Kem chromatography yog cov txheej txheem siv los cais thiab ntxuav cov tebchaw raws li lawv cov polarity thiab kev cuam tshuam nrog cov theem nyob ruaj khov thiab txawb. Nov yog cov lus qhia ib kauj ruam ntawm kev npaj thiab ua kab chromatography:
Cov ntaub ntawv xav tau:
Kem: iav lossis yas kem nrog lub stopcock.
Nyob ruaj ruaj theem: Silica gel los yog alumina (adsorbent).
Mobile theem: hnyav los yog hnyav sib tov (eluent).
Qauv: Qhov sib tov yuav tsum tau muab cais.
Cov khoom ntim (yeem): paj rwb lossis iav ntaub plaub.
Beaker lossis collection vials: Los sau cov feem ntau.
Khoom siv: Clamp thiab sawv ntsug, pipette, funnel, spatula.
Cov kauj ruam los npaj Kem Chromatography:
Xaiv cov Adsorbent:
Xaiv silica gel lossis alumina nyob ntawm qhov polarity ntawm cov tebchaw yuav tsum tau sib cais.
Qhov loj me ntawm cov adsorbent (40-63 µm) cuam tshuam rau kev sib cais.
Npaj Kem:
Txuas hauv qab ntawm kem nrog paj rwb los yog iav ntaub plaub kom tiv thaiv adsorbent los ntawm to.
Clamp lub kem vertically rau ib qho chaw muag khoom nrog lub stopcock kaw.
Wet Packing (Xav tau rau cov txiaj ntsig zoo tshaj):
Ntxiv ib qho me me ntawm cov kuab tshuaj (theem mobile) rau hauv kem.
Npaj ib qho slurry ntawm cov adsorbent nyob rau hauv cov kuab tshuaj (siv 1.5-2 zaus ntawm kab ntawv ntim ntawm adsorbent).
Maj mam ncuav lub slurry rau hauv kem siv ib tug funnel.
Tso cai rau cov adsorbent los daws los ntawm maj mam tapping kab los yog cia nws so.
Qhib lub stopcock luv luv kom ntws cov kuab tshuaj ntau dhau kom txog rau thaum lub adsorbent tsim ib lub txaj compact thiab qib.
Qhuav Packing (Lwm txoj kev):
Ntxiv cov qhuav adsorbent ncaj qha rau hauv kem.
Maj mam coj mus rhaub rau sab kom paub meej tias txawm ntim.
Ua tib zoo ntxiv cov kuab tshuaj raws phab ntsa kom tsis txhob cuam tshuam cov ntim adsorbent.
Qhov xwm txheej ntawm kab:
Thaum ntim, pre-elute lub kem nrog hnyav kom tshem tawm cov hnab tshos huab cua thiab sib npaug ntawm cov adsorbent.
Load tus qauv:
Dissolve tus qauv nyob rau hauv ib tug me me ntawm lub mobile theem.
Maj mam muab cov qauv tso rau sab saum toj ntawm cov khoom ntim adsorbent siv lub pipette lossis syringe.
Cia tus qauv kom adsorb mus rau theem nyob ruaj khov.
Ntxiv Phase Txawb:
Ua tib zoo ntxiv cov kuab tshuaj rau sab saum toj ntawm kem yam tsis muaj kev cuam tshuam lub txaj adsorbent.
Qhib lub stopcock kom pib elution, tswj tus nqi tsis tu ncua.
Sau Fractions:
Sau cov kuab tshuaj eluted nyob rau hauv sequential fractions siv cov raj los yog vials.
Saib xyuas cov feem uas siv UV teeb, TLC (thin-txheej chromatography), los yog lwm yam kev tsom xam.
Optimize lub Solvent System:
Pib nrog ib qho hnyav hnyav lossis ib qho hnyav-polarity tsawg.
Maj mam nce lub polarity yog cov tebchaw tsis elute.
Tswv yim rau kev vam meej:
Tsis txhob muaj npuas npuas: xyuas kom lub txaj adsorbent tau ntim nruj thiab sib npaug.
Tswj Flow Rate: Tus nqi ntws tsis tu ncua thiab nruab nrab muab kev sib cais zoo dua.
Gradient Elution: Siv cov kuab tshuaj sib tov nrog maj mam nce polarity kom daws tau qhov zoo dua.
Saib Xyuas Kev Txhim Kho: Siv TLC los xyuas qhov sib cais thiab txheeb xyuas cov feem.
Los ntawm kev ua raws li cov kauj ruam no, koj tuaj yeem sib cais thiab ntxuav koj cov tebchaw siv kab chromatography.