Nghim Th Hng v cs

Tp ch KHOA HC & CNG NGH

65(03): 103 - 107

MT S THNH PHN HA HC C TRONG CY HN THE (DESMDIUM HETEROPHYLUM) H CNH BM (PAPILIONACEAE)

Nghim Th Hng, Phm Vn Thnh*

Trng i hc S phm - H Thi Nguyn TM TT Cy hn the c Y hc dn tc dng lm thuc Nam cha cc bnh cm st, si thn, tiu vim rt hiu qu. Nghin cu v ho hc thc vt hn the (Desmodium heterophyllum) cho thy trong cy hn the cha 7 nhm h p cht thin nhin c hot tnh sinh h c cao l: cc ng kh, ancaloit, steroit, flavonoit, cumarin, poliphenol v cc saponin. T dch chit bng n-hexan tch c steroit v glucozit ca n l stigmatsterol v 3-O--D- glucopyranozyl stigmasterol, cn t dch chit bng CHCl3 tch c -sitosterol-3--D-glucopyranozit. Trong dch chit bng etyl axetat tch c mt flavonoit. Cu trc ho hc ca n c xc nh bng cc phng php ph hin i nh IR, MS, 1D v 2D-NMR cho bit flavonoit y chnh l 8-C--Dglucopyranozyl apigenin (vitexin). Vitexin l cht c hot tnh sinh hc cao c s dng lm thuc chng vim v hn ch s pht trin ca cc khi u. T kho: Cy hn the, Desmodium heterophyllum, steroit, flavonoit, vitex.

M U Cy hn the l thu c loi cy c mc hoang khp ni, c tn khoa hc l Desmdium heterophylum thuc h Cnh bm Papilionaceae. Trong Y hc dn tc, cy hn the c nhn dn dng nh cy thuc cha nhiu loi bnh: cha bnh c m st nng, ho c m, tiu sng, tiu vim, cha cc bnh i but, si thn, si mt, thng hi, li tiu, ton cy gi nt p ngoi vt thng [1]. C nhiu ng dng nh vy song hu nh nc ta cn cha c cng trnh no nghin cu thnh ph n ho hc c trong cy hn the. Trong khun kh bi bo ny chng ti gii thiu mt s kt qu nghin cu thnh phn ho hc cy hn the . NGUYN LIU, THIT B NGHIN CU Cy hn the thu ton cy cc bi hoang trong khu vc trng i hc S ph m Thi Nguyn vo thng 2 nm 2009 v c B mn phn loi thc vt khoa Sinh xc nh tn khoa hc l Desmdium heterophylum. Thit b nghin cu: im chy c o trn my Electrothermal IA-9200, Ph IR c ghi trn my IMPACT-410 dng vin nn tinh th KBr. Ph MS ghi trn my HP-1100 LS/MS. Trap. Ph 1H-NMR (500 MHz) v 13 C-NMR (125 MHz) c ghi trn my Bruker AM 500 FT-NMR v TMS c dng

lm cht chun ni. Sc k lp mng trng sn trn nhm DC-Alufolien Kieselgel 60 F254. Sc k ct s dng cht nhi Silica gel (Kieselgel 63-230 mesh) PHNG PHP NGHIN CU Mu thc vt ti c dit men bng cch sy nng 1100C trong 10 pht sau sy kh nhit 600C n khi lng khng i. Mu kh c nghin nh thnh bt, ngm chit phn lp bng cc dung mi vi phn cc khc nhau. Cc cht tinh khit c phn lp bng phng php sc k ct vi cc h dung mi thch hp v lm sch bng phng php kt tinh phn on lp li nhiu ln theo s 1. KT QU V THO LUN Trn c s phn tch nh tnh bng cc phn ng mu nh n dng cc nhm hp cht thin nhin c hot tnh sinh hc cao, pht hin th y trong cy hn the c cha 7 nhm cht c hot tnh sinh hc, kt qu c nu ra bng 1.

