Sc k ct l phng php dng tch cc cht (cu t) ra khi hn hp da vo tnh phn cc ca tng cht.

. Dng c: ct sc k (ging ging nh ci buret, gm 1 ng thy tinh v 1 ci kha, nhng khng cn vch chia , i khi bn cng khng cn ci kha ny na, kch thc ct c ln, c nh, t b t chng nh ci ct nh n b t to nh ci ng tim) cht nhi ct (pha tnh, stationary phase, thng dng silicagel, c 2 loi l silicagel pha thun v silicagel pha o, silicagel pha thun thng dng hn, gm Si c nh cc nhm OH, silicagel pha o gm Si c nh cc dy alkan R, ngoi ra cn dng alumin) Cht nhi ct quyt nh qu trnh sc k ca bn. Nu dng pha thun, silicagel c gn nhm OH th n s gi cht cc cht phn cc ca bn hn, cc cht khng phn cc s lin kt vi silicagel yu hn nn s ra trc, cc cht phn cc hn s ra sau. Ngoi ra ty thuc vo lng mu ca bn c m bn s dng lng silicagel bao nhiu, t bn s chn kch c ct sc k ca bn nh th no. Mt yu t na cng quyt nh n lng silicagel l bn mng ca bn, ty thuc cht bn tch gn nhau hay xa nhau m bn dng lng silicagel gp 20, 30 hay 50 ln lng mu. Ch lng silicagel phi n khong gp 10 ln ng knh ct th qu trnh tch s din ra tt. Nu trn bn mng lng cht ca bn gn nhau th lng silicagel dng phi gp 50 ln lng mu, cn xa nhau th ch cn gp 20 ln l . Dung mi: thng thng bn chn dung mi ph thuc vo cht nhi ct. Nu dng silicagel pha thng (phn cc) th chn dung mi i t khng phn cc n phn cc (v d t chloroform m methanol), thng thng dng hn hp dung mi, cch chn dung mi nh th no l do bn quyt nh da vo thng tin trn bn mng. Ch dung mi chy sc k ct phi phn cc hn mt cht so vi dung mi chy bn mng v ht silicagel dng cho ct sc k s to hn ht trn bn mng nn kh nng tch s km hn mt cht. (sc k ct l phng php tch cht sau khi c tt c thng tin trn bn mng, TLC, thin layer chromatography, l bin php u tin dng "m mm" thng tin v hn hp ca mnh cng nh bin php tch cht cn ly). Sau khi c tt c thng tin ri th bn tin hnh chy ct. Thc hin Bc 1 l nhi ct. Sau khi chn ct, lm kh v cn silicagel cn dng, pha dung mi chy h ri th ha tan silicagel vo dung mi trong mt erlen chng hn. Ly mt ming nh bng gn nhi di y ct chn silicagel li, nu ct

qu di th dng 1 dy km tht bng gn xung, ch dng t bng gn thi. Vi nhng ct c sn ming xp hay lc thy tinh chn silicagel ri th khng cn lm vic ny, tuy nhin ming xp c nhc im l sau khi chy ct xong ta phi ra li nhiu ln cho sch, rt mt thi gian so vi vic nhi bng gn, khi chy ct xong ch cn vt bng gn i l xong. Sau khi nhi cht bng gn th tin hnh khuy silicagel trong dung mi (khng phn cc) pha sn ri rt nh nhng vo ct. Trong lc rt th nn m kha silicagel lng u, nh 1 erlen bn di thu hi dung mi, tip tc lm nh vy n khi cho ht lng silicagel vo, tin hnh rt thm dung mi (t erlen bn di) n nh h, ch i ch li nhiu ln n khi h n nh, ct khng b nt hay gy th xem nh vic nhi ct hon thnh. Kha ct li. Bc 2: np mu cht vo. C 2 loi np mu l np mu kh v np mu t. Np mu t l hn hp ca bn cng tan trong dung mi chy ct nn ch cn cho mu vo l c. Cn nu mu ca bn khng tan trong dung mi chy ct th bn phi ha tan mu vo dung mi gin tip trong 1 erlen chng hn. Sau dng lng t silicagel cho vo erlen hp thu mu, sau cho ht vo bnh c quay c quay ui dung mi i thu c silicagel kh c cha mu. Lc ny np ht silicagel vo ct v cho dung mi chy ct vo. Ci ny gi l np mu kh. Khi c quay th nh lt ming bng gn ming bnh c quay trnh silicagel bay ln. C quay gi nm na l chng ct trong p sut km (chn khng). Bc 3: sau khi hon tt vic np mu ri th lt 1 ming bng gn bn trn mu cht n nh h ri tip tc chm dung mi vo, t t thay i phn cc ca h, ch khng c thay i t ngt cng nh chuyn t khng phn cc sang phn cc ri li quay li khng phn cc, lm nh vy s gy ct v phi nhi li t u, mt ht cht. Bc 4: m kha, lc ny ct bt u tch cht, hng lng dung mi chy ra c km theo cht tch c bng h bi, mi ln hng khong 1/5 h (mi h bi cha c khong 50-200mL ty kch thc ln nh). Sau em chm bn cc h bi, nhng h c vt tng t nhau s c gom li, l 1 cht. Tip tc nh vy th cui cng ta s tch c cc cht mong mun. Ghi ch Mi ln tch cn khong 50-200 h bi, thi gian tch t 2 n 7 ngy. Dng kha iu chnh tc dng chy dung mi, ra chm hay nhanh, vic ny quyt nh qu trnh tch ca bn c tt hay khng. Nu thn trng qu cho chy chm th bn phi ch lu, nu nn nng qu cho chy nhanh th silicagel ca bn

