'"<ap chi Khoa hgc Truang Dgi hgc Can Tha Phdn .i. Khoa hgc Tu nhien. Cong nglie vd Mdi Iruang: 31 (2014): 81-84
Tap chi Khoa hoc Tru'dng Dai hpc Can Thtf website: sj.ctu.edu.vn
HQP C H A T FLAVONOIT TU* L A CAY M U O N G HOANG Y E N (CASSIA FISTULA L.) Pham Thi Nhat Trinh*, Ddng Thi Ckm Nhung' va Le Tiin Nhung^
' Tnrdng Dgi hgc Tien Giang
^ Vi4n Cdng ngh^ Hda hgc Thdng tin chung:
Ngdy nhgn: 02/10/2013 Ngdy chip nlign: 28/04/2014 Tale:
Flavonoids from the leaves of Cassia fistula L.
Tu khoa:
Mudng hodng yin, flavonoit Keywords:
Cassia fistula, flavonoid
ABSTRACT
Two flavonoids. kaempferol (1) and liquiritigenin (2) were extracted and isolated from the ethyl acetate fraction of the leaves of Cassia fistula. Their structures were elucidated by ID and 2D NMR spectral analysis and comparison with published data.
Hai hgp clidt flavonoit, kaempferol (I) vd liquiritigenin (2) da dugc phdn Igp tir dich chiit etyl axetat Id mudng hodng yin. Cdu trite cdc hgp chdt dugc xdc dinh dua vdo phd cgng hudng tir hgt nhdn 1 chiiu. 2 chiiu vd kit hgp vol tdi lieu tham klido.
1 MODAU
Cay Mudng faoang yin cd tin khoa hgc li Cassia fistula L., thudc hp Vang (Caesalpiniaeeae).
Hi|n nay, cdy dugc trong rdt nfaiiu dl ldm canh d cac tinh, tfadnh pfad ldn nfau Hi Npi, tfaanh pfad Hd Cfai Minh, Ddng Nai... TTieo y hpc ddn gian, nudc sdc ciia Id ciy Mudng hodng yin cd tic dung nfauin trang, dugc dimg dl cfaiia cfaung kfad tiiu, dau lung, benfa ki sinh triing dudng nipt va cdc benfa ngoii da [1]. Cic nghiin cuu fren thi gidi cho tfady li Mudng hodng yen cd thdnfa phan cfainh Ii flavonoid vi antraquuion, cd tdc dung khdng oxy hod. khing vilra, khdng ndm, khang kfauin [2], Ngfailn ciiu dugc tinfa lira sing cua li cay len mgt sd dgng vit cfao tfaay nfaiiu kit qui quan frgng, dac bi|t Id kfad ndng bdo ve gan [4] vi lam Iinfa vit tfauong [3].
d nudc ta, ciy Mudng facing yin vin cfaua dugc nghien ciiu ve ragt hod hpc ciing nhu hoat tinh sulfa hgc. Bii bio nay tfaong bdo nfaiing kit qui ndiien cuu ban dau ve tiianfa pfadn hod fagc ciia cay Mudng faodng yen. Bing cdc phuong pfadp sic ky.
faai fagp cfadt kaempferol (1) va liquiritigenin (2) da dugc phdn Igp tii dich chiet etyl ^lxetat ciia la cdy Mudng hodng yin.
2 PHlTOfNG PHAP NGHIEN CUtJ 2.1 Phinmg phap nghiin ciru
Sdc ky Idp mdng (TLC) dugc thyc hiln frln bdng radng fring sdn DC-AIufolien 60 F254 (Merck), RPig Fi^is (Merck). Phdt faien cfait bing den tu ngogi d hai budc sdng 254 nm vd 365 nm hogc diing thudc thijr la dung dich 10%
H2S04/Et0H faay 1% FeCls/EtOH dugc phun diu len bin mdng, sdy khd rdi ha ndng frln bip dien tiJr til din kfai hi|n mdu.
s i c ky cOt (CC) dugc tiln hdnh vdi chit hip phu la silica gel pha thudng vd pha ddo. Silica gel pha tiiudng cd cd hat Id 0,40-0,063 mra (240-430 mesfa). Silica gel pha ddo RP-I8 (30-50 pm). Ngodi ra cdn su dung gel sepfaadex LH-20.
Diim ndng cfady (mp) dugc do frln mdy Elecfrotfaermal IA seriee, dimg raao qudn kfadng failu chinfa ciia Vien Cdng nghe Hoa hpc.
