Y Hpc TP. H o Chi M i n h * Tgp 20 * So 6 * 2016 N g h i e n curu Y hpc
KHAO SAT DAC DIEM VI HOC VA HQP CHAT ALKALOID CUA CAY TO XANH (CASSYTHA^FIUFORAaS L.)
BMt The Vinh*, Trdn Cong Ludn**
T O M T A T
MifC tieu nghien cdu: Gdp pUdn Hiu chuan hda dugc lieu Ta xanh (Cassytha filiformis L.), chdng nham lan v&i Ta hong (Cuscuta sinensis Lamk.), nghiin cieu ndy Hin hdnh khdo sdt dqc dian vi hoc vd hda hgc, dqc biet la nhdm aporphinoid cua Ta xanh.
Doi tugng nghiai cdu: C&y To xanh dugc thu hdi tai tinh Long An, dugc djnh danh b&i phdng Tdi nguyin vd Dugc lieu, Trung tdm Sdm vd Dugc lieu Tp. HCM. Khdo sdt dqc dian thuc v&t hgc bSng kinh hien vi; phan lap vd xdc dinh cau true bang ky thuqt sac kyvdky thudt pho
Kei qud: Tie m&u thuc vdt tuoi cua To xanh, da khdo sdt dugc dqc dian vi hgc, vi phdu; qua sac ky cot silicagel pha thudn da phdn lap vd loc dinh dugc cdu true cita 1 aporphine alkaloid Id cassythin.
Kei lugn: Khdo sdt dugc dqc diem vi hgc vd nh&m diat alkaloid cua To xanh (Cassytha filiformis L.), phan lap va xdc dinh dugc edit trdc ciia 1 aporphin alkaloid Id cassythin.
Titkhoa: To xanh, aporphin alkaloid, cassythin.
ABSTRACT
STUDIES ON MICROSCOPIC CHARACTERISTICS AND THE ALKALOID OF CASSYTHA FILIFORMIS L.
Bui The Vinh, Tran Cong Luan * Y Hoc TP. Ho Chi Minh * Vol. 20 - No 6 - 2016:191 -196 Objectives. This study was developed to reveal a standardizaHon of this plant
Materials and methods: Phytology research was performed by determinaHon the microscopic charaderistics; isolaHon and structural eluddation based on chromatography and spectrum technique.
Results: The botany characteristics, microscopic charaderistics and the strutural eluddation of one aporphine alkaloid were reveal.
Conclusions: The botany characteristics, microscopic charaderisHcs and the strutural eluddation of one aporphine alkaloid named cassythine.
Keywords: Cassytha filifrrmis L, cassythine, aporphinoid alkaloid
DAT VAN DJI duong, mat mo, chan tay yeii moi''''^'. Tuy nhien, ciing co nhiing duoe lieu voi hinh thai To xanh (Cassytha filiformis L.) la mpt cay ^g^^j ^ ^ ^ ^ ^ ^ ^^^ ^^_^g ^ ^ ^-^ ^^^^
thuoc dan gian da dupe dung tir lau va pho ^ ^ ^ j ^ ^ ^ ^ ^ ^ j ^ - ^ . ^^ ^^ ^ ^ ^ ^^ 1^ T^
bien tai nhieu nuoc tren the giai nhu a j^^^g iCuscuta sinensis Lamk.). Vdi muc tieu Indonesia va Chau Phi dimg chua giun san, ky ^^^ ^^^ ^^^^ ^^^^ j^^3 ^^^^ ^^^ j ^ ^ ^ ^ ^ sinh taomg, Trung Quoc dimg chiia vang da a ^^^^^ ^ ^ l | „ ^ ^ To hong; nghien ciiu nay tre em. Nuoc ta. To xanh riing dixoc xem nhu ^^^^ ^anh khao sat dae diem vi hoc va hoa hoc, mot vi thuoc dan gian rat co ich, nuoc sac To ^-^ ^..^ j - ^ ^ ^ aporphinoid a i a To xanh.
xanh dimg lam thuoc bo, chua than hu, liet Trung tam S3m va Dugt lidu, " Truong Dai hoc Tay Do
Tdc gid lien lac: DS. Bui The Vinh DT. 0918 0918 64 Email: buithe vinh@
Nghien cmi Y hpc Y Hpc TP. Ho Chi Minh * T?p 20 * So 6 * 2016 NGUYENIJEU-PHUtlNGHIAPNGHIENajtJ
Nguyen lieu nghien ciru
Cay To xanh (than ban ky sinh tren cay chii) dupe thu hai tai tinh Long An, dupe dinh danh boi phong Tai nguyen va Dupe lieu, Trung tam Sam va Dupe lieu Tp. Ho Chi Minh.
