1. Sc k lng

2. 1. Khi nim v k thut sc k lng- Phng php tch- Cc cu t c tch phn b giapha tnh v pha ng Mobile phase(Pha ng)Stationary phase(Pha tnh) 3. - Qu trnh tch da vo tnh cht ha hc, vtl v ha l ca cc cht.- Da trn 2 qu trnh: Hp ph Gii hp ph- Xy ra lin tc gia 2 pha:Pha tnh: cht rn hoc lngPha ng: cht lng (1 cht hoc hn hp nhiu cht) 4. Pha ng: ha tan v di chuyn cht phn tchPha tnh: gi cht phn tchSKL chia thnh 2 nhm- SK lng p sut thng (sc k c in)- SK lng p sut cao (SKL hiu nng cao: HPLC) (High Performance Liquid Chromatography) 5. Da vo bn cht ca qu trnh sc k, HPLC:- SK phn b- SK pha thng (normal phase chromatography- SK pha o (reversed phase chromatography)- Sk trao i ion (ion exchange chromatography)- SK ghp cp ion (ion pair chromatography) 6. - Da vo trng thi pha tnhPha ng: Lng SK lng lng (LLC)Pha tnh: Lng(Liquid liquid chromatography)Pha ng: lng SK lng rn (LSC)Pha tnh: Rn (Liquid solid chromatography) 7. Khi ni vi u do (detector), HPLC cho php:- nh tnh: da vo thi gian lu- nh lng: da vo chiu cao hoc din tch peak 8. 2. Nguyn tc cu to ca h thng myHPLC 0 123450: Ngun cung cp pha ng (mobile phase)- Bnh cha pha ng 9. 1: Bm cao p (h thng cung cp dungmi)- Bm pha ng vo ct tch- iu khin tc dng, p sut ca phang 10. 2. Van bm mu (Injection valve):- Bm mu PT vo ct tch theo nhng lngmu nht nhTim mu bng tay Tim mu t ng (Auto sampler) 11. 3: Ct tch (Column)- Ct cha pha tnh- Yu t quyt nh qu trnh tch sc k - Ct tch c kch c khc nhau - Chiu di: 10 25cm - ng knh: 2 5mm 12. 4: u d (detector)- Thit b pht hin cht phn tch (nh tnhv nh lng)- C nhiu loi khc nhau ty mc ch phntch: UV-VIS, Huynh Quang, oDan, ien Hoa, Khoi Pho, 13. 5. H thng ghi nhn v x l tn hiu:- Thu thp v x l kt qu- Recorder, Computer + printer, software 14. InjectorH thng HPLC n ginColumn Mobile phaseDetector tM tR1 tR2 tR3 tR4 Start 15. Detector UV-Vis: detector ph hp thu phn tXv nh cc cht c kh nng hp thu quang Hunh quang (Fluorescence detector): xc nh cc cht c kh nng pht hunh quang- Alflatoxin, Mycotoxin, Amino Acid, thuc tr suh Carbamate,.udchskhc x (Refractive Index Detector: RI) 16. u d dn (Conductivity detector):Xc nh cc ion v c, hu c u d khi ph (MS: mass spectrometry)Xc nh phn ln cc cht hu c 17. 3. Cc qu trnh tch trong sc k lng- Qu trnh quan trng nht trong phng phpsc k- Nhng cn bng ng xy ra gia pha tnh vpha ng trong ct sc k- L s vn chuyn v phn b lin tc ca chtPT t u ct n cui ct 18. - Cht phn tch lun phn b gia 2 pha,trong pha ng lun chy qua ct tch vimt tc nht nh hoc gradient- Hiu qu ca qu trnh tch ph thuc rtnhiu vo tng tc gia cc cht trong phatnh v pha ng- Mc ch chnh ca sc k l tch v nhtnh cc cht trong hn hp cht phc tp 19. - Thi gian cht PT b pha tnh lu gi (thigian lu) quyt nh bi: Bn cht ca pha tnh, cu trc v tnh cht ca cht PTBn cht v thnh phn ca pha ng dng ra gii cht PT ra khi ct sc k (pha tnh) 20. - Ghi li ton b qu trnh tch sc k ca hnhp cht PT sc k gm nhiu peak.- c im ca peak PT: Cc peak c th tch ri nhau hon ton Chp nhau mt phn Chp nhau hon ton 21. - Sc k phn nh qu trnh tch sc k trong ct tt hay khng tt.- Tch tt: hn hp c bao nhiu cht c by nhiu peak ring bit khng chp nhau- Cht no b lu gi mnh s c ra gii ra sau cng, cht lu gi km s ra trc 22. 