Tap chi Khoa hgc Trudng Dgi hgc Cdn Tha Phdn A: Khoa hgc Tv nhiin, Cdng nghf vd Mdi truang: 31 (2014): 81-84 Phi cgng faudng tir fagt nfadn (NMR): 'H-NMR
(500 MHz) vi '^C-NMR (125 MHz) dugc do frln may Bruker AM500 FT-NMR Specfrometer, Vien Hod fapc, Vien Hin ldm Kfaoa fapc va Cdng nghe Viet Nara.
2.2 Nguyen li|u
Mau thyc vdt dugc dung frong nghiin cuu Id Ja cay Mudng hodng yin (Cassia fistula L.). Mdu dugc thu hii tai thdnfa phi Hd Chi Minh vao thdng 6 ndm 2010.
2.3 Phan l^p cic chat
Nguyin li|u siy kho, nghiin thdnh bgt (2,9 kg) dugc ngdm chiet bdng etanol. Djcfa chiit dugc cd dudi dung mdi d ap suit kem tfau cao etanol (142,6 g). Phin bd lgi cao etanol bdng sic ky cpt nfaanfa silica gel vdi eac dung moi n-faexan, clorofom, etyl axetat vi metanol. Cd cgn cdc pfadn dogn cfao cdc cao tuong ung cd kfadi lugng ldn lugt Ii 15,8; 32,1;
41,4 vd 25,9 g. Tii cao etyl axetat (41,4 g), su dyng sic ky cpt silica gel pha thudng nhilu lin kit hgp vdi sic k^ cpt silica gel pha dao Rp-I8 vi sac k^
lgc gel sephadex LH-20 thu dugc hgp chdt 1 (5 mg) vd 2 (2 mg).
Hgp chit I: dgng bpt miu vdng, nhiet dp n6iig chay 276-278''C. 'H-NMR (500 MHz, CD3COCD3), 6 (ppm), J (Hz): 12,16 (IH, s, 5-OH); 9,76 (s, OH); 9,09 (s, OH) 8,15 (2H, d, 9Hz, H-2V H-6'); 7,02 (2H, d, 9Hz, H-3'/H-5');
6,53 (IH, s, H-8); 6,26 (IH, s, H-6). '^C-NMR (125 MHz, CD3COCD3 ), S (ppm): 176,6 (C-4);
165,0 (C-7); 162,3 (C-5); 160,1 (C-4'); 157,8 (C- 9); 147,0 (C-2); 136,6 (C-3); 130,4 (C-2V C-6');
123,3 (C-I'); 116,3 (C-37 C-5'); 104,1 (C-10);
99,2 (C-6); 94,5 (C-8).
Hgp chat 2: tinh the tring, nhipt dp ndng chay 209-211<<;. 'H-NMR (500 MHz. CD3COCD3), 5 (ppm), J (Hz): 7,73 (IH, d, 8,5Hz, H-5); 7,40 (2H, d, 9,0Hz, H-2'/H-6'); 6,.90 (2H, d, 8,0Hz, H- 37 H-5'); 6,57 (IH, dd, 8,0Hz; 2,5Hz, H-6); 6,42 ( IH, 4 2,5Hz, H-8); 5,46 (IH, dd, 13,0Hz; 3,0Hz, H-2); 3,06 (IH, dd, I6,5Hz; 13Hz, H-3e); 2,69 (IH, dd, I6,5Hz; 3,0Hz, H-3a). '^C-NMR (125 MHz, CD3COCD3), 5 (ppm): 190,4 ( C-4); 165,3 (C-7); 164,5 (C-9); 158,6 (C-4'); 131,3 (C-l');
129,5 (C-5); 128,9 (C-2'/C-6'); 116,1 (C-3'/C-5');
115,2 (C-IO); 111,2 (C-6); 103,7 (C-8); 80,5 (C-2);
44,6 (C-3).
3 KET QUA VA THAO LUAN Hgp chdt 1 cd Igp dugc d dang bdt miu ving cd nfailt dp ndng chiy 276-278"C dung moi acetone.
Hgp chit niy cho phan ung duang tinh vdi tiiudc tiiu FeCb/EiOH, cfaung td diy li mdt fapp cfait phenol.
