NGHIEN CLTU THANH PHAN HOA HQC PHAN DOAN W-HEXANE CUA CAY AN B\tU NON HEDYOTIS PiluUFERA (PIT.) T.N.NINH - RUBIACEAE
Li Thi Thanh Phuang', Nguyin Khdnh Thdy Linh^
(1) Lap Dugc 5. Trudng Dgi hgc YDugc Hui (2) Khoa Dugc, Tne&ng Dgi hgc YDugc Hui
TomtAt
Dat van dl: Loli An dien ndn dupe ngudi din viing bao ton Khe Trin dung de chDa cdc bpnh vl dudng tieu hda va dau nhirc xuang khdp. Tuy nhien, cho dSn nay chua thay cd cdng bo khoa hpc nio vl thinh phan hda hpc hay tic dyng sinh hpc cua loli nly. Do do, vipc nghiSn ciiu thinh phin hda bpc vl hogt tinh sinh hpc ciia loli nly 11 can thiet nhim cung cap ca sd khoa hgc, djnh hudng vipc bio t^n vl phit triln cly thuoc phyc vy cdng tic chira bpnh. Muc tiSu: Chiet tach phan Igp mdt so hpp chit tinh khiet tir phin doan n-hexane. Xac djnh cau tnic hda hpc cic hpp chit phan Igp dupe. D6i tiring nghiSn ciftii: Phan tren mgt dat ciia cay An dien ndn dugc thu tgt huypn VTnh Linh, tinh QuIng Trj. Phirang phap nghien cu-u: Chilt xuit bing phuangphap chilt ldng - long, rin - ldng. Phan l|p chit tinh khiet bang cac phuong phap sac ky. Xic djnh cau triic bang phuang phap ph^. Kit qua: Phan Igp dupe 02 hgp chat rotungenic acid (1) v l a-tocopherolquinone (2). Hai hgp chit nly lan diu tiSn dupe phin lap tu cay An dien ndn Hedyotis pilulifera (Pit.) T.N.Ninh cung nhu chi Hedyotis L. Hgp chat 1 cd hogt tinh khang khuan vl iirc che protein tyrosine phosphatase IB trong dilu trj bpnh dai thao dudng. Hgp chat 2 the hipn hogt tinh chong ung thu tren nhiSu ddng tl bao. Ket lu^n: Tir cao chiet phan dogn /i-hexane ciia cly An dien ndn (Hedyotis pilulifera (Pit.) T.N.Ninh, hg Rubiaceae) da phin lip dugc 02 hgp chit bao g6m rotungenic acid (1), a-tocopherolquinone (2) cd gll trj hoat tinh sinh hgc cao.
Tie khod: Hedyotis pilulifera, rotungenic acid, a-tocopherolquinone.
Abstract
STUDY ON CHEMICAL CONSTITUENTS FROM /V-HEXANE FRACTION OB HEDYOTIS PILULIFERA (PIT.) T.NJSINH- RUBIACEAE
Le Ihi Thanh Phuong', Nguyen Khanh Thuy Linf^
(1) Student of Faculty of Pharmacy. Hue University of Medicine and Pharmacy (2) Faculty of Pharmacy. Hue University of Medicine and Pharmacy Background: Hedyotis pilulifera is used for treating rheumatism, bowel complaints by the locals in Khe Tran, Quang Tri province. Therefore, the study of the chemical comppsition and biological activity of this species is necessary to provide a scientific basis, the conservation- oriented development and service of medicinal plants healing. Purposes: Extraction and isolation of pure compounds from n-hexane fraction. Determination of chemical structures of compounds isolated. Materials: The aerial parts of Hedyotis pilulifera (Pit.) T.N.Ninh were collected in Vinh Linh, Quang Tri province. Method: Extraction methods: liquid-liquid extraction, solid-liquid extraction. Isolated by column chromatography and thin-layer chromatography. Determining structure by spectroscopic methods. Results: Two compounds including: rotungenic acid (1) vl a-tocopherolquinone (2). The two compounds were first isolated from Hedyotis L. as well as
- Dfa cht liin h^: Nguyin Khdnh Thiiy Linh. emad [email protected] i' •..
