Njengethuluzi elinemba okuphezulu, i-HPLC ingaholela kalula ezinkingeni ezincane ezikhathazayo uma ingasetshenziswa ngendlela efanele ngesikhathi sokusetshenziswa.Enye yezinkinga ezivame kakhulu inkinga yokucindezela ikholomu.Uyixazulula kanjani ngokushesha i-chromatograph enephutha.Uhlelo lwe-HPLC ikakhulukazi luqukethe ibhodlela le-reservoir, iphampu, i-injector, ikholomu, igumbi lekholomu lokushisa, umtshina kanye nesistimu yokucubungula idatha.Kuwo wonke uhlelo, izinsika, amaphampu kanye nemishini yokuhlola yizici ezibalulekile kanye nezindawo eziyinhloko ezivame ukuba nezinkinga.
Isihluthulelo sokucindezela kwekholomu indawo edinga ukunakwa kakhulu uma usebenzisa i-HPLC.Ukuzinza kwengcindezi yekholomu kuhlobene eduze nokuma kwesiqongo sechromatographic, ukusebenza kahle kwekholomu, ukusebenza kahle kokuhlukaniswa kanye nesikhathi sokugcina.Ukuzinza kokucindezela kwekholomu akusho ukuthi inani lokucindezela lizinzile kunani elizinzile, kodwa kunalokho ububanzi bokuguquguquka kokucindezela buphakathi kuka-345kPa noma 50PSI (okuvumela ukusetshenziswa kokukhanya kwegradient lapho ingcindezi yekholomu izinzile futhi ishintsha kancane).Ukucindezela okuphezulu kakhulu noma okuphansi kakhulu kuyinkinga yokucindezela kwekholomu.
Okuvame kakhulu ukwehluleka kwe-HPLC nezisombululo
1, ukucindezela okuphezulu kuyinkinga evame kakhulu ekusetshenzisweni kwe-HPLC.Lokhu kusho ukunyuka kwengcindezi kungazelelwe.Ngokuvamile, kunezizathu ezilandelayo: (1) Ngokuvamile, lokhu kungenxa yokuvaleka kwesiteshi sokugeleza.Kuleli qophelo, kufanele sikuhlole ngokucophelela.a.Okokuqala, nqamula i-inlet yepompo ye-vacuum.Kuleli qophelo, ishubhu ye-PEEK yayigcwele uketshezi ukuze ishubhu le-PEEK libe lincane kunebhodlela le-solvent ukuze libone ukuthi uketshezi lwaluconsa ngokuthanda kwakhe.Uma uketshezi lungaconzi noma lungaconsi kancane, ikhanda lesihlungi se-solvent liyavinjelwa.Ukwelashwa: Gxila ku-30% we-nitric acid isigamu sehora bese ugeza ngamanzi ahlanzekile.Uma uketshezi luconsa ngokungahleliwe, ikhanda lesihlungi se-solvent livamile futhi liyahlolwa;b.Vula i-valve ye-Purge ukuze isigaba seselula singadluli kukholomu, futhi uma ukucindezela kungancishiswa kakhulu, ikhanda elimhlophe lokuhlunga livinjiwe.Ukwelashwa: Amakhanda amhlophe ahlungiwe asusiwe futhi ahlanganiswa ne-10% isopropanol isigamu sehora.Ngokucabanga ukuthi ingcindezi yehla ngaphansi kwe-100PSI, ikhanda elimhlophe elihlungiwe lijwayelekile futhi liyahlolwa;c.Susa ukuphela kokuphuma kwekholomu, uma ukucindezela kunganciphi, ikholomu ivinjiwe.Ukwelashwa: Uma kuyisivimbeli sikasawoti esivimbayo, hlambulula ngo-95% kuze kube yilapho ukucindezela sekuvamile.Uma ukuvinjelwa kubangelwe enye into egcinwe kakhulu, ukugeleza okunamandla kunesigaba samanje seselula kufanele kusetshenziselwe ukuphuthuma kungcindezi evamile.Uma ingcindezi yokuhlanza isikhathi eside inganciphisi ngokwendlela engenhla, i-inlet kanye ne-outlet yekholomu ingabhekwa njengexhunywe kuthuluzi ngokuphambene nalokho, futhi ikholomu ingahlanzwa ngesigaba seselula.Ngalesi sikhathi, uma ingcindezi yekholomu ingakancishiswa, ipuleti ye-sieve yekholomu ingashintshwa kuphela, kodwa uma ukusebenza kungalungile, kulula ukuholela ekunciphiseni umphumela wekholomu, ngakho-ke zama ukusebenzisa kancane.Ngezinkinga ezinzima, ukushintshwa kwekholomu kungase kucatshangelwe.