cha kp tch phi cho ra cht, nh vy cng khng tch c. C nhng h phi tch rt lu, cho d m kha ht c vn khng chy xung nhanh th bn dng my y (my sc oxi dng nui c). N s sc khng kh vo tng tc qu trnh y dung mi xung (ch dng khi tc dng di khong 3 git/pht). Ch cht ca bn khng d b oxi ha th mi dng phng php ny c. Nu khng oxi khng kh s oxi ha ht cht ca bn. Nu dng silicagel pha o th ct khng cn kha v pha o chy rt chm, nhng nhc im l bn phi ngi canh n lin tc, tc l nu tch 3 ngy th bn phi thc trng 2 m canh ct! CH khi chy pha o th khng c dng my y, v bn cht pha o l chm (ging nh chy ngc chiu s cng an thi th ai cng phi chy chm, s b tng na). Nu dng my y th nh ni trn, silicagel khng tch c, qu trnh tch ca bn hoi cng. Khi chy ct th iu ti k l kha ct li qu lu. D nhin chy ct 2-3 ngy th bn phi kha ct li cui ngy i v, nhng hm sau phi ln lm ngay, nu qu lu th cht b gi li trong ct lu s kh tch ra hn. Kin thc cn hn hp, ni hi kh hiu nhng hy vng ng gp c cht t. Thn!

Shadow 09-22-2008, 09:23 AM Hi ! Cm n tie.pok rt nhiu, ng l mnh lm cng 3, 4 ti ri nhng cha trch cao bao gi :24h_104: nhng bn khng hiu cu hi ca mnh ri ! Khi c mt mu cao trn tay, lm sao bit c l nhng cht khng phn cc hay l phn cc ? ( nhng phn on ang c lp lm sao bit nhng cht trn ! ). Cn vn thang chia phn cc, thc ra n ch bit ci no ln hn, ci no nh hn thi, v d nh nhm flavone gi l phn cc hn nhm triterpen, nhng flavon l phn cc hay khng phn cc, hay gi l phn cc trung bnh ? Khng c thang chia no r rng c :24h_093: Cn v qu trnh trch cao, quy trnh ca bn a ra rt chi tit nhng khng bit bn c lm cha hay ch nghe ni :hutthuoc( v c mt s im l lm !

1. Mi ln ngm cao khng ai ngm 10,20 g c ! lm nh vy chng no mi xong :24h_125: v cng khng ai sc k lp mng cao MeOH ht -> v trong cao MeOH c rt nhiu cht trong -> 100% khng th thy tch g c !!! 2. V khng th tch ra c nn ngi ta mi t cao MeOH chia thnh nhiu phn on c phn cc khc nhau, bng cch trch lng-lng vi tng loi dung mi c phn cc khc nhau, hay dng cch trch rn lng nh bn ni cng c ( cch ny c nghe ni nhng cha thy bao gi ) -> sau mi thc hin sc k lp mng vi cc phn on thu c -> thng th chng ta hay la loi cao no tch p, d lm trc ( ng ra l khng phi la chn nh vy, phi th hot tnh tng phn on ri la chn loi cao c hot tnh cao kho st ) 3- C ch tch trn ct HPLC v sc k ct thng thng l ging nhau, bn thy khc nhau v bn cha chy sc k ct pha o bng C18 thi. HPLC tch tt hn v ht nhi trong ct nh hn ht nhi trong sc k ct rt nhiu ( nu bn dng loi ht nh ny nhi trong sc k ct th cng c nhng dung mi khng th chy qua ! in hnh cho trng hp ny l sc k lp mng thng tch tt hn sc k ct, ti sao n tch tt hn :24h_125: -> n gin v ht ca sc k lp mng nh hn ht ca sc k ct thi. Mnh bit ch c vy :018: c g ng gp nha :24h_057: tie.pok 09-23-2008, 09:20 PM Hello shadow Cm n bn trao i. Mnh c mt s kin nh th ny: 1. Cc flavonoid th ai cng bit l chng thuc lp cht phn cc (bn cht do s chuyn dch in t trong vng thm v nhm C=O cng c ng gp cho s phn cc ca flavonoid). Cng chnh v n phn cc nn gn y c lng mi kho st cc flavonoid trong HCTN (cc cht km phn cc c cc tin bi kho st ht ri v cho bit rng chng chng c hot tnh j hay ho), m khi lm loi ny, ngi ta hay vt ngay i ci phn chit ban u t nHexan. Theo kinh nghim th flavonoids thng xut hin phn chit Etyl acetat v Metanol.