- Ngdy nhgn bdi: 25/3/2016 * Ngdy dongy ddng 15/4/2016* Ngdy xudt ban: 29J4/2016 14 Tap chf Y Duvc hpc - TViriyng Dgi hpc Y Dirffc Hui - Si D|c bi^t
Hedyotis pilulifera (Pit.) T.N.Ninh. Compound 1 has and inhibitory activity of protein tyrosine phosphatase 1B in the treatment of diabetes. Compound 2 shows anticancer activity across multiple cell lines. Conclusion: From the extract of n-hexane fraction has isolated 2 compounds including : rotungenic acid (1) va a-tocopherolquinone (2), have the value of high biological activity.
Key words: Hedyotis pilulifera, rotungenic acid, a-tocopherolquinone.
L D ^ T V A N D t
Nhieu nghien cihi tren thi gidi dl cho thiy djch chiet cic phan dogn v l cac hgp chit phin l§p tir cic loai thudc chi Hedyotis L. cd nhieu hogt tinh sinh hpc tot nhu hogt tinh khing khuin, chdng oxy hda, gly dgc te bio, bio vS than kinh... [8]. Cly An diln ndn - Hedyotis pilulifera (Pit.) T.N.Ninh II loli thu0c chi Hedyotis L., theo dilu tra vl da dgng sinh hgc va kinh nghipm lim thuoc cua ngudi dan d viing Bio ton Khe Trin thi day la loli dugc lipu dupe sir dyng lam thuoc [I]. Cho den nay, theo tong quan tli lipu cho thiy chua cd nghiSn ciru nao ve thinh phan hda hpc cua loli nly d ViSt Nam. Vi the, chiing tdi lya chpn cly An dien ndn Um doi tugng nghien c^u vdi mong mu6n cd nhihig bilu biet ve mgt hda hpc ciia cay Hedyotis pilulifera (Pit.) T.N.Ninh, lim ting gii trj irng dyng ciia cly vio thyc te cupc song cung nhu gdp phan lam phong phii han nhihig hiiu biit vl cic cly thudc chi Hedyotis L.
D I tli "Nghiin citu thdnh phan hda hgc phdn dogn n-hexane cua cdy An diin non Hedyotis pilulifera (Pit.) T.N.Ninh - Rubiaceae" vdi hai
myc tieu sau:
1. Chiit tdch phdn ldp mgi s6 hgp chai linh khiet tuphdn dogn n-hexane.
2. Xdc dfnh cdu trdc hda hgc cdc hgp chdl phdn Igp dugc
2. DOI TUQfNG VA P H U O N G PHAP NGHIEN c t r u
2.1, B^i tupng nghiSn ciru
Phin trSn m|t dit ciia cly An diln ndn, mlu dugc thu t^i huySn VTnh Linh, tinh QuIng Trj vao thang 8 nlm 2014. Ten khoa hpc dugc xac djnh la Hedyotis pilulifera (Pit.) T.N.Ninh - Hp Cl phe (Rubiaceae) bdi TS. Nguyin ThI Cudng - Vipn Sinh thii vl Tli nguyen sinh vgt - ViSn Hin lam Khoa hpc vl Cdng nghp Vipt Nam.
2.2. Phinrag phap nghien ciru
2.2.1. XA ly mau, chiit xu6t cao todn phan vd cao cdc phdn dogn.
Dugc li^u dugc phai say khd, xay nhd thinh bpt mjn. Bgt dupe liSu dugc ngam chiet 3 lan d nhiSt dd phdng, mdi lan 7 ngay vdi 10 lit methanol/lin, ggp cac djch chiet, cat thu hoi dung mdi dudi ap suat giim cho den can thu dugc
• cao toln phin.
Liy cao cin toan phin phan tin vao nude roi chiet xuat phan dogn ttlng cic phuang phap chiet long-ldng vdi cic dung mdi n-hexane, chloroform, ethyl acetate, thu dupe djch chiet d c phan doan va phan nude cdn Igi.
