7 A r r
cm, rdng l-3cm, day khoang 3mm, phii ldng dang vay mau nau den nau tdi. Mat tren va hai ben than cd cac vet seo trdn ciia gdc la ldi hoac ldm, it khi cdn re hinh sgi. The nhe, gidn, de be gay, mat gay mau nau dd, cd nhihig ddm vang xep thanh mdt vdng. Miii nhe, vi nhat va hai se.
4. Dac diem vi hoc:
* Dac diem bdt dugc lieu: Bgt mau vang nau, soi dudi kinh hien vi thay: Nhung manh bieu bi mau vang sim, manh md mem thanh mdng, mach gd hinh thang.
5. Dinh tinh:
Dinh tinh flavonoid Chudn bi mdu:
- M I U thu: lg dugc lieu da xay nhd, chilt vdi 10 ml methanol, sieu am 30
phiit, gan, lgc, dich lgc thu dugc diing dl chim sic ky. Lugng chim 4 -6 |il. - Miu ddi chilu: Naringin 0,1 mg/ml methanol, lugng chim lO^il.
L&p mong: TLC silica gel 60 F254 Merck, hoat hda d 105°C trong 1 gid trudc khi
sii dung.
He dung moi sdc ky: Ethylacetat: methanol: aceton : nudc ( 1 0 : 1 : 1 : 1 ) .
Phdt hien: TT 1: nhdm chlorid 2% trong ethanol + UV366nm -^ Hinh 4.5 .A.
- TT 2: Vanillin- acid sulphuric -> Hinh 4.5.B.
Kit qud:Y[mh 4.5.{A,B). Dinh tinh hffp chdt phenolic
Chudn bi mdu:
Miu thu: lg dugc lieu da xay nhd, chilt vdi 10 ml methanol, sieu am 30 phiit, gan, lgc, dich lgc thu dugc dung dl chim sic ky. Lugng chim 4 -6 ^il.
Miu ddi chilu: Naringin 0,1 mg/ml methanol, lugng chim lOjil.
L&p mong: TLC silica gel 60 F254 Merck, hoat hda d 105^C trong 1 gid trudc khi
sii dung.
He dung moi sdc ky: Ethylacetat :acid acetic: acid formic : nudc (10:1:1:2). Phdt hien: TT fast blue salt.
IHIHHHHHI^IHB^^^^^^^^^^a ^H 1 ( t S - ^ M * - ^ 1 1-r.vj.,^^ p..^ , B i gmiii^ 0,5 Na 1 2 3 4 Na 1 2 3 4 N a l 2 3 4
Hinh 4.5. s i c ky do dinh tinh thanh phan nhom hoat chat du-gc lieu cot toai bd
{Rhizoma Drynariae).
Ky hieu: Naringin (Na); Cac miu cot toai bd nghien ciiu (vlt 1- 4).
SKD djnh tinh flavonoid va hgp chit phenolic (Hinh. A/UV366, Hinh. B/VIS va Hinh. C/VIS)
6. Dinh lirgng:
Phuong phdp Chudn bi mdu:
- Mau thii: lg dugc lieu da dugc xay nhd, chilt vdi 15ml hon hgp methanol va
nudc( 4:1 ), sieu am 20 phiit, lam nhic lai nhu vay 3 lin. Gdp dich chilt, them dung mdi viia dii 50 ml, lgc qua giiy lgc, bd 1-2 ml dich lgc diu, liy 2-3ml dich tiip theo
• » r / • de cham sac ky.
- Mau ddi chieu: Dung dich naringin 0,1 mg/ml trong methanol. (adsbygoogle = window.adsbygoogle || []).push({});
L&p mong: HPTLC Silica gel 60 F254, Merck, kich thudc kinh 20 x 10 cm, hoat
hda d 105^C trong 1 gid trudc khi sii dung.
Chdm mdu: Cham thang chuan: 0,3fil, 0,6)il x 2, 0,9)j,l, 1,2^1 va l,5|j.l; Miu thii: 8-
14^1 (chim nhac lai miu thu 2 -3 lan). Cham each mep kinh 15 mm (chieu X), 8 mm (chieu Y), chieu rdng vet (8mm), khoang each giua cac vet (10mm).
He dung moi sdc ky: Toluen: ethylacetat: acid formic: nudc (1:12:2,5:3). Phdt hien: Do TLC-scanning, huynh quang, X 366nm va kinh lgc K 400.
Cdch tiin hdnh: Kinh sau khi triin khai sic ky, liy ra dl khd 30 phiit, nhiing dung
dich nhdm chlorid 2% trong ethanol, dl khd 30 phiit, nhiing tiip dung dich PEG 400 10% trong ethanol, dl khd tiip 30 phiit. Tiln hanh do TLC scanning tren may
CAMAG.TLC. SCANNER3, huynh quang, budc sdng X 366 nm, kinh lgc K 400,
tdc do quet 20mm/giay, ehi do do remission. Chuang trinh duge thuc hien dudi su dieu khien ciia phan mem wintcats. Dgc kit qua do tren may. Tinh ham lugng % naringin cd trong dugc lieu cdt toai bd dugc tinh theo cdng thiic sau:
m.V.100.100
X% =
u.lOOO. P ( I O O - B ) Trong dd :
X% : Ham lugng % naringin trong dugc lieu khd tuyet ddi
m : Lugng miu thu do dugc ()ig) (kit qua trung binh cua 2-3 lin nhic lai)
1): Lugng miu thii cham tren may {]i\)
V: The tich dich chiet miu thii (ml) P : Khdi lugng dugc lieu dem can (g) B : Do am cua dugc lieu
Ket qud:
Ket qua dinh lugng naringin trong dugc lieu cdt toai bd dugc ghi d Bang 4.3 va hinh 4.6.
Bang 4.3. Ket qua dinh lugng naringin trong dugc lieu cdt toai bd
STT 1. 1. 2. 3. 4. Ky hieu mau CTB 201 CTB 202 CTB 203 CTB 204 Ham lucmg % naringin 0,0567 0,2046 0,5813 0,8300 Ghi chu
Mau thu hai nt nt nt
Ham lugng naringin trong dugc lieu cdt toai bd khd tuyet ddi nam trong khoang 0,0567-0,8300%.
A-.-
L ^ A-
Naringin
B
L.L
Duac lieu cot toai bo
U M 3 - IU«4M«-X >- CHMe M v . I KV
S K e » (adsbygoogle = window.adsbygoogle || []).push({});
Do thi xac dinh khoang tuyen tinh
r ^ ^ ^ ^
Hinh 4.6. Sac ky do TLC scanning va do thi xac dinh KTT Dinh lirgng naringin trong dirge lieu cot toai bo
D I E P H A C H A U D A N G {Herba phyllanthi amari)