Tp ch Khoa hc 2012:21a 134-138

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NGHIN CU THNH PHN HA HC TRONG CAO ETHYL ACETATE CA R CAU (ARECA CATECHU L.)
L Thanh Phc v Bnh Nguyn Anh Ho 1

ABSTRACT
This paper presents the results on separating, identifying the chemical composition in Areca catechu L. roots in Phong Dien district, Can Tho city. The results showed that two compounds: lamotrigine (C9H7N5Cl2) and -trehalose (C12H22O11) were isolated from the ethyl acetate extracts. The structures of these compounds have been elucidated by modern spectroscopic methods: ESI-MS, 1H-NMR, 13C-NMR, HSQC, COSY and HMBC. Keywords: Areca catechu L., components, lamotrigine,-trehalose, root Title: Study on the chemical components in ethyl acetate extract from Areca catechu L. root

TM TT
Bi bo trnh by cc kt qu nghin cu v chit, tch, nh danh thnh phn ha hc trong r Cau trng ti huyn Phong in, thnh ph Cn Th. Kt qu: hai hp cht: lamotrigine (C 9 H 7 N 5 Cl2 ) v -trehalose (C 12 H 22 O 11 ) c c lp t cao ethyl acetate. Cu trc ha hc ca cc cht ny c xc nh bng cc loi ph ESI-MS, 1 H-NMR, 13 C-NMR, HSQC, COSY v HMBC. T kha: Areca catechu L., lamotrigine (C9H7N5Cl2) v -trehalose (C12H22O11).

1 T VN Vit Nam, Cau l cy trng lu i rt quen thuc khp ni, nht l vng trung du v ng bng. Cy Cau c tn khoa hc l Areca catechu L., thuc h Cau (Arecaceae) ( Tt Li, 1995). Cc b phn ca cy Cau c dng nhiu trong y hc c truyn cha nhiu bnh nh: ht Cau dng cha bnh sn x mt, sn l, cha vim rut, v qu Cau dng tr thy thng, bng y trng, b tiu tin; r Cau ni c tc dng b dng, cha i nht ( Huy Bch, 2004). Trn th gii, Cau c kt hp mt s nguyn liu thin nhin khc to cht khng oxi ha dng lm m phm, lm thuc chng bnh trm cm, bnh cao huyt p (P. Wetwitayaklunga et al., 2006). Thnh phn ha hc trong cc phn khc nhau ca cy Cau: thn, l, tri, c nhiu tc gi trong v ngoi nc nghin cu. Tuy nhin, vic nghin cu thnh phn ha hc ca r cy Cau c rt t bi bo c cng b (o Hng Cng, 2010). Trong cc nghin cu trc y chng ti c lp v nh danh c hai hp cht: lupeol v lupeol acetate c trong dch chit petroleum ether (L Thanh Phc et al., 2011). tip tc gp phn nghin cu thnh phn ha hc ca r Cau v lm sng t thm c s khoa hc cho cc bi thuc dn gian, chng ti phn lp v xc nh cu trc mt s hp cht t cao ethyl acetate ca r Cau (Areca catechu L.) huyn Phong in, thnh ph Cn Th.
1

