• Nghien CLFU - Ky thuat
2. Mclnnis et al.: Method and apparatus for recovering zein from com, United Stale Patent (2003).
3 M. I. Beck, I. Tomka, E. Waysek: Physico - chemical characterization of zein as a film coating polymer. A direct comparison with ethyl cellulose.
International Joumal<^Phammcei^cs (1996), p.140-145.
4. Oshlack et al.: Controlled release coatings derived from aqueous dispersions of zein, United
State Pa(en((1994) p. 1 - 2 2 .
5. The United States Pharmacopeia (USP 30 NF 25), p. 461,1248.
6, Zl Yi Yang, Yan Lu, and Xing Tang:
Pseudoephedrine Hydrochloride Sustained-Release Pellets Prepared by a Combination of Hot-Melt Subcoating and Polymer Coating, Drug Development and Industrial Phanvacy (2008), 34. p. 1323-1330.
Khao sat thanh phan hoa hoc cay ray gai (Lasia spinosa L.)
NguySn Minh Duong', Nguyin T^n Pbdt^, Phdng Vfin T^u^g^ Nguyen Ng^c Hanh^
'Dgi hgc Cdn Tha ' Vi^n Cong nghg Hod hgc - Vign Khoa hgc vd Cong nghi Viet Nam
D a t v a n d e
Cdy rdy gai {Lasia spinosa L.) edn d u y c gpi Id mdp gai, son thyc gai, thupc hp rdy (Araceae). Cay thupc logi cay thao, eao 0,4-0,7m, thdn r i va cuong Id d i u cd gal. Cf n u d c ta, cay phdn b i rai rac khap cdc dia p h u a n g d viing d i n g bang, trung du vd mil thap. Bp phdn t h u d n g diing Id thdn vd re. Cay cd tfnh mdt, cd tde dyng tieu viem, giai dpc, thanh nhi§t t r d t h i p . Theo kinh nghi$m ddn gian vd y hpc c i t r u y i n rdy gai Id n g u i n d u y c li$u quan trpng dung de chua ho, dau hpng, phCi thung do suy gan, di chdng do s i t rdt'^'^'.
Trong nhung ndm g i n ddy, da cd n h i i u nghidn edu v i thdnh p h i n hda hpc vd hogt tfnh sinh hpe cdy rdy gai d Vi$t Nam cung n h u t r i n t h i gidi'^"'^'^''', Trong bdi bdo ndy, chiing tdi trinh bdy vigc phdn l$p vd xdc djnh c i u tnic ciia cde chat t d cao ethyl acetat vd cao butanol ciia cdy rdy gai.
N g u y e n l i e u Vci phu'O'ng p h d p Nguyen ligu:
Cay ray gai (Lasia spinosa I.) d u y c cung c i p bdi Cdng ty D u y c Domesco, trong tgi xd A n C u , huygn Tjnh Bien, tfnh An Giang, thu hdi p h i n tren mat d i t vdo thdng 12 nam 2 0 1 1 , cdy d u y c 3 nam tuoi.
Phipcng phdp xac djnh c i u t r u e vd chiit xuit
Diem ndng chay d u y c do tren may ELECTROTHERMAL 9100 (UK), mao qudn khdng hi^u chfnh. Cdc p h i cdng h u d n g t d hat nhdn: ' H - N M R , ' ^ C - N M R , DEPT, COSY. HSQC, HMBC d u y c ghi trdn mdy BRUKER AVANCE (500 MHz) dd djch c h u y i n hod hpc tfnh theo (ppm), h i n g s6 t u o n g tdc (J) tfnh b i n g Hz. Pho k h i i l u o n g d u y c do trdn mdy AGILENT TECHNOLOGIES 6120 (Quadrupole LC/MS).
s i c ky Idp mdng (TLC) d u y c t h y c hign tren bdn nhdm silica gel Merck-GF60F254 trang san, kich t h u d c 20 X 20 cm, dp ddy Idp h i p phu 0,2 mm eiia hang Merck, Germany. S i c ky cot trung dp diing silica gel 60, Merck, d u d n g kinh hgt 0,040- 0,063 mm.
Thdn Id cdy rdy gai khd (1,5 kg) d u y c chiit kigt b i n g c h i i t sieu dm vdi ethanol 9 6 % d 60°C.
Sau khi thu h i i dung mdi, d u y c cao ethanol (117 g), cao ndy d u y c c h i i t Idng-ldng l i n luyt vdi n-hexan, ethyl acetat vd butanol thu d u v c edc cao t u o n g ung la cao LSH 27,5 g (chiim 23,5%); cao LSE 4,5 g ( c h i i m 3,8%) vd cao L S B 1 1 g ( c h i i m 9,4%).