Fhuong phAp
Khdo sdt dqc diem thifc vat hoc
Mo ta hinh thai ngoai dac diem hoa, qua va hat tren mau tuoi. Khao sat dac diem vi phlu cat ngang 6 than tuoi vai thuoc nhupm kep do carmin va luc iod, khao sat dac diem ciia bpt dupe lieu.
Khdo sdt thdnh pftdn alkaloid
Dinh tinh nhom alkaloid bang phan iing hoa hgc va sac ky lop mong: Lam ain 10 g bpt dupe lieu bang NH40H 10%, ngam chief vcd 100 ml CHzCb X 3 Kn. Gop cac dich chief CH2CI2 co giam ap den can. Hoa 1 phan can voi luong HJS041% viia dli fhuc hien phan iing dinh tinh vai cac thuoc fhii alkaloid dac trung:
Dragendorff, Mayer, Bouchardat. Phan can eon lai hoa vol CHzCh thuc hien sac ky lop mong voi he pha dpng CH2CI2: MeOH : NH4OH ( 9 : 1 : 1 gipt), phat hien bang fhuoc fhu Dragendorff.
Chief xuat, phan lap alkaloid: 4,2 kg Bpt To xanh dupe kiem hoa bang dung dich N H 4 0 H 15% den p H 10. Them CH2C12 den khi ngap be mat bpt To xanh. Ngam trong khoang 12 gio, thinh thoang co khuay trpn, Ipe thu dich chiet. BCiem tra sir hien dien ciia alkaloid trong dieh chief bang phucmg phap sac ky lop mong, sau do tien hanh co quay giam ap thu eao chiet, dung moi CH2CI2 thu hoi dupe sii dung de chief tiep ba dupe lieu. Quy trinh lap lai cho den khi dich chief khong con alkaloid.
Cao alkaloid tong ttiu dupe la 380 g. Dung 10 g cao tien hanh sSc ky qua nhieu cot sihca gel pha fhuan vai he dung moi CH2CI2 : MeOH vai dp phan cue tang dan (100:0; 99:1; 98:2 den 95:5). Thu nhan dupe 27 mg chat A ket tinh hinh kim, mau cam nhat.
KET QUA VA BAN LU A N Khao sat dac d i ^ thuc vat hpc Thdn
Dai 1,5 - 2 m hay han, day leo, xoan, than hoi CO ranh dpc, co long nau min, it khi nhin.
Mau vang lue den xanh luc, tiet dien tron, it khi dep, duang kinh trung binh 1,5 - 2 mm, khoang each giua cac long 7 -9 cm, cac nhanh thudng ben mpc fhang goc voi true chinh.
Khong ttiay nhanh cap 2.
Ld
Tieu giam fhanh vay, vay hinh tam giac dai khoang l-l,5mm. Noi than tiep xiic voi cae bo phan cay chii hay vdi cfiinh than cua no co cac giac miit hinh bau due kieh fhudc 1-1,5 mm.
Hoa
Cum hoa dang gie, mpc tren nhanh ben.
Tnic mang hoa dai 1,5-5 cm, mau xanh, eo long min va cac not bi khong, mang khoang 8 hoa, hoa khong cuong. Hoa luong tinh, mau trang, khi chua no co hinh tning co 1,5-2 mm.
Bdi
Co 3 dai phu, 3 la dai mau xanh co it long nau. La dai dinh 6 day, cung ton tai vdi vanh hoa den khi ra qua. Vanh: Co 3 vanh hoa mau trang. Dai va vanh dinh d day. Nhuy due: x ^ fhanh 4 luan sinh, moi luan sinh gom 3 tieu nhuy. Luan sinh ngoai cimg dinh vao earth hoa, luan sinh trong cimg lep. Tieu nhuy khai bang 2 nap. Nhuy cai: Bau noan thupng, mot bau nhuy, mpt voi, d chfnh giiia hoa.
Qud
Gia qua, do dai va vanh khong ning ma dong trudng ciing qua tao thanh bao day, mpng, mau xanh, cd chat nhay. Gia qua hinh cau, kich thudc 5-8 mm, tron nhan. Bao lay qua be ben trong, hinh tning nhpn mpt dau, khi chin mau nau den, vd sii si, kieh ttiuoc khoang 3-4 mm.
