hesperidin trong huylt tuang chupt sau khi diing ndy cho cac nghien ciiu tilp theo v l dupc dpng thuoc dudng udng tgi cac thai diem khac nhau, hpc ciia thudc, dupc lieu hay cdc bdi thuoc y hpc dong thdi qua dd xay dyng dudng cong nong dp- co truyen nhdm ddng gdp cac thdng tin quan thdi gian vd ddnh gia dupc cdc thdng s6 dupc trpng cho viee nghien ciiu ve phdn lilu, bao ehi dpng hpc ciia hesperidin dudng udng tren ddi vd cdch su dpng dupc lieu hay thudc y hpc ed tupng chupt nhdt trdng trudmg thanh. Nghien ciiu truyin an toan, hpp ly va hieu qud.
Tk\ li|u tham khdo
1. BO Y tl (2010), Duipc diin Vi^t Nam IV. NXB Y hpc. 2. Phirong Thi|n Thuong (2012). Nghien cuu qui trinh cong nghe chiet xudt citrojlavonoid ttr vo qud ciia mpt so lodi thupc chi Citrus, hq Rutaceae. BSo cao t6ng k^t d i Iii cip So Y Khoa hgc vi Cflng nghe H i Ngi. 3. Rizza S et al (2011), Citrus polyphenol hesperidin stimulates production of nitric oxide in endothelial cells while improving endothelial fiinction and reducing inflammatory markers in patients with metabolic syndrome. Clinical Endocrinol Metabolism Journal 96(5), 782-792. 4. Anand AZ et al (2014), Polyphenols in Human Health and Disease: Chapter 76 - Cardiovascular Effects of hesperidin: A Flavanone Glycoside. Academic Press 2, 989 - 992. 5.
Leandro CS et al (2013), Evidence for the involvement of the serotonergic 5-HTIA receptors in the antidepressant-like effect caused by hesperidin in mice. Progress in Neuro-Psychopharmacology and Biological Psychiatry 40, 103 - 109. 6.
Milenkovic D et al (2011), Hesperidin displays relevant role in the nutrigcnomic effect of orange juice on blood leukocytes in human volunteers: a randomized controlled cross-over study. /'LoSOA'£6(ll), e2666. 7, Morand C ef fl/(2011), Hesperidin contributes to the vascular protective effects of orange juice: a randomized crossover study in healthy volunteers. American Journal of Clinical Nutrition 93, 73 - 80. 8. Tong L. et al (2012), Simultaneous determination of naringin, hesperidin, neohesperidin, naringenin and hesperetin of Fruclus auranlii extract in rat plasma by liquid chromatography tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis 58, 58 - 64. 9. Sirlak M et al (2010), Micronized purified flavonoid fraction in pretreating CABG patients. Texas Heart Institute Journal 2il{2), 1 7 2 - 177. 10. Sampath Cera/(2012), Pharmacokinetics of valerenic acid in rats after intravenous and oral administrations. Planta Medica 78, 575-581.11. BQ Y tfi (2011), Duvc ldm sdng. NXB Y hpc, trang 2 4 - 3 5 . 12. T? Thj Thao (2010), Gido trinh mon hoc thong ke trong hda phdn tich. Tru&ng D^i hgc Khoa hgc Ty nhien - D^i hgc QGHN H^ NOi. 13. Xiao J et al (2011), Molecular propeny-binding affinity relationship of flavonoids for common rat plasma proteins in vitro Biochimie 93(2), 134 - 140. 14. Fang T et al (2006), A rapid LC/MS/MS quantitation assay for naringin and its two metabolites in rats plasma. Journal of Pharmaceutical and Biomedical Analysis 40, 454 - 459.
Tgp chi Dirge li^u, tap 19, s6 4/2014 (Trdng 231 - 234)
KHAO SAT THANH PHAN HOA HOC TREN CAO CHIET CLOROFORM -
C A Y BORRERIA ALATA (AUBL.) DC.
To Cdm Loan', Phgm Nguyin Kim Tuyin\ Nguyin Kim Phi Phun^\ Tir Biic Dun^' ' Dgi hQc An Giang, ^Dgi hgc Sdi Gon.