(2) Ukulungiselelwa kwezinga lokugeleza okungalungile: Izinga lokugeleza elilungile lingasethwa kabusha.
(3) Isilinganiso sokugeleza esingalungile: inkomba ye-viscosity yezilinganiso ezihlukene zokugeleza ihlukile, futhi ingcindezi yesistimu ehambisanayo yokugeleza nge-viscosity ephakeme nayo inkulu.Uma kungenzeka, izincibilikisi ze-viscosity eziphansi zingashintshwa noma zisethwe kabusha futhi zilungiswe.
(4) Umfutho wesistimu uziro drift: lungisa uziro wenzwa yezinga loketshezi.
2, ingcindezi iphansi kakhulu (1) ngokuvamile ibangelwa ukuvuza kwesistimu.Okufanele ukwenze: Thola ukuxhumana ngakunye, ikakhulukazi isixhumi esibonakalayo kuzo zombili iziphetho zekholomu, bese uqinisa indawo evuzayo.Susa okuthunyelwe bese uqinisa noma ulayinise ifilimu ye-PTFE ngamandla afanele.
(2) Igesi ingena empompini, kodwa ingcindezi ngokuvamile ayizinzile ngalesi sikhathi, iphezulu futhi iphansi.Okubi nakakhulu, ipompo ngeke ikwazi ukumunca uketshezi.Indlela yokwelapha: vula i-valve yokuhlanza futhi uhlanze ngesilinganiso sokugeleza kwe-3 ~ 5ml / min.Uma kungenjalo, amabhamuza omoya ayeshiswa kuvalvu yokukhipha umoya kusetshenziswa ishubhu yenaliti elizinikele.
(3) Akukho ukuphuma kwesigaba esihambayo: hlola ukuthi ingabe sikhona yini isigaba esihambayo ebhodleleni lamanzi, ukuthi usinki ucwiliswe esigabeni esihambayo, kanye nokuthi iphampu iyasebenza yini.
(4) I-valve yereferensi ayivaliwe: i-valve yesithenjwa ivaliwe ngemva kokwehla.Ngokuvamile yehlela ku-0.1.~ 0.2mL/ min ngemva kokuvala ivalvu yereferensi.
Isifinyezo:
Kuleli phepha, kuhlaziywa kuphela izinkinga ezijwayelekile ku-liquid chromatography.Yebo, ekusebenziseni kwethu okungokoqobo, sizobhekana nezinye izinkinga ezengeziwe.Ekuphatheni amaphutha, kufanele silandele izimiso ezilandelayo: shintsha kuphela isici esisodwa ngesikhathi ukuze sinqume ubudlelwano phakathi kwesici esicatshangelwayo nenkinga;Ngokujwayelekile, lapho sishintsha izingxenye zokuxazulula inkinga, kufanele sinake ukubuyisela izingxenye ezihlakaziwe ezingashintshiwe ukuze kuvinjelwe imfucuza;Ukwakha umkhuba omuhle wokuqopha kuyisihluthulelo sempumelelo yokusingatha amaphutha.Sengiphetha, lapho usebenzisa i-HPLC, kubalulekile ukunaka isampula yokwelashwa kwangaphambili kanye nokusebenza okufanele nokugcinwa kwezinto zokusebenza.
Isikhathi sokuthumela: Sep-18-2023