T d cao LSE (4,5 g) tien hdnh sac ky cdt tmng dp vdi hg dung mdi cd dy phdn c y c tang d i n n- hexan, ethyl acetat va methanol; tgi phan dogn 5
52 T^P CHi DirOC HOC - 05/2012 fSO 433 NAM 52)
» Nghien CCFU - Ky thuat
tien hanh hanh sac ky cot trung dp vdi hg dung mdi CHCI3 MeOH (93 : 7), thu d u y c hyp c h i t (1) (5 mg); tgi phan dogn 6 tien hdnh hdnh sac ky ept trung dp vdi hg dung mdi CHCI3 MeOH HjO (8 : 2 : 0,2), thu d u y c h y p chat (2) (27 mg). T u cao LSB (11 g) tien hdnh sac ky cOt tmng dp vdi he dung mdi cd dO phan eye tang d i n n-hexan, ethyl acetat, methanol vd nudc. Tgi phdn dogn 7, thu duyc hyp c h i t (3) (30mg).
Ket qua vd thao luan
Hyp chat (1) thu d u y c d u d i dgng b i t , mau trang; d i i m ndng chdy: 298-300 C; s i c ky Idp mong (TLC) t r i i n khai bang hon hyp dung mdi CHCI3 MeOH ( 9 : 1 ) hl#n mdu b i n g dung djch H2SO4 10 trang EtOH, cho v i t trdn xanh tim cd Rf = 0,25 khi ho ndng.
Phd ^H-NMR {500 MHz, DMSO, 6 ppm) cho tin higu cda 3 proton olefin d 6 H 5 , 3 3 ( 1 H . d, J = 5, He), 5,14-5,26 (2H, m, H22, H23); 5 proton cua nhdm methin k i oxygen d 5H 3,46 (1H, m, H3), 2,90 (1H, td, J = 5 vd 9, H2), 3,14 (1H, td, J = 4 va 8,5, H3), 3,02 (1H, td, J = 4 vd 9, H4) va 3,07 (IN, dd, J = 4 vd 13,5, Hg); 2 proton ciia nhom methylen k i oxygen d 5H 3,65 (1H, ddd, J = 11, 6 va 2, Hea) vd 3,40 (1H, m, Hgt); 1 proton anomer d 5 H 4 , 2 1 (1H, d, J = 8, H i ) ; proton eua 6 nhdm methyl d 5H 0,67-1,01. Pho "C-NMR {125 MHz, DMSO, 6 ppm) k i t h y p DEPT cho tfn higu eiia 35 carbon; trong do cd 1 carbon bgc 4 mang n i l ddi d c 140,4 (C^); 3 carbon olefin d c 137,9 (C22), 129,5 (C23) va 121,0 (Cg); 1 carbon acetal d c 100,8 {Cy); 5 carbon methin ke oxygen d c 76,9 (C3), 73,5 (C2), 76,8 (Cs), 70,2 (C4) va 76,7 (Cg); 1 carbon methylen ke oxygen d c 61,1 (Ce); 6 carbon methyl; 2 carbon bac 4; 9 carbon methylen; 7 carbon methin. Chung td (1) cd khung aglycon Id stigmasterol 29 carbon, mang noi ddi d vj tri 5 vd vi trf 22 vdi phan d u d n g 6 carbon. Pho COSY vd HSQC, xdc nhgn day Id d u d n g D- glueose. Proton anomer d 5H 4,21 (1H, d, J = 8, Hi) Chung td Id d u d n g jB. Ngodi ra, proton anomer d 5 H 4 , 2 1 ( 1 H , d, J = 8, H i ) edn tuang tac vdi carbon d c 76,9 (C3), chdng td phan dudng gdn vao khung aglycon d vj tri C3.
T u d u lieu p h i ^H-NMR. "C-NMR, k i t h y p vdi pho DEPT. COSY, HSQC, HMBC; cde dgc trung vdt ly vd so sdnh vdi cac tdi li^u dd edng
T ^ CHI Dirac HOC - osmnjsO 433 NAM 52)
bi'^^, chiing toi nhdn danh hyp c h i t (1) Id: 3 - 0 - LS-D-glucopyranosyl]-stigmasta-5,22-dien.
