids from the Aerial Parts of Bidens pitosa"
Planta Med. 2; 76: 893-896.
4.Wu W. L , Chiang Y. M. et al (2004), "Polyacetyienes Function as Anti-angiogenic Agents". Pharmaceutical Research. Vol. 21, No. 11; p 2112-2119.
5. Yuan L. P., Chen F. H., Ling L , Dou P F., Bo H., Zhong M. M., Xia L J., (2008), "Protective effects of total flavonoids of Bidens pitosa L. (TFB) on animal liver injury and liver fibrosis", Joumal of Ethnopharmacology 116: p 539-546
