Mimwe misoro yeLC troubleshooting haisati yapera, sezvo paine nyaya muLC kudzidzira, kunyangwe tekinoroji yezviridzwa inovandudzwa nekufamba kwenguva.Kune nzira dzakawanda dzekuti matambudziko angamuka muLC system uye anopedzisira aita hurombo hwepamusoro chimiro.Kana nyaya dzine chekuita nepeak shape dzamuka, pfupiso pfupi yezvikonzero zvinokonzeresa izvi zvinobatsira kurerutsa ruzivo rwedu rwekugadzirisa matambudziko.
Zvanga zvichinakidza kunyora iyi "LC Troubleshooting" column uye kufunga pamusoro pemisoro mwedzi wega wega, nekuti mimwe misoro haimbobude muchimiro.Tichiri mundima yechromatography tsvakurudzo dzimwe misoro kana pfungwa dzinopera basa sezvo dzichitsiviwa nemazano matsva uye ari nani, muchikamu chekugadzirisa matambudziko, sezvo chinyorwa chekutanga chekugadzirisa matambudziko chakabuda mujenari rino (iyo LC ver 3 Journal ichiri makore gumi apfuura) , Ini ndakatarisa akati wandei LC Troubleshooting zvikamu pane zvemazuva ano maitiro anokanganisa liquid chromatography (LC) (semuenzaniso, kuenzanisa kwehukama hwekunzwisisa kwedu kwemhedzisiro yekumanikidza kuchengetedza [2] New Advances) Dudziro yedu yeLC zvabuda uye nzira yekugadzirisa nemidziyo yeLC yemazuva ano. s yeLC troubleshooting - zvinhu zvakakosha kune chero mugadziri wematambudziko akakosha, zvisinei nezera rehurongwa hwatiri kushandisa.Musoro wemusoro weiyi nhevedzano yakakosha kune yakakurumbira "LC Troubleshooting Guide" yemadziro chati yeLCGC (4) yakarembera mumarabhoritari mazhinji.Pachikamu chechitatu cheiyi nhevedzano, ndakasarudza kutarisisa nyaya dzine chekuita nepamusoro-soro yemhando yepamusoro 4 chati inokonzeresa yemhando yepamusoro. t funga nezvezvinhu zvose izvi zvakadzama munyaya imwe chete, saka muchikamu chekutanga ichi pamusoro pehurukuro, ndichaisa pfungwa pane dzimwe dzandinoona kakawanda.Ndinovimba kuti vechidiki nevakuru veLC vanoshandisa vachawana mazano anobatsira uye zviyeuchidzo panyaya iyi inokosha.
Ndinozviona ndava kuramba ndichipindura mibvunzo yekupedza dambudziko ndichiti “chero chinhu chinogoneka”. Mhinduro iyi ingaite seyakapfava kana tichifunga zvakaonekwa zvakaoma kududzira, asi ndinoona zvakakodzera.Nezvizhinji zvinokonzeresa kushata kwemamiriro ezvinhu, zvakakosha kuti tirambe takavhurika pfungwa kana tichifunga kuti dambudziko ringavei, uye kuti tikwanise kukoshesa zvingakonzeresa kutanga nhamburiko dzedu, tichitarisa pane izvo zvinonyanya kukosha, zvinokwanisika.
Nhanho inokosha mune chero maitiro ekugadzirisa matambudziko - asi imwe yandinofunga kuti yakaderedzwa - kuziva kuti pane dambudziko rinoda kugadziriswa.Kuziva kuti pane dambudziko rinowanzoreva kuziva kuti chii chinoitika kune chigadziro chakasiyana nezvatinotarisira, izvo zvinoumbwa nedzidziso, ruzivo rwemagetsi, uye ruzivo (5) .Iyo "peak shape" kwete chete, iyo inonzi peak of smooth, shapesymmetric, kwete chete pano, inoratidzika seyesymmetric. ffy, kutungamirira kumucheto, kuswededza, nezvimwewo), asiwo kuhupamhi.Zvatinotarisira kune chimiro chaicho chepamusoro chiri nyore.Chidzidzo (6) chinonyatsotsigira tarisiro yebhuku rekunyora kuti, kazhinji, mapeji echromatographic anofanira kunge akaenzana uye anoenderana nechimiro chekugoverwa kweGaussian, sezvinoratidzwa mumufananidzo 1a.Zvatinotarisira kubva pane imwe nyaya yehumwe hupamhi huchakurukurirana mune imwe nyaya yeramangwana. gure 1 rinoratidza zvimwe zvingango onekwa—nemamwe manzwi, dzimwe nzira dzingangoshata.Muchikamu chasara chechikamu chino, tichapedza nguva tichikurukura mimwe mienzaniso chaiyo yemamiriro ezvinhu anogona kutungamirira kumhando dzemaumbirwo aya.
