TAP CHI HOA HOC T. 50(2)201-204 THANG 4 NAM 2012
NGHIEN CLFU THANH P H A N HOA HOC LA CAY SON TRA POILANE {ERIOBOTRYA POILANEI J. E. VID.), HO HOA H 6 N G ( R O S A C E A E )
III - Cac ho-p c h M tritecpen axit va fiavollgnan Dinh Gia Thien', Nguyen Thi Hoang Anh^*, Tran Van Sung^
'Tnrdng THPT Le Quy Ddn. Thdnh pho Tam Ky - Tinh Qudng Nam
^ Vien Hda hgc. Vien Khoa hgc vd Cong nghe Vi^t Nam Din Tda sogn 9-11-2011
Abstract
Three compounds a fatty alcohol («-hexate[racontan-l-ol), a triterpene acid (arjunic acid) and a flavolignane (catiguanin B) were isolated from M-hexane and ethylacetate extracts of Eriobotrya poilaiiei's leaves. Their structures were elucidated by analysis of MS, NMR spectra and comparison with the published data.
Keywords: Eriobotryapodanet, triterpene and flavolignan.
1. MO DAU
Cdy Son trd Poilane {Eriobotryapodanei) la mgt trong sd 13 loai thugc chi San trd {Eriobotrya), hg Hoa hdng (Rosaceae) cd d Viet Nam [1].
Theo tra ciiu tu lieu, chua cd cdng bd nao ve thanh phdn hoa hgc ciia cay Son tra Poilane trudc nghien curu ciia chiing tdi [4, 5]. Bai bao nay tiep tyc thdng bao vipc phan lap va xdc djnh cau triic ciia 3 chat thugc cac khung khac nhau: ancol beo, tritecpen axit, flavolignan dugc tach tir dich chiet «-hexan va etyl axetat cua Id cdy Son tra Poilane. Cau triic cua cac chat do dugc xac djnh Id «-hexatetracontan-l-ol (1), axit arjunic (2) vd catiguanin B (3).
2. THUC NGHIEM 2.1. Hoa chat va thiet bi
Phd cdng hudng tir hat nhan NMR dugc ghi tren may Bruker Avance 500 MHz, phd khdi ESI-MS dugc do tren mdy AGILENT 1100 LC-MSD Trap tai Vipn Hda hoc, Vien Khoa hgc va Cdng nghe Viet Nam. Sac ky ban mdng dugc tien hdnh tren ban silica gel Merck 6OF254. Sac ky cgt sir dyng silica gel CO hat 0,04-0,063 mm.
2.1.1. Mdu thuc vgt
Mau la Son tra Poilane dugc thu hai tgi rirng qu6c gia Bi Diip, Nui Ba thugc tinh Lam Ddng vdo thang 3 nam 2009, do TS. Nguyen Tiln Hipp - Vien Sinh thai vd Tdi nguyen Sinh vgt - Vien Khoa hgc vd Cong nghe Viet Nam xac djnh ten khoa hgc.
2.1 2. Chiet mdu thuc vgt vd phdn lap cdc chdt
Mdu la (1800 g) dugc say kho, nghien nho vd ngam chiet 3 lan trong hdn hgp metanol/nudc (85/15) d nhiet do phdng. Sau khi cat loai dung moi dudi dp sudt gidm, djch nudc cdn lai dugc chiet lan luot vdi cac dung moi »-hexan, etyl axetat, //-butanol. Cat loai dung mdi dudi dp suat giam thu dugc cac can djch chiet tuong irng voi khdi lugng Id 12, llOva I30g.
Can chiet «-hexan (9,4 g) dugc sac ky tren cot silica gel, dung mdi «-hexan/EtOAc voi ti le EtOAc tang dan tir 2-100% thu dugc 12 phan dogn. Phan doan EPH5/12 (157 mg) dugc tinh che lai qua cgt silica gel vdi n-hexan/EtOAc = 25/1 thu dugc 15,2 mg chat 1 d dang ran, mau trang; Rf = 0,23 (/;- hexan/EtOAc = 10/1), FT-IR M,m (cm"'): 3423, 2953, 2832, 1115, 1023. ' H - N M R (500 MHz, CDCI3): 5H = 3,65 (2H-2, \,J= 6,6 Hz); 1,56 (2H-3, 2H-4, m); 1,26 (m) vd 0,89 (3H-46, t, 7 = 6,8 Hz).
