Muaj ntau ntau txoj hauv kev los tshuaj xyuas qhov muaj pes tsawg leeg thiab kawm txog cov khoom ntawm ntau lub tebchaw thiab cov khoom sib xyaw. Ib txoj hauv kev zoo li no yog chromatography. Lub authorship nyob rau hauv lub invention thiab daim ntawv thov ntawm txoj kev belongs rau Lavxias teb sab botanist MS Tsvet, uas nyob rau hauv thaum pib ntawm lub xyoo pua 20th ua kev sib cais ntawm cov nroj tsuag pigments.
Definition and fundamentals of method
Chromatography yog ib txoj hauv kev physicochemical rau kev sib cais cov khoom sib xyaw thiab txiav txim siab lawv cov khoom, raws li kev faib tawm ntawm lub xov tooj ntawm tes thiab nyob twj ywm theem ntawm cov tshuaj uas tsim cov khoom sib xyaw (piv txwv). Cov theem nyob ruaj khov yog cov khoom ntxeem tau - ib qho sorbent. Nws tseem tuaj yeem ua cov yeeb yaj kiab ua kua tso rau ntawm qhov chaw ruaj khov. Lub sijhawm txawb - lub eluent - yuav tsum txav mus raws theem nyob ruaj khov lossis ntws los ntawm nws, raug lim los ntawm cov sorbent.
Lub ntsiab lus ntawm chromatography yog qhov sib txawv ntawm cov khoom sib tov yog qhov tsim nyog los ntawm cov khoom sib txawv, xws li qhov hnyav molecular, solubility, adsorbability, thiab lwm yam. Yog li ntawd, tus nqi ntawm kev sib cuam tshuam ntawm lub Cheebtsam ntawm lub mobile theem - sorbates - nrog rau qhov chaw nres tshebtsis zoo ib yam. Qhov no ua rau muaj qhov sib txawv ntawm qhov nrawm ntawm cov molecules ntawm cov khoom sib xyaw nrog rau theem nyob ruaj khov, vim tias cov khoom sib cais thiab cov concentrated nyob rau hauv ntau thaj tsam ntawm cov sorbent. Ib txhia ntawm lawv tawm cov sorbent ua ke nrog rau theem mobile - cov no yog cov thiaj li hu ua unretained Cheebtsam.
Qhov tshwj xeeb zoo ntawm chromatography yog tias nws tso cai rau koj sai sai cais cov sib xyaw ua ke ntawm cov tshuaj, suav nrog cov khoom zoo sib xws.
Txoj kev faib cov hom chromatography
Txoj kev siv hauv kev tshuaj ntsuam tuaj yeem faib ua raws li ntau yam. Cov txheej txheem tseem ceeb ntawm cov qauv no yog raws li hauv qab no:
- tag nrho lub xeev ntawm qhov chaw nres tsheb thiab lub sijhawm txawb;
- lub cev thiab tshuaj lom neeg ntawm kev sib cuam tshuam ntawm sorbent thiab sorbates;
- yuav ua li cas qhia eluent thiab txav nws;
- txheej txheem ntawm kev tso kawm ntawm theem nyob ruaj khov, piv txwv li cov txheej txheem chromatography;
- chromatography lub hom phiaj.
Ntxiv rau, txoj hauv kev tuaj yeem ua raws li qhov sib txawv ntawm cov txheej txheem sorption, ntawm cov txheej txheem ntawm kev sib cais chromatographic (piv txwv li, qis lossis siab siab).
Cia peb saib ze rau ntawm cov txheej txheem tseem ceeb saum toj no thiab cov hom kev siv chromatography feem ntau cuam tshuam nrog lawv.
Eluent and sorbent state of aggregation
Rau lub hauv paus no, chromatography muab faib ua kua thiab roj. Cov npe txheej txheem qhia txog lub xeev ntawm lub sijhawm txawb.
Ua kua chromatography yog cov txheej txheem sivnyob rau hauv cov txheej txheem ntawm kev sib cais ntawm cov mixs ntawm macromolecular tebchaw, nrog rau cov tseem ceeb biologically sawv daws yuav. Nyob ntawm lub xeev ntawm kev sib sau ua ke ntawm cov sorbent, nws muab faib ua kua-kua thiab kua-kua-khoom.
