7- 10 cm. Mat tren la mau luc sam, mat dudi mau luc nhat, phien la day, nhan, mep
> y F
nguyen cd 3- 5 gan hinh cung, chay dgc theo gan chinh va chum lai ve phia dau gdc la, gan ldi ve phia mat dudi la rat ro. Cudng dai tu 5- 10cm, rdng ve phia gdc.
4. Dac diem vi hoc:
Dac diem bdt dugc lieu: Bdt mau nau nhat, vi hai dang. Soi dudi kinh hien vi thay: Manh biiu bi mang Id khi. Id khi. manh biiu bi cudng la, bd sgi, gigt dau, manh canh hoa, manh mach.
5. Dinh tinh
Dinh tinh iridoid
m
Chudn bi mdu:
Miu thii: Can Ig dugc lieu da dugc tan nhd, chilt vdi 20ml hdn hgp methanol nudc (4:1), sieu am 20 phiit, lam nhic lai 2 lin. Gdp dich chilt, lam bdc hoi dung mdi tren each thiiy din gin can, tiip tuc lam khd trong hdt. Hda cin thu dugc trong
10 ml methanol, lgc, thu dugc dung dich dk chim sic ky, lugng chim 6-12 ^1.
- Miu ddi chilu: Dung dich aucubin 0,2 mg/ml trong methanol, chim 12 ^il.
L&p mong: TLC silica gel 60 F254 Merck.
He dung moi sdc ky: Aceton: xylen: isopropanol: amoniac dam dac (10: 20: 60: 10) Phdt Men: DMAB acid hydrochloric
Kit qud: Uinh 4.14.A.
Dinh tinh ddn xudt acid caffeic Chudn bi mdu:
- Miu thii: Can lg dugc lieu da dugc tan nhd, chilt vdi 20ml hon hgp methanol
nudc (4:1), sieu am 20 phiit, lam nhic lai 2 lin. Gdp dich chilt, lam bdc hoi dung mdi tren each thuy din gin can, tiip tuc lam khd trong hdt. Hda can thu dugc trong
10 ml methanol, lgc, thu duge dung djch dl chim sic ky, lugng cham 6-12 ^il.
- Miu ddi chilu: Dung dich acid chlorogenic 0,1 mg/ml trong methanol, lugng cham 6^.1.
L&p mong: TLC silica gel 60 F254 Merck.
He dung moi sdc ky: Toluen: ethylacetat: acid formic: nudc (5 : 100 : 10 : 10) Phdt Men: Natural products - PEG 4000, UV 366nm
Hmh 4.14. Sac ky do dinh tinh thanh phan nhom hoat chat du'gc lieu ma dl
{Folium Plantaginis).
Ky hieu: Aucubin (Au), acid chlorogenic (Ch), acid ursolic ( Ur); Cac miu ma dl thu hai (vlt 1-6).
SKD dinh tinh iridoid glycoside (Hinh. A/VIS); SKD dinh tinh dan xuit acid caffeic (Hmh. B/UV366); SKD dinh tinh terpenoid (Hinh. C/VIS, Hinh. D/UV366).
Dinh tinh terpenoid
9 «rf
Chudn bi mdu:
- Miu thii: Can lg dugc lieu da dugc tan nhd, chilt vdi 20ml methanol, sieu am
20 phiit, lam nhic lai 2 lin. Gdp, lgc dich chilt, lam boc hoi dung mdi tren each (adsbygoogle = window.adsbygoogle || []).push({});
F ,
thiiy den can. Hoa can thu dugc trong vd 10ml methanol, thu dugc dung djch dk
chim sic ky, lugng chim 6-12 ^il.
- Miu ddi chilu: Dung dich acid ursolic 0,1 mg/ml trong methanol, lugng chim 10|il.
L&p mong: HPTLC RP18 , nia giai kinh vdi methanol, dl khd d nhiet do phdng.
F
He dung moi sdc ky: Methanol: nudc: tetrahydrofuran (94: 5: 1)
Phdt Men: Acid sulfuric 10% (v/v) trong ethanol, siy 105^ C trong 3-5 phiit, quan
sat/ VIS va UV 366nm.
Kit qud: Hinh4.14.(C, .D).