'^C-NMR (125MHz, CDCI3): 5c-63,1 (C-l, t), 32,8 t; 31,9 t; 29,71; 29,6 t; 29,41; 29,3 t; 25,7 t; 22,7 t va 14,1 (C-46,q).
Can chiet EtOAc (50 g) dugc dua len cot silica gel, giai hdp bdng 77-hexan/EtOAc vdi ti le EtOAc tu 50 din 100%, sau dd bdng hdn hgp EtOAc/MeOH vdi lugng MeOH tir 0 den 50%o thu dugc 20 phan doan. Tiep tyc tinh che phdn dogn 7-8/20 (2,6 g) bdng sdc ky cgt silica gel, dung mdi «-hexan/EtOAc vdi ti le EtOAc tdng dan tir 10-20%. thu dugc 5 phan dogn. Phdn doan gidu chit 2 (218 mg) dugc tiep tue ldm sach ldn lugt bdng cgt sephadex LH-20 (MeOH), silica gel (o-hexan/CHjCb/MeOH 2/3/0,3) cho 20 mg chat 2 d dgng rdn, mau trdng, Rf
TCHH, T.50(2). 2012
= 0.63 (/;-hcxan/ElOAc/Me011 = 1/4/0.2), Bi;n c.inh chat 2, tir can chiet ElOAc mdt lugng Idn axit ursolic vd pomolic da dirge phan Igp |4]. Dc thugn Igi han cho vice phan Igp cdc chat khac trong c^n chiet nay. chiing toi da tim cdch ket linh dc logi bd bdt hai a\ii neu tren (5]. Tir 29 g cgn cliict l.lOAc sau khi ket tinh Ihu dugc 19g can djch dd lo^ii bdt axil ursolic vd pomolic. Tinh che 19 g cgn do biing cgt silica gel. dung mdi »;-hexan/CI LCli/McOl I theo cac li lp cd do phan c^rc tdng dan thu dugc 17 phan dogn. Tinh che phan dtign 15/17 (1.6 g) kin lugt qua cac cgt silica gel (CINCL/l lOAc'McOlI = 3/2/0,1), sephadex I 11-20 (McOl I), thu dugc 31 mg chat 3. Rp
= 0.67 (McOII/HjO = 2/1 tren ban mdng ngirgc pha RP-18). FT-IR: v'„,„ (cm') = 3413. 1728. 3321, 3300-3100.2925.2855, 1628. 1253, 1169, 1067.
3. KET QUA vA THAO LUAN
Pho FT-IR ciia chat 1 cho dinh hiip thy ddc trung Clia nhdm h\droxy d 3423 cm"'. Phd khoi phdn gidi cao HRESI-MS cho biet pic ion phdn tir d m/z
Nguyen Thi Hodng Anh vd cgng sir 663.73831 [M+H]"*, tuang irng vol cdng thirc la (•,f,Hc,.0 (theo tinh loan la 663,73829). Sy phan mdnh trong phd khoi the hipn dac diem rat dgc trimg cho mgt hidrocacbon no qua cac ion phan manh luon mdt mgt nlidm CI I2 hay mgt so nhdm Cl Ii tuong dng vdi 14 don vj khoi lugng hodc bgi so cua 14. Vidu:
III/: 606 JM-Cllt,]' (M-56); 578 [M-CftHpf (M-84); 522 |M-C|„II;o]' (M-140),.... Su co mat nhdm h_\ilroxy trong phdn tir ciing dugc the hipn vk rd qua ion phdn mdnh tai ;;;/- 646 [M-OH+H]' (Ni- le). Pho '11- va '-'C-NMR khdng djnh chat 1 Id ancol beo mgch cldi, the hipn qua pic ciia rdt nhicu nhom CH: cgng hudng chong chgp aSu = 1,26; (5c= 29,63;
mgt nhdm mctyl cua mgch hidrocacbon no d S\\ = 0.89 (\.3\\).Sc= 14,12; mgt nhdm CILOII d tS,, = 3.65 (t, 211), Sc = 63.13; hai nhdm metylen ben canh nhom CII2OII df),i= 1,56 (4H), 4- = 32,85; 31,94.