Gas chromatography yog cov hauv qab no:
- Gas adsorption (gas-solid-theem), uas siv cov khoom sorbent, xws li cov thee, silica gel, zeolites lossis porous polymers. Ib qho roj inert (argon, helium), nitrogen, carbon dioxide ua raws li ib qho eluent - cov cab kuj ntawm cov sib tov sib cais. Kev sib cais ntawm cov khoom tsis hloov pauv ntawm qhov sib tov yog ua tiav vim qhov sib txawv ntawm lawv cov adsorption.
- Gas-kua. Cov theem nyob ruaj khov nyob rau hauv cov ntaub ntawv no muaj cov kua zaj duab xis tso rau ntawm ib qho khoom inert puag. Cov khoom piv txwv raug cais raws li lawv cov adsorbability lossis solubility.
Gas chromatography yog dav siv rau kev soj ntsuam ntawm kev sib xyaw ntawm cov organic tebchaw (siv lawv cov khoom decomposition lossis derivatives hauv daim ntawv gaseous).
Kev sib cuam tshuam ntawm sorbent thiab sorbates
Raws li qhov kev ntsuas no, cov hom no txawv xws li:
- Adsorption chromatography, dhau los ntawm cov khoom sib tov sib cais vim qhov sib txawv ntawm cov qib ntawm adsorption ntawm cov tshuaj los ntawm cov sorbent immobile.
- Distribution. Nrog nws cov kev pab, kev sib cais yog nqa tawm raws li qhov sib txawv solubility ntawm cov khoom sib xyaw. Kev sib cais tshwm sim nyob rau hauv lub xov tooj ntawm tes thiab nyob ruaj ruaj theem (hauv cov kua chromatography), los yog tsuas yog nyob rau hauv lub chaw nres tsheb (hauv roj-kua).chromatography).
- Tswv Ntuj. Txoj kev chromatography no yog ua raws li qhov sib txawv ntawm qhov sib txawv ntawm cov txheej txheem precipitates ntawm cov khoom sib cais.
- Kev cais tawm, lossis gel chromatography. Nws yog raws li qhov sib txawv ntawm qhov loj ntawm cov molecules, vim tias lawv muaj peev xwm nkag mus rau hauv cov pores ntawm cov sorbent, lub npe hu ua gel matrix, txawv.
- Affine. Txoj kev tshwj xeeb no, uas yog raws li hom tshwj xeeb ntawm biochemical kev sib cuam tshuam ntawm kev sib cais impurities nrog lub ligand uas tsim cov complex compound nrog ib qho inert carrier nyob rau hauv cov theem nyob ruaj khov. Txoj kev no muaj txiaj ntsig zoo hauv kev sib cais ntawm cov protein-enzymes thiab muaj ntau hauv biochemistry.
- Ion pauv. Raws li cov qauv sib cais, txoj kev no siv qhov sib txawv ntawm qhov muaj peev xwm ntawm cov khoom sib xyaw rau ion txauv nrog rau theem nyob ruaj khov (ion exchanger). Thaum lub sij hawm tus txheej txheem, cov ions ntawm cov theem nyob ruaj ruaj yog hloov los ntawm ions ntawm cov khoom nyob rau hauv lub muaj pes tsawg leeg ntawm lub eluent, thaum lub sij hawm vim qhov sib txawv affinity ntawm lub tom kawg mus rau lub ion exchanger, ib tug sib txawv tshwm sim nyob rau hauv lub ceev ntawm lawv lub zog, thiab yog li lub sib tov sib cais. Rau cov theem nyob ruaj ruaj, ion pauv resins feem ntau yog siv - tshwj xeeb hluavtaws polymers.
Ion-exchange chromatography muaj ob txoj kev xaiv - anionic (khaws cov ions tsis zoo) thiab cationic (khaws cov ions zoo, raws li). Txoj kev no yog siv dav heev: nyob rau hauv kev sib cais ntawm electrolytes, tsis tshua muaj lub ntiaj teb thiab transuranium ntsiab, nyob rau hauv dej purification, nyob rau hauv kev tshuaj ntsuam tshuaj.
