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 zitigera ziva muburyo.Mu gihe mubijyanye nubushakashatsi bwa chromatografiya 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 kibazo (LC Ikinyamakuru icyo gihe cyashize)) ku mpinduka zigezweho zigira ingaruka kuri chromatografiya (LC) (kurugero, kugereranya ugereranije no gusobanukirwa ningaruka zumuvuduko mukugumana [2] Iterambere Rishya) Ibisobanuro byacu bya LC nuburyo bwo gukemura ibibazo hamwe nibikoresho bigezweho bya LC.Mu gice cyuku kwezi, ndakomeza urukurikirane rwanjye (3), rwatangiye mu Kuboza 2021, rwibanze ku ngingo zimwe na zimwe z '"ubuzima n' urupfu" ni ibibazo byingenzi bikemura ibibazo bya LC. y'uruhererekane rufite akamaro kanini cyane ku mbonerahamwe ya LCGC izwi cyane ya "LC Troubleshooting Guide" yerekana urukuta (4) yimanitse muri laboratoire nyinshi.Ku gice cya gatatu cy'uruhererekane, nahisemo kwibanda ku bibazo bijyanye n'imiterere y'impinga cyangwa ibiranga impinga. Igitangaje ni uko imbonerahamwe y'urukuta igaragaza urutonde 44 rutandukanye rushobora gutera ibibazo mu buryo budasubirwaho mu ngingo imwe, bityo rero muri iki gice cya mbere nzabibanda ku ngingo za kera, kandi ndizera ko muri iki gice cya mbere. ers 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 nkeka 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, ibishushanyo, byerekana ko ari impuzamugambi, ibishushanyo mbonera, byerekana ko ari byiza. ku bugari. Ibyo dutegereje kumiterere yimpinga nyayo biroroshye. Igitekerezo (6) gishyigikira neza igitabo cyateganijwe ko, mubihe byinshi, impinga ya chromatografique igomba kuba ihuye kandi igahuza nuburyo bwo gukwirakwiza Gaussiya, nkuko bigaragara ku gishushanyo cya 1a.Ibyo dutegereje mubugari bwimisozi ni ikibazo kitoroshye, kandi tuzaganira kuri iyi ngingo mu buryo buzaza mu zindi ngingo. inder yiki 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.
Ubudozi bwo hejuru, impinga imbere, 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 impinga zitandukanye muri chromatogramu, urashobora kubona ibimenyetso byingenzi aribyo nyirabayazana. ni imiti.
Impamvu ziterwa na chimique zitera umurizo ziragoye cyane kubiganiraho muri make hano.Umusomyi ushimishijwe yerekejwe kukibazo giheruka cya "LC Troubleshooting" kugirango baganire ku buryo bwimbitse (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 "massage overtable". kugirango imiterere yimpinga nziza iboneke nubwo hamwe nimbaraga nini zatewe.
Hariho nimpamvu nyinshi zishobora kuba zifatika zo kudoda 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 mumashusho 1d, twashizwemo na feri mucyumweru gishize twashizemo na feri ya feri, kandi twashizeho muri feri ya feri, twashizemo inshinge nkeya twashizeho. itegeko ryari ryarahinduwe kuri capillary idafite ingese.Nyuma yubushakashatsi bwambere bwo gukemura ibibazo, twabonye ko ubujyakuzimu bwicyambu muri stator ya valve yatewe inshinge zimbitse cyane kuruta uko twari dusanzwe tubimenyereye, bikavamo umubare munini wapfuye munsi yicyambu. Iki kibazo cyakemutse byoroshye mugusimbuza uruzitiro rwinshyi hamwe nubundi buryo, dushobora guhindura ferrule mukibanza gikwiye kugirango tumenye icyuma cyapfuye.
