ngoai mau vang nhat din vang nau(mau ho phach), trong, md, sang bong hoac ed van dgc sau hoac ndng khdng diu, cd khi edn sdt lai vd ngoai mau nau xam. Chit Cling, dai, cd chat nhay dinh, mat cit dang chit simg, tru giira mau tring nga. Mui nhe, vi nggt, hai ding.
4. Dac diem vi hoc:
• •
Dac diim bdt dugc lieu: Bdt mau vang tring, soi dudi kinh hiin vi thiy: Manh md mim, manh mach, tl bao cOmg, tinh thi calxi oxalat hinh kim.
5. Djnh tinh
Dinh tinh terpenoid
Chudn bi mdu:
- Mau thir: Can 2 g dugc lieu da tan nhd, ngam vdi 5ml nudc dl yen 1 gid cho
"^ F •\ ^
truang nd, nghien nhd trong cdi, dua dugc lieu da nghien nhd vao binh ndn bang 20ml methanol, sieu am 30 phiit, tiip tuc dun tren each thiiy 30 phiit, gan liy phin dich chiet, phan cdn lai dugc chiet nhic lai lan 2 vdi 20 ml methanol. Gdp dich
F F \ F
chiet, lam bay hai dung mdi tren each thuy den gan can. Hda can thu dugc vdi 5ml
F \ t F F F
nudc, lac vdi 20ml chloroform x 2 lan, de lang, gan lay phan doan chloroform, bdc
F F t F F
hoi dung mdi den can. Hda can thu dugc trong 2ml methanol de cham sac ky Lugng cham 8 - lOnl.
- Miu ddi chilu: Dung dich P- sitosterol 0,1 mg/ml trong methanol, lugng cham 6 |il.
L&p mong: TLC silica gel 60 F254. Hoat hoa kinh d nhiet do 105° C trong 1 gid
trudc khi sii dung.
He dung moi sdc ky: n.Hexan: ethylacetat (2: 1) D I bao hoa dung mdi sic ky 20
phiit
Phdt Men: Liebermann Burchard (TT), siy kinh d nhiet do 105° C trong 2-3 phiit
quan sat VIS va UV366nm
Kit qud: Hinh 4.21.A., .B.
Dinh tinh acid amin
Chudn bi mdu:
- Miu thii: Can 0,5g dugc lieu, them 3 ml nude, dl yen 1 gid, nghiln thanh
nhac lai them 2 lin vdi 20 ml hdn hgp methanol va nudc 4:1. Gdp dich chilt them dung mdi vira dii 50ml. Lgc bd 1-2 ml dieh lgc diu, liy 3 ml dich tiip theo dl chim sac ky, lugng chim 2 - 4 j^l.
• Mau ddi chilu: Dung djch asparagin 0,05 mg/ml trong methanol, luong
cham 6 \x\.
L&p mong: HPTLC cellulose, Merck, nia giai methanol, dk khd d nhiet dg phdng He dung moi sdc ky: n. Propanol: acid acetic: nudc (3: 1: 1), dl bao hoa dung mdi (adsbygoogle = window.adsbygoogle || []).push({});
sac ky 1 gid
Phdt Men: Ninhydrin (TT) siy kinh d nhiet do 100 °C trong 5 phut Kit qud: Hinh 4.21.C.
Si 1
Inh 4.21. s i c ky do dinh tinh thanh phan nhdm hoat chit dirge lieu thien mdn ddng
(Radix Asparagi).
hieu: p sitosterol (Si); Asparagin (As); Cac mau thien mdn ddng nghien ciiu (vlt 1-4).
D dinh tinh terpenoid (Hinh A/ VIS, Hinh. B/UV366); SKD dinh tinh acid amin (Hinh. VIS).
6. Djnh lu'gng
Phuffng phdp
Chudn bi mdu :
- Miu thu: 0,5g dugc lieu , them 3 ml nude, dl yen 1 gid, nghiln thanh bdt
min, them 12ml methanol, chilt sieu am 20 phut, tiip tuc chilt hdi luu tren each thuy 30 phut, gan liy phin dich chilt, chilt nhic lai 2 lin vdi 20 ml hdn hgp methanol va nudc 4:1. Gdp djch chilt, them dung mdi vira du 100ml. Lgc bd 1 ml dich lgc dau, lay 3 ml djch tiep theo, pha loang x 5 lin vdi hdn hgp dung mdi tren, thu dugc dung djch dl chim sic ky
- Mau ddi chilu: Dung dich asparagin 0,01 mg/ml trong hdn hgp methanol nude (4:1).
L&p mong: HPTLC Cellulose Merck, kich thudc kinh 20 x 10 cm, nia giai
methanol, de khd d nhiet do phdng.
Chdm mdu: Cham thang chuin: 4(il x 2, 8|il x 2 , 12|LI1 va 16|LI1. Miu thu : 4 - 16 |il, chim nhac lai miu thOr 2 - 3 lin. Chim each mep kinh 15 mm (chilu X), 8 mm (chieu Y), chieu rdng vet (8mm). khoang each giira cae vet (10mm).
He dung moi sdc k}'-: n Propanol: acid acetic: nudc (3: 1: 1), chay binh cao 20cm,
bao hoa dung mdi 1 gid.
Phdt Men: Dung dich ninhydrin 0,3% trong ethanol cd chiia 3% acid acetic, do hap
thu X 61 Onm
Cdch tiin hdnh: Kinh sau khi da triin khai sac ky, de khd 1 gid, nhiing thude thu, de
yen 30 phut. Siy kinh d nhiet do 100 °C trong 10 phut, do TLC scanning tren may CAMAG.TLC. SCANNER3, budc sdng X 610 nm, toe do quet 20mm/giay, chi do do remission. Chuang trinh dugc thuc hien dudi sir diiu khiln ciia phin mim wintcats. Dgc kit qua do tren may. Tinh ham lugng % asparagin va acid amin (tinh theo asparagin) 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 (^g) (kit qua tmng binh ciia 2-3 lin nhic lai) u : Lugng miu thii chim tren may ((il)
V: Thi tich dich chilt miu thu (ml) P : Khdi lugng dugc lieu dem can (g) B : Do i m cua dugc lieu
Kit qud
Ket qua dinh lugng asparagin va acid amin tdng sd trong dugc lieu thien mdn ddng duge ghi d Bang 4.11 va Hinh 4.22. (adsbygoogle = window.adsbygoogle || []).push({});
Bang 4.11. Ket qua dinh lugng asparagin va acid amin tdng sd trong dugc lieu thien
mdn ddng. STT 1. 2. 3. 4. 5. Ky hieu mdu TMD201 TMD202 TMD203 TMD204 TMD205 Hdm lugfng Vo asparagin 2,07 1,53 1,18 1.53 1,16 Hdm lugfng acid amin .X. A tong so 8,89 7,18 5,94 5,53 4,55 Ghi chit
Mau thu hai nt nt nt nt