H^t
Trong hat ed 2 la mam to, mau xanh luc nh^f, CO 2 ldp vd, bao ngoai la ldp vd qua ciing
Y Hpc TP. Ho Chi Minh * T^p 20 * Sfi 6 * 2016 Nghien cum Y hpc va den nau.
Dac diem cay To xanh qua khao sat d nhieu mlu tfli cd dae tinh cau tao ehung nhu tren, phii hpp vdi cac tai tieu da cong bof^-^'. Tuy nhien, do To xanh la loai song ban ki sinh, do do cd the tuy vao cay chii ma dae diem sinh hpe eiia nd ed the khac di mpt it ve kich thudc va mau sac.
Vi phdu thdn
Than tiet dien tron, ngoai cimg la ldp bieu bi kich thude deu ed cutin kha day, mang nhreu
long che ehd don bao. Md day phim deu 1 - 2 ldp, vimg md mem vd dao chiem khoang 4-6 ldp, trong rai rac cd cac hat luc lap. Npi bi khdng rd, spi tru bi xep thanh cum lip tren cac bd hbe 1, khoang 20-26 bd libe gd xep tren mpt vdng, cae bd go 1 nhd, 3-4 mach phan hda li tam ro, ben cgnh go 1 quanh mo mem tiiy cd cac mach gd hau mpc rat to. Md mem tiiy trong eung kich thudc to, dang gan trdn xep chira cac dao nhd.
J'^^^'^^'^
Hinh 2. Vi phan thdn Ta xanh
193
N ^ i e n cmi Y hpc Y Hpc TP. Hb Chi Minh * Tap 20 * So 6 * 2016
thom (1608,73; 1516,53; 1410,51 cm-'); C-O-C (1249,08 cm-'); O-CH2-O (1054,86; 869,21 cm').
Hinh 3 Cac cm tir cua bot duac head Long che cho 2 Manh mach dian 3 Biaibivalokhi 4: Bd sgi mo cieng; 5: Mdnh mqch xoan)
Bgt dugc lieu
Bpt mau nau xam, miii cay ed dae trung.
Trong bpt tim thay nhieu nha't cac caii hi: Long che chd gap hoac diit gay, manh bi&i bi vdi nhieu Id khi kieu song bao xep thanh day, manh mach xoan, mach diem, cac bd sai mo cung, cac khoi chat nhua mau cam.
Khao sat thanh phan alkaloid Dinh iinh alkaloid bang phdn iing hoa hgc
Mayer
Kgt qua djnh tinh
+: Phan ung dyong tinh +++. Phan ling dipang tinh rat ro Ket qua dinh tinh bang phan iing hda hpc va sic ky ldp mdng cho fhay mau chief cd su hien dien eiia nhdm alkaloid rat rd. Tren sac ky do, cd the thay it nhat 6 vet chat bat mau duong tmh vdi fhuoc thii Dragendorff, cho thay trong To xanh ed nhieu ve ham lupng va cac loai alkaloid.
Phan lap va bien giai cau true cua alkaloid da phan l^p
A la chat ket tinh hinh kim, mau nau, tan tot trong cae dung mdi phan cvec nhu methanol, khong tan trong nude. Pho UV cho 3 dinh hap ttiu 220 nm, 283 nm, 303 nm. Pho IR cho tin hieu ciia cac nhdm chiic OH (3416,63 em-^); C=C nhan
Hinh 4: Djnh tinh alkaloid bang sac ky l&p mdng he dung mdi: CHCb: MeOH: NH4OH (9:1:0,5); mh:
Alkaloid todn plmn; phdt hiin bang thuo'c thu Dragendorff
Pho 'H-NMR cho tin hieu cpng hudng iing vdi si^ hien dien hai nhdm proton methoxy 5H [4,06 (3H; s; 3-OMe); 3,87 (3H; s; 10-OMe)], bdn nhdm proton mettiylen trong dd cd 1 nhdm d vimg trudng thap [5H 6,12; 5H 6,01 (O- CH2-O, A; O-CH2-O, B)], so nhdm edn lai d vung frudng cao [2,96; 2,98 (H-4A; H-4B); 3,73;
3,35 (H-5A, H-5B); 3,02; 2,84 (H-7A, H-7B)], ba nhdm proton methin 5H [4,36 (lH;H-6a), 6,75 (IH; H-8), 7,63 ( l H ; H - n ) ] .
