than va nggn cd nu hoa ve phoi hoac say khd.
3. Md ta: Than non vudng. nhe, xdp, dai 20- 50 cm, dudng kinh 0,1-0,3 cm, mat
' f F F
ngoai than mau nau tim, cd ranh dgc va cd long trang nhd. La mgc ddi, cudng ngan, phien hinh mui mac dai tir 4- 6cm, rdng 2-3 cm, mep cd r^ng cua, dau la thudn nhgn, hai mat la mau luc sam. phan gan chinh cd phii long min. Cum hoa xim co d nggn than va d dau canh. Dugc lieu cd miii hang, vi hoi man va sau hai dang.
4. Dac diem vi hoc:
• •
* Dac diem vi phau la.
Bieu bi tren va dudi mang ldng che chd va long tiet. Ldng che chd cd nhieu d phan gan giua, gdm 2 6 tl bao, mat long phu cutin ldm chdm. Ldng tilt cd chan ngan 1 - 2 tl bao, diu 2 - 4 tl bao, chan ngin mdt tl bao nim sau trong ldp biiu bi. Hai dam md day nim sat bieu bi tren va dudi d phan gan chinh. Bd libe - gd xep hinh vdng cung tach ddi nim giua phin md mim ciia gan chinh. Phin phiin la cd md mim giau gdm mdt hang tl bao hinh chir nhat xip thing diing dudi ldp biiu bi tren. Md mim khuyit gdm 4 - 6 hang tl bao hinh trdn.
Cac ldng che chd nhiiu loai: Ngin hinh ndn, dan bao hoac dai da bao mdt day. Cac ldng tilt nim d chd ldm cua biiu bi, mdt sd thi khdng cudng diu da bao, mdt sd khae thi diu 1-2 tl bao nim tren mdt cudng ngin. Chi cd mdt hang te bao hinh dau d phien la.
* Dac diim bdt duoc lieu: Manh biiu bi va Id khi kiiu true bao, ldng che chd va
• • ' ' F
ldng tilt cd dang nhu da md ta d phin vi phiu. Manh mach vach, mach xoan, mach mang.
5. Dinh tinh
Dinh tinh flavonoid Chudn bi mdu
Miu thu: 0,5g dugc lieu da xay nhd, chilt vdi 10 ml methanol, sieu am 30 phiit, gan, lgc, dich lgc thu dugc dung dl cham sac ky.
- Miu ddi chilu: Dung djch sinensetin 0,025 mg/ml trong methanol.
L&p mong: TLC silica gel 60 F254 Merck. Hoat hoa kinh d nhiet do 105° C trong 1
gid trudc khi sii dung.
Kitqud:mnh4.n.A.
ffinh 4.18. s i c ky do dinh tinh thanh phan nhom hoat chat dirge heu
rau meo (Herba Orthosiphonis spiralis).
Ky hieu: Sinensetin (Si); Acid rosmarinic (Ro); Acid ursolic (Ur); Cac mau rau meo
nghien ciiu (vet 1-5). . WTT. i I-./T7TO
SKD djnh tinh flavonoid (Hinh. AAJV366); SKD dinh tinh terpenoid (Hmh. B/VIS, Hinh. B1/UV366); SKD dinh tinh din xuit acid caffeic (Hinh. C/UV254, Hmh.Cl/ UV366 va Hinh.C2/UV366)
Dinh tinh terpenoid
Chudn bi mdu
- Miu thii: 0,5g dugc lieu da xay nhd, chilt vdi 10 ml methanol, sieu am 30
phiit, gan, lgc, dich lgc thu dugc dung dk chim sic ky.
- Miu ddi chilu: Dung dich acid ursolic 0,25mg/ml trong methanol.
L&p mong: TLC silica gel 60 F254 Merck. Hoat hoa kinh d nhiet do 105° C trong 1
gid trudc khi sii dung.
Luang chdm: 8 |nl mau ddi chieu acid ursolic va lOfil miu thii He dung moi sdc ky: Chloroform : ethylacetat (5 : 2)
Phdt Men: Acid sulphuric 10% trong ethanol, siy 105°C din khi xuit hien vlt, quan
sat dudi anh sang thudng va dudi UV366nm Kit qua : Hinh 4.18.Bva .Bl.
Dinh tinh dan xudt acid caffeic
Chudn bi mdu
- Miu thii: 0,5g dugc lieu da xay nhd, chilt vdi 10 ml methanol, sieu am 30
phiit, gan, lgc, dich lgc thu dugc dimg de cham sac ky.
- Mau ddi chieu: Dung dich acid rosmarinic 0,4mg/ml trong methanol.
L&p mong: TLC silica gel 60 F254 Merck.
Luang chdm: 6}il mau ddi chieu acid rosmarinic va lOfil mau thii
F
He dung moi sdc ky: HDM 3: Toluen : ethylacetat: aceton : acid formic ( 5 : 2 : 2 :
1)
Phdt Men:
- Quan sat UV 254 va UV366 - NP/PEG (TT)/ UV366nm
Kit qud: Hinh4.18.C, .CI va .C2.