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Nghim Th Hng v cs

Tp ch KHOA HC & CNG NGH

65(03): 100 - 104

Bng 1: Nhn dng cc nhm cht c hot tnh sinh hc trong cy hn the.
TT 1 2 3 4 5 6 7 8 Nhm cht ng kh Ancaloit Steroit Flavonoit Poliphenol Cumarin Glicozit tim Saponin Thuc th Felinh Dragendooc Libecman - Bocsa Xianiin NH3 c FeCl3 Axit Kim Kelle -Kiliani To bt Hin tng Cho kt ta mu gach Mu vng da cam Mu hng n xanh l cy Mu hng Vng m Mu xanh en Kt ta bng Khng c hin tng gi Bt bn trong mi trng kim Kt qu ++ + ++ ++ ++ ++ ++ ++

Ghi ch: Du (+) l cho phn ng dng tnh, du (++) l phn ng dng tnh rt r, du (-) l khng c phn ng

Cht PH1 Thu c t cn n-hexan ca cy hn the l cht rn dng tinh th hnh kim, khng mu, khi lng 25 mg. Nhit nng chy o c l 156-1580C, Rf = 0,8 trong h dung mi n-hexan etylaxetat (85:15). Ph FT-IR cho php khng nh s c mt ca nhm OH
1

l khc ca ch t PH1 vi stigmasterol ca phng th nghi m [2] chng ti nh n th y chng hon ton tng t nhau. V v y c th quy kt ch t PH1 l stigmasterol (hay stigmast-5,22-dien-24R-3 -ol).

max =

3406 cm-1, cn ph

H-NMR cho H 3 = 3,58 ppm. C 2 ni i

khng lin hp vi

max = 1621 cm-1, cn

Stigmast-5,22-dien-24R-3 -ol (stigmasterol)

trong ph 1H-NMR c 3H cc nhm metin (C-H) lin kt vi cacbon cha no cc ni i trn l H-6 = 5,37ppm v H-22 = 5,15 ppm, J = 15HZ v 7 HZ v H-23 = 5,02 ppm, J = 15 v 7 HZ). So snh cc d liu ph FT-IR, ph 1HNMR, nhi t nng ch y, cc h ng s v t
104

Cht PH2: Ch t PH2 l cht rn v nh hnh, tan c trong etylaxetat, trong metanol, d tan trong hn hp dung mi nhexan - etyl axetat (20:80) c nhi t nng ch y 288-2900C.

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Nghim Th Hng v cs

Tp ch KHOA HC & CNG NGH

65(03): 100 - 104

Thu phn cht PH2 trong dung dch HCl 2N thu c stigmasterol, ph 1H-NMR ca sn ph m sau thu phn khng cn nhng tn hiu ca phn ng v hon ton tng t nh ph ca stigmasterol. Phn dung dch c phn tch bng sc k, so snh vi cc ng mu chun ca ca phng th nghim cho thy tng ng vi ng glucoz. Phn tch ph 1H-NMR ca cht PH2 cng cho thy cc phn r r t tng ng vi ph 1 H-NMR ca stigmasterol v phn ng tng ng vi cc chuyn dch ho hc ca cc H trong phn ng ( H 3 t 3,24 n 4,5 ppm). T cc d kin thc nghim trn chng ti cho rng cht PH2 l 3-O--Dglucopyranozyl stigmasterol Cht PC1: Hp cht PC1 c phn lp trong cn dch clorofom bng sc k ct nhi silicagen, ra gii bng h dung mi etylaxetat : metanol (70-30), thu c 10 mg tinh th dng bt mu xm. Rf = 0,65 trong h dung mi clorofom metanol (30:10), Rf = 0,45 trong h dung mi n-butanol-axit axetic-nc (4:1:5). Nhit nng chy o c l 276-2800C. Phn tch ph 1H-NMR ca cht PC1 thy rt ging vi ph ca cht PH2 tc l ging vi ph ca stigmasterolglucozit ngo i tr khng pht hin th y H =5,15 v H =5,03 ca cc proton lin kt C-H t i cc v tr C 22 v C23, iu ny gi cho bit c th cht PC1 l glycozit ca -sitosterol. Khi thu phn cht PC1 bng dung dch HCl 2N ta thu c cht kt ta m c Rf ln ph 1H-NMR ca n u trng lp vi ph v Rf ca -sitosterol [2]. Phn dung dch thu phn c nhn dng ng glucoz bng phng php sc k giy c so snh vi glucoz ca phng th nghim th y cho cc s liu trng nhau. T kt qu trn c th quy kt cht PC1 l 3-O-D-glucopyranozyt sitosterol. Cht PE1: Hp ch t PE1 thu c di d ng ch t r n v nh hnh mu vng, kh i l ng 12 mg, thu c c n dch etylaxetat khi r a gi i h dung mi etylaxetat: metanol
105