Hyp chat (2) thu d u y c d u d i dang b i t , mdu vdng; d i i m nong chdy: 269-270 C; sdc k f Idp mdng (TLC) t r i i n khai bdng h i n hyp dung mdi CHCI3 MeOH H2O ( 7 : 3 : 0,5) hign mdu b i n g dung djch H2SO4 10 trong EtOH, cho v i t trdn mdu vdng ed Rf = 0,5 khi ho ndng.
P h i k h i i luong MS cho pic ion gia phdn t d vdi m/z [M-H]" = 431 tuong dng vdi cdng thue phdn tuC2iH!oOio.
Phd ^H-NMR (500 MHz, DMSO, 5 ppm) (Bdng 1) cho tin higu cua cde proton vdng thom d 5H 6,27-8,01. Trong dd 1 proton eiia vdng thom bt t h i 5 lan d 6H 6,27 (1H, s. He) vd 4 proton eiia vong thom the doi xung 1,4 d 5H 6,88 ( I N , d, J = 8,5, H2', Hg') vd 6H 8,01 (1H, d, J
= 8,5, Ha', H5'); 1 proton ciia nhdm hydroxyl k i i m n i l d 5H 13,17 (1H, s, OH5). Ngoai ra, cdn ed proton anomer d 5H 4,68 (1H, d, J = 9,5, Hi") va proton methin k i oxygen d 6H 3,22-3,84.
Pho "C-NMR (125 MHz, DMSO, 6 ppm) k i t hyp vdi DEPT eho tin higu eiia 21 carbon; trong do ed 5 carbon bac 4 vong tham ke oxygen d c 163,9 (C2), 160,3 (Cs), 162,6 (C7), 156,0 (Cg) vd 161,1(C4'); 3 carbon bdc 4 vong tham d c 104,6 (Cs), 104,0 (Co) vd 121,6 (Ci'); 1 carbon carbonyl d c 182,0 (C4); 6 carbon methin vdng thom d c 102,4 (C3), 98,2 (Ce), 128,9 (Cj', Ce') va 115,8 (C3', Cs'); 5 carbon methin ke oxygen vd 1 nhdm methyl ke oxygen d c61,3 (Ce") eho thay hyp chat (2) Id flavonoid glucoside vdi p h i n dudng 6 carbon. Pho COSY va HSQC xde nhgn dan vj dudng Id D-glueose. Hang so ghep cgp J ciia proton anomer d 6H 4,68 (1H, d, J = 9,5, Hi"), eho biet day Id dudng ^-D-glueose.
Phd HMBC cho t h i y proton anomer d 5H 4,68 (1H, d, J = 9,5, Hi") tuang tac vdi carbon bdc 4 vdng tham d c 104,6 (Cg), vay p h i n dudng phai gdn vao khung aglycon d vj tri Ce;
ngodi ra H i " cung cho tuong tdc vdi carbon bgc 4 vdng thom d c 162,6 (C7) vd 156,0 (C9) do dd proton duy n h i t eiia vdng A d 5H 6,27 (1H, s.
He). Dieu nay cung d u o c chdng minh, proton d 5H 6,27 (1H, s. He) tuang tdc vdi carbon bdc 4 vdng tham d c 104 (Cio); 104,6 (Ca); 160,3 (Cg) va 162,6 (C7). Ngodi ra, tren pho HMBC cung cho t h i y proton d 5H 6,78 (1H, s) tuang tdc vdi carbon carbonyl d c 182,0 (C4) va carbon bgc 4 vdng thom ke oxygen d -- 163,9 (Cz), nen proton ndy Id H3.
• Ngiiien cii'u - Ky tiiuat
Tif dO li$u ph6 H-NMR, "C-NMR, kit hi?p vdi ph6 DEPT, COSY, HSQC, HMBC, cio ajc truTig vjt ly; so s4nh vdi ti\ li^u dS c6ng b6"' vS ket qua cua nli6m t4c %\i NguySn Ttii H6ng van aa c6 Mp \i! cay rSy gai ttiu hSi b Hi NOi, ctiOng tSi nhSn danli ti^p ch4t (2) \i: Vitexin.
Hop cti4t (3) tiiu iuqic A\i6\ dsng bOt, mau vang sang; diim n6ng ctidy: 213-215 C; sic ky i*p mdng (TLC) triin khai bing h6n hyp dung m6i CHCij IWeOH H2O ( 7 : 3 : 0,5) iiiin mau bang dung dich HzS04 10 trong EtOH, oho vit trdn mau vSng cd Rf = 0,25 khi ho ndng.