6. Dinh lirmig
Phuffng phdp
Chudn bi mdu::
Miu thu: Can 0,5 g dugc lieu da xay nhd, chilt hdi luu tren each thuy 45 phiit vdi 20ml chloroform x 3 lin. Gan, gdp djch chilt, lam bay hoi dung mdi tren each thuy din can. Hoa cin thu dugc trong vira dii 10ml methanol, lie diu, lgc qua giiy lgc, bd 0,5-lml dich lgc diu, liy l-2ml dich lgc tiip theo diing dl chim sac ky.
- Miu ddi chilu: Dung dich sinensetin 0,025 mg/ml trong methanol; Dung djch acid ursolic 0,25mg/ml trong methanol.
L&p mong: HPTLC Silica gel 60 F254 Merck, kich thudc kinh 20 x 10 cm.
Chdm mdu: Chim thang chuin: 2 ^ 1 , 4^1 x 2, 6^1, 8^il va lO^il cua mdi dung dieh
He dung moi sdc ky: Dichloromethan: ethylacetat ( 2:1) de binh sac ky bao hda
dung mdi 30 phiit.
Phdt Men: Do TLC scanning, huynh quang. A, 366nm, kinh lgc K400 (dinh lugng
sinensetin). Do TLC scanning, hap thu, X 520nm, sau khi tao san pham dan xuat vdi
TT acid sulphuric (dinh lugng acid ursolic).
Cdch tien hdnh: Kinh sau khi da trien khai vdi he dung mdi neu tren, eao 85mm, lay
ra, de bay hoi het dung mdi trong hdt , thdi gian 30 phiit. Do TLC scanning tren
may CAMAG.TLC. SCANNER3, huynh quang, budc sdng X 366nm, kinh lgc
K400, tdc do quet 20mm/giay, ehe do do remission. Chuang trinh dugc thuc hien dudi su dieu khien ciia phan mem wintcats. Dgc ket qua do tren may, ghi ket qua do dinh lugng sinensetin thu dugc. Tiep tuc tien hanh cae budc tiep theo de dinh lugng acid ursolic: Nhiing ban mdng trong dung djch acid sulphuric 10% trong ethanol,
siy d nhiet do 110°C trong 5 phiit. Do scanning, budc sdng X 520 nm, chi do do
nhu tren. Dgc kit qua do tren may. Tinh ham lugng % sinenserin, % acid ursolic cd trong dugc lieu rau meo theo cdng thiic sau:
m.V.100.100 X % =
U.lOOO. P ( 1 0 0 - B ) Trong dd :
X% : Ham lugng % hoat chit trong dugc lieu khd tuyet ddi
m : Lugng miu thir do dugc (|ig) (kit qua trung binh ciia 2-3 lin nhac lai) u : Lugng miu thir chim tren may (^il)
V: Thi tich dich chilt miu thir (ml) P : Khdi lugng dugc lieu dem can (g) B : Do i m ciia dugc lieu
Kit qud
Kit qua dinh lugng sinensetin va acid ursolic trong dugc lieu rau meo dugc ghi d Bang 4.10 va Hinh 4.19, Hinh 4.20..
Bang 4.10. Ket qua dinh lugng sinensetin va acid ursolic trong cac mau dugc lieu rau meo TT 1. 2. 3. 4. 5. 6. Ten duffc lieu RM201 RM202 RM203 RM204 RM205 RM206 Hdm luong Xsinensetin 0,020 0,038 0,021 0,045 0,029 0,018 Hdm lugfng % Acid ursolic 0,229 0,303 0,232 0,393 0,317 0,206 Ghi cha
Mau thu hai nt nt nt nt nt
Ham lugng sinensetin trong cac mau dugc lieu rau meo nam trong khoang tir 0,018 - 0,045%, ham lugng acid ursolic nam trong khoang tir 0,206 - 0,393%.
A . -- - F 1 ^' •^' •»> •^' . V . ... , v . ;^ 7^~ I Sinensetin B - 1 ! 1 • . i J , .A «M *•* • ! • Ai) « ^ ( t e , i , .VJ ,•„
Dugc lieu rau meo
• V - 1?0T SIO . n »73 • X r - 0 •>«»• M . - 2 tOX V ^ 2 9 S M - , : MP aoooo- t e a e o - ymec- 9000- « M X y^ X ^ M ' OD IM M 1W DO 30cf.oa a » 00 w e 00 : "•
Dd thi xac dinh khoang tuyen tinh
Hinh 4.19. Sic ky dd TLC scanning va do thi xac d|nh KTT Dinh lirgng sinensetin trong dirge lieu rau meo
Acid ursolic B
7^ Tfj Tn r« ; c ~
Duac lieu rau meo
. « t i U*-Xt'0 tt512 Mv • * *1«
000 OSO !0i T 00 2.00 2.M
Do thi xac dinh khoang tuyen tinh
Hinh 4.20. Sic ky dd TLC scanning va do thi xac dinh KTT Dinh lirgng acid ursolic trong dirge lieu ran meo
T H I E N M O N D O N G (Radix Asparagi)