Ingingo zimwe na zimwe za LC zo gukemura ibibazo ntizigera zishaje, kuko hariho ibibazo mubikorwa bya LC, nubwo tekinoroji yibikoresho igenda itera imbere mugihe.Hariho inzira nyinshi zishobora kuvuka ibibazo muri sisitemu ya LC bikarangira bitameze neza.
Byarashimishije kwandika iyi nkingi ya "LC Troubleshooting" no gutekereza ku ngingo buri kwezi, kubera ko ingingo zimwe na zimwe zitajya ziva muburyo.Mu gihe mubijyanye na chromatografiya ubushakashatsi ingingo zimwe cyangwa ibitekerezo biba bishaje kuko bisimbuwe nibitekerezo bishya kandi byiza, mubijyanye no gukemura ibibazo, kubera ko ingingo ya mbere yo gukemura ibibazo yagaragaye muri iki kinyamakuru (LC Ikinyamakuru muri kiriya gihe) kuva ku ngingo zimwe na zimwe za LC (icyo gihe LC) icyo gihe). Gukemura ibibazo kubigezweho bigezweho bigira ingaruka kumazi ya chromatografiya (LC) (kurugero, kugereranya ugereranije no gusobanukirwa kwingaruka zumuvuduko mukugumana [2] Iterambere Rishya) Ibisobanuro byacu bya LC nuburyo bwo gukemura ibibazo hamwe nibikoresho bya LC bigezweho. Mugice cyuku kwezi, ndakomeza urukurikirane rwanjye (3), rwatangiye mukibazo cyikibazo cyikibazo cyikibazo cyikibazo cyibibazo bya LC. ya sisitemu dukoresha. Ingingo yibanze yuruhererekane irafitanye isano cyane na LCGC izwi cyane ya "LC Troubleshooting Guide" igicapo cyurukuta (4) kimanitse muri laboratoire nyinshi.Ku gice cya gatatu cyuruhererekane, nahisemo kwibanda kubibazo bijyanye nimiterere yimpinga cyangwa ibiranga impinga. Ntibisanzwe, imbonerahamwe yurukuta irerekana ingingo zose zishobora guterwa muri iyi ngingo muburyo bwa mbere, nzashyiraho ingingo zose mubisobanuro birambuye kuri iyi ngingo. kenshi.Ndizera ko abakoresha bato n'abakuru LC bazabona inama zingirakamaro nibutsa kuriyi ngingo yingenzi.
Njye mbona ndushijeho gusubiza ibibazo byo gukemura ibibazo hamwe n "" byose birashoboka ".Iki gisubizo gishobora gusa nkicyoroshye mugihe urebye kwitegereza bigoye kubisobanura, ariko ndabona akenshi bikwiye. Hamwe nimpamvu nyinshi zishoboka zitera imiterere mibi yimisozi mibi, ni ngombwa guhora ufunguye ibitekerezo mugihe usuzumye ikibazo gishobora kuba, kandi ukabasha gushyira imbere ibitera imbaraga kugirango utangire imbaraga zacu zo gukemura ibibazo, kwibanda kubintu bishoboka cyane.
Intambwe y'ingenzi mu myitozo iyo ari yo yose yo gukemura ibibazo - ariko imwe nibaza ko idashyigikiwe - ni ukumenya ko hari ikibazo kigomba gukemurwa.Kumenya ko hari ikibazo akenshi bisobanura kumenya ko ibiba ku gikoresho bitandukanye nibyo dutegereje, bigizwe na théorie, ubumenyi bufatika, hamwe n'uburambe (5). "Imiterere y'impinga" ivugwa hano ntabwo isobanura gusa imiterere, impuzamugambi, impuzamugambi, yerekana neza. ) inzira ibintu bishobora kugenda nabi.Mu gice gisigaye cyiki gice, tuzamara umwanya tuganira kuburorero bumwebumwe bwibihe bishobora kuganisha kuri ubu bwoko.
