Nkhani zina za LC zothetsa mavuto sizikhala zachikale, chifukwa pali zovuta muzochita za LC, ngakhale ukadaulo wa zida ukuyenda bwino pakapita nthawi. Pali njira zambiri zomwe mavuto angabwere mu dongosolo la LC ndikutha ndi mawonekedwe owopsa.
Zakhala zosangalatsa kulemba ndime iyi ya "LC Troubleshooting" ndikuganizira za mitu mwezi uliwonse, chifukwa mitu ina simachoka m'kalembedwe. Pamene tikuyang'ana kafukufuku wa chromatography mitu ina kapena mfundo zina zimakhala zosagwiritsidwa ntchito pamene zimachotsedwa ndi malingaliro atsopano komanso abwino, m'munda wa kuthetsa mavuto, kuyambira pamene nkhani yoyamba yothetsa mavuto idawonekera m'magazini ino (zolemba za LC 9 zaka zapitazo) zidakalipo zaka zitatu zapitazo. , Ndayang'ana kwambiri magawo angapo a LC Troubleshootings pazochitika zamakono zomwe zimakhudza chromatography yamadzi (LC) (mwachitsanzo, kufananitsa kwachibale kwa kumvetsetsa kwathu za zotsatira za kukakamizidwa kusunga [2] New Advances) Kutanthauzira kwathu kwa zotsatira za LC ndi momwe tingathetsere mavuto ndi zida zamakono za LC. s of LC troubleshooting — zinthu zomwe zili zabwino kwa aliyense wothetsa mavuto ndi zofunika, ziribe kanthu msinkhu wa dongosolo ife ntchito. Mutu waukulu wa mndandanda wa kwambiri zogwirizana ndi LCGC wotchuka "LC Troubleshooting Guide" khoma tchati (4) atapachikidwa mu laboratories ambiri.Pa gawo lachitatu la mndandanda, Ndinasankha kuganizira nkhani zokhudzana ndi nsonga za nsonga za 4 mawonekedwe a khoma kapena nsonga zapamwamba za tchati cha khoma. t kuganizira zonsezi mwatsatanetsatane m'nkhani imodzi, kotero mu gawo loyamba ili pa mutuwo, ndidzayang'ana pa zina zomwe ndimaziwona nthawi zambiri.Ndikuyembekeza kuti achinyamata ndi achikulire omwe amagwiritsa ntchito LC adzapeza malangizo ndi zikumbutso zothandiza pa mutu wofunikawu.
Ndimadzipeza ndikumayankha mafunso othetsa mavuto ndi "chilichonse ndi chotheka". Yankho ili limatha kuwoneka losavuta ndikaganizira zomwe ndizovuta kuzimasulira, koma ndimawona kuti ndizoyenera. Ndizifukwa zambiri zomwe zimapangitsa kuti pakhale zovuta, ndikofunikira kukhala ndi malingaliro omasuka poganizira zomwe vuto lingakhale, ndikuyika patsogolo zomwe zingayambitse kuti tiyambitse zovuta zathu, kuyang'ana kwambiri zomwe zingatheke.
Chinthu chofunika kwambiri pazochitika zilizonse zothetsera mavuto - koma zomwe ndikuganiza kuti ndizochepa - ndikuzindikira kuti pali vuto lomwe liyenera kuthetsedwa.Kuzindikira kuti pali vuto nthawi zambiri kumatanthauza kuzindikira kuti zomwe zimachitika ku chida ndizosiyana ndi zomwe timayembekezera, zomwe zimapangidwira ndi chiphunzitso, chidziwitso champhamvu, ndi zochitika (5) . ffy, kutsogolera m'mphepete, mchira, etc.), komanso m'lifupi.Zoyembekeza zathu za mawonekedwe enieni apamwamba ndi ophweka.Lingaliro (6) limathandizira bwino chiyembekezero cha bukuli kuti, nthawi zambiri, nsonga za chromatographic ziyenera kukhala zofanana ndikugwirizana ndi mawonekedwe a kugawa kwa Gaussian, monga momwe tawonetsera mu Chithunzi 1a. Zomwe tikuyembekezera pamutuwu ndizovuta kwambiri, zomwe tikuyembekezera pamutuwu ndizovuta kwambiri. chithunzi 1 chikuwonetsa zina mwa zina zomwe zingathe kuwonedwa-mwa kuyankhula kwina, njira zina zomwe zingawonongeke.
Nthawi zina nsonga sizimawonedwa konse mu chromatogram momwe zimayembekezeredwa kuchotsedwa.Tchati yapakhoma yomwe ili pamwambayi ikuwonetsa kuti kusakhalapo pachimake (poganiza kuti chitsanzocho chili ndi chowunikira chomwe chikuyenera kupangitsa kuti chowunikiracho chikhale chokwanira kuti chiziwone pamwamba pa phokoso) nthawi zambiri chimagwirizana ndi vuto la chida kapena mikhalidwe yolakwika ya gawo la foni (ngati iwonedwa konse).nsonga, nthawi zambiri "zofooka") .Mndandanda waufupi wamavuto omwe angakhalepo ndi mayankho mgululi akupezeka mu Gulu 1.
