BCDUOCLIEU20thang9

GVHD: Ths. Vn Mi*

I. TNG QUAN V SC K LP MNG

- Sc k lp mng (thin layer chromatography - TLC) l mt k thut sc k c dng tch cc cht trong hn hp.- Phng php sc k lp mng bao gm: Pha tnh l mt lp mng cc cht hp ph, thng l silica gel, aluminium oxide, hoc cellulose c ph trn mt mt phng cht tr. Pha ng bao gm dung dch cn phn tch c ha tan trongmt dung mi thch hp v c ht ln bn sc k bi mao dn, tch dung dch th nghim da trn tnh phn cc ca cc thnh phn trong dung dch.*


*H s di chuyn Rf l i lng c trng quan trng v mc tch. H s di chuyn Rf c tnh theo cng thc:

Trong :- a l khong cch t im xut pht n tm ca vt mu th, tnh bng cm.- b l khong cch t im xut pht n mc dung mi o trn cng ng i ca vt, tnh bng cm.- Rf: Ch c gi tr t 0 n l.

I. TNG QUAN V SC K LP MNGKhi Rf = 0 th cht tan hon ton khng di chuyn, cn khi Rf = 1 th cht tan di chuyn bng tc ca dung mi. C nhiu yu t nh hng n Rf nhng quan trng l: Cht lng v hot tnh cht hp th. B dy ca lp mng. Cht lng v tinh khit ca pha ng.

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Mt s ci tin c th kt hp phng php truyn thng t ng ha mt vi bc, lm tng dung gii ca sc k lp mng v cho s liu chnh xc hn.

Phng php ny c gi l sc k lp mng hiu nng cao (high performance TLC - HPTLC).*

II. Cc bc chun b sc k lp mng

Chn cht hp ph: cng tng t nh sc k ct c th dng cc cht hp ph l silicagel, nhm oxit, bt cellulose, tinh bt, sephadex...

- cho cht hp ph bm chc vo phin knh, ngi ta cho thm cht dnh vo cht hp ph. Cht dnh thng l CaSO4 vi t l khong 10%.*


- Chun b phin knh v lp mng cht hp ph trn knh:Cc phin knh thng c c 20x5 cm, 20x20 cm, 7,5x2,6 cm. Ra sch, sy kh trc khi ri. Cn 5g bt silicagel cho vo ci nghin u vi 10ml nc ct, lng ny c th ri trn 2 phin knh c 20x5cm. Sau ri ngay ln phin knh bng tay hoc bng my.

kh nhit phng sau sy 1000C t 30 - 120 pht tu theo yu cu. dy chun ca lp mng l 0,25 - 0,30 mm.*


Chun b mu th: cng ging nh sc k giy, cc mu th cn c cch chun b ring loi b tp cht v c c mu. Dng ng vi qun chm mu cch b di phin knh khong 1-2 cm. Vt chm phi nh, ng knh 2 - 5 mm cch xa nhau t nht 1 cm.*

Mt s dng c cho SKMLBnh trin khai*

Mt s dng c cho SKMLBng mng trng sn*

Mt s dng c cho SKML*Uv lamps cabinet 365, 254nmMao qunMy sy bng mng

III. Mt vi h dung mi dng khai trin trn cc bn mng silicagel-G:

Saponin triterpenoid:Chloroform methanol nc (65:35:10)

b) Ethyl acetat acid acetic nc (8:2:1)

c) n-butanol ethanol*


2. Saponin nhm spirostan:Chloroform methanol nc (65:35:10)

b) Chloroform methanol (8:2)

c) n-butanol bo ha nc*


3. Saponin kim: Chloroform ethanol dung dch amoniac 1% trong nc (2:2:1)

b) Ethanol pyridin nc (3:1:3)*

Dung mi phi l loi tinh khit, thng dng hn hp 2-3 cc dung mi. Cc dung mi xp theo th t mnh dn (sc y, phn hp ph): hexan, heptan, cyclohexan, carbon tetraclorid, benzen, cloroform, butyl acetat, ether, ethyl acetat, pyridin, aceton, ethanol, methanol, nc.*

IV. Tin hnh SKLMChit xut v tinh ch s b Saponin.Chm dung dch trn bn mng.Trin khai SKLM.Cch pht hin.

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Chm mu thK mt vch thng nm ngang bng bt ch, cch mp di ca bn mng 1cm lm vch xut pht.Dng mao qun hay micropipet chm cc vt dung dch th v dung dch chun ln . Cc vt phi cch nhau v cch mp bn mng t nht 1 cm.