Phd '-^C-NMR cua 1 xuit hiln 13 tin failu, frong do 2 tin hieu tai 8c 130,4 vd 116,3 cd cudng dp cacbon gdp doi. V§y cdng thurc phan tii ciia 1 cd 15 cacbon, gom I cacbon cacbonyl [5c 176,6 (C-4)], 12 cacbon vdng thom [5c 165,0 (C-7); 162,3 (C-5);
157,8 (C-9); 130,4 (C-2' vd C-6'); 123,3 (C-I');
116,3 (C-3' vi C-5'); 104,1 (C-10); 99,2 (C-6) vi 94,5 (C-8)] vi 2 cacbon olefm mang oxygen [5c 147,0 (C-2) vd 136,6 (C-3)]. Tii da lieu phi 'H NMR, '^C NMR vd DEPT eho dy doin 1 cd khung flavone.
Phd 'H NMR cua 1 cdn sdu proton vdng thom, frong dd, hai tin hi|u proton vdng thom xudt hi?n vdi cudng dp proton gip doi [SH 8,15 (2H, d) vd 7,02 (2H, d)], faing sd ghep J = 8,5 cfao tfaiy day Id 2 cip proton tuong duong ndra d vi fri orto vdi nhau. Tin faieu proton t^ 5H 7,02 cho tuong quan vdi tin faieu cacbon tgi 5c 116,3 vd tin faieu proton tai 5H 8,15 cho tuong quan vdi cacbon tai 5c 130,4 frln phi HSQC vd tuong quan qua lai lln nhau frln phd HMBC. Dilu ndy chung td bdn proton nay Cling ndm trin vdng B cd tirtii dil xung.
Tin hi?u kiim nii tai 5H 12,16 fren phd ^H NMR khing djnh nhdm -OH phdi gan vio C-5 khung flavon. Tren phi HMBC, proton cua -OH kiim nil tuong quan vdi m^t cacbon vong tham mang oxygen 5c 162,3, mdt cacbon vdng thara tii cap 5c 104,1 vd mgt cacbon vdng tham mang hydrogen Sc 99,2 khing dinh ba tin failu cacbon nay lin Iu0 Ii C-5, C-IO vd C-6. Proton H-6 [SH 6,26] gfalp meta vdi proton SH 6,53, nin proton nay cfai cd the la H-8. Mdt kfaac, hai proton H-6 vd H-8 Cling tuong quan vdi carbon 5c 165,0 frln pho HMBC, xac nhgn cacbon ndy phdi Id C-7. Cacbon 5c 157,8 cung cho tuong quan vdi vdi proton H-8 frln phi HMBC nin cacbon ndy Id C-9.
Xet vdng B, tren phd HMBC, tin faieu proton SH 8,15 cho tuang quan vdi hai carbon sp^ mang oxygen [Sc 160,1 va 147,0] vi mdt carbon sp^ tii cip [Sc 123,3]. Tin hi^u proton 5H 7,02 cung cho tucmg quan vdl hai frong ba carbon frln la Sc 160,1 va Sc 123,3. Do dd cd thi suy ra tin hi|u proton SH 8,15 ling vdi H-2' vd H-6', tin faipu proton tai SH 7,02 u:ng vdi H-3' vi H-5'; ba carbon sp^ Sc 130,4;
147,0 v i 123,3 lin lugt Ii C-6', C-2 vd C-l'. Nhu viy, frong 13 tin failu carbon ciia khung flavon frln phd "C NMR chi cdn tin hi|u carbon sp^ Sc 136,6 nen carbon niy phdi Id C-3.
Tgp chi Khoa hgc Truong Dgi hgc Can Tho Phdn A Khoa hgc Tv nhien. Cong nghf vd Mdi iru&ng: 31 (2014): 81-84 Tii cdc biln ludn frln, kit h ^ vdi tii lieu
tfaam kfaao [5], cfaung tdi kit ludn fagp cfait 1 Id kaempferol.
Kaempferol cd ldc dung ti'ch cyc frong vile
Hgp cfadt 2 thu dugc dang tinfa tfae mau trang.
Nhi?t dp ndng chdy 209-21 I^C dung mdi acetone, 2 cfao pfadn ling duong tinfa vdi tiiudc thu FeCb/EtOH, chiing td day Id radt dan xuit phenol.