Phi khii tu'ong cho pic ion gia phfin tu' vdi m/z [IVI-H]" = 593 tu'ong ij'ng vdi cdng thifc phan tl> C2,H3oO,5.
Tu-ong ty chit (3) cQng cd dO li#u phi gin nhu chit (2) (Bang 1), gim 1 khung flavonoid vi 2 San vi du'dng. Phi COSy va HSQC xac nh^n don v]
du*ng thd hai ia D-giucose. Hang s i ghdp cap J cua proton anomer d 6H 3,91 (1H, d, J = 7,5, Hi-), cho biit day la au*ng /3. Phi HMBC cho thiy sg- ttjong tdc giO'a proton anomer cua du'dng thir hai d SH 3,91 (1H, d, J = 7,5 Hz, H,-) tuong tdc vdi 6c 81,1 (Cr), v^y du'dng thCr 2 gan vdo phin du'dng thir nhit tai vj tri Cy.
Td dO' li#u phi 'H-NMR, "C-NMR, kit hop vdi phi DEPT, COSY, HSCK, HMBC, dc dac trung v^t 1^; so sdnh vdi tdi iidu da cdng bi'^vd kit qud ciJa nhdm tac gid Nguyin Thj Hing Vdn da c i tap ti> cdy rdy gai thu hai d Hd Nii, chung tdi nhdn danh hop chit (3) ta: Vitexin-2'-0-/3-D- giucopyranosid.
3-0-[jS-D-giucopyranosyi]-stigmasta- Vitexin (2) 5,22-dien (1)
Bang 1: DiJ' lieu ph6 'H, "C-NMR va HMBC cua chit (2) vi (3)
Vitexin-2"-O^D- glucopyranoside (3)
"C NMR Vj tri ppm
2 3 4 5 6 7 8 9 10
r
2' 3' 4' 5' 6' 1"
2- (2) 163,9 102.4 182.0 160.3 98.2 162.6 104.6 156.0 104,0 121,6 128,9 115,8 161,1 115,8 128,9 73,3 70,8
(3) 163,7 102,6 182,0 160.6 98.1 162.6 103.6 156,2 103,8 121,7 128.8 115,8 161,0 115,8 128,8 71,5 81,1
'HNMR ppm, J = (2)
6,78 (IH.s) OHs 13.17 (1H.S)
6.27 (IH.s)
8.01 (1H. d. J = 9) 6,88(1H. d. J = 8.5) 6.88(1H. d. J - 8 , 5 ) 8,01 (1H,d,J = 9) 4,68(1H, d, J = 9,5) 3,84(1H,t.J = 9)
Hz (3)
6.74 ( I H . s ) OHs 13,15 (IN, s)
6,23 (1H,s)
799(1H, d, J = 8.5) 6.89(1H. d. J = 8.5) 6,89(1H, d.J = 8.5) 7.99(1H, d. J = 8.5) 4,79(1H.d.J = 9,5) 4,07(1H, t, J = 9,5)
HMBC 'H "C (2) Cl ,C2,C4,Cl0
CsiC7,C8,Cl0
C2,C4',C6*
Cr,C4',C5' Cr,C3',C4' C2,C2',C4"
C7.C8, Ca, C2",C3',C4"
CB,CI",C3"
(3) Cv,C2,Cio
C8,CiD 1 ^.:f'
>«:»i-
Cz,C4',C6' Cr,C4',C5' Cr,C3',C4' C2,Cz',C4"
C7,Ce, _ C9.C2",C3' C,',C3- T*p CHi Dirac HOC - 05/2012 (s6 4.33 NAIM 52).
• Ngiiien ciJU - Ky tiiuat
VJIri 3- 4"
5"
6- 1 "
2"
3"
4 "
5"
6"
"C NMR ppm (2) 78.6 81.7 70,6 61,3
(3) 78.4 70.1 81,7 60,9 " » • 105,1
74,3 76,2 75,9 69.5 60.4
(2) 3.27 (2H.