Rimwe na rimwe, impinga ntizigaragara na gato muri chromatogramu aho ziteganijwe gutorwa.Imbonerahamwe yo hejuru y'urukuta yerekana ko kuba nta mpinga (tuvuge ko icyitegererezo kirimo analyte igamije kwibanda ku kintu cyagombye gutuma igisubizo cya detector gihagije kugira ngo kibone hejuru y'urusaku) ubusanzwe gifitanye isano n'ibibazo bimwe na bimwe by'ibikoresho cyangwa niba ibyiciro bya mobile bitari byo (niba bigaragara kuri byose). impinga, mubisanzwe "intege nke") .Urutonde rugufi rwibibazo nibisubizo muriki cyiciro urashobora kubisanga mumeza I.
Nkuko byavuzwe haruguru, ikibazo cyukuntu kwaguka kwimpinga bigomba kwihanganira mbere yo kubyitondera no kugerageza kugikemura ni ingingo igoye nzabiganiraho mu kiganiro kizaza. Ubunararibonye bwanjye nuko kwaguka kwimpinga gukunze guherekezwa nimpinduka zikomeye kumiterere yimpinga, kandi umurizo wimisozi ukunze kugaragara kuruta mbere yimpinga cyangwa gutandukana.Nyamara, impinga zisa nizina nazo ziraguka, zishobora guterwa nimpamvu nke zitandukanye: zishobora guterwa nimpamvu nke:
Buri kimwe muri ibyo bibazo cyaganiriweho ku buryo burambuye mu bibazo byabanjirije ikibazo cyo gukemura ibibazo LC, kandi abasomyi bashishikajwe n’izi ngingo barashobora kwifashisha izi ngingo zabanjirije iyi kugira ngo bamenye amakuru ku ntandaro n’ibisubizo byakemuka kuri ibyo bibazo. Ibisobanuro birambuye.
Umurizo wimpinga, impinga imbere, hamwe no gutandukana byose birashobora guterwa nibintu byimiti cyangwa umubiri, kandi urutonde rwibisubizo byibyo bibazo biratandukanye cyane, ukurikije niba dukemura ikibazo cyimiti cyangwa umubiri. Mugihe, ugereranije nimpinga zitandukanye muri chromatogramu, urashobora kubona ibimenyetso byingenzi aribyo nyirabayazana. reba neza, igitera birashoboka cyane mumiti.
Impamvu ziterwa na chimique zitera umurizo ziragoye cyane kubiganiraho muri make hano.Umusomyi ushimishijwe yoherezwa kubibazo biherutse gusohoka bya "LC Troubleshooting" kugirango biganirweho byimbitse (10) .Nyamara, ikintu cyoroshye kugerageza nukugabanya imbaga ya analyte yatewe hanyuma ukareba niba imiterere yimpanuka itera imbere.Niba aribyo, noneho ibi nibimenyetso byiza byerekana ko ikibazo ari "imbogamizi zirenze urugero". imiterere ya chromatografi igomba guhinduka kugirango imiterere yimpinga nziza iboneke nubwo haterwa imbaga nini.
Hariho nimpamvu nyinshi zishobora kuba zifatika zitera umurizo.Abasomyi bashishikajwe no kuganira birambuye kubishoboka boherezwa kukindi kibazo giherutse kuba cyitwa "LC Troubleshooting" (11) .Imwe mumpamvu zikunze kugaragara kumubiri zitera umurizo ni ihuriro ribi mugihe kiri hagati yuwatewe inshinge na detekeri (12) .Urugero rukabije rwerekanwe mubishusho 1d, twabibonye muri sisitemu yanjye hashize ibyumweru bike twashizeho. Gutera inshinge hamwe na ferrule yari yarabumbabumbwe kuri capillary idafite ingese.Nyuma yubushakashatsi bwambere bwo gukemura ibibazo, twabonye ko ubujyakuzimu bwicyambu muri stator ya valve yatewe bwimbitse cyane kuruta uko twari dusanzwe tubimenyereye, bikavamo ubwinshi bwapfuye munsi yicyambu. Iki kibazo cyakemutse byoroshye mugusimbuza uruzitiro rwumubyimba mukibanza gikwiye kugirango duhindure ferrule kugirango tuyikureho feri kugirango tuyikureho.