^ Dgi hpc Khoa hpc tu nhien, Dgi hgc Qudc gia TP HCM
*Email: [email protected] hodc [email protected] (Nhan bdi ngdy 20 thdng 5 nam 2014)
T6m ik
Borreria alata (Aubl.) DC. (hay Spermacoce alata Aubt., Borreria latifolia K. Schum.) la cay pho bien d viing nhi?t ddi v4 cfin nhi$I Chau MJ, ChSu Phi, Chau Au v^ Chau A. CJ Vi^t Nam, cay Borreria alata chi dir<?c diing lim cay phu dat \i chira c6 nghien ciiii nSo v^ thinh ph4n hda hgc ciJng nhu ho^t tinh sinh hgc. Tit dich chiel cloroform ciia cSy Borreria alata, chiing tdi dfl ph9n ISp dupc 3 h(Fp ch^t: Acid ursolic (1), acid olcanolic (2) va kaempfcrol (3). Cau injc cua cic hgp ch4t dugc xdc dinh thdng qua phi 'H-, '^C-NMR vft so s ^ h vdri cic tii Ii?u tham khao.
Til' khfla: Rubiaceac. Borreria alata. Acid ursolic, Acid oleanolic. Kaempferol.
Tgp chi Dugc liiu. tdp 19, sd'4/2014 231
Summary
Chemical Constituents of Chloroform Extract of Bon-eriai)toa(AubL) DC.
Borreria alata (Aubl.) DC. (or Spermacoce alata Aubl., Borreria latifolia K. Schum.) is a species widely distributed in tropical and subtropical regions around the world. In Viemam, it is only planted for ground cover, and no chemical and biological studies on this plant have been reported so far. A phytochemical investigation of chloroform soluble extract of A alata ted to isolate three known compounds, including ursolic acid 1, oleanolic acid 2 and kaempferol 3. Their structures were identified by comparing their NMR data with those in literatures.
Keywords: Rubiaceae, Borreria alata, Oleanolic acid. Ursolic acid. Kaempferol.
1. Dat van de
Chi Borreria thupc hp Cd phe vd cd khodng 150 loai mpc phd biln d cdc vung nhi?t ddi vd can nhipt. Cac nghien ciiu trudc day da xdc nhan rdng djch chiet tii cdc loai thupc chi Borreria cung nhu hpp chdt phdn lap ciia nd cd ho^it tinh sinh hpc da dang, bao gdm chdng viem, khdng u, khang khudn, chdng oxy hda, chdng loet...[8]
O Nepal, re cdy Borreria alata dupc dung de chiia b?nh sdt ret.[10] Tren the gidi chi mdi cd cay Borreria alata thu hai d Indonesia da dupc nghien ciiu, phdn lap dupc 7 hpp chat iridoid glycosid va mpt hpp chdt diterpenoid [7]. 0 Vi?t Nam, Borreria alata dupc xem la mpt loai cd dai va dupc dung de lam phan xanh, thiic an gia sue va chua co nghien ciiu ndo ve thanh phan hda hpc ciing nhu dupc tinh.
Do dd, cdy B. alata (Aubl.) DC, dupc thu hai tai huypn Di Linh, tinh Ldm Dong, da dupc chpn de lam ddi tupng nghien ciiu. Trong bai viet nay, chiing toi thdng bdo phan l$p va xae djnh cdu tnic
Hinh I. Cay Borreria alata (Aubl.) DC dirge thu hii tai Di Linh -Lam Dflng
hda hpc cua 3 hpp chat: Acid ursolic 1, Acid oleanolic 2 va kaempferol 3. Day la nhung hpp chat cd boat tinh sinh hpc cao dang dupc cac nha khoa hpc nghien cuu iing dung trong ITnh vyc y hpc vd dupc hpc [3],[4],[5],[6]. Cdu true ciia cac hop chat 1-3 dupc xdc djnh thdng qua pho NMR va so sdnh vdi eac tai lieu da cdng bd.
2. Doi tuong va phirong phap nghien ciru 2.1. Doi tU0ig
Cay Borreria alata, dupc djnh lodi bdi nha thuc vat hpc Vo Vdn Chi, co ten khoa hpc la Borreria alata (Aubl.) EX:., ho Ca phe (Rubiaceae).
Mdt mau cay ep khd ky hipu sd US-C031, dupc lim trong quyin luu giii tieu ban thyc vdt t?i b?
mdn Hda hiru co, Trudng Dai hpc Khoa hpc tv nhien, Thanh Phd Hd Chi Minh.
