xam din vang nau, su si, vdi nhung dudng van mit dgc, khdng diu, dl bdc; mat trong miu vang nhat din vang xam, tuong ddi nhin cd van dgc nhd. Chit nhe va gidn, dl be giy, mat giy khdng phing, ldp ngoai mau vang nau, ldp trong mau trang xam. Miii nhe, vi hai nggt sau ding.
4. Dac diem vi hoc:
* Dac diim vi phiu: Mat cit ngang dugc lieu la dai hinh cung. Dudi kinh hiin vi nhin tii ngoai vao trong thiy: Ldp bin gdm 6-7 hang tl bao xip ldn xdn. Md mim vd la nhung te bao thanh mdng hinh da giac xip ldn xdn. Bd libe cip hai chiim khoang 2/3 vi phiu cd hinh bd thap. Tia rugt gdm 2-3 hang tl bao. Cac tl bao chiia day tinh the calci oxalat dang cat. Tang phat sinh libe-gd gdm 2-3 hang tl bao diu dan.
* Dac diem bdt duge lieu: Bdt vd re dia cdt bi mau vang nau, miii nhe, vi dac biet.
t F > > / ^
Soi dudi kinh hien vi thay: Manh ban, manh md mem. Rat nhieu hat tinh bdt hinh
F > trdn tap trung thanh tiing khdi trong cac manh md mem hay diing rieng le, dudng trdn tap trung thanh tiing khdi trong cac manh md mem hay diing rieng le, dudng kinh hat tmh bgt 0,01-0,02nim. Tinh thi calci oxalat dang cat khd nhin dudi kinh hien vi thudng, de nhin han dudi kinh hien vi phan cue.
5. Dinh tinh:
Dinh tinh coumarin Chudn bi mdu:
- Miu thii: 2g dugc lieu da cit nhd, chilt vdi 10ml methanol, sieu am 30 phiit,
gan, lgc liy phin dich chilt, dung dl chim sic ky. Lugng chim 8-16 ^il.
Miu ddi chilu: Dung dich scopoletin 0,04mg/ml trong methanol. Lugng chim 6nl.
L&p mong: TLC silica gel 60 F254 Merck, hoat hda ban mdng d 105°C trong 1 gid
trudc khi su dung.
He dung moi sdc ky: Ethylacetat: methanol: nudc (10: 1:1) Phdt Men: - Quan sat UV 366nm -^ Hinh 4.9.A.
- Dung dich KOH/ethanol, UV366nm-^ Hinh 4.9. B
Sc 1 2 3 4 5 Sc 1 2 3 4 5
A1 ^^.
i
B e l 2 3 4 5 6 Si 1 2 3 4 5
r '\ ^ r F
Hinh 4.9. Sac ky do dinh tinh thanh phan nhom hoat chat dirge lieu dja cot bi
{Cortex Lycii).
Ky hieu: Scopoletin (Sc); Betain (Be); (3 sitosterol (Si); Cac mau dugc lieu dia cdt bi thu hai (vet 1- 5); Huang gia bi (vet 6).
SKD dinh tinh coumarin (Hinh. A/UV366, Hinh. B/UV366); SKD djnh tinh alcaloid (Hinh.C/VIS); SKD dinh tinh terpenoid (Hinh. D/ VIS); SKD TLC
scarming / scopoletin va DL dia cdt bi, huynh quang X 366nm / K 540(Hinh. Alva
Dinh tinh alcaloid
Chudn bi mdu:
- Mau thu: 2g dugc lieu da duge cit nhd, chilt vdi 30ml hdn hgp methanol va
nude (4:1), sieu am 30 phiit, tiip tuc dun hdi luu tren each thuy 30 phiit. Dl ngudi, gan lay phan dich chilt, lam bay hoi dung mdi din can. Hda cin thu dugc trong 5ml methanol cd chiia 0,5% acid hydrochloric, lgc qua giiy lgc, bd 1ml dich lgc diu, liy 3ml dich tiep theo dung dl chim sic ky. Lugng chim 10 ul.
Mau ddi chieu: Dung dich betain 4 mg/ml trong methanol. Lugng chim 5^1.
L&p mong: TLC silica gel 60 F254 Merck, ban nhira.