194
Y Hpc TP. Hb Chi Minh * T?p 20 * So 6 * 2016 Nghien cuu Y hpc Cac dii lieu phci "C-NMR, DEPT cho fhay Al
cd tong so 19 carbon eho tin hieu pho tu 20 ppm dwi 150 ppm. Trong dd cd 10 carbon bae IV (C), 3 carbon bac m (CH), 4 carbon bac II (CH2) va 2 carbon blie I (CHa). Tat ea 10 carbon bac IV deu dich chuyen d vimg trudng thap (&c tii 115 ppm den 150 ppm). Trong dd cd 4 carbon mang di td oxy (6c 145,73; 137,66; 148,48; 147,41 ppm), cac carbon con lai deu la ciia ndi ddi.
Trong so 3 carbon bae IH cd 1 carbon d vimg trudng cao (6c 54,52 ppm; &H 4,36 ppm), 2 carbon cdn lai d vung trudng ttiap (110 -115 ppm).
Cd 4 carbon bac U, trong dd cd 1 carbon bac n xuat hien vimg trudng fhap [5c 102,73 ppm, &H (6,12; 6,01) ppm]. Day la d|ic trung eiia nhdm methylendioxid ttiudng gap trong cae alkaloid eiia chi Stephania. Cac carbon bac II edn lai la cac carbon mang noi don xuat hien d vimg trudng cao [6c 42,55 ppm; 6H (3,73; 3,31) ppm], [6c 33,67 ppm; 6H(3,02; 2,84) ppm], [&c 21,47 ppm; 6c (2,96;
2,98) ppm].
Trong nhdm methyl, cd 2 nhdm cd dp ddi hda hpc dac trung ciia nhdm methoxy [6c 60,12 ppm; 6H (4,06; 3H) ppm] va [6c 56,66 ppm; 6H (3,87; 3H) ppm].
Vdi cac dli lieu phd NMR nhu tren ta cd the du doan Al la mot alkaloid cd mpt nhdm the meftiylen dioxid, hai nhdm methoxy, hai nhdm methylen, mpt nhdm NH va mdt nhdm -OH.
Tham khao cac tai lieu cdng bd d|c,die'm pho NMR cd the d u doan dupe Al la khung aporphin.
Dua vao phd HMBC (PL 6.4.5), hai proton cua nhdm methylen dioxid (6,12 va 6,01) ed tuong tac vdi ea hai carbon bgc IV 6c (145,73 va 137,66, cd the la C-1 va C-2). Them vao dd proton ciia nhdm methoxy 6H (4,06) cd xuat hien tuong Bdng 1. Kei qud phd'NMR ciui A so sdnh v&i Cassythine.
tac vdi ca C-1, C-2 va mdt Ctubon bac IV (6c 140,61) do dd carbon bac IV nay cd the la C-3 hoae C-3^. Ca hai proton methylen {6H 2,96; 6H 2,96) va hai proton methylen (6H 3,73; 6H 3,35) deu eho tuang tac vdi mot carbon bac IV (6c 117,54) nen carbon bac IV (6c 140,61) phai la vi tri C-3 va carbon bac IV (6c 117,54) phai la vi hi C- 3a. Them vao do hai proton methylen (6H 2,96; 6H 2,96) ed tuong tac vdi C-3, mot carbon bgc IV (6c 122,13) va carbon mettiylen (6c 42,55), nen carbon bae IV (6c 122,13) phai la vi tri C-lb, carbon methylen (6c 42,55) la v; tri C-5 va hai proton methylen (6H 2,96; bn 2,98) gan vdi carbon (6c21,47) d vi tri C-4. Hon niia H-5A, H-5B deu tuong quan vdi C-3a, C-4 va mpt carbon b | c m (6c 54,52) nen carbon bae DI nay phai la C-6a.
Dua vao phd COSY (PL 6.4.6), H-6a ghep cap vdi hai proton methylen (6H 3,02; 6H 2,84), nen hai proton nay la H-7a, H-7b. Hai proton H-7a, H-7b lai gan vdi carbon (6c 33,67) do dd carbon nay la C-7.
Phd HMBC cho Ihay H-6a cd tuang quan vdi carbon bac IV (6c 112,25) ma khdng tuong quan vdi carbon bae IV (6c 125,78) cdn H-7 ffii nguoc lai, nen (6c 112,25) la v; tri C-la, (6c 125,78) la vi tri C-lla. Ngoai ra, H-6- va H-7 cimg tuong quan vdi carbon bac IV (6c 123,23), nen (6c 123,23) la vi tri C-7a. H-7a tuong quan vdi ca hai carbon metfiine (6c 111,89; 6c 116,16) cdn proton methin (6H 6,75) thi khdng tuong quan vdi carbon methin (6c 111,89), nen (6c 111,89) la vi tri C-11 va (6c 116,16) la vi tri C-8. Ca hai H-7a va H-8 deu cd tuang quan vdi carbon bae IV (6c 148,48), nen (6c 148,48) la vi tri C-9. Cdn lai la vi tri C-10.
