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 haimbobuda muchimiro. Patiri mundima yechromatography tsvakurudzo dzimwe misoro kana pfungwa dzinopera basa sezvo dzichitsiviwa nemazano matsva uye ari nani, mundima yekugadzirisa matambudziko, sezvo chinyorwa chekutanga chekugadzirisa matambudziko chakabuda mujenari rino (nenguva iyo LC Journal ichiri kushanda) 1983 (1) . Mumakore mashoma apfuura, ndakatarisa akati wandei LC Troubleshooting zvikamu pamazuva ano maitiro anokanganisa liquid chromatography (LC) (somuenzaniso, kuenzanisa kuenzanirana kwekunzwisisa kwedu kwemhedzisiro yekumanikidza kuchengetedza [2] New Advances) Dudziro yedu yemhedzisiro yeLC uye maitiro ekugadzirisa matambudziko nemwedzi uno weLC'm inoenderera mberi, mumwedzi waZvita 3 'zvishandiso zvakatanga. 2021, iyo yakanangana nedzimwe nyaya dze "hupenyu nerufu" dzeLC troubleshooting - zvinhu zvakakosha kune chero mugadziri wematambudziko akakosha, zvisinei nezera rehurongwa hwatiri kushandisa.Musoro wemusoro weiyi nhevedzano unoenderana zvakanyanya neLCGC yakakurumbira "LC Troubleshooting Guide" wall chart (4) yakarembera mumarabhoritari akawanda. maitiro.Zvinoshamisa, chati yemadziro inonyora 44 zvikonzero zvakasiyana-siyana zvinogona kukonzera urombo hwepamusoro chimiro!Hatigoni kufunga nezvese nyaya idzi zvakadzama munyaya imwe chete, saka muchikamu chino chekutanga pamusoro pehurukuro, ndichaisa pfungwa pane dzimwe dzandinoona kakawanda.Ndinovimba kuti vashandisi veLC vaduku nevakuru 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) . "Peak shape" kwete chete pano, peak yereferensi yezvinyorwa ( shapesymmetric) kwete chete pano, peak of referendum. asymmetrical, smooth, fluffy, lead edge, tailing, etc.), asiwo kuhupamhi.Zvatinotarisira kune chaiyo peak shape zviri nyore.Theory (6) inotsigira zvakanaka tarisiro yebhuku rekunyora kuti, kazhinji, chromatographic peaks inofanira kunge yakaenzana uye inopindirana nechimiro chekugovera kweGaussian, sezvinoratidzwa mumufananidzo uyu, isu tinotarisira kubva kune imwe nyaya yakaoma. musoro munyaya yeramangwana.Mamwe maumbirwo epamusoro-soro muMufananidzo 1 anoratidza zvimwe zvingangoonekwa-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. nhongonya imwe chete kana kuti shoma inobatwa, asi zvimwe zvacho zvinotaridzika zvakanaka, chikonzero chinogona kunge chiri chemakemikari.
Zvikonzero zvekemikari zvepeak tailing zvakanyanya kuoma kuti zvikurukure muchidimbu pano.Muverengi anofarira anotaurirwa kunyaya ichangoburwa ye "LC Troubleshooting" kuti iwedzere kukurukurirana kwakadzika (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, diki diki diki. analyte masses, kana mamiriro echromatographic anofanirwa kuchinjwa 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 ndeyekusabatana kwakashata pane imwe nzvimbo pakati peinjector uye detector (12) .Muenzaniso wakanyanyisa unoratidzwa muFigure 1d iyi jekiseni ine jekiseni idzva rakagadzirwa mavhiki mashomanana apfuura muvhavha yangu. iyo yatakanga tisina kushandisa kare, uye yakaisa diki vhoriyamu jekiseni loop ine ferrule yakanga yakaumbwa pasimbi isina tsvina capillary.Mushure meimwe miedzo yepakutanga yekugadzirisa dambudziko, takaona kuti kudzika kwechiteshi mujekiseni yevhavha stator kwakadzika zvakanyanya kudarika zvatakajaira, zvichiita kuti ivhu guru rakafa pasi pechiteshi. bvisa vhoriyamu yakafa pasi pechiteshi.