Phi 'H-NMR (aceton-rfe) cho thdy sy hien diln cua 7 proton vdng thom, frong do cd 2 cip proton ddi xiing fren vong benzen SH [7,40 (2H; d; 9 Hz)]
vd SH [6,90 (2H, d; 9 Hz)] vd 3 proton nfadn tiiom SH [7,73 (IH; d; 8,5 Hz)]; 6,57 (IH; dd; 8,5 Hz, 2,5 Hz); 6,42 (IH; d; 2,5 Hz)], 2 proton ciia nhdm raetfaylen vd I proton ciia carbon sp^ 6H 5,46 (IH, dd, 13,0Hz;3,0Hz).
Phi ''C-NMR (aceton-de) kit hgp vdi ky tiiuit DEPT cbo biit fagp chit 2 cd 13 tin failu. Trong dd, moi tin fai|u a Sc 128,9 vi 5c 116,1 cd cudng do cacbon gip ddi. Vdy frong hgp chit 2 cd 15 cacbon, gdm 1 cacbon cacbonyl [5c 190,4 (C-4)], 12 cacbon vdng tiiam [5c 165,3 (C-7); 129,4 (C-5);
164,5 (C-9); 128,9 (C-2' vi C-6'); 131,3 (C-l');
116,1 (C-3'viC-5'); 115,2 (C-10); 111,2 (C-6) va 103,7 (C-8)] vd 1 cacbon sp' mang oxygen 6c 80,5 (C-2) va 1 cacbon metylen Sc 44,6 (C-3).
Du lieu phd NMR cfao dy doan day la rapt flavanon faai proton ciia nhdm metfaylen 6H [3.06 (IH; dd;" l6.5Hz; 13 Hz); 2.69 (IH; dd; 16.5 Hz;
3.0Hz)] tuong ling li H-3e vi H-3a.
Phi HSQC cfao biit C-3 cd dp dich cfauyen hda hoc 6c 44.6. Proton H-3 cfao tuong quan HMBC vdi mdt cacbon bgc 3 ndi oxy 6c 80,5 it bin Id C-2, vay proton tgi 5H5,46 la H-2.
Trin HMBC. proton [5H 7.73; d; 8.5 Hz] frrong quan vdi cacbon cacbonyl xdc nhdn proton nay la H-5, suy ^ proton ghep orto vdi H-5 Id H-6 [6H 6 57' (IH; dd; 8,0 Hz; 2.5 Hz)]; proton ^ e p meta
chdng ung thu va binh tim mgch. Nfaiiu ngfaien Clhi ciing cfai ra mdt sd tdc dung chiJa benfa eiia kaempferol nfau cfaong ddng kinfa, cfadng viem, kfadng kfauin, cfadng oxy fada, chdng co that, tri ddi thdo dudng, gidm dau vd ldm giim ho [7].
vdi H-6 Id H-8 [5H 6,42; d; 2,5 Hz], Nhu viy nhdra till -OH hdn tgi C-7. Proton H-8 vi H- 5 cho tuong quan vdi hai cacbon bic bon mang oxy 5c [165,3;
164,5], nen hai tin hieu ndy ldn lugt Id C-7 va C-9.
Hai tin hi|u proton gfaep orto tai SH [7,40; 2H;
d; 9Hz] vd 6H [6,90; 2H; d; 8,5Hz] chiing to cfaung thudc vdng benzen the 1,4- cd ciu tnic ddi xiing.
Vdy diy Id nh&ng tin hieu cua vdng B vd vdng B mang mgt nhdra thi -OH tai C-4".
Pfadn tich tin hieu proton cua nhdm metylen frln C-3, proton SH [3m06 (IH; dd; 16 Hz; 13 Hz)]
Id proton H-3a, proton SH [2,69 (IH; dd; 16,5Hz;
3,0Hz)] Id proton H-3e, nhu viy vdi hdng si gfaep J=I3,0 (ghep aa) cua H-3a vd J=3,0 (^ep ae) cua H-3e nin H-2 chdc cfadn phdi ndm d vj fri tryc vd vong benzen ndm d vi tri xich dgo. Nhu vdy cau hinh cua C-2 laS.
J™,=16-5 Su gbep spin giira cac proton trong vdng C ciia
fagp chat 2
Tu cic diJ lieu pfad \a tdi lieu tham khao (Bing 1), hgp chit 2 thu dugc la liquiritigenin.
Liquiritigenin cd tic dyng cfadng lai sy nfaiem ddc kim loai nang frong raiu. Ngodi ra, faquiritigenin cdn cd khd ning bio vl gan, ngin chin ung tiiu gan [7].
