Lihlooho tse ling tsa LC tsa ho rarolla mathata ha ho mohla li siiloeng ke nako, kaha ho na le litaba tsa ts'ebetso ea LC, joalo ka ha theknoloji ea lisebelisoa e ntlafala ha nako e ntse e feta.Ho na le litsela tse ngata tseo ka tsona mathata a ka hlahang tsamaisong ea LC mme a qetella a le boemong bo bobe ba tlhoro.
Ho bile monate ho ngola kholomo ena ea "LC Troubleshooting" le ho nahana ka lihlooho khoeli le khoeli, hobane lihlooho tse ling ha li tsoe ka mokhoa oa mokhoa.Ha re ntse re le lefapheng la lipatlisiso tsa chromatography lihlooho tse itseng kapa mehopolo e felloa ke nako kaha e nkeloa sebaka ke mehopolo e mecha le e betere, lebaleng la ho rarolla mathata, kaha sengoloa sa pele sa ho rarolla mathata se hlahile koranteng ena (the LC 3 years are still related in the LC Journal of the past 8 years at the past 8) , Ke tsepamisitse maikutlo a 'maloa a LC Troubleshooting likarolo tse mabapi le mekhoa ea kajeno e amang liquid chromatography (LC) (mohlala, papiso e lekanyelitsoeng ea kutloisiso ea rona ea phello ea khatello ea ho boloka [2] New Advances) Tlhaloso ea rona ea liphetho tsa LC le mokhoa oa ho rarolla mathata ka lisebelisoa tsa morao-rao tsa LC. s of LC troubleshooting — elements that are great for any troubleshoor are important, ho sa tsotellehe lilemo tsa tsamaiso eo re e sebelisang.Sehlooho sa mantlha sa letoto lena se bohlokoa haholo ho chate ea lebota e tummeng ea “LC Troubleshooting Guide” (4) e leketlileng lilaboratoring tse ngata t ho nahana ka lintlha tsena kaofela ka botlalo sehloohong se le seng, kahoo karolong ena ea pele ea sehlooho, ke tla tsepamisa maikutlo ho tse ling tseo ke li bonang hangata.Ke tšepa hore basebelisi ba LC ba bacha le ba baholo ba tla fumana malebela le likhopotso tse molemo tabeng ena ea bohlokoa.
Ke iphumana ke ntse ke tsoela pele ho araba lipotso tsa ho rarolla bothata ka "eng kapa eng e ka khoneha". Karabo ena e kanna ea utloahala e le bonolo ha ke nahana ka lintlha tseo ho leng thata ho li hlalosa, empa hangata ke fumana ho nepahetse. Ka lisosa tse ngata tse ka bang teng tsa boemo bo bobe ba maemo a holimo, ho bohlokoa ho ba le kelello e bulehileng ha re nahana hore na bothata e ka ba eng, le ho khona ho etelletsa pele lisosa tse ka bang teng ho qala ts'ebetso ea rona ea ho rarolla mathata, ho tsepamisa maikutlo ho tsona ke lintlha tsa bohlokoa haholo.
Mohato oa bohlokoa mosebetsing ofe kapa ofe oa ho rarolla mathata - empa eo ke nahanang hore e tlase - ke ho hlokomela hore ho na le bothata bo lokelang ho rarolloa.Ho lemoha hore bothata bo teng hangata ho bolela ho lemoha hore se etsahalang ka sesebelisoa se fapane le litebello tsa rona, tse bōpiloeng ke khopolo, tsebo ea matla, le phihlelo (5). ffy, mohatla o etellang pele, mohatla, joalo-joalo), empa hape le bophara.Litebello tsa rona bakeng sa sebopeho sa tlhōrō ea sebele li bonolo.Khopolo (6) e tšehetsa hantle tebello ea buka ea hore, maemong a mangata, litlhōrō tsa chromatographic li lokela ho lekana le ho lumellana le sebopeho sa kabo ea Gaussian, joalokaha ho bontšitsoe setšoantšong sa 1a. gure 1 bontša tse ling tsa menyetla e meng e neng e ka hlokomeloa-ka mantsoe a mang, tse ling tsa litsela tseo lintho li ka senyehang ka tsona.Karolong e setseng ea karolo ena, re tla qeta nako re buisana ka mehlala e itseng e tobileng ea maemo a ka lebisang mefuteng ena ea sebōpeho.
Ka linako tse ling litlhōrō ha li hlokomeloe ho hang ka chromatogram moo ho lebelletsoeng hore li tlosoelitlhōrō, hangata le tsona li “fokola”).Lethathamo le lekhutšoane la mathata a ka bang teng le tharollo sehlopheng sena le ka fumanoa ho Lethathamo la I.
Joalokaha ho boletsoe ka holimo, potso ea hore na ho atolosoa ha tlhōrō e kae ho lokela ho mamelloa pele ho ela hloko le ho leka ho e lokisa ke taba e rarahaneng eo ke tla e tšohla sehloohong se tlang. Phihlelo ea ka ke hore ho atolosoa ha tlhōrō ea bohlokoa hangata ho tsamaisana le phetoho e kholo ea sebopeho sa tlhōrō, 'me peak tailing e tloaelehile ho feta pele ho tlhōrō kapa ho arohana.
