Kāore ētahi kaupapa rapurongoā mō ngā taputapu LC e tawhito, nā te mea he raruraru kei roto i te mahi LC, ahakoa kei te whakapai ake te hangarau taputapu i te roanga o te wā. He maha ngā huarahi ka puta ai ngā raruraru i roto i te pūnaha LC, ā, ka mutu ki te āhua kino o te tihi. Ina puta mai ngā take e pā ana ki te āhua o te tihi, ka āwhina he rārangi poto o ngā take pea mō ēnei hua ki te whakahaere i tā mātou wheako rapurongoā.
He mea ngahau te tuhi i tēnei pou "LC Troubleshooting" me te whakaaro ki ngā kaupapa i ia marama, nā te mea kāore ētahi kaupapa e tawhito. I a mātou e rangahau ana i te chromatography, ka tawhito ētahi kaupapa, whakaaro rānei i te mea kua whakakapia e ngā whakaaro hou ake, pai ake, i roto i te mara o te rapu raruraru, mai i te putanga mai o te tuhinga rapu raruraru tuatahi i roto i tēnei hautaka (te LC Journal i taua wā) nā te mea kei te whai tikanga tonu ētahi kaupapa) i te tau 1983(1). I roto i ngā tau kua pahure ake nei, kua arotahi ahau ki ētahi wāhanga LC Troubleshooting ki ngā au o nāianei e pā ana ki te chromatography wai (LC) (hei tauira, te whakataurite whanaunga o tō mātou māramatanga ki te pānga o te pēhanga ki te pupuri [2] New Advances) Tā mātou whakamārama i ngā hua LC me pēhea te rapu raruraru me ngā taputapu LC hou. I roto i te wāhanga o tēnei marama, kei te haere tonu ahau i taku raupapa (3), i tīmata i te Hakihea 2021, i arotahi ki ētahi o ngā kaupapa "ora me te mate" o te rapu raruraru LC - he mea nui ngā huānga pai mō tētahi kaiwhakatau raruraru, ahakoa te pakeke o te pūnaha e whakamahia ana e mātou. Ko te kaupapa matua o tēnei raupapa he tino whai tikanga ki te rongonui o te LCGC "Aratohu Whakatau Rapanga LC" tūtohi pakitara (4) e iri ana i roto i ngā taiwhanga maha. Mō te wāhanga tuatoru o tēnei raupapa, i whiriwhiria e au te arotahi ki ngā take e pā ana ki te āhua o te tihi, ki ngā āhuatanga rānei o te tihi. He mea whakamiharo, e 44 ngā take pea o te āhua kino o te tihi e rārangi ana i roto i te tūtohi pakitara! Kāore e taea e tātou te whakaaro taipitopito i ēnei take katoa i roto i te tuhinga kotahi, nō reira i tēnei wāhanga tuatahi mō te kaupapa, ka arotahi ahau ki ētahi o ngā mea e kitea pinepine ana e au. Ko taku tumanako ka kitea e ngā kaiwhakamahi LC taitamariki me ngā kaumātua ētahi tohutohu me ngā whakamaumahara āwhina mō tēnei kaupapa nui.
Kei te piki haere taku whakautu i ngā pātai rapurongoā me te kī, "ka taea ngā mea katoa". Tērā pea he ngāwari tēnei whakautu ina whakaarohia ngā kitenga he uaua ki te whakamārama, engari he maha ngā wā ka kitea e au he tika. He maha ngā take pea o te āhua kino o te tihi, he mea nui kia tuwhera te hinengaro ina whakaarohia he aha te raruraru, kia taea ai te whakarite i ngā take pea hei tīmata i ā tātou mahi rapurongoā, me te arotahi ki ngā mea tino taea, he mea tino nui tēnei. Ka taea.