Monga tafotokozera pamwambapa, funso la kuchuluka kwa nsonga kuyenera kulekerera musanayambe kulabadira ndikuyesera kukonza ndi nkhani yovuta yomwe ndidzakambirana m'nkhani yamtsogolo. Zomwe ndikukumana nazo ndikuti kufalikira kwapamwamba kwambiri nthawi zambiri kumatsagana ndi kusintha kwakukulu kwa mawonekedwe apamwamba, ndipo nsonga zamtunduwu zimakhala zofala kwambiri kusiyana ndi chisanadze kapena kugawanika.
Nkhani zonsezi zakambidwa mwatsatanetsatane m'nkhani zam'mbuyo za Kuthetsa Mavuto a LC, ndipo owerenga omwe ali ndi chidwi pamituyi akhoza kutchula nkhani zam'mbuyomu kuti mudziwe zomwe zimayambitsa komanso njira zothetsera vutoli.Zambiri.
Kuthamanga kwapamwamba, kutsogolo kwapamwamba, ndi kugawanika kungayambitsidwe ndi zochitika za mankhwala kapena zakuthupi, ndipo mndandanda wa njira zothetsera mavutowa zimasiyana mosiyanasiyana, malingana ndi momwe tikulimbana ndi vuto la mankhwala kapena lakuthupi. nsonga zimakhudzidwa, koma zina zonse zimawoneka bwino, chifukwa chake ndi mankhwala.
Zomwe zimayambitsa nsonga zam'mwamba zimakhala zovuta kwambiri kuti tikambirane mwachidule apa.Wowerenga chidwi amatchulidwa ku nkhani yaposachedwa ya "LC Troubleshooting" kuti tikambirane mozama (10). kapena mawonekedwe a chromatographic ayenera kusinthidwa kuti mawonekedwe abwino apamwamba athe kupezeka ngakhale atabayidwa unyinji wokulirapo.
Palinso zifukwa zambiri zakuthupi zomwe zimapangitsa kuti pakhale nsonga zapamwamba.Owerenga omwe ali ndi chidwi chokambirana mwatsatanetsatane za zotheka amatumizidwa ku nkhani ina yaposachedwa ya "LC Troubleshooting" (11) . Chimodzi mwa zifukwa zomwe zimawonekera kwambiri za nsonga zapamwamba ndi kugwirizana kosauka pakati pa jekeseni ndi detector (12) . kale, ndipo anaika yaing'ono voliyumu jekeseni kuzungulira ndi ferrule kuti anali kuumbidwa pa zitsulo zosapanga dzimbiri capillary.After zina zoyesa zothetsa mavuto koyamba, tinazindikira kuti doko kuya mu jekeseni valavu stator anali zakuya kwambiri kuposa ife anazolowera, chifukwa chachikulu akufa buku pansi pa doko. .
Zomwe zimawonekera pachimake ngati zomwe zikuwonetsedwa mu Chithunzi 1e zitha kuyambitsidwanso ndi zovuta zakuthupi kapena zamankhwala. Chomwe chimayambitsa nsonga yotsogola ndikuti bedi la tinthu tating'onoting'ono silinapakidwe bwino, kapena kuti tinthu tating'onoting'ono takonzedwanso pakapita nthawi. Monga momwe zimakhalira pachimake chifukwa cha chodabwitsa ichi, njira yabwino yothetsera izi ndikusintha chigawocho ndikupitabe patsogolo ndi zomwe timayambitsa, zomwe zimayambira nthawi zambiri. Pansi pamikhalidwe yabwino (mizere), kuchuluka kwa analyte komwe kumasungidwa ndi gawo loyima (motero, chinthu chosungira) chimagwirizana molingana ndi kuchuluka kwa analyte mu column. Kuonjezera apo, mawonekedwe osakhala amtundu amatsimikizira mawonekedwe a nsonga za chromatographic, zomwe zimapangitsa kuti pakhale nsonga zotsogola kapena zotsatizana.Monga momwe zimakhalira ndi kuchulukana kwakukulu komwe kumayambitsa nsonga yapamwamba (10), nsonga yotsogola yomwe imayambitsidwa ndi kusungidwa kopanda malire imathanso kupezeka mwa kuchepetsa jekeseni wa analyte mass. khalidwe.
Nthawi zina timawona zomwe zikuwoneka ngati "kugawanika" pachimake, monga momwe tawonetsera mu Chithunzi 1f. Njira yoyamba yothetsera vutoli ndiyo kudziwa ngati mawonekedwe apamwamba amachokera ku mgwirizano wapang'ono (ie, kukhalapo kwa mitundu iwiri yosiyana koma yoyandikana kwambiri). nsonga zimagwirizana ndi thupi Magwiridwe alibe kanthu kochita ndi mzati wokha.Nthawi zambiri, chidziwitso chofunikira kwambiri pa chisankho ichi ndi chakuti nsonga zonse za chromatogram zimasonyeza mawonekedwe ogawanika, kapena chimodzi kapena ziwiri.ngati nsonga zonse zagawanika, mwina ndi nkhani yakuthupi, yokhudzana ndi gawo lomwelo.