Lm thao tc chit lng lng v bn cht ngha v mc ch cng ch l c lp cc lp cht khng phn cc, phn cc trung bnh v phn cc. 2. Trong bi vit trn ca mnh, ngm 10 - 20g mu trong MeOH l bc kho st ban u ( chm bn mng tm h dung mi, s b nh gi tng quan hm lng gia cht trong mu, v a i th hot tnh khng nm khng khun ca mu tng). Ch khng phi ch ngm tng y lm hm b lng cc bc sau ri a lln ct. Mun lm a ln ct, t nht cng fi c 200g n... rt nhiu g mu kh. M nh bn ni, c rt nhiu cht trong mu chit MeOH nn ngi ta mi phi th vi SKBM vi c chc h dm khc nhau. Khi SKBM dng h dm khng phn cc th cc cht khng phn cc s chy ln cn cc cht phn cc vn nm nguyn vt gc, sau SKBM vi cc h dm phn cc tng dn cho n khi trn bn mng th hin cc cht khng phn cc v phn cc va nm ht tin tuyn dm cn cc cht phn cc c tch bn di vi cc Rf khc nhau cn vt gc ch cn m m l ok. Tt nhin y ch l bc kho st c bn ban u nn vic tch r rng cc cht ra khi nhau l 1 iu khng tng. 3. ng nh bn ni vic tch trn CT ca SK ct (tr ct Sephadex) v tch trn CT ca HPLC l hon ton ging nhau v nguyn tc v c ch tch. Mnh bit l bn lm nhiu v HPLC nn bn hiu. Nhng iu mnh mun ni l c ch hot ng ca HPLC li khc nhiu v c c cc iu kin tuyt vi nh p sut, gradient nng ,... 4. Bn nhc n loi ct C18, c phi ci m thin h lu nay vn gi l ct pha o khng? Vt liu nhi trong ct l silica gel c nh vi gc Hydrocacbon no R (chui 18 C) chnh thng R ny lm cho loi silica gel ny tr nn rt khng phn cc. Khi chy ct C18 ngi ta chy bng H2O, H2O:MeOH, MeOH. M loi ny c ci hay rt hp vi tnh tit kim ca ngi Vit l khi dng xong c th ra i v ln sau lm th nhi ct dng li. Uh, nu ng l n th mnh cng c may mn dng ht 1 thng ca Meck ri. thuhanoi2304 11-06-2008, 11:26 PM

anh Tie.pok mi dng ht 1 thng thi ah?thn tng ca t y.ci ny t khng ni TungPham1986 03-18-2009, 02:55 PM Thy mi ngi bn v HCTN mnh cng xin gp mt s kin th ny. Vic chn h dung mi chy sc k ct da vo kt qu sc k lp mng. Sau khi c c cc cn chit t dch cn ( Cn n-hexan, iclometan, Etyl Axetat, Metanol), sc k lp mng c tin hnh vi cc h dung mi vi phn cc tng dn, mnh thng lm vi N-hexan ( ete du ha)->iclometan->etylaxetat>metanol. Cc h dung mi c kho st vi t l tng dn cht phn cc hn. Cn chn c h dung mi ph hp th ty vo kinh nghim ca tng ngi v ty vo mch ch phn tch. Trong qu trnh SKLM th c dng thm thuc th nh FeCl3 hay Vanidin/H2SO4 th nh tnh mt s cht c trong cn chit. Cn vic chy SKLM sau khi thu c cc phn on ch yu l gom cc phn on ging nhau li ( c th l n cht hay hn hp) nu l n cht th kt tinh v chp ph l OK. Cn nu l hn hp th tin hnh sc k ct ln hai( mnh thng lm trong cng t ht) s thu c cht tinh khit. TungPham1986 03-18-2009, 02:59 PM anh Tie.pok mi dng ht 1 thng thi ah?thn tng ca t y.ci ny t khng ni HUyn phi ko? Anh kia tht l i gia chi hn mt thng ha cht Merck. Chc l lm trong vin no c ti ln, hay l lm nc ngoi. Ch VN th ha cht chai Tu l i gia lm ri. Dn HCTN ton mua dung mi cng nghip v ct li ch cha thy ai mua ha cht Merck bao gi.