3,27 (2H, 3,40 (1H, ,3,76(1H. dd. J
H,'b 3,62 (1 m) m) m)
'HNMR ppm, J = Hz
= 4.5 vd 10,5) H, m)
<3) 3,41-3,46 (3H,m) 3,41-3,46 (3H, m) 3,24-3,27 (2H, m) H,-. 3,74-3.77 (IN. dd.J = 5,5vS10,5)
H."B 3,41-3,46 (3H,m) 3,91(1H,d, J = 7,5)
2,77(1H, t, J = 8) 2,88-2.94 (2H. m) 2.46 (IH.m) 2,88-2.94 (2H. m) H6-.3.16(1H.dd.J = 3vS11) Hm3.05(1H.dd.J = 2,5vd11)
HMBC 'H "C (2) Cs- Cs- C3-.C4"
(3) C2-.C4-
Cs-
C2-,C3- C3- C«-.C5-
Cs".C4-
Kit luan
T u phan tren mgt dat ciia cay rdy gai d u y c 3 nam tuoi trong tgi An Giang, chdng tdi da phan Igp va nhdn danh cdu tnJc ba hyp chat Id: 3-0-
^D-glueopyranosyl]-stigmasta-5,25(27)-dien (1), vitexin (2) va vftexin-2"-0-p-D-glucopyranosid (3).
Trang do (1) Id lan ddu tien d u y c tim thay trong cay ray gai so vdi nhung nghien c u u trudc ddy.
Summary
Of Lasia spinosa L. cultivated in An Giang province, three compounds were isolated, 3-O-^D- giucopyranosyl]-^gmasta-5,22-dien (1), Vitexin (2) from Uie ethylacetate extracts and Vitexin-2"-0-P-D- glucopyranoside (3) ^om the butand ones. Their stnictures were elucidated by NMR (1D and 2D - NMR) in reference to the literature data.
Keywords: Lasia spinosa L, 3-O^-D- glucopyranosyl]-^gmasta-5,22-dien (1), Vitexin (2) and Vftexin-2"-0-^D-glu<x>pymnoside (3).
Tai lieu tham khao 1. Do Huy Bfch, Dgng Quang Chung, Bill Xudn Chuo-ng, Nguyen Thuyng Dong, Pham Van Hi^u, Vu Nggc L0, Pham Duy Mai, Phgm Kim Man, Dodn Thi Thu, Nguyin T i p , Tran Todn: Cdy thudc va dOng vat ISm thu6c & Viit Nam, Nha xuit biin Khoa hQC va Ky thu$t, Vi|n duyc ligu, (2003) tap II, 617-618.
2. D 6 Tat LyI: Nhung cay thu6c vd vi thudc Viet Nam, NXB YhQC. (2004), 757-758.
3. Nguyen Thj Hong Van, Chdu Van Minh, Pham Quoc Long, Braca Alessandra, Morelli Ivano, Nguyin The DOng: Budc ddu nghien cdu thanh phan hPa hoc
ciia cdy rSy gai, T?p chi Dugc li$u, (2005) t§p 10, s6 2, trang 45-49.
4. NguySn Thj Hfing VSn, Chau Van Minh, Phgm Qu6c Long, Braca Alessandra, Morelli Ivano, Nguyin ThS Dung: Three flavonoid glucoside and two phenylpropane derivatives of lasia spinosa L., Biodjemiscat SystemaScs and Ecology. (2006), 34, p. 882-884.
5. W. Suthikrai, R. Jintana, S. Sophon. R.
Hengtakulsin, V. Usawang, M. Kamonpatana: Effects of Lasia spinosa Thw. on growth rate and reproductive hormone of weaned Swamp buffalo and Murrah X Swamp buffalo calves, Italian Journal of Animal Science, (2007), vol. 6, p. 532-535.
6. FaknjI Alam, Mozammel Haque, Md. Hossain Sohrab, Mohammad A. Monsur, Choudhury M. Hasan and Nuruddin Ahmed: Antimicrobial and Cytotoxic activity from Lasia spinosa and isolated Lignan, Latin American Journal of Ptiarmacy, (2011), p. 550-553.
7. Temjenmongla and Arun K. Yadav: Anticestodal efficacy of Lasia spinosa extract agaisnt experimental hymenolepis diminuta infections in rats, PhamiaceuScal Biology. (2006), vol. 44, No. 7, p. 499-502.
8. M. Sanwar Alam, Neeraj Chopra, Mohammed All, Masatake Nlwa: Oleanen and stigmasterol derivatives from Ambroma Augusfa, Phytochemistry, (1996) Vol. 41, No. 4, p. 1197-1200.
9. Erika Vdzquez, Estaban M.Martinez, Juan Antonio Cogordan, Guillemio Delgado. Triterpenes, phenols, and other constituents from the leaves of Ochroma pyramidale (Balsa wood, Bombacaceae).
Prefened confomiations of 8-C-p-D-glucopyranosyl- apigenin (vitexin), Joumal of the Mexican chemical Society, julio-septiembre, ano/vol. 46, nCimero 003 Sociedad Qufmica de Mexico Mexico Mexico, (2002), p. 254-258.
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