Ko tētahi taahiraa matua i roto i tētahi mahi rapurongoā - engari ki taku whakaaro he iti te aro ki tēnei - ko te mōhio kei reira tētahi raruraru me whakaoti. Ko te mōhio kei reira tētahi raruraru he tikanga maha te mōhio he rerekē te mea e pā ana ki te taputapu i ō tātou tumanakohanga, e hangaia ana e te ariā, te mōhiotanga ā-wheako, me te wheako (5). Ko te "āhua tihi" e kōrerohia ana i konei ehara i te mea ko te āhua o te tihi anake (he ōrite, he kore ōrite, he maeneene, he pupuhi, he taha ārahi, he hiku, me ētahi atu), engari ko te whānui hoki. He māmā noa iho ā tātou tumanakohanga mō te āhua tihi tuturu. E tautoko pai ana te ariā (6) i te tumanako a te pukapuka ako, i te nuinga o ngā wā, me ōrite ngā tihi chromatographic me te rite ki te āhua o te tohatoha Gaussian, e whakaaturia ana i te Pikitia 1a. Ko tā tātou e tumanako ana mai i ngā whānui tihi he take uaua ake, ā, ka matapakihia e tātou tēnei kaupapa i roto i tētahi tuhinga a muri ake nei. Ko ētahi atu āhua tihi i roto i te Pikitia 1 e whakaatu ana i ētahi atu mea ka taea te kite - arā, ko ētahi o ngā huarahi ka hē ai ngā mea. I te toenga o tēnei wāhanga, ka pau te wā ki te matapaki i ētahi tauira motuhake o ngā āhuatanga ka arahi ki ēnei momo āhua.
I ētahi wā, kāore e kitea ngā tihi i roto i te chromatogram e tika ana kia tangohia. E whakaatu ana te tūtohi pakitara i runga ake nei ko te ngaro o te tihi (mehemea kei roto i te tauira te analyte ūnga i te kukū e tika ana kia nui ai te urupare a te pūoko ki te kite i runga ake i te haruru) he mea e pā ana ki tētahi raruraru taputapu, ki ngā āhuatanga wāhanga neke hē rānei (mēnā ka kitea). ngā tihi, he "ngoikore" rawa te nuinga o te wā). Ka kitea he rārangi poto o ngā raruraru me ngā otinga pea i roto i tēnei kāwai i te Ripanga I.
E ai ki te kōrero i runga ake nei, ko te pātai mō te nui o te whānui o te tihi me manawanui i mua i te aro atu me te ngana ki te whakatika he kaupapa uaua ka matapakihia e au i roto i tētahi tuhinga a muri ake nei. Ko taku wheako he maha ngā wā ka haere tahi te whānui nui o te tihi me te huringa nui o te āhua o te tihi, ā, he nui ake te kitea o te hiku o te tihi i te wā i mua i te tihi, i te wā rānei e wehea ana. Heoi, ka whānui ake hoki ngā tihi ōrite, ka puta pea i ētahi take rerekē:
Kua matapakihia katoatia ēnei take i roto i ngā putanga o mua o te Troubleshooting LC, ā, ka taea e ngā kaipānui e hiahia ana ki ēnei kaupapa te tirotiro i ēnei tuhinga o mua mō ngā mōhiohio e pā ana ki ngā pūtake me ngā otinga pea mō ēnei take. Ētahi atu taipitopito.
Ka taea te puta te piko o te tihi, te tihi mua, me te wehewehe i ngā āhuatanga matū, ā, he rerekē te rārangi o ngā otinga pea mō ēnei raruraru, i runga i te mea kei te aro atu tātou ki tētahi raruraru matū, ā-tinana rānei. He maha ngā wā, mā te whakataurite i ngā tihi rerekē i roto i tētahi chromatogram, ka kitea e koe ngā tohu nui mō te take. Mena he rite ngā āhua o ngā tihi katoa i roto i tētahi chromatogram, ko te take pea ehara i te mea ā-tinana. Mena kotahi, e toru rānei ngā tihi e pāngia ana, engari he pai te āhua o ētahi atu, ko te take pea he matū.
