* Dinh lirgng acid chlorogenic
Phuffng phdp:
Chudn bi mdu:
- Mau thii: Can 0,5g dugc lieu da xay nhd, chiet sieu am 20 phiit vdi 50ml hdn
hgp methanol va nudc ti le 4:1, lap lai lan 2 vdi 30ml dung mdi. Gan dich chiet vao binh dinh miic 100ml, them dung mdi vira dii 100ml. Lgc qua giiy lgc, bd 1-2 ml dich lgc diu, liy 15ml dich tiep theo. Hiit chinh xac 10ml dich lgc, them methanol viia dii 25ml, thu dugc dung dich dl chim sac ky.
- Miu ddi chilu: Dung dich acid chlorogenic 0,1 mg/ml trong dung mdi methanol.
L&p mdng: HPTLC Silica gel 60 F254 Merck, kich thudc kinh 20 x 10 cm.
Chdm mdu: Chim thang chuin: 3^1, 6|LI1 X 2, 9|al X 2 , va 12^1; Miu thii : 8- 16[il
(chim nhic lai miu thii 2 - 3 lin). Chim each mep kinh 15 mm (chilu X), 8 mm (chilu Y), chilu rdng vlt (8mm), khoang each giua cac vlt (10mm).
He dung moi sdc ky: Ethylacetat: acid formic : nudc (8: 1 :1), dl binh sic ky bao
hoa dung mdi 20 phiit.
Phdt Men: Do TLC scanning, ?imax 325 nm.
Cdch tien hdnh: Kinh sau khi da triin khai vdi he dung mdi neu tren, cao 85mm, liy
ra, de bay hai hit dung mdi trong hdt, thdi gian 30 phiit. Tiln hanh do TLC
scanning tren may CAMAG.TLC. SCANNER3, budc sdng X 325nm, tdc do quet
20mm/giay, che do do remission. Chuang trinh dugc thuc hien dudi sir diiu khiln ciia phan mim wintcats. Dgc kit qua do tren may. Tinh ham lugng % acid chlorogenic 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 % acid chlorogenic trong dugc lieu khd tuyet ddi
m : Lugng miu thii do dugc (jig) (kit qua trung binh cua 2-3 lin nhic lai) u : Lugng miu thii chim tren may (fil)
V: Thi tich dich chilt miu thii (ml)
F
P : Khdi lugng dugc lieu dem can (g)
t
B : Do am ciia duge lieu
r
Kit qud
F
Ket qua dinh lugng acid chlorogenic trong dugc lieu kim ngan hoa dugc ghi d Bang 4.6 va Hmh 4.12.
Bang 4.6. Kit qua dinh lugng acid chlorogenic trong dugc lieu kim ngan hoa
TT 3. Ten duac lieu/ ky hieu KN201 KN202 KN203 Hdm luong % acid chlorogenic/duffc lieu 2,1600 2,0300 5,5720 Ghi chd
Mau thu hai nt
nt
Ham lugng acid chlorogenic trong dugc lieu kim ngan hoa dat tur 2,0300 - 5,5720%.
Acid chlorogenic
B „ ^
S Tn S 75-
Dugc lieu kim ngan hoa
•*-3aiVB*iimrM»-Xtmojmn» Mr-»47%
OM OJO CM tM %X 1.40
Do thj xac dinh khoang tuyen tinh
Hmh 4.12. SSc ky dd TLC scanning va dd thi xac dinh khoang tuyen tinh D|nh lirgng acid chlorogenic trong dirge lieu kim ngan hoa
Dinh lu'gng p sitosterol
Phuffng phdp
Chudn bi mdu:
Mau thu: Can 0,5g dugc lieu da xay nhd, chilt sieu am 20 phiit vdi 20ml methanol, chilt lap lai lin 2 vdi 20ml dung mdi. Gan djch chilt cho vao binh dinh miic 50ml, them dung mdi vua du. Lgc qua giiy lgc, bd 1-2 ml djch lgc diu, liy 2-3 ml dich lgc tiep theo, thu dugc dung dich de chim sic ky.
1^ F F
- Mau ddi chieu: Dung dich p- sitosterol 0,1 mg/ml trong methanol. L&p mong: TLC Silica gel 60 G F254 Merck, kich thudc kinh 20 x 10 em.
Chdm mdu: Chim thang chuin: 2|LI1 , 4|LI1 X 2, 6^1, 8|il, va lOjul; Miu thii : 8- lO^il,
chim nhic lai miu thii 2 - 3 lin. Chim each mep kinh 15 mm (chilu X), 8 mm (chilu Y), chilu rdng vlt (8mm), khoang c^ch giu-a cac vlt (10mm).
F ^ ^
He dung moi sdc ky: n.Hexan: ethylacetat (2: 1), de binh sac ky bao hoa dung mdi
20 phiit.
Phdt Men: Do TLC scarming, X 570 nm.
F t f
Cdch tien hdnh: Kinh sau khi da trien khai vdi he dung mdi neu tren, cao 85mm, lay
t F F F
ra, de bay hai het dung mdi trong hdt, thdi gian. Nhiing vdi thudc thu acid sulphuric
10%/ ethanol (V/V), ^^ khd, siy d 105 V trong 10 phiit. Do TLC scanning tren may CAMAG.TLC. SCANNER3, budc sdng X 570 nm, tdc do quet 20mm/giay, chi do
F
do remission, thdi gian do trong khoang 15- 30 phiit sau khi say kinh. Chuang trinh dugc thuc hien dudi su dieu khien ciia phan mem wintcats. Dgc ket qua do tren may. Tinh ham lugng % P- sitosterol cd trong dugc lieu dugc tinh theo edng thiie sau:
m,V.100.100 X % =
U.lOOO. P ( 1 0 0 - B ) Trong do :
X% : Ham lugng % p- sitosterol trong dugc lieu khd tuyet ddi
m : Lugng miu thii do dugc (^ig) (kit qua trung binh ciia 2-3 lin nhic lai) u : Lugng miu thii chim tren may (|al)
V: Thi tich dich chilt miu thii (ml) P : Khdi lugng dugc lieu dem can (g) B : Dd im ciia duac lieu
Kit qud
Ket qua dinh lugng p sitosterol trong dugc lieu kim ngan hoa dugc ghi d Bang 4.7 va Hinh 4.13
F
Bang 4.7. Ket qua dinh lugng p sitosterol trong duge lieu kim ngan hoa
TT 4. 4. 5. 6. Ten dugfc lieu/ ky hieu KN201 KN202 KN203 Hdm lugng % p- sitosterol /dugfc li^u 0,0507 0,0793 0,0955 Ghi chii
Mau thu hai nt
nt
. v ^
'^ SS s Si iE ni—re-
Beta sitosterol
B .;-
Dugc lieu kim ngan hoa
c r