E 'ngoe le e' ngoe ea litaba tsena e 'nile ea tšohloa ka ho qaqileng litabeng tse fetileng tsa Troubleshooting LC,' me babali ba thahasellang lihlooho tsena ba ka sheba lihlooho tsena tse fetileng bakeng sa tlhahisoleseding mabapi le lisosa le tharollo e ka 'nang ea e-ba teng litabeng tsena.Lintlha tse ling.
Peak mohatla, peak fronting, le petsoha kaofela li ka bakoa ke lik'hemik'hale kapa lintho tse etsahalang 'meleng,' me lethathamo la tharollo e ka bang teng mathateng ana le fapana haholo, ho itšetlehile ka hore na re sebetsana le bothata ba lik'hemik'hale kapa tsa 'mele.Hangata, ka ho bapisa litlhōrō tse fapaneng tsa chromatogram, u ka fumana lintlha tsa bohlokoa mabapi le hore na molato ke ofe. litlhōrō li ameha, empa tse ling kaofela li shebahala li le ntle, sesosa se ka etsahala haholo ke lik'hemik'hale.
Lisosa tsa lik'hemik'hale tsa peak tailing li rarahane haholo hore li ka tšohloa ka bokhutšoanyane mona.'Mali ea thahasellang o fetisetsoa khatisong ea morao-rao ea "LC Troubleshooting" bakeng sa puisano e tebileng haholoanyane (10). Leha ho le joalo, ntho e bonolo ho leka ke ho fokotsa boima ba mohlahlobi o kentsoeng le ho bona hore na sebopeho sa tlhōrō se ntlafala. kapa maemo a chromatographic a tlameha ho fetoloa e le hore libopeho tse ntle tsa tlhoro li ka fumanoa le ka bongata bo boholo bo kentsoeng.
Hape ho na le mabaka a mangata a 'mele a ka bakang mohatla oa tlhōrō.Babali ba thahasellang puisano e qaqileng ea menyetla ba fetisetsoa khatisong e' ngoe ea morao-rao ea "LC Troubleshooting" (11) .E 'ngoe ea lisosa tse tloaelehileng haholo tsa' mele tsa peak mohatla ke khokahanyo e mpe ka ntlha e 'ngoe pakeng tsa injector le detector (12).Mohlala o feteletseng o bontšoa ho Figure 1d, re ile ra sebelisa mochine o mocha libeke tse seng kae tse fetileng. pele, 'me a kenya e nyenyane bophahamo ba modumo ente lupu le ferrule e neng e bōpehile holim'a tšepe e sa hloekang capillary.Ka mor'a liteko tse ling tsa pele tsa ho rarolla mathata, re ile ra hlokomela hore botebo boema-kepe ka stator belofo ente e ne e tebile haholo ho feta re ne re tloaetse ho, ho fella ka e khōlō bafu bophahamo ba modumo botlaaseng ba koung. .
Litlhōrō tse kang tse bontšitsoeng setšoantšong sa 1 li ka boela tsa bakoa ke mathata a 'mele kapa a lik'hemik'hale.Sesosa se tloaelehileng sa' mele se lebisang pele ke hore karoloana ea karoloana ha e tletse hantle, kapa hore likaroloana li hlophisitsoe bocha ha nako e ntse e ea. Joalo ka peak tailing e bakiloeng ke ketsahalo ena ea 'mele, tsela e molemo ka ho fetisisa ea ho lokisa sena ke ho nkela kholumo sebaka le ho tsoela pele. tion maemo. Tlas'a maemo a loketseng (linear), palo ea analyte e bolokoang ke mokhahlelo o emeng (ka hona, ntlha ea ho boloka) e amana ka mokhoa o lumellanang le mahloriso a mohlahlobi kholumong.Chromatographically, sena se bolela hore ha bongata ba mohlahlobi bo kentsoeng kholomong bo ntse bo eketseha, tlhoro e ba telele, empa e se bophara. Ho phaella moo, libopeho tse se nang moeli li khetholla sebopeho sa litlhōrō tsa chromatographic, e leng se etsang hore ho be le litlhōrō tse etellang pele kapa tse latellanang. Joalo ka boima bo bongata bo bakang peak tailing (10), tlhōrō e etellang pele e bakoang ke ho boloka nonlinear e ka boela ea fumanoa ka ho fokotsa boima ba analyte e entsoeng. boitšoaro.