6. Djnh lirgng
Phuffng phdp
Chudn bi mdu
- Mau thii: Can lg dugc lieu da dugc tan nhd, chiet vdi 20ml hdn hgp
' > F F
methanol nudc (4:1), sieu am 20 phiit, lam nhac lai 2 lan. Gdp dich chiet, lam bdc
F '\ F F F
hoi dung mdi tren each thiiy den gan can, tiep tuc lam khd trong hdt. Hda can thu dugc trong vira dii 25 ml methanol, lgc bd 2ml dich lgc dau, lay 5 ml dich tiep theo
t F F
de cham sac ky.
- Miu ddi chilu: Dung dich aucubin 0,1 mg/ml trong methanol.
L&p mong: TLC Silica gel 60 F254 Merck, kich thudc kinh 20 x 10 cm, nia giai
kinh bing methanol, dl khd kinh trong hiit am.
Chdm mdu: Chim thang chuin: 5^1, lO^il x 2, 15^1 x 2 , va 20^1; Miu thu : 4- lO^il
(chim nhic lai miu thii 2 - 3 lin). Chim each mep kinh 15 mm (chieu X), 8 mm (chilu Y), chilu rdng vlt (8mm), khoang each giua cac vlt (lOmm).
He dung moi sdc ky: Aceton : xylen : isopropanol : amoniac dam dac (10: 30: 50:
10), bao hoa dung mdi sic ky 30 phiit.
Phdt Men: Thudc thii DMAB, do TLC scanning, >i 590 nm.
Cdch tiin hdnh: Kinh sau khi da triin khai vdi he dung mdi neu tren, cao 85mm, liy
ra, dl bay hai hit dung mdi trong hdt, nhudm thudc thii DMAB (hdn hgp ti le 2:1 ciia dung dich 4. dimethylaminobenzaldehyd 1% trong ethanol va acid hydrochloric 36%), dl yen 15 phiit va tiip tuc lam khd trong hdt, thdi gian Ih 45 (adsbygoogle = window.adsbygoogle || []).push({});
phiit. Do TLC scanning tren may CAMAG.TLC. SCANNER3, budc sdng X 590
nm, tdc do quet 20mm/giay, chi do do remission. Chuang trinh dugc thuc hien dudi sir diiu khiln cua phin mim wintcats. Dgc kit qua do tren may. Tinh ham lugng %
m.V.100.100
X% =
U.lOOO. P ( 1 0 0 - B )
Trong do :
X% : Ham lugng % aucubin trong dugc lieu khd tuyet ddi
m : Lugng miu thu do dugc (|ag) (kit qua trung binh cua 2-3 lin nhic lai) u : Lugng miu thu chim tren may (|il)
V: Thi tich dich chilt miu thii (ml) P : Khdi lugng dugc lieu dem can (g) B : Do am ciia dugc lieu
Kit qud:
Kit qua dinh lugng aucubin trong dugc lieu ma dk dugc ghi d Bang 4.8 va Hinh
4.15
Bang 4.8. Ket qua dinh lugng aucubin trong dugc lieu
•WW •\ t . /» y ma de TT 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Ky hieu mdu MD201 MD202 MD203 MD204 MD205 MD206 MD207 MD208 MD209 MD210 Hdm luffng % aucubin 0,5160 0,6740 0,7480 0,6340 0,6220 0,2790 0,2970 0,4750 0,6670 0,2800 Ghi chd
Mau thu hai nt nt nt nt nt nt nt nt nt
A -. - - r - " - ( ^ ^ • ^ • ' - - ' V . A- . . - - A - ^ ~ ' - • k'- • ' - - • ',!, Aucubin B , - . i •"' r ; * • " • « -vi •-! .Si ,V: .V, ; t — Dugc lieu ma de g~^ » * * « • - « SMiM»ai 1 « E « e - • • • r « > - > - - * * • j w » . MM *** MOC I K C . • «000- < • j ' l a i - s w » i •* r. cMme m«t • i jjx y : y^ --^ f : e o : CM toa t . » j e s
Do thi xac dinh khoang tuyen tinh
Hmh 4.15. Sac ky do TLC scanning va do thi xac dinh KTT Dinh lirong aucubin trong dirac lieu ma de
N G U V I T U {Fructus Schisandrae)