2.2.2. Phiegngphdp nghien dku vi hod hgc Phan Igp cic chat tinh khilt bing sic ky cpt silica gel pha tiiudng (Silica gel 60 0,040-0,063mm (230- 400 mesh ASTM), Merck); silica gel pha dao YMC (30-50 (im, Fuji Silysia Chemical Ltd.); Sephadex LH-20. Tbeo ddi cic phin dogn bing sic 1^ ldp mdng pha thudng, pha ^ o (TLC-Silicagel 60 F254 Merck). Phat hipn vet chat bing d^n tli ngogi d hai budc sdng 254 nm va 366 nm ho$c dimg thuoc thti 11 dung djch H^SO^ 10% phun diu ISn bin mdng, say khd roi ha ndng tir tir din khi hipn mlu.
Xic djnh cau tnic cac hgp chat phan lap dupe dya tren cic phuang phip pho bao gom: phd cgng hudng t i hgt nhin mdt chieu ('H-NMR, '^C-NMR) va hai chieu (HMBC, HSQC) do tien may Bruker Avance AM500 FT-NMR tgi Vipn Hoi hgc, Vipn Han lam Khoa hpc v l Cdng nghp ViSt Nam.
S.KtTQUA
3.1. K4t qua nghien cuu ve hda hpc 2,4 kg dugc lipu da xay thinh bdt dupe ngam chiet trong methanol (10 lit x 3), djch chilt sau dd dugc cd khd d ap suat thap, thu dugc cao cin.
cin methanol dugc phan bo vio nude roi lin lugt chilt vdi n-hexane, chloroform, ethyl acetate, cho cic phan dogn chilt n-hexane (H), chloroform (C), ethyl acetate (E) vl djch nude cdn Igi. Djch nude dugc tich phan dogn bing cgt DianJon HP-20 (W) (Hinh 3.1).
Tgp chf Y Dirprc hpc - Trirfrng Dai hpc Y Duprc Hue - Sh D?c bipt 15
Nhi#t dO
1 n-hexane j - HPH 1 1 (40,6 B) 1
1 Ch! j l p h s T ^
c
ftps
B $ t dugrc l i f u ( 2 , 4 k g ) j
1 N e S m i) nhS^t d& p h i n g / m e t h a n o l j Djch c h i ^ 1
kt gidm 1 c a q u a y 1 can (100.5 8> 1 1 Phan tAn 1 Djch nude 1
1 l.ac phOn dotin 1 Chloroform HPC 1
< 1 9 , 2 B ) 1
n-hexane: acetone
1 100:I 1 1 Hi 1 1 H2
30-I
1 Ethyl acetate 1 HPE 1 1 (lO.Sg) 1
i ..
1 H3 1 1 H4 1 1 <**e) 1 1 (20E> 1
1 Nude HPW 1
<20,Sg) 1
H 5
Hinh 3.1. Sa do chilt xuit cic phin dogn tii cay An dien ndn Phin dogn n-hexane dugc tich tren cdt silicagel,
nia giii bing n-hexane/acetone lan lupt vdi ty Ip 100:1, 50:1, 5:1, 1:1, 0:1 tiiu dupe 5 phan dogn (Hl-5). cin ciia hai phin doan H3 vl H4 dugc ggp chung (Ky hipu H34). Phan dogn H34 dugc hda tan holn toln vio lupng n-hexane t6i thiSu vl triln khai tren cpt silicagel pha thudng nia giai bing hdn hpp n-hexane - acetone 3:1 thu dugc 2 phan dogn H34A-B. Phan dogn H34A dugc tach trdn cdt silicagel pha thudng, nia giii bing h$
dung mdi chloroform - methanol 50:1 thu dugc 3 phin doan H34A1-3. Phin dogn H34A2 tiep tyc tien hanh sic k}^ trgn cdt Sephadex LH-20 vdi hp dung mdi methanol - nude 1:1 thu dugc 2 phin doan H34A2A-B. H34A2A dugc phan Igp bing
cpt silicagel pha dao vdi hp methanol - nude 15:1 thu dugc 2 phan dogn H34A2A1-2. H34A2A2 du^c tinh chi bing SKC pha thudng nia giii hlng n-hexane - acetone vdi tJ Ip ban dau 10:1 sau dd doi thanh ^ Ip 6:1 cho din khi qui trinh nia gill kit tiiiic. Thu dugc chit sgch so 1 la H34A1A2I.