Peak fronts seaya anoratidzwa muFigure 1e anogonawo kukonzerwa nematambudziko emuviri kana kemikari.Chikonzero chinowanzoita chechikamu chinotungamira ndechekuti iyo particle bed ye column haina kunyatso kurongedza, kana kuti zvidimbu zvakagadziridzwa nekufamba kwenguva.Sezvo nepeak tailing inokonzerwa nechiitiko ichi chemuviri, nzira yakanakisisa yekugadzirisa iyi ndeyekutsiva mbiru uye kuramba tichienda. Mamiriro ekuchengetedza "asina mutsara".Pasi pemamiriro ezvinhu akanaka (mutsara), huwandu hweanalyte hunochengetedzwa nechikamu chakamira (saka, chinhu chekuchengetedza) chinoenderana nemutsara weanoongorora mukoromo.Chromatographically, izvi zvinoreva kuti sezvo kuwanda kweanalyte injected mu column inowedzera, iyo peak inova kureba, asi hukama hwekuchengetedza hunova hurefu, asi hukama hwekuchengetedza hunova hurefu uye husina hukama hwakakura. mapeaks haangove akareba chete asiwo akafara sezvo huwandu huzhinji huri jekiseni.Mukuwedzera, maumbirwo asina mutsara anotarisa chimiro chekromatographic peaks, zvichiita kuti itungamirire kana kuti inotevedza mipendero.Sezvo nekuremerwa kwehuwandu hunokonzera peak tailing (10), peak inotungamira inokonzerwa nekuchengetedzwa kusina mutsara inogonawo kuongororwa nekuderedza injected analyte nzira inokonzera kuti iwedzere majekiseni emhando, kana peak analyte nzira igadziriswe. inotungamira, kana mamiriro echromatographic anofanirwa kuchinjwa kuderedza maitiro aya.
Dzimwe nguva tinocherechedza izvo zvinoratidzika 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) . "Kutsemuka" nhongonya dzinechekuita nemuviri Kuita hakunei nechekuita nekoramu pachayo.Kazhinji, chinonyanya kukosha chezano kune iyi sarudzo ndeyekuti peaks ese muchromatogram anoratidza maumbirwo akapatsanurwa, 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 chakasvika kumagumo ekupedzisira ekupindura.Kune optical absorbance detectors (UV-vis munyaya iyi), apo kuongorora kweanalyte kwakanyanya kwazvo, analyte inobata zvizhinji zvechiedza chinopfuura kuburikidza ne detector flow cell, zvichisiya zvishoma zvishoma zvinotarisa magetsi kubva pamagetsi. inopesvedzerwa zvakanyanya neakasiyana siyana eruzha, senge mwenje wekurasika uye "rima razvino", zvichiita kuti chiratidzo chive "chisina kujeka" pakutaridzika 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.
Muchikoro chechromatography, tinoshandisa detector signal (kureva, y-axis muchromatogram) sechiratidzo chekuongorora kweanalyte mumuenzaniso.Saka zvinoratidzika zvisinganzwisisike kuona chromatogram ine chiratidzo chiri pasi pe zero, sezvo dudziro iri nyore ndeyokuti izvi zvinoratidza kukanganisa kwekuongorora - izvo zvisingagoneki mumuviri zvinogoneka. (semuenzaniso, UV-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 izvi marefu. shandisa nhongonya dzisina kunaka kugadzirira curve yecalibration uye kuwana ruzivo rwakakwana rwehuwandu, saka hapana chikonzero chakakosha chekudzivisa pase (iyi nzira dzimwe nguva inodaidzwa se "indirect UV kucherechedzwa") (13).Zvisinei, kana tichida chaizvo kudzivirira nhongonya dzisina kunaka zvachose, kana iri nyaya yekuona kunyura, mhinduro yakanaka ndeyokushandisa imwe nzira yakasiyana-siyana kuitira kuti iwedzere kushandura mawaveleng ekufambisa kwefoni, kuitira kuti iwedzere kushandura mafungu enhare. chikamu kuitira kuti Vanotora chiedza chishoma pane vanoongorora.
Negative peaks inogonawo kuoneka kana uchishandisa refractive index (RI) kuona kana refractive index yezvimwe zvikamu kunze kwe analyte mumuenzaniso, seye solvent matrix, yakasiyana nerefractive index ye mobile phase.Izvi zvinoitikawo nekuonekwa kweUV-vis, asi mhedzisiro iyi inotaridza kudzikiswa maererano neRI kuona. phase.
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 tarisiro kuri pachena.Peak shape matambudziko ane zvakawanda zvakasiyana zvinogona kukonzera (yakanyanya kupeta, kupinza, nezvimwewo). zvikonzero zvandinowanzoona.Kuziva mashoko aya kunopa nzvimbo yakanaka yekutanga kugadzirisa matambudziko, asi hakutore zvose zvingangoitika.Vaverengi vanofarira rumwe ruzivo rwakadzama rwezvikonzero uye mhinduro vanogona kureva LCGC “LC Troubleshooting Guide” wall chart.
(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.