Dya vao cdc tin hipu phd dd dugc phdn tich dtren ket hgp so sdnh vdi so lieu trong tai lipu (6), cau triic chat 1 dugc xac djnh Id H-hexatetracontan-l-ol. Ch5t nay da dugc phdn lap trudc ddy lir lodi Erylhrina sigmoidea [6].
1: H-hexatetracontan-l-ol
HO,, 2 / ^
2: axit arjunic
HO ^ " O'" "OMe OH
3: catiguanin B
Phd khdi phdn gidi cao HR-ESI-MS ciia chat 2 cd pic ion phan tir tgi m/z 511,33940 [M+Na]\
tuong irng vdi cong thiic C3oH4805Na (theo tinh toan Id 511,33994). So lipu pho khdi, ket hgp vdi phd cgng hudng lir hat nhdn cho phep du dodn chat 2 Id mgt tritecpen axit. Pho ' H - vd '^C-NMR cho bilt phdn tir chdt 2 cd chira 7 nhdm mctyl bac 3 d <Ji( = 0,79; 0,84; 0,96; 0,99; 1,02; 1,04; 1,31 (moi tin hipu 3H,s), I noi ddi dang >C=CH-[(5i[ = 5,30(lH,t,y = 3,4 (H-12); Sc = 124,73 (C-12), 144,70 (C-13)]; 1 nhdm -COOH d <5b = 182,65. Ngoai ra, sy cd mat
Clia 3 nhdm CH-OH cung dugc the hipn qua cdc tin hipu d £5ii = 3,63 (m, H-2); 3,28 (d, J = 3,7, H-19);
2,94 (d, J = 9,6, H-3) vd <5b = 69,53 (C-2); 82,48 (C;
19); 84,55 (C-3). Ben canh cac tin hipu kl tren, pho '•'C-NMR, DEPT cdn cho thay sg cd mat cua 8 nhdm CH2, 3 nhdm CH cung nhu 6 cacbon bgc 4 trong phan tir. Vdi cdc so lieu da neu tren va cac tuong tdc trong pho 2D-NMR (HSQC, HMBC COSY), ket hgp so sanh vdi cac so lipu da cong b6 [7], cdu true chat 2 dugc xdc dinh la axit 2a,3p,19o- trihydroxy-olean-12-en-28-oic hay axit arjunic. Hop 202
TCHH, T. 50(2), 2012 Nghien ciru thdnh phdn hda hgc...
Sfl«g7:Sdlieuphd 'H-va '^C-NMR ciia chdt 2,3(500 vd 125 MHz, MeOD) vd cdc so lieu trong tdi lieu Vjtri
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
2
*
3,63 (m)
-
2,94 (d. 9.6)
-
5.30 (t. 3.4)
3,28 (d, 3,7)
1,04 (s) 0,84 (s) 1,02 (s) 0,79 (s) 1,31 (s) 0,96 (s) 0,99 (s)
*
47,98 t 69,53 d 84,55d 39.39 s 56.82 d 19.70 1 33,91 t 39,39 s 47,98 d 40,50 s 24.871 124.73 d 144,69 s 42.64 s 28.64 t 24,87 t 46.71 s 45.18 d 82.48 d 36,02 s 29,42 t 34,01 t 27,10 q 17,38 q 16,95 q 17,77q 29,27 q 182,65 s 29,27 q 25,07 q
Axit arjunic 5:
(CDCI,) [7]
47,2 67,0 82.2 38.9 54,9 16,9 32,5 40,0 47,2 38,6 23.2 122.0 143.0 41.9 28,4 24.0 46.6 43,1 80,1 36,9 28,5 34,8 27,1 16,8 16,1 17,1 28,7 179,2 28,8 24,5
Vitri 2 3 4 5 6 7 8 4a Sa 1' 2' y 4' 5' 6' V 2"
y 4 "
5 "
6 "
9 10 COOMe
3 Sa 4,92 (s) 4,25 (m)
2,82 (dd, 17,0; 2,0)
2,92 (dd, 17,0,3.6)
6,12(s)
-
7.00 (d. 1.5) 6.79 (d, 8,2)
-
6,86 (dd, 1,5; 8,2)
6,47 (s)
- -
6,62 (s) 4,48 (dd, 3,0; 9,0)
2,50 (dd,9,0; 14,7)
3,60 (s)
*
80,07 d 66,83 d 29,49 t 156,49 s 95,82 d 152,82 s 104,36 s 103,05 s 153,39 s 132,08 s 115,18d 145,80 s 145,95 s 115,94 d 119,20 d 115,11 s 146,08 s 104,43 d 146,41 s 142,10 s 115,94 d 31,15d 44,78 t 174,65 s 52,03 q