Dzimwe nguva mapeaks haaonekwe zvachose muchromatogram maanotarisirwa kuvharwa.Iyo pamusoro pemadziro chati inoratidza kuti kusavapo kwepamusoro (tichifunga kuti sampuli inenge iine chinangwa chekuongorora pahutano hunofanira kuita kuti detector iite mhinduro yakakwana kuti ione iyo pamusoro pemheremhere) inowanzoenderana kune imwe nyaya yezviridzwa kana kuti mamiriro ezvinhu asina kunaka efoni (kana akacherechedza zvachose).peaks, kazhinji zvakare "vakaneta"). Rondedzero pfupi yezvingangoitika matambudziko nemhinduro muchikamu ichi inogona kuwanikwa muTafura I.
Sezvambotaurwa pamusoro apa, mubvunzo wekuti inguva yakawanda yekuwedzera kwepamusoro inofanira kushivirirwa isati yateerera uye kuedza kuigadzirisa inyaya yakaoma yandichakurukura munyaya inotevera.Chiitiko changu ndechokuti kuwedzera kukuru kwepamusoro kunowanzoperekedzwa nekushanduka kukuru kwepeak shape, uye peak tailing inonyanya kufanana kupfuura pre-peak kana kupatsanurwa.Zvisinei, iyo inonzi symmetric peaks inogona kukonzerwa nezvikonzero zvakasiyana-siyana zvinogonawo kukonzerwa:
Imwe neimwe yenyaya idzi yakakurukurwa zvakadzama muzvinyorwa zvakapfuura zve Troubleshooting LC, uye vaverengi vanofarira nyaya idzi vanogona kutarisa kune izvi zvakapfuura zvinyorwa kuti vawane ruzivo nezve midzi zvikonzero uye zvinogona kugadzirisa nyaya idzi.Mamwe mashoko.
Peak tailing, peak fronting, uye kupatsanurana zvose zvinogona kukonzerwa nekemikari kana zviitiko zvemuviri, uye urongwa hwezvinogona kugadzirisa matambudziko aya hunosiyana-siyana, zvichienderana nokuti tiri kutarisana nekemikari kana dambudziko remuviri.Kazhinji, nekuenzanisa mapeji akasiyana-siyana muchromatogram, unogona kuwana zviratidzo zvakakosha pamusoro pekuti ndeupi muparidzi.Kana iyo yakawanda inokonzera chimiro chimwe chete chinenge chisingaiti chinenge chiri muchimiro chimwe chete kana kuti chromatogram. mapeaks akanganiswa, asi mamwe ese anotaridzika zvakanaka, chikonzero chinogona kunge chiri chemakemikari.
Zvikonzero zvemakemikari zvinokonzeresa peak tailing zvakanyanya kuoma kuti zvikurukure muchidimbu pano.Muverengi anofarira anotumirwa kune ichangoburwa nyaya ye "LC Troubleshooting" kune imwe kurukurirano yakadzama (10) .Zvisinei, chinhu chiri nyore kuedza ndechekuderedza kuwanda kweiyo injected analyte uye kuona kana iyo peak chimiro ichivandudza.Kana zvakadaro, saka ichi ndicho chiziviso chakanaka chekuti dambudziko rinopfuura, mudiki massess anofanira kunge ari diki nzira. kana mamiriro echromatographic anofanirwa kushandurwa kuitira kuti maumbirwo akanaka epamusoro anogona kuwanikwa kunyangwe nemajekiseni akakura.