He uaua rawa ngā take matū o te piko o te tihi, kāore e taea te kōrero poto i konei. Ka tukuna te kaipānui hiahia ki te putanga hou o "LC Troubleshooting" mō tētahi kōrero hohonu ake (10). Heoi, he mea ngāwari ki te whakamātau ko te whakaiti i te papatipu o te analyte i werohia, me te tirotiro mēnā ka pai ake te āhua o te tihi. Mēnā koinā, he tohu pai tēnei ko te raruraru ko te "taikaha papatipu". I tēnei take, me whakawhāiti te tikanga ki te werohanga i ngā papatipu analyte iti, me whakarerekē rānei ngā āhuatanga chromatographic kia taea ai te whiwhi i ngā āhua tihi pai ahakoa he nui ake ngā papatipu i werohia.
He maha hoki ngā take ā-tinana pea mō te piko o te wai. Ko ngā kaipānui e hiahia ana ki tētahi kōrero taipitopito mō ngā mea ka taea, ka tirohia tētahi atu putanga hou o "LC Troubleshooting" (11). Ko tētahi o ngā take ā-tinana e kitea whānuitia ana mō te piko o te wai ko te hononga ngoikore i te wāhi i waenga i te werowero me te pūoko (12). Kei te whakaatuhia tētahi tauira tino nui i te Pikitia 1d, i riro mai i taku taiwhanga i ētahi wiki kua pahure. I tēnei take, i hangaia e mātou he pūnaha me tētahi takirere werohanga hou kāore anō kia whakamahia e mātou i mua, ā, i tāutahia he koropiko werohanga iti me tētahi ferrule i hangaia ki runga i tētahi capillary kowiri tira. Whai muri i ētahi whakamātautau rapurongoā tuatahi, i kite mātou he hohonu ake te hohonutanga o te tauranga i roto i te stator takirere werohanga i tā mātou i whakamahi ai, ka hua ake he nui te rōrahi mate i raro i te tauranga. Ka taea te whakatau ngāwari i tēnei raruraru mā te whakakapi i te koropiko werohanga ki tētahi atu ngongo, ka taea e mātou te whakatika i te ferrule ki te tūranga tika hei whakakore i te rōrahi mate i raro i te tauranga.
Ko ngā mata tihi pērā i ngā mea e whakaaturia ana i te Pikitia 1e ka puta mai anō hoki i ngā raruraru ā-tinana, ā-matū rānei. Ko tētahi take ā-tinana noa o te taha ārahi ko te kore e pai te tākai o te moenga matūriki o te pou, kua whakaritea anō rānei ngā matūriki i roto i te wā. Pērā i te hiku tihi i puta mai i tēnei āhuatanga ā-tinana, ko te huarahi pai ki te whakatika i tēnei ko te whakakapi i te pou me te haere tonu. Ko te tikanga, ko ngā āhua tihi taha ārahi me te takenga matū ka puta mai i ngā mea e kiia nei e tātou he "kore-raina". I raro i ngā āhuatanga pai (raina), ko te nui o te analyte i puritia e te wāhanga tūmau (nō reira, ko te tauwehe pupuri) e pā ana ki te kukū o te analyte i roto i te pou. I roto i te chromatographic, ko te tikanga o tēnei, i te mea ka piki ake te papatipu o te analyte i werohia ki roto i te pou, ka teitei ake te tihi, engari kāore i te whānui ake. Ka pakaru tēnei whanaungatanga ina kore-raina te whanonga pupuri, ā, kāore ngā tihi e teitei ake engari ka whānui ake i te mea ka nui ake te papatipu e werohia ana. Hei tāpiri, ko ngā āhua kore-raina te whakatau i te āhua o ngā tihi chromatographic, ka hua ake he taha ārahi, he taha whai muri rānei. Pērā i te taumaha nui e puta ai te hiku tihi (10), Ka taea hoki te tātaritanga i te tihi ārahi i puta mai i te pupuri kore-raina mā te whakaiti i te papatipu analyte i werohia. Mena ka pai ake te āhua o te tihi, me whakarerekē te tikanga kia kaua e neke ake i te kounga werohanga e puta ai te mata ārahi, me whakarerekē rānei ngā āhuatanga chromatography hei whakaiti i tēnei whanonga.