4. u im ca phng php sc k- C th phn tch ng thi nhiu hp cht- Khng cn lm bay hi mu- phn gii cao nh qu trnh tch trn ct- nhy cao (ppm-ppb) nh u d- Th tch mu phn tch nh (1-100L) 23. Hn hp cht tch khi nhau th no ?Flow Pha tnh 24. Ti sao li c s khc nhau?MnhYu Do lc tng tc khc nhau 25. Cc cn bng trong ct HPLCMu PT i qua ct: tn ti ng thi 3 thnh phn:- SP khng chuyn ng- Cht PT va chuyn ng t u ct n cuict, ng thi phn b li gia SP v MP: B SP hp ph, ri ra gii bi MP- MP: dung mi ra gii v chuyn ng Tng tc vi cc cht, li ko v mang cht PT ra khi ct 26. 3 tng tc chnh:- S tng tc v cn bng ca cht PT vi SP- S tng tc v cn bng ca cht PT vi MP- S tng tc ca SP v MP 27. PTFbFa FcSPMP 28. Ftot = Fa + Fb + Fc- Cht no c lc tng tc ln nht s bgi li lu trn ct- Cht no c F nh b ra gii u tin- Ftot khc nhau nhiu th qu trnh tch tt 29. Qu trnh tch din ra trong ct sc kcolumn Vt liu nhi ct 3- 5m 30. Qu trnh tchmixed sampleMobile phase column 31. 5. Sc k lng pha thng v pha oPha thngPha oPha tnh Phn cc Khng phn ccPha ngKhng phn Phn cc cc 32. Ct nhi trong sc k pha thng- Ct silica trung tnh: -B mt c cha nhmHH H phn cc (a nc): -OHOO O -Xc nh cc cht khngSiSi Si phn cc hoc t phnO OOO ccSiSi 33. - Ct silica trn nn mch carbon: -Si-CH2CH2CH2CN -Si-CH2CH2CH2NH2 -Si-CH2CH2CH2OCH(OH)-CH2(OH)+ Ct cyano: a mc ch+ Ct amino: phn tch ng+ Ct diol: protein 34. - Dung mi ch yu: khng phn cc+ Hydrocarbon: hexan, pentan, octan+ Aromatic hydrocarbon: benzen, toluen, xylen,+ Chloroform CHCl3+ Methylene chloride CH2Cl2- Dung mi ph: phn cc hoc hi phn cc+ Ethanol, methanol,. 35. Lin kt hydrogen v thi gian lu HO 1MnhPhn SiOHcc2 1SiOHYuRt yuOH Rt 2 36. Lin kt hydrogen Mu phn tch c:COOH: nhm carboxyl Lin ktOH: nhm hydroxylhydrogen mnhNH2 : nhm amino Mu phn tch c nhm tert-butyl hoc ccnhm khng phn cc ln LK hydrogen s yu 37. Ct nhi trong sc k pha o- Ct: khng phn cc- Pha ng: phn cc- Ct silica alkyl ha cc nhm OH trnb mt silica trung tnh: Ct C18 (ODS)B mt khng phn cc Ct C8 (octyl) Ct C4 (butyl)hay t phn ccCt phenyl (k nc) 38. Xc nh cht phn cc, khng phn cc v t phn cc 39. Thi gian lu v lin kt k nc HO1 C18 (ODS) YuMnhOH 12 2 40. Dung mi trong HPLC pha o- Dung mi phn cc: Methanol(CH3OH: MEOH), acetonitrile (CH3CN: ACN)- Dung dch m Ti u ha t l dd m/ dung mi rt quan trng qu trnh tch tt 41. So snh pha thng v pha o Thng sNormal Phase Reversed Phase phn cc ca ct CaoThp phn cc ca dung Thp Caomi Th t ra gii Cht km phnCht phn cc racc ra trc trcTng phn cc Ra gii nhanhRa gii chmdung mihnhn 42. ng dng ca HPLC- Ch yu xc nh cc hp cht hu c khbay hi trong nhiu i tng khc nhau:+ Amino acid+ Acid hu c+ Thuc tr su+ . 43. Sc k ion (Ion Chromatography)Ion Exchange R Lc ion N+ RSample R + + + + + + - Sample+SO3+ + + + + + 44. - Phn tch cc hp cht ion- C 2 loi ct:+ Ct trao i cation -Strong cation exchange : (R-SO3 )-Weak cation exchange : (R-COO )+ Ct trao i anion:Strong anion exchange : (R4N+)Weak anion exchange :diethyl aminoethyl) 45. Pha ng: thng l dung dch m trong dung mi ncAnion: Carbonat/bicarbonate (Na2CO3/NaHCO3) Potassium hydroxide (KOH)Cation: Nitric acid Tartaric acid Tartaric acid/dipicolinic acid Tartaric acid/citric acid 46. n tp1. Khi nim v phn tch nh lng2. Tnh ton v x l s liu phn tch3. Phng php phn tch dng c- PP ph hp th phn t- PP ph nguyn t- PP in ha- PP sc k lng4. Cc bi thc tp, tnh ton s liu t s othc nghim