Khoa Khoa hc T nhin, Trng i hc Cn Th

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2 PHNG TIN V PHNG PHP 2.1 Nguyn liu R Cau non thu hi ti huyn Phong in, thnh ph Cn Th. R Cau c ra sch, loi b phn su, sy kh nhit 55C n khi lng khng i v xay nh trc khi s dng. m trung bnh ca mu r Cau l 8.8%. R Cau c nh danh khoa hc l r ca Areca catechu L. bi Ths. Ng Thanh Phong, B Mn Sinh, Khoa Khoa hc T Nhin, i Hc Cn Th. 2.2 Phng php Chit hot cht: bt r Cau (3500 g) c chit ngm kit vi ethanol 96 (EtOH) trong 7 ngy, tch ly phn lng em c quay di p sut km thu cao EtOH th (305 g). Cho phn cao th ha tan trong mt lng nc ct nht nh, sau chit lng lng ln lt vi cc dung mi petroleum ether (PE), ethyl acetate (EtOAc), n-butanol (BuOH). Thu gom cc dch trch v sau khi loi dung mi di p sut km thu c cc cao PE (59 g), cao EtOAc (49 g) v cao BuOH (56.5 g), tng ng. Phn lp cc cht t cao ethyl acetate: thc hin sc k ct, cht hp ph l silica gel, s dng nhng h dung mi gii ly gm PE, EtOAc v methanol (MeOH) c phn cc tng dn. Theo di qu trnh sc k ct bng sc k bn mng vi h dung mi gii ly l chloroform:methanol (CHCl3:MeOH), thuc th hin vt l dung dch acid sulfuric 10% trong ethanol hoc dung dch KMnO4. Cc phn on th hin Rf ging nhau trn sc k bn mng c gp li. Tin hnh sc k ct ln 2 vi cc phn on ging nhau c lp c cht sch. Xc nh cu trc ca cc cht c lp c bng cch s dng cc phng php ph nghim: MS, 1H-NMR, 13C-NMR, COSY, HMBC. Ph NMR c o trn my Bruker Advance 500 MHz (Vin Cng Ngh, 18 Hong Quc Vit, Cu Giy, H Ni). Silica gel 60 F254 (0.04-0.05 mm), bn mng TLC ca hng Merck, c. Cc ha cht tinh khit khc c xut x t Trung Quc. 3 KT QU V THO LUN 3.1 Kt qu sc k ct T cao EtOAc (9.023 g), tin hnh sc k ct thng vi h dung mi gii ly PE:EtOAc v EtOAc:MeOH c phn cc tng dn. Kim tra cc phn on bng TLC. Cc phn on c Rf ging nhau c gom chung. Kt qu phn on EtOAc:MeOH = 95:5 thu c 50 mg cht sch di dng tinh th mu trng, TLC gii ly bng h CHCl3:MeOH:NH4OH 85:10:5 hin hnh bng thuc th KMnO4 cho vt trn mu trng c Rf 0.63 k hiu tinh th ny l PhuocHao4H. phn on EtOAc:MeOH = 1:2 thu c 25 mg cht sch di dng tinh th mu trng, TLC gii ly bng h CHCl3:MeOH= 4:6 hin hnh bng thuc th

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H2SO4 10% trong EtOH cho vt trn mu c Rf 0.48 k hiu tinh th ny l Phuoc-Hao6. 3.2 Kt qu d liu ph Hp cht Phuoc-Hao4H Ph 1H-NMR (500 Mz, DMSO, ppm, J = Hz): trong phn t Phuoc-Hao4H c 7 tn hiu proton, trong c 3 tn hiu ca cc proton gn trn vng benzen cc v tr 7.71 (dd, J = 8; 1.5, Ha), 7.44 (t, J = 8; 7.5, Hb), 7.36 (dd, J = 7.5; 1.5, Hc). Tn hiu cng hng 7.44 ppm dng triplet nhng li c hai gi tr J=8 v 7.5 Hz l v hai proton Ha v Hc khng tng ng v mt ha hc. T hng s ghp cp J, ta thy Ha v Hb v tr ortho vi nhau, Hb v Hc v tr ortho vi nhau, Ha v Hc v tr meta vi nhau. Bn tn hiu proton 6.90 (2H, br s, -NH2), 6.42 (2H, br s, -NH2). Ph 13C-NMR (125.8 MHz, DMSO, ppm) kt hp DEPT cho thy trong phn t Phuoc-Hao4H c 9 carbon trong c 6 carbon t cp. Ph HMBC cho thy s tng quan gia proton Ha vi tn hiu C 131.58, proton Hb vi tn hiu C 136.76, 131.99, proton Hc vi tn hiu C 138.29, 131.58.

NH2 Hc
6

H
1'

N
2'

Cl

3'

Ha

Cl

Hnh 1: Tng quan HMBC ca hp cht Phuoc-Hao4H

Ph ESI-MS: cho bit [M+H]+ = 257.1 amu tng ng vi phn t c khi lng phn t l 256.1 (C9H7N5Cl2). T nhng d kin trn Phuoc-Hao4H c nhn danh l lamotrigine hay cn gi l 6-(2,3-dichlorophenyl)-1,2,4-triazine-3,5-diamine. Kt qu ny ph hp vi kt qu ca Guntoori et al. 2003.. Kt qu o GC-MS dch chit alkaloid t r Cau ca PGS.TS. o Hng Cng, i Hc Nng (2010) cng cho thy trong r Cau c lamotrigine. Lamotrigine c tc dng tr ng kinh, bnh parkinson v tc dng ln knh sodium (Na+) (N. R. Mirza et al., 2005).
4

H 2N

5 6

NH2
6' 1' 5' 4' 3'

N
2

N
1

Cl

2'

