Iyo mobile phase yakaenzana nechikamu chemvura yeropa, uye pane zvinhu zvakasiyana-siyana zvekuteerera panguva yekushandiswa. Pakati pazvo, pane mamwe "misungo" inofanira kutariswa.
01. Edza iyo pH ye mobile phase mushure mekuwedzera organic solvent
Kana iwe ukayera pH neiyo organic additive, iyo pH yaunowana ichave yakasiyana pane usati wawedzera iyo organic solvent. Zvisinei, chinhu chinonyanya kukosha ndechekuenderana. Kana iwe uchigara uchiyera pH mushure mekuwedzera iyo organic solvent, ita shuwa yekutaura nhanho dzako munzira yaunoshandisa kuitira kuti vamwe vatevedze nzira imwechete. Iyi nzira haisi 100% yakarurama, asi zvishoma ichaita kuti nzira yacho ienderane. Izvi zvingave zvakanyanya kukosha pane kuwana chaiyo pH kukosha.
02. Hapana buffer yakashandiswa
Chinangwa chebhafa ndechekudzora pH uye kuidzivirira kubva pakuchinja. Dzimwe nzira dzakawanda dzinoshandura pH yechikamu chenhare, izvo zvinogona kukonzera shanduko munguva yekuchengetedza, chimiro chepamusoro, uye mhinduro yepamusoro.
Formic acid, TFA, nezvimwewo hazvisi zvibhafa
03. Kusashandisa buffer mukati meyakajairika pH renji
Imwe neimwe buffer ine 2 pH unit hupamhi hwehupamhi, mukati mayo inopa yakanyanya kugadzikana pH. Mabhafa kunze kwehwindo iri haape zvinobudirira kuramba pH shanduko. Shandisa buffer muchikamu chakakodzera, kana sarudza bhafa inovhara pH renji yaunoda.
04. Wedzera buffer kune organic solution
Kusanganisa buffer solution ne organic phase kungango konzera kuti buffer iwedzere. Muzviitiko zvakawanda, kunyange kana kunaya kwakaitika, kuchiri kunetsa kuona. Rangarira kugara uchiwedzera iyo organic solution kuchikamu chemvura, izvo zvinogona kuderedza zvakanyanya mukana wekunaya kwebuffer.
05. Sanganisa kunyura kubva pa0% nepombi
Mapombi aripo nhasi anogona kunyatso kusanganisa mobile phases uye degas inline, asi havasi vese vari kushandisa nzira yako vachave nemhando yepamusoro pombi. Sanganisa A uye B mune imwe mhinduro woimhanyisa 100% mukati.
Semuenzaniso, 950 ml ye organic kutanga musanganiswa inogona kugadzirwa nekusanganiswa ne50 ml yemvura. Chakanakira izvi ndechekuti inogona kuderedza kusiyanisa pakati peHPLC uye kuderedza mukana wemabhuru uye kunaya kwemvura muhurongwa. Zvakakosha kuziva kuti chiyero chemusanganiswa wepombi ndeye 95: 5, izvo hazvirevi kuti nguva yakasanganiswa yekuchengetedza mubhodhoro zvakare 95: 5.
06. Kusashandisa iyo chaiyo yakagadziridzwa asidhi (base) kushandura buffer
Shandisa asidhi kana base inoumba buffer salt yauri kushandisa. Semuenzaniso, sodium phosphate buffer inofanirwa kugadzirwa ine phosphoric acid chete kana sodium hydroxide.
07. Kwete kutaura ruzivo rwese nezve buffer munzira, sekuwedzera 5g yesodium phosphate kusvika 1000ml yemvura.
Mhando yebuffer inosarudza iyo pH renji inogona kuvharwa. Iko kusungirirwa kunodiwa kunogadzirisa simba rebhafa. 5 magiramu kana anhydrous sodium phosphate uye 5 magiramu e monosodium phosphate monohydrate ane akasiyana buffer masimba.
08. Kuwedzera organic solvents usati watarisa
Kana nzira yapfuura yakashandisa gadziriso yebhafa yekutanga B, uye nzira yako ichishandisa organic mhinduro yehwaro B, unogona netarisiro kugadzirisa bhafa mupombi tubing uye pombi musoro.
09. Simudza bhodhoro uye dururira donhwe rekupedzisira
Pane mukana wakanaka wekuti iwe hauzove neyakakwana mobile phase yekupedzisa kumhanya kwese uye sampuro yako ichaputa. Kunze kwemukana wekupisa pombi system uye koramu, nharembozha inobuda zvachose uye chikamu chenhare chiri pamusoro pebhodhoro chinoshanduka.
10. Shandisa ultrasonic degassing mobile phase
Chinhu chakanyanya kukosha ndechekuita shuwa kuti ese mabuffer salt anyungudika, asi iyi ndiyo nzira yakaipisisa ye degas uye inokurumidza kupisa chikamu chenhare, zvichiita kuti organic zvikamu zvinyunguduke. Kuti uchengetedze dambudziko risingakoshe gare gare, tora maminetsi mashanu kuti uvhare sefa yako mobile phase.
Nguva yekutumira: Aug-27-2024