Catiguanin B
ft-
(MeOD) [9]
80,1 66,9 29,5 156,5 95,8 152,9 104.5 103,0 153,4 132,1 115,1 146,1 146,0 116,0 119,2 115,9 146,4 104,4 145,8 142,1 115,2 31,2 44,8 174,7 52,0
chdt nay da dugc phan lap trudc day tir cdy Terminalia arjuna vd the hipn hogt tinh ddc vdi cdc dong te bao ung thu d ngudi: bieu mo (KB), buong trirng (PA-1) va ung thu gan (HepG-2 vd WRL-68), dac bipt la ddng HepG-2 vdi IC50 = 0,7 ^ig/mL [7], Dong phdn lap the cua axil arjunic (axit 2a,3a,19a- trihydroxy-oiean-12-en-28-oic) dd dugc tim thay trong loai Eriobotrya deflexa [8].
Pho IR Clia chat 3 cho cdc dinh hap thu d 3413 (hydroxyl), 1728 cm"' (cacbonyl). Cdng thirc phdn tir Id C25H22O10 dugc nit ra tiir pic ion phan tu d m/z = 505,11054 [M+Na]^ (CisHzjOjoNa, theo tinh todn Id 505,11107) trong pho khdi phdn gidi cao HR-ESIMS.
Pho 'H-; '^C-NMR va DEPT cho thay chdt 3 cd chira mdt dan vj epicatechin the hien qua cdc tin hieu ciia vdng C, gdm cd 2 nhdm metin gdn vdi oxi d r^i = 4,92 (IH, s, H-2); 4,25 (IH, m, H-3) vd ^ = 80,07 (C-2);
66,83 (C-3); tin hieu cua nhdm CH2-4 d ^ = 2,82;
2,92 vd ^ = 29,49. Ben cgnh dd, cdc proton thom tuong tac kilu ABX ciia vdng B cung dugc tim thdy trong phd 'H-NMR, gdm cd hai doublet b S^ = 6,79 vd 7,00 vdi hdng sd tuang tac lan lugt Id 8,2 vd 1,5 Hz;
mgt doublet kep d ^, = 6,86 ( 7 - 8 , 2 & 1,5 Hz). Do djch chuyen hda hgc cua 15 nguyen tii cacbon gan gidng epicatechin cimg vdi cac tuong tdc quan sdt thay giua H-2 vdi C- r (132,08 ppm), C-2' (115,18 ppm) va 203
TCHH, T. 50(2), 2012
C-6' (119,20 ppm) trong pho IIMBC khdng dinh sy cd mat cua don vi epicatechin trong phdn lii 3. Ngoai ra, phd NMR cdn cho thdy cac tin hipu ciia mgt nhom metin d<5ii=^ 4.48 (HI. dd. J = 3.0 & 9.0 Hz) va ,i_ "- 31,15, mgt nhdm metylen d <^i = 2.50 (211, dd../= 9,0
& 14.7 Hz) vd ^ = 44.78. mgt \diig benzen bj Ihl 4 lan trong dd cd 2 nhdm Ihl la hydroxy the hipn qua tin hieu singlet ciia 2 proton tlKmi d <^i = 6.47 vd 6.62 va 6 cacbon thorn trong dd cd 2 cacbon djch chuyin vc vung trudng thap d ("v =-- 142.10 \ii 116.41 Tuong tdc trong phd HMBC giiJa 11-9 \oi C-7, C-8. C-l", C-2"
\a C=0 chi ra rang C-9 licn kc! vdi C-8 ciia vdng A va C-l" Clia vdng thom bj the 4 lan. Tuong tdc ciia mgt trong hai proton cu.i nhdm CIL-10 a 2.98 ppm vdi l l - : \ H-6' vd H-6" trong phd ROF.SY chimg minh hda lap the cii.i chat 3 nhu trong hinh \c. Cudi cimg cau triic cli.it 3 dugc xac djnh la catiguanin B khi so sdnh vdi so lieu phd trong tdi lieu [9]. Catiguanin B, mgt dan xuat phenylpropanoit ciia epicatechin mdi dugc phdt hipn trong vd than cay Trlchilia caiigua vdo nam 2007 va nd the hipn hogt tinh chdng oxi hda vdi giatrilCso-2.49fiM[9].