Dua tren ket qua phd ket hpp so sanh tai lieu tham khao <^' '^i f^', cd the nhan dinh chat A la mot aporphin alkaloid ten gpi la cassythin.
Vi tri 1 1a 1b 2
'H(a)
Alkaloid A l
" C ( a | 145.73 112,25 122,13 137,66
HiVIBC COSY
Cassythine p ] 'H(b) "C(b)
143 111,1 128,8 135,1
N g h i e n c u u Y hgc Y Hpc TP. Hb Chi M i n h * Tap 20 * S£» 6 * 2016
Vi HI 3 3a 4A 4B 5A 58 6a 7A 7B 7a 8 9 10 11 11a 3-OMe 10-OMe OCHiJDA
B
AiltaioidAI 'H(al
2,96 m 2,98 m 3,73 dd 3.31m 4.36 (dd)
3.02 m 2.841
6,75
7.63
4,06 3,87 6,12 6,01
"C(a) 140.61 117.54 21.47 21.47 42.55 42.55 54.52 33.67 33.67 123.23 116.16 147.41 148.48 111.89 125,78 60,12 56,66 102,73 102,73
HiMBC
Cib, Ca, Caa. Cs
Caa, C4, Caa Clb, C7, C7a Cib, Csa. Cya. Ca, Cila
C7, Cza, Ca. Cila
CiB, Cva. Ca. Cio, Cila
Ci. Ca. Ca Ca, Cio Ci, Ca
COSY
Ha
H, H, He.
Cassythine [3]
'H(b)
2,68-2,74 m 2,6&-2,74 m 3,39 m 2,94 m 3,87 dd 2.78 2,65 m
6.79 s
7.59 s
4.01s 3.92 s 6,1 5.9
" C (b) 139.8 119.5 23.9 23.9 43.1 43.1 53,8 36,5 36,5 128.5 114,2 144.6 145.4 109.7 123.3 59.4 56.2 100.5 100.6 (a) Phff A do trong MeOH, 'H-NMR 500 MHz vi
(b) PhoCassylhme do trong CDCh, ^H-NMR 400, OMe
Hinh 5. Cong thiec cua cassythin
K E T L U A N
Nghien ciiu nay da khao sat dupe dac diem vi hpc va nhdm chat alkaloid a i a To xanh (Cassytha filifrrmis L.), phan lap va xac dinh dupe cau tnic ciia 1 aporphine alkaloid la cassjihin.
Ket qua nghien ciiu gdp phan xay d u n g tieu i"C-NA4RI25MHz.
13 MHz vd 'K-NMR 100.62 MHz.
chuaji dupe Ueu nay, dong thai ed ttie dimg Cass3rfhine lam chat ehuah doi chieu danh gia chat lupng ham lupng ciia dupe Ueu nay, chong nham l i n vdi cay To hong.
TAI LIEU T H A M KHAO
1 Chang FR, Chao YC, Teng CM and Wu YC (1998). Chemical constituents fe>m Cassytha fHifbrmK I_ loumal of Natund produds, 61(7), 863-866.
2 Hoet S, Stevigny C, Blodc S, Opperdoes S, Colson P.
Baldeyiou B, Lansiaux A, Bailly C a n d Quetin-Ledetcq J (2004). Alkaloid from Cassy^ filiprmis a n d Related Aporphines: Antitrypanosomal A c b v i ^ , Cytotodxia^, and Interaction with D N A and Topoisomeiases Planta Med., 70, 407-413
3 Stevigny Q Block S, De Pauw-Ollet M C De Hoffmann E Uabres G, Adjatddje V and Quetm-Leclercq J(2002). Cytotoxic Aporphine Alkaloid from aissytiui JUifarmis. Ptania Med., 68, 1042-1044.
4 Vien d u v c lieu (2003). CSy thuot va dgng v6t lam (Jiwfir 17 Vi^
Nam (tUfC). NXB Khoa hpc ki thuat, 978.
5 V6 Van Chi (1997) Tu dieh cSy OiuSc Vi& Nam. NXB Y hgft 1236.
Ngay nhan bm bdo' Ngay phdn biin nhan .xet bm bdo.
Ngay Mi bdo dugc dang:
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