Phan dogn H34A1 tiep tyc dugc phan Igp tr6n cpt silicagel pha thudng vdi hp dung mdi n-hexane - chloroform - ethylacetate vdi ty Ip ban dau 5:1:0,1 sau dd doi thanh ty Ip 3:1:0,1 cho den khi qua trinh riia giii kit thuc, thu dugc 2 phin dogn H34A1A-B. Tilp tyc tinh che H34A1A bing cdt silicagel pha dao, nia giai bing methanol - nude 1:1 thu dupe chat sgch so 2 11 H34A1A1I (Hinh 3.2).
I H 3 4 < 3 6 g > I
SKC pha Ihudns [ n-hexane — acetone (3:1)
c
^ <9e) ID
J
SKC pha thirfmgI H 3 4 A I < 6 0 0 m e ) ~ | j H 3 4 A 2 (Sg) j
methanol (SO:l)
— : — I
. ^ ^
I H 3 4 A 1 A ( 5 2 m g ) j SKC pha di I H 3 4 A 1 A 1 I ( S m g ) I
SKC pha thujms chlorofbrm — ethyl acetate ( S : l : 0 , l ^ - 3 ! l ; 0 , I )
SKC Sephadex LH-ZO methanol S O ^
H 3 4 A 2 A ( 2 . 3 8 ) | j H 3 4 A 2 B | I H 3 4 A 1 B I S K C p h a d A o methanol — nu*o (15:1)
-^ i _ ' = i
H 3 4 A 2 A 1 ( 1 , 9 s ) H 3 4 A 2 A 2 I SKC pha thu£msI H : 3 4 A 2 A 1 I (POOmg) I
EQnh 3.2. Sa do phin lap hgp chit H34A2A1I va H34A1 AH tif phin dogn H34.
Tgp chi Y Dir^rc hpc - Tririmg Dgi hpc Y Dirgrc Hui - S6 D|c bi|t
3.2. XAe djnh cau true cdc chit dS phSn l^p durgv.
Thong qua vlpe phan tIch dO' kipn ph6 ID, 2D - NMR, elc chit s^ch 1-2 dugc xic djnh 11 rotungenic acid (1), a-tocopherol (2).
H ^ chdt 1: Bgt mlu b ^ g . PhS 'H-NMR chi ra cae tin hipu d$c trung ciia 1 proton olefin tgi 5^^
5,62; 2 proton ciia nhdm oxymethylene tgi S^3,68 (d, / = 10,5 Hz), 4,51 (d, y = 10,5 Hz); 1 proton ciia nhdm carbinol tgi 5^3,64 (dd, J= 11,5, 3,5 Hz, H-3); 7 nhdm metiiyl t^i 5„ 0,90 (3H, s), 1,09 (3H,s), l,14(3H,d,/=6,5Hz), 1,47(3H,s), 1.56 (3H, s), 1,74 (3H, s). Hlng so tuong tic cua H-3 cho phep xic n h ^ ciu hinh a ciia proton nly. Ph6 '^C-NMR'cho thay phan tir cd 30 nguyen hi carbon vdi cac tin hipu d$c trung: 1 nhdm carboxylic tgi S^
181,2 (C-28), 2 carbon n6i ddi tgi S^. 128,4 (C-12), 140,4 (C-13), 1 nhdm oxymetiiine tgi 5^ 80,7 (C- 3), 1 nguydn tii carbon bgc b6n gin oxy tgi 5(.73,2 (C-19)va 1 nhdm oxymethylene tgi 5(, 65,1.
Cac kit qui thu dugc cho thiy hpp chit 1 11 mdt triterpene khung ursane. D6i chilu vdi tai li$u tham khio cho ph^p xic djnh hpp chit 1 la 3b,19a,24-trihydroxy-urs-12-en-28-oic acid hay cdn gpi Ik rotungenic acid (Hinh 3.3). Hpp
chat nly dupe phan l§p lan dau tiSn tii loli Hex rotunda [5].