Theo tra ciiu tai lipu ciia chimg tdi thi day la lan dau tien cac chdt ;i-hexatetraconlan-l-ol. axit arjunic va catiguanin B dugc tim thdy trong chi Eriobotrya.
Loi cam on: Cong trinh dugc hodn thdnh vdi sir tdi trg kinh phi cua Quy Phdt irien Khoa hgc vd Cdng nghe Qudc gia cho De tdi nghien ci'nt ca bdn, ma so 104.01.128.09.
TAI LIEU THAM K H A O
1. Pham Hoang Hg Cdy co Viet Nam. Nxb. Tre, quyen 1,776(1999).
^. Nunziatina De Tommasi, Francesco De Simone.
Nguyen Thi Hodng Anh vd cgng sir Cosimo Pizza. Constituents of Eriobotrya japonica.
A study of their antiviral properlies, Joumal of Natural Products, 55(8), 1067-1073 (1992).
Yan Huang, Jun Li, Qi cao, Shi-Chun Yu, Xiong- Wcn Lv, Yong Jin. Anti-oxidalive effect of triterpene acids of Eriobotrya japonica (Thunb) Lindl. leaf in chronic bronchitis rats. Life Sciences, 78(23), 2749- 2757 (2005).
Dinh Gia Thi§n, Triln Van Chien, Nguyen Thj Hoang Anh. Tr^n VSn Sung, Nghien ciru thdnh phan hoa hgc Id cdy San Ird Poilane (Eriobotrya poilanei J E.
i nl). hg Hoa hdng (Rosaceae) - Phdn I. Triterpen khung ursan. Tgp chi I Ida hgc, 49(2), 223-226 (2011).
Dinh Gia Thi^n, Nguyen Thj Hodng Anh. Trin Van Sung 201 I. Nghien cdv thdnh phdn hoa hgc Id cdy San trd Podane (Eriobotrya poilanei J.E. Vid.), hif lloa hdng (Rosaceae) ~ Phdn II Triterpen khung ursan vd otean. Tap chi H6a hgc, 49(3), 362-366 ((2011).
J. C. Ndom, Kouam, J. C. Vardamides, J. D. Wansi, A. W. Kamdem, J. T. Mbafor, Z T. Fomum.
Constituents of Erylhrina sigmoidea. Bull. Chem.
Soc, Ethiop., 15, 151 (2001).
Mohil Saxcna, Uzma Fandi, Rupal Mishra, M. M.
Gupta, M. P. Darokar, S. K. Srivastava, Digvijai Singh, Suaib Luqman. S P. SKhanuja. Cytotoxic Agents from Terminalia arjuna. Plania Medica, 73, 1486-1490(2007).
By Lee, Tzong-Huei, Lee, Shoei-Sheng, Kuo, Yuh- Chi, Chou, Chang-Hung. Monoterpene Glycosides and Triterpene Acids from Eriobotrya deflexa. J. Nat.
Prod, 64(7), 865-869(2001).
Wanxia Tang, Hideaki Hioki, Kenichi Harada, Miwa Kubo, Yoshiyasu Fukuyama. Antioxidant Phcnvlpropanoid-Substiluted Epicatechins from Tncinlia caligua. J. Nat. Prod., 70, 2010-2013 (2007).
Lien h?: Nguyen Thi Hoang Anh Vipn Hda hgc
Vipn Khoa hgc vd Cdng nghp Viet Nam 18 Hoang Qudc Viet, Cau Gidy, Hd Ngi Email: hoanganhvhh(ggmail.com
204