Kunewo zvakawanda zvinogona zvikonzero zvemuviri zvepeak tailing.Vaverengi vanofarira kukurukurirana kwakadzama kwezvingabvira vanotumirwa kune imwe nyaya ichangoburwa ye "LC Troubleshooting" (11) .Chimwe chezvikonzero zvinowanzoitika zvemuviri zvepeak tailing ndiko kusabatana kwakashata pane imwe nzvimbo pakati pejekiseni uye detector (12) .Muenzaniso wakanyanyisa unoratidzwa muFigure 1b, takashandisa majekiseni matsva masvondo adarika, takashandisa majekiseni mashomanana apfuura. isati yasvika, uye yakaisa chidimbu chejekiseni chevhoriyamu ine ferrule yakanga yakaumbwa pasimbi isina tsvina capillary.Mushure memamwe maitiro ekutanga ekugadzirisa matambudziko, takaona kuti kudzika kwechiteshi mujekiseni rejekiseni stator kwakadzika zvakanyanya kudarika zvatakajaira, zvichiita kuti ivhu guru rakafa pasi pechiteshi. .
Peak fronts seaya anoratidzwa muFigure 1e anogonawo kukonzerwa nedambudziko remuviri kana kemikari.Chikonzero chinowanzoita chechikamu chinotungamira ndechekuti iyo particle bed yembiru haina kunyatso kurongedza, kana kuti zvimedu zvakagadziridzwa nekufamba kwenguva.Sezvo nepeak tailing inokonzerwa nechiitiko ichi chemuviri, nzira yakanakisa yekugadzirisa izvi ndeyekutsiva iyo column uye kuramba tichienda. tion conditions.Pasi pezvakanakira (linear) mamiriro, huwandu hweanalyte hunochengetwa nechikamu chakamira (saka, the retention factor) inofananidzwa nemutsara weconcentration yeanalyte mukoramu.Chromatographically, izvi zvinoreva kuti sezvo huwandu hweanalyte hunopinzwa mukoramu hunowedzera, peak inova kureba, asi kwete yakafara.Uhu hukama hunoputsika uye husina hurefu hunove husingarebe kana hurefu hunova huzhinji husingaverengeki. inoiswa jekiseni.Mukuwedzera, maumbirwo asina mutsara anotarisa chimiro chechromatographic peaks, zvichiita kuti kutungamirira kana kutevedza mipendero.Sezvinei nehuwandu hunorema hunoita kuti peak tailing (10), peak inotungamira inokonzerwa nekuchengetwa kusina mutsara inogonawo kuongororwa nekuderedza injected analyte mass.Kana peak shape inovandudza, nzira yacho inofanira kuchinjwa kuti iite majekiseni anotungamira kuti asapfuure ematorographic maitiro, iyo inokonzera kuchinjwa kwehutano hunoita kuti isapfuure ematorographic. maitiro.
Dzimwe nguva tinocherechedza izvo zvinoita se "kuparadzaniswa" kwepamusoro, sezvinoratidzwa mumufananidzo 1f. Danho rokutanga pakugadzirisa dambudziko iri nderokuona kana chimiro chepamusoro chinokonzerwa nechikamu chekubatana (kureva, kuvapo kwezvikamu zviviri zvakasiyana asi zvakanyatsojeka). nhongonya dzine chekuita nemuviri Kuita hakunei nechekuita nembiru pachayo.Kazhinji, chinonyanya kukosha chekuratidza sarudzo iyi ndechekuti nhongonya dzese dziri muchromatogram dzinoratidza zvimiro zvakapatsanurwa, kana imwe chete kana maviri.Kana iri imwe chete kana maviri, ingangove inyaya yekubatanidzwa;kana nhongonya dzese dzakapatsanurwa, ingangove inyaya yemuviri, ingangove ine chekuita nekoramu pachayo.
Kupatsanurwa kwepamusoro kunoenderana nemaitiro emuviri wekoramu pachayo kunowanzo kukonzerwa nechikamu chakavharwa chinopinda kana chinobuda frits, kana kurongeka patsva kwezvikamu mukoramu, zvichibvumira kuti mobile phase ifambe nekukurumidza kupfuura iyo mobile phase mune dzimwe nzvimbo dze column channel formation .mune mamwe matunhu (11) .zvisinei, mune zvakaitika kwandiri, izvi zvinowanzova zvenguva pfupi pane kugadzirisa kwenguva refu.Izvi zvinowanzouraya nembiru dzemazuva ano kana zvikamu zvibatanidza zvakare mukati membiru.Panguva iyi, zvakanakisisa kutsiva mbiru uye kuenderera mberi.
