Qhov sib txawv tseem ceeb ntawm RP HPLC thiab HIC yog tias RP HPLC siv lub zog ntawm lub xov tooj ntawm tes ntau dua thiab qis dua polar nyob ruaj khov. Whereas, HIC siv lub sijhawm hydrophobic nyob ruaj ruaj uas tso cai rau cov hydrophobic molecules txuas rau nws.
Chromatography yog cov txheej txheem tshuaj ntsuam xyuas uas pab cais cov sib tov rau hauv nws cov khoom. Ua ntej, peb yuav tsum yaj cov qauv kom muab cais ua ib qho kev daws teeb meem. Qhov kev daws teeb meem-cov khoom sib tov no yog hu ua theem mobile. Tom qab ntawd lub sijhawm txawb tau dhau los ntawm lwm cov khoom siv hu ua lub sijhawm nyob ruaj khov. Cov theem nyob ruaj khov txiav txim siab qhov kev sib cais ntawm cov khoom. Kev sib cais tshwm sim vim yog kev faib khoom ntawm cov khoom siv ntawm cov theem mobile thiab cov theem nyob ruaj khov.
RP HPLC yog dab tsi?
Lo lus RP HPLC sawv cev rau Reverse Phase High-Performance Liquid Chromatography. Nws koom nrog kev sib cais ntawm cov khoom sib xyaw ua ke, raws li hydrophobicity. Lub hydrophobic Cheebtsam nyob rau hauv lub mobile theem txuas nrog lub immobilized hydrophobic ligands nyob rau hauv lub nyob ruaj khov theem, thaum hydrophilic Cheebtsam elute los ntawm cov theem nyob ruaj ruaj yam tsis tau txuas mus rau saum npoo ntawm cov theem nyob ruaj ruaj.
Daim duab 01: HPLC Apparatus
Ntxiv mus, txoj kev no muaj kev tsim tawm ntau dua piv rau lwm cov txheej txheem chromatographic, thiab nws qhia tau hais tias siv dav dav thiab. Yog li, peb siv txoj kev no hauv chav kuaj rau ntau dua 75% ntawm tag nrho HPLC txoj kev. Feem ntau ntawm lub sijhawm, peb siv cov dej sib xyaw ua ke ntawm cov dej nrog miscible polar organic hnyav raws li lub sijhawm txawb. Yog li, nws yuav xyuas kom meej qhov txuas ntawm hydrophobic Cheebtsam hauv kev daws rau saum npoo ntawm qhov chaw nyob ruaj khov.
HIC yog dab tsi?
Lub sij hawm HIC sawv rau Hydrophobic Interaction Chromatography. Nws yog ib hom kev rov qab theem HPLC, thiab txoj kev no yog siv feem ntau rau kev sib cais ntawm cov biomolecules loj xws li cov proteins. Hauv txoj kev no, peb yuav tsum tau siv qhov nruab nrab aqueous los ua cov qauv ntawm biomolecule. Nws yog vim hais tias cov kuab tshuaj organic tuaj yeem ua rau denaturing ntawm cov proteins. Ntxiv mus, cov txheej txheem no siv kev sib cuam tshuam ntawm cov molecules loj nrog me ntsis hydrophobic nto ntawm qhov chaw nyob ruaj khov. Tsis tas li ntawd, peb yuav tsum siv cov ntsev ntau hauv cov qauv vim tias nws txhawb nqa cov protein kom khaws cia ntawm qhov ntim; txheej txheem no hu ua s alting tawm. Los ntawm kev txo qis ntsev concentration maj mam, peb tuaj yeem ua rau biomolecules elute cais raws li lawv cov hydrophobicity.
Feem ntau, txoj kev no siv cov xwm txheej tawm tsam ntawm ion-exchange chromatography. Hauv cov txheej txheem no, ua ntej, peb yuav tsum dhau ib qho kev daws teeb meem los ntawm kem txhawm rau txhawm rau txo cov kev daws teeb meem hauv cov qauv. Nws ua rau thaj tsam hydrophobic ntawm cov protein kom nthuav tawm. Txawm li cas los xij, thaum lub molecule yog ntau hydrophobic, tsawg ntsev yuav tsum tau los txhawb txoj kev khi. Ntawm no, qhov tsawg hydrophobic solutes yog eluted ua ntej, thiab raws li qhov kev hloov ntawm ntsev concentration, ntau hydrophobic solutes yog eluted kawg.
Qhov txawv ntawm RP HPLC thiab HIC yog dab tsi?
Chromatography yog cov txheej txheem tshuaj ntsuam xyuas rau kev sib cais ntawm cov khoom sib xyaw. RP HPLC thiab HIC yog ob txoj kev tshwj xeeb chromatographic. Qhov sib txawv tseem ceeb ntawm RP HPLC thiab HIC yog tias RP HPLC siv lub zog ntawm lub xov tooj ntawm tes ntau dua thiab tsawg dua polar nyob ruaj khov, hos HIC siv cov theem hydrophobic nyob ruaj ruaj, uas tso cai rau cov hydrophobic molecules txuas rau nws.
Cov hauv qab no qhia txog qhov sib txawv ntawm RP HPLC thiab HIC.
Summary – RP HPLC vs. HIC
Chromatography yog cov txheej txheem tshuaj ntsuam xyuas rau kev sib cais ntawm cov khoom sib xyaw. RP HPLC thiab HIC yog ob txoj kev tshwj xeeb chromatographic. Qhov sib txawv tseem ceeb ntawm RP HPLC thiab HIC yog tias RP HPLC siv lub zog ntawm lub xov tooj ntawm tes ntau dua thiab qis dua polar nyob ruaj khov theem, hos HIC siv lub hydrophobic nyob ruaj ruaj, uas tso cai rau cov hydrophobic molecules txuas rau nws.