2.2. Phiromg phap nghien cuu
2.2.1. Phuang phap phan lap cdc hpp chat; Sit dyng cac phuang phap nhu phuang phap ngam ddm, trich tdng-ldng, sdc ki cpt silica gel, sdc 1^
Idp mdng... vdi cdc dung mdi: n-hexan, chlorofofffi,
232 Tgp chi Dugc lieu, tap 19, so 4/2014
ethyl acetat, methanol...
2.2.2. Phuong phdp xae dinh cdu true cac hgp chat: Pho ' H - va "C-NMR duoc ghi tren may cpng hucmg tir hat nhan Brulcer Avance 500, tSn so 500 MHz doi vcri pha ' H - N M R va 125 MHz doi vcri pho "C-NMR. Nhiet dp n6ng chay duoc ghi tren may Polythemi A Heiztisch Mikioskop (Truong Dai hoc Khoa hgc Tu nhien Tp. HCM).
2.3. Chiit xudt vaphdn lap
Cay tuoi sau khi thu hai dugc loai bo cac la sau b^nh, vang ua roi dem rira sach, phot kho va xay nhuyin thanh bpt. B^t kho ciia cay (6,0 kg) dugc trich ki?t bSng phucmg phap ngam d ^ vai dung moi metanol a nhiet dg SO^C (2 ngay/lan x 10 lan). Loc phan dich trich, co quay duoi ap suat th4p d 40 °C va thu hoi dung moi. Thuc hien nhieu lin, thu dugc cao metanol (290,0 g). Cao metanol dugc trich long-long bang binh long vai cac don dung moi vdi dg phan cue tang dan: n- hexan, chloroform, ethyl acetat va methanol.
Dung djch giai li dugc co quay thu hoi dung moi duoi ap suat thap, ket qua thu dugc 4 loai cao phan do^n tuong irng: cao n-hexan (120,5 g), cao cloroform (15,3 g), cao ethyl acetat (20,0 g) va cao nu6c (120,5 g). Cao chloroform dugc sac ky cgt v6i he dung moi tang dSn n-hexan : ethyl acetat (10:0 den 0:10) thu dugc cac phan doan Cl (2,5 g), 02 (3,7 g), C3 (3,2 g), C4 (2,0 g) ya CS (2,1 g). Phan doan C l dugc sdc ky cgt nhi6u lan v6i he dung moi cloroform : metanol (100:0 den 99:1) thu dugc cac hgp chat 1 (10,0 mg) va 2 (15,0 mg). Phan doan C5 dugc sac ky cgt nhieu lan voi he dung moi cloroform : metanol (100:0 din 90:10) thu dugc hgp chit 3 (3,0 mg) (Hinh 1)
3. Ket qua nghien ciiu
Wxff chat 1 (acid ursolic). Chat ran, dang bgt, miu trjng, dilm nong chay 288 °C (ket tinh trong methanol). 'H-NMR (500 MHz, DMSO-rf^): 6„
ppm 3,00 (IH; dd; 10,0; 5,0; H-3 ); 5,09 (IH; t;
3,0; H-12); 2,12 (IH; d; 8,5; H-18); 0,90 (3H; s:
H-23); 0,67 (3H; s; H-24); 0,86 (3H; s; H-25);
0,75 (3H; s; H-26); 1,03 (3H; s; H-27); 0,80 (3H;
d; 6,5; H-29); 0,90 (3H; d; 6,5; H-30), "C-NMR (125 MHz, DMSO-ds): 5c ppm 38,3 (C-l); 27,0 (C-2); 76,8 (C-3); 38,3(C-4); 54,6 (C-5); 18,0 (C- 6); 30,4 (C-7); 39,1 (C-8); 47,1 (C-9); 36,5 (C- 10); 24,0 (C-11); 124,1 (C-12); 138,7 (C-13);
41,7 (C-14); 32,8 (C-15); 22,9 (C-16); 46,8 (C- 17); 52,6 (C-18); 38,4 (C-19); 38,6 (C-20); 27,7 (C-21); 36,6 (C-22); 28,3 (C-23); 17,1 (C-24);
16.1 (C-25); 15,2 (C-26); 23,3 (C-27); 178,9 (C- 28); 17,1 (C-29); 21,2 (C-30).