Tap chi Khoa hgc Truang Dgi hgc Cdn Tho Phdn A • Khoa hgc Tv nhien, Cong nghi vd Mdi tru&ng: 31 (2014): 81-84
Bang 1: So sinh so lieu phi NMR ciia 2 vdi phi NMR cua liaquiritigenin [6]
Vj trf 2 3 4 5 6 7 8 9 10
] '
2' 3' 4' 5' 6'
2 (Acetone-dfi) 5H ppm (J, Hz)
500 MHz 5.46 (d, 13-3) 3.06(dd,I6-I3) 2.69(dd;16.5-3) 7.73 (d; 8.5) 6.57 (dd; 8-2) 6.42 (d; 2.5)
7.40 (d; 9) 6.90 (d; 8.5) 6.90 (d; 8.5) 7.40 (d; 9)
5c ppm HMBC 125 MHz (H -> C)
80.54 C-2' 44.69 C-2; C-4 C-2' 190.48
129.45 C-2; C-9 C-4 111.23C-8;C-10 165.37
103.70C-6;C-9 C-10 164.54
115.20 131.31
128.92 C-2; C-4' C-6' lI6.15C-r;C-4' C-5' 158.62
116.15C-I';C-4' C-3' 128.92 C-2; C-4' C-2'
Liquiritigenin (Acetone-d6) ba ppm (/, Hz)
500 MHz 5.36 (dd;12.9-3.0) 3.0I(dd; 16.8-12.) 2.67 (dd; 16.8-3.0) 7.69 (d; 8.8) 6.43 (dd; 8.8-2.2) 6.25 (d; 2.2)
7.33(d; 8.5) 6.83 (d; 8.5) 6.83 (d; 8.5) 7.33 (d; 8.5)
5c ppm 125 MHz
81.0 44.9 193.5 129.8 111.8 166.9 103.45 165.5 114.9 131.3 129.0 116.3 159.0 116.3 129.0 4 KETLU^N
Tu dich chiit phan dogn etyl axetat cua la Mudng hoang yen, bang cdc phucmg phdp sac ky da phdn ldp dugc hai hgp chat flavonoit:
kaempferol vi liquiritigenm. Cau tnic hoa hgc cua cdc hgp chat ndy dugc xac dinh bang phd cdng hudng tii hat nhan NMR (ID, 2D).
TAI LifU THAM KHAO 1. Dd Huy Bich, Dang Quang Chung, Bill
Xuan Chucmg, Nguyin Thugng Dong, Dd Trung Dam, Phgm Van Hiin, Vu Nggc Lg, Pham Duy Mai, Phgra Kim Man, Doan Thi Nhu, Nguyin Tap, Trin Toan (2004), Cay thuic va dpng vat lira thudc d Viet Nam, tgp 1, Nha xudt bdn Khoa hgc vi kT tiiudt Ha N6i,ti^g215-217.
2. Mohd. Danish, Pradeep Singh, Garima Mishra, Shrutl Srivastava, K.K. Jha, R.L.Khosa (2011), "Cassia fisfrila Linn.
(Amulthus)- An important medicinal plant:
A review of its fraditional uses, phytochemistry and pharmacological properties", J. Nat. Prod. Plant Resour, 1 (1), 101-118.
Muthusaray Senthil Kumar, M.Sc, Ramasamy Sripriya, Ph.D., Harinarayanan Vijaya Raghavan, M.Sc, and Praveen Kumar Sehgal, Ph.D. (2006), "Wound Healing Potential of Cassia fistula on Infected Albino Rat Model", Joumal of Surgical Research, 131,283-289.
T. Bhakta, S. Banerjee, Subhash C. Mandal, Tapan K. Maity, B. P. Saha and M. Pal (2001), "Hepatoprotective activity of Cassia fistula leaf exfract", Phytomedicine, 8 (3), 220-224.
Deepa T. Vasudevan, Kavitha R. Dinesh, S.
Gopalakrishnan, S.K. Sreekanth And Sonal Shekar (2009), "the potential of aqueous and isolated fraction from leaves of Cassia fistula linn as antibacterial agent". Int. J.
Chem. Sci., 7(4), 2363-2367.
Le Tien Dung (2008), "Nghien ciiu hod hpc va boat tinh sinh hpc ciia ciy Ban nhit (Hypericum japonicum)", Luin an tiin sT hoi hpc, Viin hoa hpc cdc fagp chdt thiin nliiln.
Dictionary of Natural Prduct on DVD, 2012, CRC press, Taylor and Francis group.
