Isigaba esihambayo silingana nesigaba soketshezi segazi, futhi kunezinto ezihlukahlukene okufanele uzinake ngesikhathi sokusetshenziswa. Phakathi kwazo, kukhona “izihibe” okumelwe zinakwe.
01. Linganisa i-pH yesigaba seselula ngemva kokwengeza i-solvent ephilayo
Uma ukala i-pH ngesithako se-organic, i-pH oyitholayo izohluka kunangaphambi kokwengeza i-solvent ephilayo. Nokho, okubaluleke kakhulu ukungaguquguquki. Uma njalo ukala i-pH ngemva kokwengeza i-solvent ephilayo, qiniseka ukuthi usho izinyathelo zakho endleleni oyisebenzisayo ukuze abanye balandele indlela efanayo. Le ndlela ayinembile ngo-100%, kodwa okungenani izogcina indlela ihambisana. Lokhu kungase kubaluleke kakhulu kunokuthola ivelu ye-pH enembile.
02. Asikho isilondolozi esisetshenzisiwe
Inhloso ye-buffer ukulawula i-pH futhi iyivimbele ukuthi ingashintshi. Ezinye izindlela eziningi zishintsha i-pH yesigaba seselula, okungabangela ukushintsha kwesikhathi sokugcina, ukuma okuphezulu, nokuphendula okuphezulu.
I-Formic acid, i-TFA, njll. akuzona izibhafa
03. Ukungasebenzisi isigcinalwazi ngaphakathi kwebanga le-pH elivamile
Ibhafa ngayinye inobubanzi beyunithi ye-pH engu-2, ngaphakathi kwayo ihlinzeka ngokuzinza okungcono kakhulu kwe-pH. Amabhafa angaphandle kwaleli windi ngeke anikeze ukumelana okusebenzayo nezinguquko ze-pH. Sebenzisa ibhafa ebangeni elilungile, noma khetha isigcinalwazi esimboza ibanga le-pH oludingayo.
04. Engeza isilondolozi kwisixazululo sezinto eziphilayo
Ukuxuba isisombululo se-buffer nesigaba se-organic cishe kuzodala ukuthi isigcinalwazi sidume. Ezimweni eziningi, noma ngabe imvula ike yaba khona, kusenzima ukuyibona. Khumbula njalo ukwengeza ikhambi lezinto eziphilayo esigabeni esinamanzi, okunganciphisa kakhulu ithuba lokuna kwebhafa.
05. Hlanganisa i-concentration gradient kusuka ku-0% ngepompo
Amaphampu atholakalayo namuhla angaxuba ngempumelelo izigaba zeselula kanye ne-degas emgqeni, kodwa akuwona wonke umuntu osebenzisa indlela yakho ozoba nepompo yekhwalithi ephezulu. Xuba u-A no-B kwisixazululo esisodwa bese usisebenzisa ngo-100% emgqeni.
Isibonelo, i-950 ml yengxube yokuqala ye-organic ingalungiswa ngokuxuba no-50 ml wamanzi. Inzuzo yalokhu ukuthi inganciphisa ukuhluka phakathi kwama-HPLC futhi inciphise amathuba okuba namabhamuza kanye nemvula ohlelweni. Kuyaphawuleka ukuthi isilinganiso senhlanganisela yepompo singu-95: 5, akusho ukuthi isikhathi sokugcina esixutshwe ngaphambili ebhodleleni siphinde sibe ngu-95: 5.
06. Ukungasebenzisi i-asidi eguquliwe efanele (isisekelo) ukushintsha isigcinalwazi
Sebenzisa kuphela i-asidi noma isisekelo esakha isigcinalwazi sikasawoti osisebenzisayo. Isibonelo, i-sodium phosphate buffer kufanele ilungiswe nge-phosphoric acid noma i-sodium hydroxide kuphela.
07. Ukungasho lonke ulwazi mayelana nesigcinalwazi endleleni, njengokwengeza u-5g wei-sodium phosphate ku-1000ml wamanzi.
Uhlobo lwebhafa lunquma ububanzi be-pH obungafakwa kubhafa. Ukugxila okudingekayo kunquma amandla ebhafa. Amagremu ama-5 noma i-anhydrous sodium phosphate kanye namagremu angu-5 e-monosodium phosphate monohydrate anamandla okuvimba ahlukene.
08. Ukwengeza izincibilikisi zemvelo ngaphambi kokuhlola
Uma indlela yangaphambilini ibisebenzisa isixazululo se-buffer sesisekelo B, futhi indlela yakho isebenzisa isixazululo se-organic sesisekelo B, ngethemba ungakwazi ukuxazulula isivanzi kushubhu wepompo kanye nekhanda lokumpompa.
09. Phakamisa ibhodlela bese uthulula iconsi lokugcina
Kunethuba elihle lokuthi ngeke ube nesigaba seselula esanele ukuze uqedele konke ukugijima futhi isampula yakho izobhema. Ngaphandle kwamathuba okushisa isistimu yepompo kanye nekholomu, isigaba seselula sizoshabalala ngokuphelele futhi isigaba seselula phezulu ebhodleleni sizoshintsha.
10. Sebenzisa isigaba seselula se-ultrasonic degassing
Iphuzu elibaluleke kakhulu ukwenza isiqiniseko sokuthi wonke usawoti we-buffer uyancibilika, kodwa lena indlela embi kakhulu ye-degas futhi izoshisa ngokushesha isigaba seselula, okubangele ukuthi izakhi ze-organic zihwamuke. Ukuze ulondoloze izinkinga ezingadingekile kamuva, thatha imizuzu emihlanu ukuhlanza isigaba sakho seselula.
Isikhathi sokuthumela: Aug-27-2024