Kugawanika nsonga zokhudzana ndi maonekedwe a thupi la gawolo nthawi zambiri zimakhala chifukwa cha kutsekedwa pang'ono kolowera kapena kutuluka kwa frits, kapena kukonzanso kwa tinthu tating'onoting'ono tating'onoting'ono tating'onoting'ono, kulola kuti gawo la mafoni liziyenda mofulumira kusiyana ndi gawo la mafoni m'madera ena a mapangidwe a ndondomeko .m'madera ena (11) .komabe, muzochitika zanga, izi nthawi zambiri zimakhala zaufupi kusiyana ndi nthawi yayitali.Izi nthawi zambiri zimapha ndi mizati yamakono ngati tinthu tating'ono timagwirizanitsanso mkati mwa chigawocho.Panthawiyi, ndibwino kuti musinthe ndondomekoyi ndikupitirizabe.
Pachimake mu Chithunzi 1g, komanso kuchokera ku zochitika zaposachedwa mu labu yanga, nthawi zambiri zimasonyeza kuti chizindikirocho ndi chapamwamba kwambiri moti chafika kumapeto kwa chiwerengero choyankha. magwero osiyanasiyana a phokoso, monga kuwala kosokera ndi "mdima wapano", kupangitsa chizindikirocho kukhala "chosamveka" pamawonekedwe komanso osadalira ndende ya analyte.Izi zikachitika, vutoli likhoza kuthetsedwa mosavuta pochepetsa kuchuluka kwa jekeseni wa analyte-kuchepetsa mphamvu ya jekeseni, kuchepetsa chitsanzo, kapena zonse ziwiri.
Mu sukulu ya chromatography, timagwiritsa ntchito siginecha (mwachitsanzo, y-axis mu chromatogram) ngati chizindikiritso cha ziwonetserozo. ).
Pachifukwa ichi, nsonga yoyipa imangotanthauza kuti mamolekyu omwe amatuluka kuchokera pamzati amamwa kuwala kochepa kusiyana ndi gawo la foni lomwe nthawi yomweyo isanayambe komanso itatha chiwombankhanga. Izi zikhoza kuchitika, mwachitsanzo, pogwiritsa ntchito mafunde otsika kwambiri (<230 nm) ndi zowonjezera zowonjezera zomwe zimayamwa kuwala kwambiri pa mafundewa. Kukonzekera kokhotakhota ndikupeza zidziwitso zolondola za kuchuluka kwake, kotero palibe chifukwa chofunikira chopewera chilichonse (njira imeneyi nthawi zina imatchedwa "kuzindikira kwa UV kosalunjika") (13). tes.
Nsonga zopanda pake zimatha kuwonekeranso mukamagwiritsa ntchito kuzindikira kwa refractive index (RI) pomwe index ya refractive ya zigawo zina kupatula ma analyte mu zitsanzo, monga zosungunulira matrix, ndizosiyana ndi refractive index of the mobile phase. Izi zimachitikanso ndi kuzindikira kwa UV-vis, koma izi zimakhala zocheperako poyerekeza ndi kuzindikira kwa RI.
Mu gawo lachitatu pa mutu waukulu wa LC troubleshooting, Ndinakambirana zinthu zimene anaonedwa pachimake mawonekedwe amasiyana kuyembekezera kapena wamba pachimake mawonekedwe.Kuthetsa mavuto mogwira mtima wa mavuto amenewa kumayamba ndi chidziwitso cha kuyembekezera akalumikidzidwa pachimake (kutengera chiphunzitso kapena zinachitikira m'mbuyomu ndi njira zomwe zilipo), kotero kupatuka kwa ziyembekezo izi n'zoonekeratu. Mavuto mawonekedwe pachimake ali ndi zifukwa zambiri zosiyana, kutsogolera m'mbali zambiri, Ndikambitsirana mwatsatanetsatane, kufotokoza mwatsatanetsatane, ndi zina zotero. kawirikawiri.Kudziwa zambiri izi kumapereka malo abwino oyambira kuthetsa mavuto, koma sikujambula zonse zomwe zingatheke.Owerenga omwe ali ndi chidwi ndi mndandanda wozama wazomwe zimayambitsa ndi njira zothetsera mavuto angatchule tchati chapakhoma cha LCGC "LC Troubleshooting Guide".
(4) LCGC "LC Troubleshooting Guide" wall chart.https://www.chromatographyonline.com/view/troubleshooting-wallchart (2021).
(6) A. Felinger, Data Analysis ndi Signal Processing mu Chromatography (Elsevier, New York, NY, 1998), pp. 43-96.
(8) Wahab MF, Dasgupta PK, Kadjo AF ndi Armstrong DW, Anal.Chim.Journal.Rev.907, 31–44 (2016).https://doi.org/10.1016/j.aca.2015.11.043.