He dung moi sdc ky: n.Butanol : acid hydrochlorid : nudc (8:2:1) Phdt Men: Dragendoff cd acid h\'drochlorid
Kit qud: Hinh 4.9.C.
Dinh tinh terpenoid
Chudn bi mdu:
- Mau thu: 2g dugc lieu da cit nhd, chilt vdi 10ml methanol, sieu am 30 phiit, gan, lgc liy phin dieh chilt, dung dl chim sic ky. Lugng chim 8-16 ^il.
Mau ddi chilu: Dung dich p sitosterol 0,lmg/ ml. Lugng chim 5^1.
L&p mong: TLC silica gel 60 F254 Merck, hoat hda ban mdng d 105V trong 1 gid. He dung moi sdc ky: n.Hexan : ethylacetat (2:1)
Phdt Men: Liebermann- Burchard (TT), quan sat / VIS
^eV^M^:mnh4.9.D. 6. D|nh lirgng:
Phuffng phdp
Chudn bi mdu:
- Miu thii: Can 1 g dugc lieu da dugc xay nhd, them 20 ml methanol, sieu am
30 phiit, tiep tuc chilt ndng tren each thiiy d 60°C trong 30 phiit x 2 lin. Gdp dich
' ' r F
chiet, cd tren each thuy den can. Hda can thu dugc trong vua dii 5ml methanol, lgc bd 1ml dich lgc dau, lay 2ml dich lgc tiep theo de cham sac ky.
rv F F
- Mau ddi chieu: Dung dich p sitosterol 0,1 mg/ml trong methanol.
L&p mong: HPTLC Silica gel 60 F254 Merck, kich thudc kinh 20 x 10 cm, hoat hda
ban mdng d 105°C trong 1 gid trudc khi sii dung.
Chdm mdu: Cham thang chuan: 2\A , 4fil x 2, 6p.l, 8|LI1, va 10|j.l; mau thii 4 6|il (cham nhac lai mau thii 2 - 3 lan). Cham each mep kinh 15 mm (chieu X), 8 mm (chieu Y), chilu rdng vlt (8mm), khoang each gifl-a cac vet (10mm).
He dung moi sdc ky: n.Hexan : ethylacetat (2: 1), de bao hoa dung mdi sac ky 20
phiit.
F
sulphuric 10%/ ethanol (VA^), dl khd, siy d 1 0 5 ^ trong 10 phiit. Tiln hanh do
TLC scanning tren may CAMAG.TLC. SCANNER3, budc sdng X 570 nm, tdc do
quet 20mm/giay, chi do do remission, thdi gian do trong khoang 15- 30 phiit sau khi say kinh. Chuang trinh dugc thuc hien dudi sir diiu khiln ciia phin mim wintcats. Dgc kit qua do tren may. Tinh ham lugng % p sitosterol cd trong dugc lieu dugc tinh 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 thii do dugc (|ug) (kit qua trung binh ciia 2-3 lin nhic lai)
u : Lugng miu thii chim tren may {\i\)
V: Thi tich djch chilt miu thii (ml) P : Khdi lugng dugc lieu dem can (g) B : Do am ciia dugc lieu
Kit qud:
Kit qua dinh lugng p sitosterol trong dugc lieu dia cdt bi dugc ghi d Bang 4.5 va Hinh 4.10.
F ^
Bang 4.5. Ket qua dinh lugng P- sitosterol trong dugc lieu dia cdt bi miu thu hai va
mau mua tren thi trudng.
STT 1. 2. 3. 4. 5. Tin mau DCB 201 DCB 202 DCB 203 DCB 204 DCB 205 Hdm luong % fi- sitosterol 0,0205 0,0270 0,0368 0,0306 0,0245 Ghi chd
Mau thu hai nt nt nt nt
Ham lugng p sitosterol trong dugc lieu dia cdt bi khd tuyet ddi nam trong khoang 0,0205-0,0368%.
if
m tiZ t'3
Dugc lieu dia c6t bi
CM 0 30
• » - - , . . . , Do thi xac dinh khoang tuyen tinh Do thi xac dinh khoang tuyen tinh
Hinh 4.10. Sac ky do TLC scanning va do thi xac dinh KTT. Dinh lirong B- sitosterol trong du'o'c lieu dia cot bi
K I M N G A N H O A {Flos Lonicerae)