(90:10). Nhit nng chy o c l 271-2740C.

PH2 l 3-O--D- glucopyranozyl stigmasterol.

PC1 l -sitosterol-3--D-glucopyrannozit Ph UV v ph NMR ca PE1 c dng ph ca hp cht flavonoit glycozit. C tn hiu rng singlet cng hng vng trng rt thp ( = 13,15 ppm) gi v s c mt ca nhm OH v tr C-5 (vng A) v tham gia lin kt hidro vi oxi trong nhm C=O v tr C-4 (vng C). Hai tn hiu mnh singlet khc = 6,76 c trng cho proton ni i ti C-3 ca cc flavon. vng B ch c 1 nhm OH v tr C-4 cng c minh chng trong ph 1H-NMR vi tn hiu doublet = 8,0; J = 8,1 Hz (2H c trng cho cc H-2 v H-6). Mt tn hiu doublet na = 6,9 vi J = 8,2 c trng cho proton H-3 v H-5. Proton c tn hiu = 6,26 l ca proton ca H-6. Cc proton ca phn ng c pht hin trong vng tn hiu = 3,4- 4,96. ng ch l proton cacbon anome (C-1) c gi tr cao hn c = 4,68 (J = 0,5 Hz) so vi cc gi tr tng ng trong trng hp O-glycozit. iu ny cho php ngh n lin kt glycozit hp cht PE1 l lin kt C-glycozit. Ph 13C-NMR xut hin tn hiu tng ng ca 15 C thuc khung flavon v 6 cacbon ca

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Nghim Th Hng v cs

Tp ch KHOA HC & CNG NGH

65(03): 100 - 104

mch ng. Nhm cacbonyl c nh n d ng bi tn hiu = 181,88. Vng B c 2 tn hi u cao gp i cc tn hiu khc = 128,69 v = 115,71. chnh l cc tn hiu ca C-2, C-6 v ca C-3, C-5. V tr C-4, c nh n ra vi tn hiu = 160,98. Cn tn hiu ca C-1 = 121,51. Cc gi tr C ca phn t ng l 61,19 (C6); 70,53 (C-4); 70,84 (C-2); 73,38 (C-1); 78,5 (C-3) v 81,62 (C-5) l hon ton ph hp vi cc tn hiu ca ng glucopyranozit c lin kt C-C [4]. Khi so snh cc gi tr ph v tnh cht vt l khc ca cht PE1 vi cht 8-C--Dglucopyranozyl apigenin (Vitexin) ca cc tc gi khc cho kt qu rt ph hp nhau [3,4]
Bng 2: S liu ph 13C-NMR ca PE1
V tr 2 3 4 5 6 7 8 9 10 c (ppm) 163,6 102,8 182,0 160,7 98,09 163,3 104,4 156,3 103,5 s d s s s s d s s H (J,Hz)

Trn ph HSQC v HMBC ca cht PE1 cng cho php quy kt dch chuyn ho hc ca cc v tr C v H trong cng thc cu to gi nh ca cht PE1. Tng tc HMBC ca H-1 vi C-2, C-3, C5, C-6, C-7, C-8 khng nhng khng nh thm cc gi tr dch chuyn ho hc ca cc v tr trn m cn khng nh s to lin kt C-glycozit v tr C-8. Tng tc HMBC ca H-6 ( = 6,26) vi C-5 v C-7; tng tc ca H-3 vi C-4 v C-2 cng cho bit hai nhm OH phi v tr C-5 v C-7 v ni i C-2 vi C-3 T cc lp lun trn v kt qu ca cc ph NMR c th quy kt cho cc v tr nh bng 2