Qhov txawv ntawm cov txheej txheem
Muaj ob txoj hauv kev uas tus qauv txav mus rau qhov chaw nyob ruaj khov:
- Kem chromatography nqa tawm cov txheej txheem sib cais hauv ib qho khoom tshwj xeeb - kab chromatographic - ib lub raj, nyob rau hauv lub puab kab noj hniav uas ib tug immovable sorbent yog muab tso rau. Raws li txoj kev sau, cov kab tau muab faib ua ob hom: ntim (lub thiaj li hu ua "packed") thiab capillary, nyob rau hauv uas ib txheej ntawm cov khoom sorbent los yog kua zaj duab xis ntawm cov theem nyob ruaj ruaj yog siv rau saum npoo ntawm phab ntsa sab hauv. Cov kab ntim khoom tuaj yeem muaj cov duab sib txawv: ncaj, U-puab, kauv. Capillary kab yog helical.
- Planar (planar) chromatography. Hauv qhov no, daim ntawv tshwj xeeb lossis phaj (hlau, iav, lossis yas) tuaj yeem siv los ua tus nqa khoom rau lub sijhawm nyob ruaj khov, uas muaj txheej nyias nyias ntawm sorbent. Hauv qhov no, txoj kev chromatography yog hu ua ntawv lossis nyias txheej chromatography, feem.
Tsis zoo li kab ke kab ke, qhov twg cov kab chromatographic siv ntau zaus, hauv cov qauv chromatography, txhua tus neeg nqa khoom nrog cov txheej txheem sorbent tuaj yeem siv ib zaug xwb. Txoj kev sib cais tshwm sim thaum lub phaj lossis daim ntawv tso rau hauv lub thawv nrog eluent.
Taw qhia thiab hloov pauv ntawm eluent
Qhov no yuav txiav txim siab qhov xwm txheej ntawm kev txav mus los ntawm thaj chaw chromatographic raws cov txheej txheem sorbent, uas tau tsim thaum lub sijhawm sib cais ntawm qhov sib tov. Muaj cov txheej txheem xa khoom hauv qab no:
- Pem hauv ntej. Txoj kev no yog qhov yooj yim tshaj plawstxheej txheem ua. Lub sijhawm txawb yog ncaj qha tus qauv nws tus kheej, uas yog txuas mus ntxiv rau hauv kem uas muaj cov sorbent. Nyob rau hauv cov ntaub ntawv no, qhov tsawg tshaj plaws khaws cia tivthaiv, adsorbed phem dua lwm tus, txav raws lub sorbent sai dua lwm tus. Yog li ntawd, tsuas yog thawj cov khoom no tuaj yeem raug cais tawm hauv daim ntawv ntshiab, ua raws cov cheeb tsam uas muaj cov khoom sib xyaw. Tus qauv faib zoo li no: A; A + B; A + B + C thiab lwm yam. Frontal chromatography yog li ntawd tsis muaj txiaj ntsig rau kev sib cais cov khoom sib xyaw, tab sis nws muaj txiaj ntsig zoo hauv ntau yam kev ua kom huv huv, muab cov tshuaj uas yuav tsum tau cais tawm tau qis qis.
- Txoj kev hloov pauv txawv ntawm qhov uas tom qab nkag mus rau qhov sib tov kom sib cais, ib qho eluent nrog ib qho tshwj xeeb displacer yog pub rau hauv kem - ib yam khoom uas muaj sorbability ntau dua li ib qho ntawm cov khoom sib xyaw. Nws displaces lub feem ntau khaws cia, uas displaces lub tom ntej no, thiab hais txog. Cov qauv txav mus raws kab ntawm qhov ceev ntawm cov displacer thiab tsim cov cheeb tsam nyob ib sab ntawm cov concentration. Nrog rau hom chromatography no, txhua qhov khoom tuaj yeem tau txais ib tus zuj zus hauv daim ntawv ua kua ntawm qhov hluav taws xob ntawm kab.