Hpp chat 2: Dau mlu vang. PhS 'H-NMR chi ra tin hl^u tetra-alkyl benzoquinone. Ba nhdm methyl singlet tr^n vdng quinone xuat hi§n t^i 5„
2,01 (3H), 1,98 (6H). Tin hi§u smglet cdn Igi tgi 5jj 1,21 thupe ve nhdm methyl d vj tri germinal vdi nhdm hydroxyl. Ph6 "C-NMR chi ra cac tin hipu cua vdng quinone the va mgch aliphatic logi phytol vdi 1 nhdm hydro^o-l tgi C-3'. Cau tnic ciia vdng quinone thi dugc xic nhgn bing d c tuong tic HMBC giua 2-Me (5j, 1,98) va C-1 (S^.187,33)/C-2 (5(.140,52)/C-3 (5(,140,63);
giita 3-Me (5j, 1,98) va C-2 (5(.140,52)/C-3 (5cl40,63)/C-4 (5^.187,77); giSa 5-Me (5„ 2,01) vl C-4 (6^.187,77)/C-5 (5^,140,57)/C-6 (5^.144,57).
Mgch nhinh aliphatic tgi C-6 dugc thiet Igp thdng qua tuang tic gi&a H-1' (5^ 2,52) va C-1 (5(.187,77)/C-5 (5j.l40,57)/C-6 (5^.144,57). Cac d5 ki^n ph6 NMR ciia hgp chit 2 cho phep xac djnh hpp chit nay la (2^,5-trimethyI-6-(3 '-hydroxy)-phytyl-l,4-benzoquinone) hay cdn ggi la a-tocopherolquinone-vdi cic gia trj pho holn toan phii hgp vdi tli li^u tham khao (Hinh 3.4) [11].
Hinh 3 J . Ciu triic hda hgc cua hgp chit (1)
Hinh 3.4. Ciu tiiic hda hgc cua hpp chat (2) Tgp chf Y Dirp-c hpc - Trirfrng Dgi hpc Y Dirge Hui - S6 D$c bi^t
4 . B A N L U ^
Day 11 lan diu tien hai hgp chat rotungenic acid vl a-tocophHolquinone dugc phin l|p tir chi Hedyotis L. Hgp chit rotungenic acid tiiugc nhdm tiiterpenoid mang khung ursane. Dly 11 nhdm hgp chat cd nhieu hogt tinh sinh hgc nhu: ch6ng ung thu, bio v§ gan, ch6ng vidm, khing khuin, vims.
Mdt vii hgp chit cua nhdm nhu axit corosoHc (b6 sung che dd In u6ng chong Igi b^nh tiiu dudng) II ffi cd trSn thj trudng vl mdt so ngudi khac dang dugc thir nghiem lam sang ho§c sin sing dl dugc tung ra tren thj trudng. Betulin 11 mdt trong nhihig san phim ty nhi€n dugc cd 1 ^ diu tidn nlm 1788, cd hogt tinh chong khoi u vl khang virus. Axit Betulmtc 11 phan tii hogt tinh sinh bpc cao nhit ciia nhdm. Nd da dupe chiing minh la mgt chit ure che chpn lgc khoi u Ic tinh ciia con ngudi ml hogt ddng bing clm iing ciia apoptosis. D$c bipt, cd hogt tinh chong HTV bing cIch src chi sy trudng thanh cua virus [3]. Ursolic acid (UA) 11 mpt hprp chit ty nhien du<^ dinh gii cao vdi cac nghiSn ciiu ve hogt tinh cd gii trj nhu chong tmg thu, chong viem, bio v$ gan, giam dau, khang khuan, ch6ng nim, chong virus, dieu hda miin djch; hogt tinh chong Igi 6 chiing vi khuin, kit qui tdt nhit 11 doi vdi S. aureus vdi gii trj MIC = 32mg/ mL; hi^u qui d6i vdi E. coli, K pneumoniae vl S. flexneri vdi cimg gii trj MIC = 64 mg/mL; ure che sy ting trudng cua vi khuin lao Afycobaclerium Tuberculosis H37Rv vdi MIC = 41,9 (tg/mL. Ben cgnh dd, hogt ttnh chong virus HIV ciia UA dl dugc xic djnh vdi gii trj ECj„ - 4,4 pM [2], [4], [6].