V tr 1 2 3 4 5 6 1 2 3 4 5 6

c (ppm) 121,5 128,7 115,7 160,9 115,7 128,6 73,38 70,8 78,59 70,53 81,62 61,19 s d d d d d

H (J,Hz)

6,78 s 13,155 s (OH) 6,26 s

7,93 (8,3) 6,9 (8,2) 6,9 (8,2) 8,0 (8,2) 4,68 ( 0,5) 3,84 t 3,3 3,4 3,25 3,53 t (5,4) 3,75 d (9,5)

OH OH H H H H OH OH H O

HO

OH

OH

Cht PE1: 8-C--D-glucopyranozyl apigenin (Vitexin)

KT LUN Ln u tin, cy hn the (Desmodium heterophyllum - Papilionaceac), mc hoang

106

ti Thi Nguyn c nghin cu sng lc ha thc vt, thit lp c quy trnh ngm chit mu hp l thu c 4 dch chit chn

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Nghim Th Hng v cs

Tp ch KHOA HC & CNG NGH

65(03): 100 - 104

lc. T ton b phn trn mt t (thn v l) cy hn the (Desmodium heterophyllum Papilionaceac), bng phng php phn tch nh tnh khng nh trong cy hn the t nht c cha ti 7 nhm hp cht thin nhin quan trng l: ng kh, ankaloit, steroit, flavonoit, poliphenol, cumarin v saponin. Ln u tin, phn lp v bng cc phng php quang ph c phn gii cao xc nh c cu trc ha hc ca 4 hp cht c trong cy hn the l Stigmast -5,22-dien24R-3-ol; 3-O--Dglucopyranozyl stigmasterol;-sitosterol-3--Dglucopyranozit v 8-C--D-glucopyranozyl apigenin .

TI LIU THAM KHO

[1] Tt Li (1999), Nhng cy thuc v v thuc Vit Nam, Nxb Y hc, H Ni 1999 [2] Phm Vn Thnh, Bi Vn Bnh - Nghin cu ho hc cy ngn mc hoang ti t i Thi Nguyn, Tp ch Khoa h c v Cng ngh - H Thi Nguyn 2008 (46) ,71-75. [3] Nguyn Th Hng Vn, Phan Vn Kin, Chu Vn Minh... , Cc hp cht flavonoit phn lp t l cy b kt, Tp ch Dc hc s 379 (11/2007) tr. 36-39 [4] Yin.F., Hu.L., Pan.R. novel dammarane-type glycoside from Gynostemma pentaphyllum. Chem. Pharm. Bull. (2004) Vol.52 p.1440 1444.

SOME CHEMICAL COMPOSITION OF DESMDIUM HETEROPHYLUM - PAPILIONACEAE

Nghiem Thi Huong, Pham Van Thinh1

College of Education - Thai Nguyen University

SUMMARY
Desmodium heterophyllum was very effectively treated fever, kidney stones inflammation targets in the medical tradition. Chemical components of the Desmodium heterophyllum shows containing the seven groups of natural compounds with high biological activity: reducing sugar, alkaloids, steroids, flavonoids, coumarin, polyphenol and saponin. From n-Hexane extraction were isolated steroid, stigmatsterol glucosid and 3-O--D-glucopyranozyl stigmasterol, also from CHCl3 extraction were isolated -sitosterol-3--D-glucopyranosid. From ethyl acetate extraction were isolated a flavonoid. Its chemical structure was determined by the modern methods as IR, MS, 1D and 2D-NMR and structure of extracted flavonoid is 8-C--D-glucopyranozyl apigenin (vitexin). Vitexin is biologically active substances have been used as a high anti-inflammatory drugs and limit the growth of tumors. Key words: Welding tree, Desmodium heterophyllum, steroids, flavonoid, vitexin.

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