- Txoj kev eluent (tsim) txoj kev yog feem ntau. Nyob rau hauv sib piv rau txoj kev xaav, lub eluent (carrier) nyob rau hauv cov ntaub ntawv no muaj ib tug tsawg sorbability tshaj cov piv txwv Cheebtsam. Nws tsis tu ncua dhau los ntawm txheej sorbent, ntxuav nws. Periodically, nyob rau hauv feem (pulses), qhov sib tov yuav tsum tau muab cais mus rau hauv lub eluent txaus, tom qab uas cov ntshiab eluent yog noj dua. Thaum ntxuav tawm (elution), cov khoom sib cais,Ntxiv mus, lawv cov cheeb tsam concentration tau sib cais los ntawm cov cheeb tsam eluent.
Eluent chromatography ua rau nws muaj peev xwm yuav luag tag nrho cov kev soj ntsuam sib xyaw ntawm cov tshuaj, thiab sib tov tuaj yeem ua tau ntau yam. Tsis tas li ntawd, qhov zoo ntawm txoj kev no yog kev sib cais ntawm cov khoom ntawm ib leeg thiab qhov yooj yim ntawm kev txheeb xyuas qhov sib xyaw ntawm cov khoom sib xyaw. Qhov tsis zoo muaj xws li kev noj ntau ntawm eluent thiab qis qis ntawm cov qauv piv txwv hauv nws tom qab sib cais ntawm qhov hluav taws xob sab. Txoj kev eluent yog dav siv nyob rau hauv ob qho tib si roj thiab kua chromatography.
Chromatographic txheej txheem nyob ntawm lub hom phiaj
Qhov txawv ntawm cov hom phiaj chromatography ua rau nws muaj peev xwm paub qhov txawv ntawm txoj hauv kev xws li kev txheeb xyuas, kev npaj thiab kev lag luam.
Los ntawm kev tshuaj ntsuam chromatography, kev soj ntsuam zoo thiab ntau yam ntawm cov khoom sib xyaw ua tiav. Thaum txheeb xyuas cov qauv piv txwv, thaum tawm hauv kem ntawm chromatograph, lawv mus rau lub ntes - ib qho cuab yeej rhiab heev rau kev hloov pauv hauv cov ntsiab lus ntawm cov khoom hauv eluent. Lub sijhawm dhau los ntawm lub sijhawm tus qauv coj mus rau hauv kem kom txog rau thaum qhov siab tshaj plaws ntawm cov khoom ntawm lub ntsuas ntsuas hu ua lub sijhawm khaws cia. Muab hais tias qhov kub thiab txias ntawm cov eluent tus nqi yog tas li, tus nqi no yog tas mus li rau txhua yam khoom thiab ua lub hauv paus rau kev soj ntsuam zoo ntawm qhov sib tov. Kev tsom xam ntau yog ua los ntawm kev ntsuas thaj tsam ntawm tus kheej peaks hauv chromatogram. Raws li txoj cai, txoj kev eluent yog siv nyob rau hauv analytical chromatography.
Npaj chromatography aims kom cais cov khoom ntshiab los ntawm kev sib tov. Cov kab kev npaj muaj qhov loj duatxoj kab uas hla tshaj analytical.
Kev lag luam chromatography yog siv, ua ntej, kom tau txais cov khoom ntshiab uas xav tau hauv ib qho kev tsim khoom. Thib ob, nws yog ib feem tseem ceeb ntawm kev tswj xyuas niaj hnub thiab kev tswj hwm cov txheej txheem thev naus laus zis.
Kev lag luam chromatograph muaj qhov ntsuas ntawm qhov ntsuas ntawm ib lossis lwm yam khoom thiab tau nruab nrog lub sensor, nrog rau kev tswj thiab kev sau npe. Cov qauv raug xa mus rau cov chromatographs zoo li no nrog qee zaus.
Ntau yam khoom siv Chromatography
Niaj hnub nimno chromatographs yog cov cuab yeej siv thev naus laus zis uas muaj peev xwm siv tau rau ntau qhov chaw thiab rau ntau lub hom phiaj. Cov cuab yeej no ua rau nws muaj peev xwm txheeb xyuas cov txheej txheem sib xyaw ua ke. Lawv tau nruab nrog ntau yam kev ntsuas: thermal conductometric, kho qhov muag, ionization, huab hwm coj spectrometric thiab lwm yam.