Khi tiln hinh thir hogt tinh khang khuin eiia rotungenic acid tren 6 chiing vi khuin.
Ste^hylococcus aureus (NBRC 100910), Bacillus subtilis (NBRC 13719), Mycobacterium smegmatis (NBRC 13167), Klebsiellapneumoniae (NBRC 14940), Escherichia coli (NBKC102203), Pseudomonas aeruginosa (NBRC 106052). Kit qui thu dugc cho thiy rotungenic cd hogt tinh ire chi rd r$t tiin cic chiing 11 Staphylococcus, Bacillus va Mycobacterium vdi MIC (ng/mL) lin lugt II 2.5,2.5,1.25 so vdi ampicillin tuang iing 11 5.0, 5.0, 5.0 [7]. Trong dd, Staphylococcus aureus 11 tic nhfin hlng diu trong nhiem khuin bpnh vipn.
gfiy ra hdi chiing soc nhiem ddc, ngd ddo tiiyc phim. Mycobacterium tuberculosis lim 8,7 tripu ngudi mic b^nh lao do vi khuan, 1,3 tri^u bpnh nhan tii vong tren till gidi (2012) [12]. Day II tien dl cho djnh hudng thn kilm cic tiiudc khing sinh thyc vgt mdi tiong khi tinh ti^g de khing khing sinh dang gia ting.
Ngoli ra, rotungenic d3 dugc phin l§p tir 11 cOa loli Diospyros kaki hay cdn ggi II cfiy Hong, 11 mdt logi thu6c cd truyen Trung Quoc, da dugc sii dyng trong dieu hi dpt quy vi tiiuoc hg huyet ^ . Cac triterpene acid 5 vdng cd 1^ tir 11 hdng chtnh la tiilnh phin cd hogt tinh chu ylu ciia lolL 14 hpp chat tiTterpene phan l§p dugc tit Z>tav;j)7tw tefe'tiong do cd rotung^uc acid dl du^c kilm tia hogt tinh iic che PTPIB (protein tyrosine phosphatase) vdi gii trj ICDjo la 10,9±lpM. Ma cic chit lie chi PTPIB co gia trj trong dilu trj bpnh dai thio dudng [9].
Vdi hai hpp chit phan l&p dugc, a-tocopherolquinone II mgt chat chuyen hda ciia a-tocopherol (Vitamin E). Sung J.H. va cic cdng sy da phan Igp dugc a- tocopherolqumone tir loai Solidago virga-aurea va kilm tra hogt ttnh ddc tl bio trfin 5 ddng A-549 (Ung thu phdi d ngudi), SK-OV-3 (nhin ung fliu tuyen budng trung), SK- MEL-2 (melanoma), XF 498 (ung thu bilu md thin kinh trung uong c i ^ con ngudi), vi HCTIS (adenocarcinoma rudt ket). Thu dugc kit qui ECjo (^im) lan lugt la 8.7,22.0, 8.2,5.9,6.4 so vdi chit Doxorubicm tuong iing la 0.8, 0.9, 1.4, 1.43, 2.4 [11]. Nlm 2010, Shi gao Yang v l cic cdng sy dl c h ^ g minh a-tocopherol quinone (Alpha-TQ) dc chi tdng hgp beta-amyloid (Ab) vi hogt tinh ddc t l bio, phan tach thinh tnrdc S(;;ri vl lam giim sin lugng ciia oxy phin iing, NO vi cytokine vifim, myc tidu dilu trj b§nh Alzheimer [10]. B^nh Alzheimer (AD) li m$t phiic tgp, b ^ tiioii hda than kinh da yeu td. V i ^ tdng hgp beta-amyloid (Ab) vio ling dgng s ^ nhd ngogi bio li mdt dau hi^u b§nh IJ ciia AD. Cic Ab tdng hpp giy ddc thin kinh, phin iing viem v i stress oxy hda dugc liin kit mgnh mg vdi cic nguySn nhan cila AD.