I ētahi wā ka kite tātou i te mea e āhua ana he tihi "wehewehe", e whakaaturia ana i te Pikitia 1f. Ko te taahiraa tuatahi ki te whakaoti i tēnei raruraru ko te whakatau mēnā ko te āhua o te tihi nā te whakakotahitanga wāhanga (arā, te aroaro o ngā matū e rua e wehewehea ana engari e tata ana te whakakotahitanga). Mēnā he rua ngā tātaritanga rerekē e wehewehea ana tetahi ki tetahi, ko te mea nui ko te whakapai ake i tō rātou whakataunga (hei tauira, mā te whakanui ake i te whiriwhiri, te pupuri, te tatau pereti rānei), ā, ko ngā tihi "wehewehe" e kitea ana e pā ana ki te āhua tinana. Kāore he pānga o te mahi ki te pou tonu. He maha ngā wā, ko te tohu tino nui ki tēnei whakatau ko te mea kei te whakaatu ngā tihi katoa i roto i te chromatogram i ngā āhua wehewehe, kotahi, e rua rānei. Mēnā he kotahi, e rua rānei, he take whakakotahitanga pea; ki te wehewehea ngā tihi katoa, he take tinana pea, he mea e pā ana ki te pou tonu.
Ko ngā tihi wehewehe e pā ana ki ngā āhuatanga ā-tinana o te pou tonu, he mea nā te aukati ā-wāhanga o ngā frit tomo, ā-wāhanga rānei o te putanga, te whakaritenga anō rānei o ngā matūriki i roto i te pou, e tuku ana i te wāhanga neke kia tere ake te rere i te wāhanga neke i ētahi wāhi o te hanganga awa pou. i ētahi atu rohe (11). I ētahi wā, ka taea te whakakore i te frit kua aukatia mā te huri i te rere i roto i te pou; heoi, i roto i taku wheako, he otinga poto noa iho tēnei, kaua ki te otinga roa. He maha ngā wā ka mate tēnei i roto i ngā pou hou mēnā ka honoa anō ngā matūriki i roto i te pou. I tēnei wā, he pai ake te whakakapi i te pou me te haere tonu.
Ko te tihi i te Pikitia 1g, nō tētahi tauira tata nei anō hoki i roto i taku ake taiwhanga, e tohu ana he tino teitei te tohu kua tae ki te pito teitei o te awhe urupare. Mō ngā pūoko mimiti whatu (UV-vis i tēnei take), ina tino teitei te kukū o te analyte, ka mimiti te analyte i te nuinga o te mārama e tika ana mā roto i te pūtau rere o te pūoko, ā, he iti noa iho te mārama e toe ana hei kitea. I raro i ēnei āhuatanga, ka tino pāngia te tohu hiko mai i te pūoko whakaahua e ngā momo pūtake haruru, pērā i te mārama kotiti me te "au pōuri", ka tino "pōuriuri" te āhua o te tohu, ā, kāore e pā ki te kukū o te analyte. Ina tupu tēnei, ka taea te whakatau ngāwari i te raruraru mā te whakaiti i te rōrahi werohanga o te analyte—te whakaiti i te rōrahi werohanga, te whakawaiwai i te tauira, te rua rānei.
I roto i te kura chromatography, ka whakamahia e mātou te tohu pūoko (arā, te tuaka-y i roto i te chromatogram) hei tohu mō te kukū o te analyte i roto i te tauira. Nō reira, he āhua rerekē te kite i tētahi chromatogram me te tohu i raro i te kore, nā te mea ko te whakamaoritanga ngāwari ko tēnei e tohu ana i te kukū kino o te analyte – he mea kāore e taea te mahi ā-tinana. I roto i aku wheako, ka kitea ngā tihi kino i te nuinga o te wā ina whakamahia ngā pūoko mimiti whatu (hei tauira, UV-vis).