Iyo yepamusoro muFigure 1g, zvakare kubva kune imwe nguva ichangobva kuitika mu laboratori yangu, kazhinji inoratidza kuti chiratidzo chacho chakakwirira zvokuti chasvika kumagumo ekupedzisira ekupindura.Kune optical absorbance detectors (UV-vis munyaya iyi), apo kuongorora kweanalyte kwakakwirira kwazvo, muongorori anotora chiedza chinopfuura nepakati pe detector flow cell, achisiya zvishoma zvishoma zvinoonekwa kubva kune magetsi emagetsi. kwakasiyana siyana kweruzha, senge rakarasika mwenje uye "rima razvino", zvichiita kuti chiratidzo chive "chisina kujeka" muchitarisiko uye chakazvimiririra kubva kune analyte concentration.Kana izvi zvikaitika, dambudziko rinogona kugadziriswa nyore nyore nekudzikisa huwandu hwejekiseni reanalyte-kudzikisa huwandu hwejekiseni, kudimburira sampuli, kana zvese zviri zviviri.
Mu chromatography chikoro, ie, iyo y-axis muChrromato of ChromatoGound -vis).
Muchiitiko ichi, peak yakashata inongoreva kuti mamorekuru ari kubuda kubva mukoramu anotora chiedza chiduku pane chikamu chefoni pachayo pakarepo uye mushure mepamusoro.Izvi zvinogona kuitika, semuenzaniso, kana uchishandisa zvishoma zvishoma zvekuona wavelengths (<230 nm) uye mobile phase additives iyo inotora yakawanda yechiedza pane idzi wavelengths.Zviwedzerero izvi zvinogona kuva mobile phase solvent components kana kuti negative components se-peaktatethanometer inogona kushandiswa se-negative metaket-se-peak-medium-medium-sex. kugadzirira calibration curve uye kuwana ruzivo rwakarurama rwehuwandu, saka hapana chikonzero chakakosha chekuvadzivisa pase (iyi nzira dzimwe nguva inonzi "indirect UV kuonekwa") (13) .Zvisinei, kana tichida chaizvo kudzivisa nhongonya dzisina kunaka zvachose, munyaya yekuonekwa kwekunyura, mhinduro yakanakisisa ndeyokushandisa zvakasiyana-siyana zvekuongorora mazamu, kuitira kuti chiedza chinotora zvishoma pane chigadziko chekufamba, kuitira kuti chigadzirise chikamu chemobile tes.
Negative peaks inogonawo kuoneka kana uchishandisa refractive index (RI) kuona kana refractive index yezvikamu zvisiri zve analyte mumuenzaniso, seye solvent matrix, yakasiyana nerefractive index ye mobile phase.Izvi zvinoitikawo nekuonekwa kweUV-vis, asi mhedzisiro iyi inoita kunge yakadzikiswa maererano neRI kuona.
Muchikamu chetatu pamusoro pemusoro wenyaya weLC troubleshooting, ndakakurukura mamiriro ezvinhu umo iyo yakacherechedzwa chimiro chepamusoro inosiyana kubva kune inotarisirwa kana yakajairika peak shape.Kubudirira kugadzirisa matambudziko akadaro kunotanga neruzivo rwezvinotarisirwa peak maumbirwo (zvichienderana nedzidziso kana ruzivo rwekare nenzira dziripo), saka kutsauka kubva kune izvi zvinotarisirwa zviri pachena.Peak shape matambudziko ane akawanda akasiyana anogona kukonzera zvikonzero, zvinotungamira mune izvi zvakanyanya, ndinoona zvikonzero zvakawanda, nezvimwe. kazhinji.Kuziva izvi zvinopa nzvimbo yakanaka yekutanga kugadzirisa matambudziko, asi hazvitore zvese zvingangoitika.Vaverengi vanofarira rondedzero yakadzama yezvikonzero nemhinduro vanogona kureva LCGC “LC Troubleshooting Guide” pamadziro chati.
(4) LCGC "LC Troubleshooting Guide" chati yemadziro.https://www.chromatographyonline.com/view/troubleshooting-wallchart (2021).
(6) A. Felinger, Data Analysis uye Signal Processing muChromatography (Elsevier, New York, NY, 1998), mapeji 43-96.
(8) Wahab MF, Dasgupta PK, Kadjo AF uye Armstrong DW, Anal.Chim.Journal.Rev.907, 31–44 (2016).https://doi.org/10.1016/j.aca.2015.11.043.