Cic tiiudc hi§n cd khdng dii de dilu tii holn toin cin nguySn AD. Do dd, vi§c tim kiem cic hgp chit mdi cd till dilu trj nhilu myc tiSu cilng mdt liic dang tiiu hiit chii J nhilu hon.
18 Tgp chi Y Duvc hpc - IVirfrng Dgi hpc V Dirffc Hui - S6 D|c bi$t
5. Ktr L U ^ dgt dugc md ra triln vgng mdi v i cin tiiilt cho TCr cao chiet phin dogn n-hexane ciia cly An cic nghiSn curu tiep theo vl thanh phan hoa hgc diln ndn (Hedyotis pilulifera, hg Rubiaceae) dl vi tic dyng dugc IJ cua loli nly Ak cd thi umg phin l^p dugc 02 hgp chit bao gdm rotungenic dyng trong vipc tim kilm cic hogt chit cd tic dyng acid (I), a-tocopherolqumone (2). Cic kit qui phdng dilu trj b§nh cho con ngudi.
TAI LI$U . Bossi^re M., Basuki I., Koponen P., Wan M. and SheilD. (2006), "Biodiversity and Local Perceptions on the Edge of a Conservation Area, Khe Tran Village, Viemam ", Inti Prima Karya, Jakarta, ISBN 979-24-4642-7.
. Cantrell C.L., Franzbau S.G. and Fischer N,H.
(2001), "Antimycobacterial plant terpenoids", Planta Med 67, pp. 685-694.
, Jifi PatoCfea (2003), "Biologically active pentacyclic triterpenes and their current medicine signification ", Journal of Applied Biomedicine 1, pp. 7-12.
. Kashiwada Y., Wang H.K., Nagao T , Kitanaka S., Yasuda I. and Fujioka T. (1998), "Anti-HIV activity of oleanic acid, pomilic acid, and stmcturally related triterpenoids",A^or./'rad.61(9), pp. 1090-1095.
. Kayoko Amimoto, Kazuko Yoshikawa and Shigenobu Afiihara (1993), "Triterpenes and Triterpene glycosides from the leaves of Hex rotunda", Phytochemistry. 33(6), pp. 1475-1480.
, Neto Catherine C. (2007), "Cranbeny and its phytochemicals: A Review of invitro anticancer studies ^JVwWrion. 137,pp. 186-193.
. Nguyen Thi Hoai (2014), "Studies on antibacterial agents from Vietnam plants based on the FtsZ's uihibitory activity ", Research Repori.
. Nordin H. J.L. and Rohaya A. (2006), THAMKHAO
"Phytochemical Studies and Pharmacological Activities of Plants m Genus Hedyotis/
Oldenlandia", Studies in Natural Produds Chemistry. 33, pp. 1057-1090.
9. Phuong Thien Thuong, Chul Ho Lee, Trong Tuan Dao, Phi Hung Nguyen, Wan Gi Kim, Sang Jun Lee and Won Keun Oh (2008), "Triterpenoids from the Leaves of Diospyros kaki (Persimmon) and Their Inhibitory Effects on Proteui Tyrosme Phosphatase m", J Nat. Prod. 11, pp. 1775-1778.
10. Shi-gao Yang, Wei-yun Wang, Tie-jun Ling, Ying Feng, Xue-ting Du, Xi Zhang, Xlao-xia Sun, Min Zhao, Di Xue A, Yang Yang and Rui-tian Liu (2010), "Alpha-tocopherol quinone inhibits beta- amyloid aggregation and cytotoxicity, disaggregates preformed fibrils and decreases the production of reactive oxygen species, NO and inflammatory cytokines", Neurochemistry International. 57(8), pp. 914-922.
11. Sung LH., Lee J.O., Son J.K., Park N.S., Kim M.R., Kim J.G. and Moon D.C. (1999), "Cytotoxic constituents from Solidago virga-aurea var. gigantea MIQ ", Arch Pharm Res. 22, pp. 633-634.
12. World Healtii Organization (2014), Antimicrobial resistance: global report on surveillance: France Worid.
Tgp chf Y Dugrc hpc - Trirfrng Dgi hpc Y Dirpc H a l - S6 D$c bift 19