Hgp chat 2 (acid oleanolic). Chat ran, dang bgt, mau tring, dilm nong chay 310 °C (kit tinh trong methanol). 'H-NMR (500 MHz, CDClj) 5„
ppm 3,22 (IH; dd; 11,0; 4,0; H-3 ); 5,28 (IH; /;
3,5; H-12); 2,82 (IH; dd; 13,5; 4,5; H-18); 0,99 (3H; s; H-23); 0,76 (3H; s; H-24); 0,91 (3H; s; H- 25) 0,78 (3H; s; H-26); 1,13 (3H; s; H-27); 0,93 (3H; s; H-29); 0,90 (3H; s; H-30). "C-NMR (125 MHz, CDClj) 5c ppm 38,6 (C-l); 27,4 (C-2);
79.2 (C-3); 38,9 (C-4); 55,4 (C-5); 18,5 (C-6);
32,9 (C-7); 39,5 (C-8); 47,8 (C-9); 37,3 (C-10);
23,2 (C-11); 122,9 (C-12); 143,8 (C-13); 41,8 (C- 14); 27,9 (C-15); 23,6 (C-16); 46,7 (C-17); 41,2 (C-18); 46,1 (C-19); 30,8 (C-20); 34,0 (C-21);
32,7 (C-22); 28,3 (C-23); 15,7 (C-24); 15,5 (C- 25); 17,3 (C-26); 26,1 (C-27); 182,9 (C-28); 33,2 (C-29); 23,7 (C-30).
Hgp chat 3 (kaempferol). Chit rin, dang bgt, mau vang, diem nong chay 274 "C (ket tinh trong chloroform). 'H-NMR (500 MHz, CD3OD) 6„
ppm 6,19 (IH; d; 2,0; H-6); 6,40 (IH; d: 2,0; H- 8); 8,09 (2H; d; 9,0; H-27H-6'); 6,91 (2H; d; 9,0;
H-37H-5'). "C-NMR (125 MHz, melanol-rf.) 6c ppm 148,3 (C-2); 137,0 (C-3); 177,4 (C-4); 162,3 (C-5); 99,3 (C-6); 165,3 (C-7); 94,5 (C-8); 158,2 (C-9); 104,5 (C-10); 123,7 (C-l'); 130,7 (C-2'/C- 6'); 116,3 (C-3'/C-5'); 160,4 (C-4').
4. Ban luan
Hop cbat 1 (acid ursolic): Phi "C-NMR xuit hien 30 tin hieu nguyen tir carbon. Trong d6, mgt carbon carboxyl xuit hien viing tir trudng thip 5c 178,9 (-COOH) va cap tin hieu djc trung
Tfp chi Duac lieu, tdp 19, so 4/2014 233
cua ndi ddi dang >C=CH- cua hpp chat cd khung ursan xuat hien d 5c 138,7 (C-13) va 124,1 (C- 12). So sanh dG li?u phd cua 1 vdi dii Ii8u phd cua ursolic acid [9] thdy cd su tucmg dong. Vay, hop chdt 1 dupc xdc dinh la acid ursolic.
Hgp chat 2 (acid oleanolic): Tuang tu nhu 1, phd '^C-NMR cua 2 ciing xudt hien 30 tin hieu nguyen tCr carbon. Trong dd, mpt carbon carboxyl xudt hien vimg trudng thap 8c 182,9 (-COOH) va cap tin hieu dac trung cua ndi ddi dang >C=CH- cua hpp chat cd khung olean xudt hien d 5c 143,8 (C-13) va 122,9 (C-12). So sanh dCr lieu phd cua 2 vdi dtr lieu phd cua acid oleanolic [2] thdy cd su tuang ddng. Chinh vi the, hpp chdt 2 dupc xae djnh la acid oleanolic.
Hop chat 3 (kaempferol): Phd '^C-NMR chi ra cdc tin hieu dac trung cua khung flavonol.
Trong dd, mdt carbon carbonyl xudt hien tai 5c 177,4 (C-4). Mdt khdc, phd ' H - N M R tdn tai 2 tin hieu proton nhdn tham vi tri meta- a 5H 6,40 (1H;
d; 2,0 Hz; H-8); 6,19 (IH; d; 2,0 Hz; H-6) va 2 cap tin hipu proton nhan tham vi tri orto- a 5H 8,09 (2H; d; 9,0 Hz; H-2V6'); 6,91 (2H; d; 9,0 Hz; H-3V5'). So sdnh dii li?u phd cua 3 vdi dir lipu pho cua kaempferol [11] thay cd sy tuong ddng. Vdy, hpp chdt 3 dupc xdc dinh Id
kaempferol.