Ka linako tse ling re bona seo ho bonahalang eka ke tlhōrō ea "ho arohana", joalokaha ho bontšitsoe setšoantšong sa 1f. Mohato oa pele oa ho rarolla bothata bona ke ho fumana hore na sebopeho sa tlhōrō se bakoa ke ho kopana ha karolo e itseng (ke hore, ho ba teng ha metsoako e 'meli e sa tšoaneng empa e haufi-ufi eluting). litlhōrō li amana le 'mele Ts'ebetso ha e amane le kholomo ka boeona.Hangata, leseli la bohlokoa ka ho fetisisa la qeto ena ke hore na litlhōrō tsohle tsa chromatogram li bonts'a libopeho tse arohaneng, kapa tse peli feela.Haeba e le 'ngoe kapa tse peli feela, mohlomong ke taba e kopanetsoeng;haeba litlhōrō tsohle li arohane, mohlomong ke taba ea 'mele, mohlomong e amana le kholomo ka boeona.
Litlhōrō tse arohaneng tse amanang le thepa ea 'mele ea kholomo ka boeona hangata li bakoa ke ho kenngoa ha karolo e itseng kapa li-outlet frits, kapa ho hlophisoa bocha ha likaroloana ka har'a kholomo, ho lumella karolo ea selefouno hore e phalle ka potlako ho feta mohato oa selefouno libakeng tse itseng tsa sebopeho sa mocha oa likholomo. libakeng tse ling (11) .leha ho le joalo, ka phihlelo ea ka, hangata sena ke nako e khutšoanyane ho e-na le tharollo ea nako e telele.Sena se atisa ho bolaea ka litšiea tsa morao-rao haeba likaroloana li kopana hape ka har'a kholomo.Nakong ena, ho molemo ho nkela kholomo sebaka ebe u tsoela pele.
Tlhōrō ho Setšoantšo sa 1g, hape ho tloha ketsahalong ea morao tjena ka laborateng ea ka, hangata e bontša hore letšoao le phahame hoo le fihlileng pheletsong e phahameng ea likarabo tsa likarabo. mehloli e fapaneng ea lerata, joalo ka khanya e lahlehileng le "lefifi la hona joale", ho etsa hore lets'oao le be le "fuzzy" haholo ka chebahalo ebile le sa itšetlehe ka mahloriso a analyte.Ha sena se etsahala, hangata bothata bo ka rarolloa habonolo ka ho fokotsa molumo oa ente ea analyte-ho fokotsa molumo oa ente, ho hlapolla sampuli, kapa ka bobeli.
Sekolong sa Chromatography, re sebelisa letšoao la Deptor (ke hore, li-chromatogram tse sa tsotelleng. .
Tabeng ena, tlhōrō e mpe e bolela feela hore limolek'hule tse tsoang ho kholomo li monya khanya e fokolang ho feta mohato oa selefouno hang-hang pele le ka mor'a tlhōrō.Sena se ka etsahala, ka mohlala, ha u sebelisa maqhubu a batlang a le tlaase (<230 nm) le li-additives tsa mekhahlelo ea selefouno tse monyang boholo ba leseli ka maqhubu ana. ho lokisa curve ea calibration le ho fumana lintlha tse nepahetseng tsa palo, kahoo ha ho na lebaka la bohlokoa la ho li qoba ka se seng (mokhoa ona ka linako tse ling o bitsoa "mokhoa o sa tobang oa UV") (13). tes.
Litlhōrō tse mpe li ka boela tsa hlaha ha u sebelisa tlhahlobo ea refractive index (RI) ha index ea refractive ea likarolo tse ling ntle le analyte ka sampuli, joalo ka solvent matrix, e fapane le index ea refractive ea mobile phase. Sena se boetse se etsahala ka ho lemoha ha UV-vis, empa phello ena e atisa ho fokotsoa ho latela RI ho fumanoa.
Karolong ea boraro ka taba ea mantlha ea ho rarolla mathata a LC, ke buile ka maemo ao ho ona sebopeho sa tlhoro se bonoang se fapaneng le sebopeho se lebelletsoeng kapa se tloaelehileng sa tlhoro.Tharollo e sebetsang ea mathata a joalo e qala ka tsebo ea libopeho tse lebelletsoeng tlhōrōng (ho ipapisitsoe le khopolo kapa boiphihlelo ba pele ka mekhoa e teng), kahoo ho kheloha ho tsoa litebellong tsena ho hlakile. Mathata a sebopeho sa tlhoro a na le lisosa tse ngata tse fapaneng, tse etellang pele ka ho hlaka, joalo-joalo. hangata.Ho tseba lintlha tsena ho fana ka sebaka se setle sa ho qala ho rarolla mathata, empa ha ho nke menyetla eohle.Babali ba nang le thahasello ea lethathamo le tebileng la lisosa le tharollo ba ka sheba chate ea lebota ea LCGC “LC Troubleshooting Guide”.
(4) LCGC "LC Troubleshooting Guide" chate ea lebota.https://www.chromatographyonline.com/view/troubleshooting-wallchart (2021).
(6) A. Felinger, Tlhahlobo ea Boitsebiso le Ts'ebetso ea Lipontšo ho Chromatography (Elsevier, New York, NY, 1998), leqepheng la 43-96.
(8) Wahab MF, Dasgupta PK, Kadjo AF le Armstrong DW, Anal.Chim.Journal.Rev.907, 31–44 (2016).https://doi.org/10.1016/j.aca.2015.11.043.