Impinga yimpande nkiyerekanwa ku gishushanyo cya 1e irashobora kandi guterwa nibibazo byumubiri cyangwa imiti. Impamvu isanzwe yumubiri itera imbere ni uko uburiri bwibice byinkingi butapakiwe neza, cyangwa se ko ibice byahinduwe neza mugihe cyagenwe.Nkuko umurizo wimisozi watewe niki kintu gifatika, inzira nziza yo gukosora ibi ni ugusimbuza inkingi kandi bikomeza kugenda. ingano ya analyte yagumishijwe nicyiciro gihagaze (niyo mpamvu, ibintu byo kugumana) ifitanye isano cyane no kwibanda kwa analyte mu nkingi. Chromatografique, ibi bivuze ko uko imbaga ya analyte yatewe mu nkingi yiyongera, impinga iba ndende, ariko ntigari yagutse. Iyi mibanire iracika iyo imyitwarire yo kugumana itari umurongo, kandi impinga ntizigizwe gusa na misa. impinga, bikavamo kuyobora cyangwa gukurikira.Nkuko hamwe nuburemere burenze urugero butera umurizo wimpinga (10), impinga yambere iterwa no kugumana umurongo utagabanije irashobora kandi gupimwa mukugabanya misa ya analyte yatewe.Niba imiterere yimpanuka igenda itera imbere, uburyo bugomba guhinduka kugirango butarenga ubwiza bwinshinge butera imbere, cyangwa imiterere ya chromatografique igomba guhinduka kugirango igabanye imyitwarire.
Rimwe na rimwe, twitegereza ibisa nk '“amacakubiri”, nkuko bigaragara ku gishushanyo cya 1f.Intambwe yambere yo gukemura iki kibazo nukumenya niba imiterere yimpanuka iterwa no gufatanya igice (nukuvuga ko hariho ibice bibiri bitandukanye ariko bivuguruzanya cyane) .Niba mubyukuri hariho analyite ebyiri zitandukanye zegerana hamwe, noneho nikibazo cyo kunoza imyanzuro yabo (urugero, nukwiyongera kubitandukanya, gukora, cyangwa gukora isahani) inkingi ubwayo. Nyuma, ibimenyetso byingenzi kuri iki cyemezo ni ukumenya niba impinga zose ziri muri chromatogramu zerekana imiterere itandukanye, cyangwa imwe cyangwa ebyiri gusa. Niba ari kimwe cyangwa bibiri gusa, birashoboka ko ari ikibazo cyo gufatanya; niba impinga zose zacitsemo ibice, birashoboka ko ari ikibazo cyumubiri, birashoboka cyane ko kijyanye ninkingi ubwayo.
Gutandukanya impinga zijyanye nimiterere yumubiri yinkingi ubwayo mubisanzwe biterwa no gufunga igice cyinjira cyangwa gusohoka, cyangwa kuvugurura uduce duto twinkingi, bigatuma icyiciro kigendanwa kigenda cyihuta kuruta icyiciro kigendanwa mubice bimwe na bimwe bigize umuyoboro winkingi .mu tundi turere (11) .Ibice bifunze igice gishobora rimwe na rimwe guhanagurwa no guhindura imigendekere yinkingi; icyakora, mubunararibonye bwanjye, mubisanzwe nibihe byigihe gito aho kuba igisubizo kirekire. Ibi bikunze guhitana inkingi zigezweho niba ibice byongeye guhurira mumurongo. Kuri iyi ngingo, nibyiza gusimbuza inkingi hanyuma ugakomeza.