Ntxiv rau, niaj hnub chromatography siv cov tshuab tswj tsis siv neeg rau kev tshuaj xyuas thiab ua cov chromatograms. Kev tswj tau tuaj yeem ua los ntawm lub computer lossis ncaj qha los ntawm lub cuab yeej.
Ib qho piv txwv ntawm cov cuab yeej zoo li no yog cov roj chromatograph multifunctional "Crystal 5000". Nws muaj cov txheej txheem ntawm plaub qhov kev hloov pauv tau, lub ntsuas kub hauv kem, hluav taws xob siab thiab ntws tswj, thiab cov roj valve tswj. Yuav kom daws tau ntau yam teeb meem, lub cuab yeej muajlub peev xwm los nruab ob qho tib si ntim thiab capillary kab.
Lub chromatograph yog tswj los ntawm kev siv cov keyboard puv thiab tswj cov zaub lossis (hauv lwm qhov kev hloov kho) los ntawm tus kheej lub computer. Cov cuab yeej tshiab no tuaj yeem siv tau zoo hauv kev tsim khoom thiab hauv ntau qhov chaw tshawb fawb: kev kho mob, forensic, ib puag ncig.
siab siab chromatography
Kev ua kua kem chromatography yog tus cwj pwm los ntawm lub sijhawm ntev ntawm cov txheej txheem. Txhawm rau ua kom nrawm nrawm ntawm cov kua eluent, cov khoom siv ntawm cov theem txawb mus rau sab hauv siab yog siv. Txoj kev niaj hnub thiab pheej hmoo heev no yog hu ua kev ua haujlwm siab ua kua chromatography (HPLC) txoj kev.
HPLC kua chromatograph lub twj tso kua mis muab eluent ntawm tus nqi tas li. Lub zog inlet tsim tuaj yeem ncav cuag 40 MPa. Kev tswj hwm lub khoos phis tawj ua rau nws tuaj yeem hloov pauv cov theem ntawm lub xov tooj ntawm tes raws li qhov kev pab cuam (txoj kev elution no hu ua gradient).
HPLC tuaj yeem siv ntau txoj hauv kev raws li qhov xwm txheej ntawm kev sib cuam tshuam ntawm sorbent thiab sorbate: kev faib tawm, adsorption, loj cais tawm, ion-exchange chromatography. Hom HPLC feem ntau yog cov txheej txheem thim rov qab, raws li kev sib cuam tshuam ntawm hydrophobic ntawm lub polar (aqueous) mobile theem thiab tsis muaj polar sorbent, xws li silica gel.
Txoj kev yog siv dav rau kev sib cais, tshuaj xyuas,Kev tswj xyuas zoo ntawm cov khoom tsis muaj hluav taws xob, thermally tsis ruaj khov uas tsis tuaj yeem hloov mus rau hauv lub xeev gaseous. Cov no yog agrochemicals, tshuaj, khoom noj khoom haus thiab lwm yam complex tshuaj.
Qhov tseem ceeb ntawm kev tshawb fawb chromatography
Ntau hom chromatography yog siv dav hauv ntau qhov chaw:
- inorganic chemistry;
- petrochemicals thiab mining;
- biochemistry;
- tshuaj thiab tshuaj;
- khoom noj khoom haus;
- ecology;
- criminology.
Daim ntawv teev npe no tsis tiav, tab sis qhia txog kev lag luam kev lag luam uas tsis tuaj yeem ua yam tsis muaj chromatographic txoj kev tsom xam, kev sib cais thiab kev huv ntawm cov tshuaj. Nyob rau hauv tag nrho cov cheeb tsam ntawm daim ntawv thov ntawm chromatography, los ntawm scientific lub chaw soj nstuam mus rau industrial ntau lawm, lub luag hauj lwm ntawm cov txheej txheem yog txawm ntau zuj zus raws li niaj hnub technologies rau cov ntaub ntawv ua, kev tswj thiab kev tswj ntawm complex txheej txheem yog qhia.