I tēnei take, ko te tikanga o te tihi kino ko ngā ngota e puta mai ana i te pou ka mimiti i te iti iho o te mārama i te wāhanga neke tonu i mua tonu me muri i te tihi. Hei tauira, ka puta tēnei ina whakamahia ngā roangaru kitenga iti (<230 nm) me ngā tāpiringa wāhanga neke e mimiti ana i te nuinga o te mārama i ēnei roangaru. Ko ēnei tāpiringa ka taea te whakamahi i ngā wāhanga whakarewa wāhanga neke pērā i te methanol, i ngā wāhanga pūmau rānei pērā i te acetate, te formate rānei. Ka taea te whakamahi i ngā tihi kino hei whakarite i tētahi pihi whakatikatika me te whiwhi mōhiohio tauanga tika, nō reira kāore he take matua hei karo i a rātou (e kiia ana tēnei tikanga ko te "kitenga UV kore tika") (13). Heoi, ki te hiahia tātou ki te karo katoa i ngā tihi kino, i roto i te take o te kitenga mimiti, ko te otinga pai ko te whakamahi i tētahi roangaru kitenga rerekē kia nui ake te mimiti a te analyte i te wāhanga neke, kia whakarerekē rānei i te hanganga o te wāhanga neke kia iti ake te mimiti o te mārama i ngā analyte.
Ka puta pea ngā tihi kino ina whakamahia te kimi i te taupū whakaata (RI) ina rerekē te taupū whakaata o ngā wāhanga ehara i te analyte i roto i te tauira, pērā i te matihiko whakarewa, i te taupū whakaata o te wāhanga neke. Ka puta anō hoki tēnei me te kimi UV-vis, engari ka ngoikore haere tēnei pānga i runga i te kitenga RI. I ngā take e rua, ka taea te whakaiti i ngā tihi kino mā te whakataurite tata atu i te hanganga o te matihiko tauira ki te hanganga o te wāhanga neke.
I te wāhanga tuatoru mō te kaupapa matua o te rapurongoā LC, i kōrero ahau mō ngā āhuatanga e rerekē ai te āhua o te tihi i kitea mai i te āhua o te tihi e tumanakohia ana, e noa ana rānei. Ko te rapurongoā whai hua o ēnei raruraru ka tīmata mā te mōhio ki ngā āhua o te tihi e tumanakohia ana (i runga i te ariā, i ngā wheako o mua rānei me ngā tikanga o nāianei), nō reira he mārama ngā rerekētanga mai i ēnei tumanakohanga. He maha ngā take pea o ngā raruraru āhua o te tihi (he whānui rawa, he hiku, he taha ārahi, me ētahi atu). I tēnei wāhanga, ka kōrerohia e au ētahi o ngā take e kitea ana e au i te nuinga o te wā. Mā te mōhio ki ēnei taipitopito ka pai te tīmatanga o te rapurongoā, engari kāore e hopu i ngā mea katoa ka taea. Ka taea e ngā kaipānui e hiahia ana ki tētahi rārangi hohonu ake o ngā take me ngā otinga te tirotiro i te tūtohi pakitara "LC Troubleshooting Guide" a te LCGC.
(4) Tūtohi pakitara "Aratohu Whakatau Raruraru LC" a te LCGC. https://www.chromatographyonline.com/view/troubleshooting-wallchart (2021).
(6) A. Felinger, Te Tātari Raraunga me te Tukatuka Tohu i roto i te Chromatography (Elsevier, New York, NY, 1998), wh. 43-96.
(8) Wahab MF, Dasgupta PK, Kadjo AF me Armstrong DW, Anal.Chim.Journal.Rev. 907, 31–44 (2016).https://doi.org/10.1016/j.aca.2015.11.043.