Impinga iri ku gishushanyo cya 1g, nayo uhereye kurugero ruheruka muri laboratoire yanjye, mubisanzwe yerekana ko ikimenyetso kiri hejuru kuburyo cyageze ku ndunduro y’ibisubizo. Kubireba optique yo kwinjiza optique (UV-vis muri uru rubanza), iyo analyte yibanda cyane, analyte ikurura urumuri rwinshi runyura mumasoko ya moteri, nkurumuri ruto cyane ruturuka kumasoko yumuriro uturuka kumasoko yumuriro ukabije. na "umwijima w'icuraburindi", bigatuma ibimenyetso "fuzzy" cyane mubigaragara kandi bidashingiye kubitekerezo bya analyte. Iyo ibi bibaye, ikibazo gishobora gukemurwa muburyo bworoshye kugabanya inshinge za analyte-kugabanya inshinge, kugabanya icyitegererezo, cyangwa byombi.
Mwishuri rya chromatografiya, dukoresha ibimenyetso bya detector (ni ukuvuga y-axis muri chromatogramu) nkikimenyetso cyerekana analyte yibanze muri sample.None rero birasa nkaho bidasanzwe kubona chromatogramu ifite ikimenyetso kiri munsi ya zeru, kuko ibisobanuro byoroshye ni uko ibyo byerekana kwibanda kuri analyte mbi - birumvikana ko bidashoboka muburyo bwa tekinike.
Muri iki gihe, impinga itari nziza isobanura gusa ko molekile ziva mu nkingi zinjiza urumuri ruto ugereranije na feri igendanwa ubwayo ako kanya mbere na nyuma yimpinga.Ibi birashobora kubaho, kurugero, mugihe ukoresheje uburebure buke bwo gutahura (<230 nm) hamwe ninyongeramusaruro ya feri igendanwa ikurura urumuri rwinshi kuri ubwo burebure. Calibration curve no kubona amakuru yukuri yuzuye, kubwibyo rero ntampamvu yingenzi yo kubyirinda kuri buri gihe (ubu buryo rimwe na rimwe bwitwa "indirect UV detection") (13) .Nyamara, niba koko dushaka kwirinda impinga mbi rwose, mugihe cyo gutahura ibyinjira, igisubizo cyiza nukwifashisha uburebure butandukanye bwo gutahura kugirango analyte ikuremo icyiciro cya mobile.
Impinga mbi irashobora kandi kugaragara mugihe ukoresheje indangagaciro (RI) itahura mugihe indangagaciro yo kwanga ibice bitari analyte murugero, nka matrike ya solvent, itandukanye nigipimo cyangirika cyicyiciro cya mobile.Ibi kandi bibaho hamwe no kumenya UV-vis, ariko izi ngaruka zikunda guhuzwa ugereranije nicyitegererezo cya RI.Mu bihe byombi, impinga mbi ishobora kugereranywa na matrike igendanwa cyane.
Igice cya gatatu kumutwe wibanze wo gukemura ibibazo bya LC, naganiriye ku bihe aho imiterere yimpinga yagaragaye itandukanijwe nuburyo buteganijwe cyangwa busanzwe busanzwe. Gukemura ibibazo nkibi bitangirana no kumenya imiterere yimpinga ziteganijwe (ukurikije inyigisho cyangwa uburambe bwambere hamwe nuburyo buriho), bityo rero gutandukana nibi byifuzo biragaragara. Ibibazo byimiterere yibitekerezo bifite impamvu nyinshi zitandukanye zishobora kubaho (cyane cyane, ndondora, ndabishyira mubisobanuro, ndabibona muburyo butandukanye.) ibi bisobanuro biratanga ahantu heza ho gutangirira gukemura ibibazo, ariko ntibifata ibishoboka byose.Abasomyi bashishikajwe nurutonde rwimbitse rwibitera nibisubizo barashobora kwifashisha imbonerahamwe ya LCGC "LC Troubleshooting Guide".
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(6) A. Felinger, Isesengura ryamakuru no gutunganya ibimenyetso muri Chromatografiya (Elsevier, New York, NY, 1998), p. 43-96.
(8) Wahab MF, Dasgupta PK, Kadjo AF na Armstrong DW, Anal.Chim.Journal.Rev. 907, 31–44 (2016) .https: //doi.org/10.1016/j.aca.2015.11.043.