Cl

Hnh 2: Cng thc cu to ha hc ca lamotrigine

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Hp cht Phuoc-Hao6 Ph ESI-MS: cho bit [M+H]+ = 361.3 amu tng ng vi phn t c khi lng phn t l 360.3 (C12H22O11.H2O). Ph 1H-NMR (500 MHz, DMSO, ppm, J = Hz): xut hin tn hiu proton anomer 4.87 (1H, d, J = 3.5, H1 v H1), v 5 tn hiu proton gn trn vng ng 3.23 (1H, m, H2 v H2), 3.54 (1H, m, H3 v H3), 3.13 (1H, m, H4 v H4), 3.64 (1H, m, H5 v H5), 3.46 (1H, m, H6a v H6a), 3.54 (1H, m, H6b v H6b). Ph 13C-NMR (125.8 MHz, DMSO, ppm) kt hp vi ph DEPT: cho 1 tn hiu carbon acetal 93.08 (C1 v C1), bn mi tn hiu carbon methin k O cc v tr 70.12 (C4 v C4), 71.6 (C2 v C2), 72.44 (C5 v C5), 72.87 (C3 v C3) v mt carbon metylen k O (-CH2-O) 60.76 (C6 v C6). Ph HSQC cho thy carbon CH2- ti C= 60.76 (C6 v C6) tng quan vi hai proton ti H = 3.46 v H = 3.54 ppm. Chng t hai proton ny ln lt l H6a v H6b. Da vo ph HSQC, ln lt xc nh c dch chuyn ha hc H3, H4, H5. Ph hai chiu HMBC (DMSO, ppm): cho thy proton ti 3.13 (1H, m, H4) tng tc vi carbon ti C = 72.44 (C5), C = 72.87 (C3), C = 60.76 (C6); proton ti 3.23 (1H, m, H2) tng tc vi carbon ti C = 72.44 (C5); proton ti 4.87 (1H, d, J = 3.5, H1) tng tc vi carbon ti C = 72.44 (C5), C = 72.87 (C3). Tn hiu H = 4.87 ppm (1H, d, J = 3.5, H1 v H1) trn ph 1H-NMR v c = 93.08 ppm trn ph 13C-NMR khng nh hp cht Phuoc-Hao6 c cu hnh -pyranoside. Proton ghp cp trc xch o vi H1 phi l H2 H= 3.23 (1H, m, H2). T d liu ph trn kt hp vi ti liu tham kho (Ivanka Jeri et al., 2006), chng ti xc nh Phuoc-Hao6 l [-D-glucose-(11)--D-glucose] hay-D-trehalose. -D-Trehalose c tc dng c ch men -glucosidase nn c ng dng rng trong iu tr bnh i tho ng (Ivanova et al., 2009).

H OH
4 6 5 3

H O
2

HO HO H

H OH 1 O HO H 1' O

H
2' 5'

H
3'

OH OH
6' 4'

H OH

Hnh 3: Cng thc cu to Phuoc-Hao6

4 KT LUN phn lp v xc nh cu trc ca hai hp cht: lamotrigine (C9H7N5Cl2) v -trehalose (C12H22O11) t r Cau. y l nhng cht u c hot tnh sinh hc

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nn gii thch c l do ti sao r Cau c dng cha mt s bnh trong y hc dn gian. TI LIU THAM KHO
o Hng Cng, Bi Ngc Phng Chu, 2010. Nghin cu chit tch, xc nh thnh phn hp cht ha hc trong ht v r cy Cau, Tuyn tp bo co hi ngh Sinh vin nghin cu khoa hc ln th 7, trang 464-468. Huy Bch, 2004. Cy thuc v ng vt lm thuc Vit Nam, NXB Khoa hc v K thut, H ni, (I), tr. 350-353. Tt Li, 1995. Nhng cy thuc v v thuc Vit Nam, NXB Khoa hc v K thut, tr. 172-174. Guntoori et al., 2003. Efficient process for the preparation of lamotrigine and related 3,5-diamino-6-substituted-1,2,4-triazines, United States Patent, US 6586593B1. Ivanka Jeric et al., 2006. Synthesis of Trehalose-centered Dipeptide Esters, Croatica Chemica Acta, 79 (2) 261-272.

Ivanova et al., 2009. Isolation and identification of alpha,alpha-trehalose and glycerol from
an arctic psychrotolerant Streptomyces sp. SB9 and their possible role in the strain's survival, Prep Biochem Biotechnol 39(1):46-56. L Thanh Phc, Bnh Nguyn Anh Ho, 2011. Gp phn kho st thnh phn ha hc ca r Cau (Areca catechu L.). Tp ch Khoa hc i hc Cn Th, 19b, 80-84. N. R. Mirza et al., 2005. Lamotrigine has an anxiolytic like profile in the rat conditioned emotional response test of anxiety: a potential role for sodium channels, Psychopharmacology 180: 159168.

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