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Trin khai SKLMQu trnh cho pha ng chy, ko mu phn tch di chuyn trn pha tnh.t bn mng vo bnh sc k bo ho hi dung mi ca pha ng, mp pha chm mu c nhng vo dung mi ng nhng khng c cho im chm mu chm trc tip vo dung mi ng.Sau khi dung mi chy c na hay 2/3 bn mng ta ly ra kh hay sy kh.

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Cch pht hinDa vo tnh ph huyt bng cch trng mt lp gelatin mu (ha tan 5 g gelatin trong 100 ml dung dch NaCl 9% 600C.Khi ngui n 400C th thm mu b loi fibrin) hoc phun dung dch treo mu 2% loi fibrin trn bn mng.

Cc vt saponin s cho vt mu nht trn nn hng .*

Cch pht hinCc thuc th di y dng cho loi saponin v sapogenin nn sau khi phun cn phi sy 10 pht 1100C ri quan st mu nh sng thng hoc nh sng t ngoi (365nm):Thuc th Carr Price (dung dch SbCl3bo ha trong chloroform).Thuc th Liebermann Burchard.Thuc th Salkowski, acid phosphomolibic 10% trong ethanol, acid phosphotungstic 20% trong ethanol, dung dch acid phosphoric 50% trong nc, dung dch vanillin sulfuric.Cc vt saponin s cho cc mu khc nhau, ty loi thuc th saponin.*

Cch pht hinSaponin nhm spirostan c th hin mu bng thuc th Sanni (dung dch A: vanillin 1% trong cn, dung dch B: anhydrid acetic + H2SO412:1).

Phun dung dch ri sy 1200trong 3 pht sau phun dung dch B, vt saponin c mu vng.

i vi nhm spirostan v nhm steroid alcaloid c th dng thuc th Carr Price phn bit cc dn cht c ni i v khng c ni i v tr C-5. Cc dn cht C5c mu 200C v tm sau khi sy 1050C.*

Cch pht hinCng c th phn bit 2 loi dn cht trn bng thuc th Marquis (0,2 ml dung dch formaldehyd 37% trong nc + 10 ml H2SO4), ch c loi dn cht ca C5 cho phn ng.

Cc saponin nhm spirosolan v solanidan c th pht hin bng thuc th Dragendorff.*

V. ng dng v cng dng ca SKLMng dng:nh tnh.nh lng (phi c cht chun).Pht hin thnh phn trong hn hp, nh tnh im ch.Theo di thnh phn cc cht.Phn lp cc cht.

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V. ng dng v cng dng ca SKLMCng dng:Xt nghim tinh khit ca cc ha cht phng x trong dc khoa.Xc nh cc sc t trong t bo thc vt.Pht hin thuc tr su, thuc dit cn trng trong thc n.Nhn bit nhng ha cht trong mt cht cho sn.Gim st cc phn ng hu c.

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VI. Cc hin tng ngoi mun thng gp trong SKLM v cch khc phc*

Ngun tham khohttp://www.duoclieu.org/2012/01/kiem-nghiem-duoc-lieu-chua-saponin-bang.htmlhttp://www.duoclieu.org/2012/07/phuong-phap-sac-ky-duoc-lieu.htmlhttp://123doc.org/document/1030224-ap-dung-phuong-phap-sac-ky-cot-va-sac-ky-lop-mong-vao-giang-day-hoa-hoc-pho-thong.htm*

Thank you*

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BCDUOCLIEU20thang9