Impinga zingana nkizerekanwa ku gishushanyo cya 1e nazo zishobora guterwa nibibazo byumubiri cyangwa imiti.Impamvu isanzwe yumubiri itera inkombe yambere ni uko uburiri bwibice byinkingi butapakiwe neza, cyangwa se ko ibice byongeye gutondekanya igihe.Nkuko umurizo wimpinga uterwa niki kintu gifatika, inzira nziza yo gukosora ibi ni ugusimbuza inkingi kandi ikomeza kugendana. yandujwe nicyiciro gihagaze (niyo mpamvu, ibintu byo kugumana) bifitanye isano neza 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 idafite umurongo, cyangwa impinga ntiziba ndende gusa. s hamwe nuburemere burenze urugero butera umurizo (10), impinga yambere iterwa no kugumana umurongo utari muto irashobora kandi gupimwa mugabanya misa ya analyte yatewe.Niba imiterere yimpanuka igenda itera imbere, uburyo bugomba guhinduka kugirango butarenga ubwiza bwinshinge butera imbere, cyangwa imiterere ya chromatografi igomba guhinduka kugirango igabanye iyi myitwarire.
Rimwe na rimwe, twitegereza ibisa nkimpinga "itandukanijwe", nkuko bigaragara ku gishushanyo cya 1f.Intambwe yambere mugukemura iki kibazo nukumenya niba imiterere yimpanuka iterwa no gufatanya igice (nukuvuga, kuba hariho ibice bibiri bitandukanye ariko bivuguruzanya cyane) .Niba mubyukuri hariho analyite ebyiri zitandukanye zegerana hamwe, noneho nikibazo cyo kunoza imyanzuro yabo, hamwe no gukora ibisa na byo bifitanye isano no gukora, en, ibimenyetso byingenzi kuri iki cyemezo nukumenya 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 ku rugero ruheruka muri laboratoire yanjye, ubusanzwe yerekana ko ikimenyetso ari kinini ku buryo cyageze ku ndunduro y’urwego rwo gusubiza. Kubireba optique yo kwinjiza optique (UV-vis muri iki gihe), iyo analyte yibanze cyane, analyte ikurura urumuri rwinshi runyura mumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo uturuka kumasoko yumucyo ukabije. , gukora ikimenyetso "fuzzy" cyane mubigaragara kandi bitagendeye kuri analyte yibanze.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 ubu burebure cyangwa ibice bya tekinike bishobora gukoreshwa nka methanol cyangwa bffer iboneka nka methanol cyangwa bffer ibona ibintu nka methanol cyangwa buffer. amakuru yuzuye yuzuye, kubwibyo rero ntampamvu yingenzi yo kubyirinda kuri buri gihe (ubu buryo rimwe na rimwe bwitwa "indirect UV detection") (13) .Nyamara, niba dushaka rwose kwirinda impinga mbi rwose, mugihe cyo gutahura ibyinjira, igisubizo cyiza nukwifashisha uburebure butandukanye bwo kumenya kugirango analyte ikure ibirenze icyiciro kigendanwa kugirango isesengure icyiciro cya mobile.
Impinga mbi irashobora kandi kugaragara mugihe ukoresheje indangagaciro zivunika (RI) mugihe indangagaciro yo kwanga ibice bitari analyte murugero, nka matrike ya solvent, itandukanye nigipimo cyo kwangirika cyicyiciro cya mobile.Ibi kandi bibaho no kumenya UV-vis, ariko izi ngaruka zikunda guhuzwa ugereranije nicyitegererezo cya RI.Mu bihe byombi, impinga mbi ihuye na matrike igendanwa ihuye na matrike ya mobile.
Igice cya gatatu kumutwe wibanze wo gukemura ibibazo bya LC, naganiriye ku bihe imiterere yimpinga yagaragaye itandukanye nuburyo buteganijwe cyangwa busanzwe bwo hejuru.Gukemura ibibazo nkibi bitangirana no kumenya imiterere yimpinga ziteganijwe (ukurikije inyigisho cyangwa uburambe bwambere hamwe nuburyo buriho), bityo rero gutandukana nibi bitekerezo biragaragara. Ibibazo byimiterere yibitekerezo bifite ibisobanuro byinshi muburyo butandukanye (ndabigaragaza cyane muburyo butandukanye. ahantu 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.