5. Ket luan
Tir todn cdy Borreria alata, thu hdi t^ii tinh Lam Ddng, 3 hpp chdt: acid ursolic I, acid oleanolic 2 va kaempferol 3 da dupc phdn lap.
Day la nhung hpp chat cd hogt tinh sinh hpc va phd bien trong hp Cd phe. Trong dd, acid ursolic cd khd nang khang viem, khang virus, chdng oxy hda, tic che su phat trien cua nhieu ddng te bao ung thu d ngudi, nhu cdc ddng te bdo ung thu gan R-HepG2, ung thu bach cdu HL-60, ung thu b^ich huyet Daudi, ung thu da day SNU-I, ung thu bi6u md A431 va HCT15 (ICso 30pM)...;
[3],[6] acid oleanolic da dupc biet den vdi nhieu ddc tinh dupc ly cd Igi cho sire khde con nguai nhu khang viem, khang khudn, chdng loet, chSng oxy hda, chong khdi u, ha dudng huyet...;[5],[6]
kaempferol cd kha ndng chdng viem, khang khudn, chdng oxy hda, chdng di ling, bao v? tim mach[4]... Tuy nhien, day la lan ddu tien 3 hpp chat nay dupc phdn lap tCr loai Borreria alata, trong dd kaempferol ldn ddu tien dupc tim thay cd su hien dien trong chi Borreria. Hi^n nay, cae nghien ciru ve thdnh phdn hda hpc vdn dang dupc ti6n hanh tren cac cao cdn Igi cua cay Borreria alata cung nhu dupc tinh cCia cay.
Thi li|u tham khio
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Secondary Metabolites from Schejflera odorata Blanco. Philippine Journal of Science. 134(1), 63-67.3. Zhang D, Tang PM, Chan JY, Lam H, Au SW, Kong S, Tsui SK, Waye MM, Mak TC, Fung K (2007). Anti-proliferative effect of ursolic acid on multidrug resistant hepatoma cells R-HepG2 by apoptosis induction. Cancer Biology & Therapy, 6 (9), 1381-1389. 4.
Calder6n-Montano JM, Burgos-Moron E, P^rez-Guerrero C, Lopez-L^aro M (2011). A Review on the dietary flavonoid kaempferol. Medicinal Chemistry, 11, 298-344. 5. Martinez-Gonzdlez J, Rodriguez-Rodriguez R, Gonz^lez-Diez M, Rodiigura C, Herrera MD, Ruiz-Gutierrez V, Badimon L (2008) Oleanolic acid induces prostacyclin release in human vascular smooth muscle cells through a Cyclooxygenase-2-dependent mechanism. Journal of Nulrilion, 138 (3), 443-448. 6. Li L, Quo W, Qlng-Yao Y. (2002). Effects of ursolic acid and oleanolic acid on human colon carcinoma cell lineHCTl. World Journal i^
Gastroenterol, 8 (3), 493-495. 7. Kamiya K, Fujita Y, Saiki Y, Hanani E, Mansur U, Satake T. (2002). Studies on the constituents of Indonesian Borreria latifolia. Helerocycles, 56, 537-544. 8. Maria Conserva M, Costa Ferreira J, Junior (2012). Borreria and Spermacoce species (Rubiaceae): A review of their ethnomedicinal properties, chemical constituents.
and biological activities. Pharmacognosy Review, 6(11), 46-55. 9. Silval MG, Vieira LG, Mendes F>J, Albuquerque IU Santos RN, Silva FO, Morals SM (2008). Variation of ursolic acid content in eight Ocimum species from Northeastern Brazil.
Molecules, 13, 2482-2487. 10. Manandlar NP (1995). An inventory of some vegetable drag resources of Makawanpw district Nepal Fitoterapia, 66, 2i\-23S. U . Panichayupakaranant P. Kaewsuwan S. (2004). Bloassay-guided isolation of ih*
antioxidant constituent from Cassia alata L. leaves. Songklanakarin Journal of Science and Technology, 26 {I), 103-107.
234 Tfp chi Duac lieu, tap 19, so 412014
