Izimiso Nezindlela Zokuhlaziya Okuningi nge-Liquid Chromatography

Indlela yokwehlukanisa ye-chromatography ewuketshezi isuselwe emehlukweni wokuhlobana kwezingxenye zengxube yezigaba ezimbili.

Ngokwezigaba ezihlukene zokuma, i-chromatography ewuketshezi ihlukaniswa nge-chromatography e-liquid-solid, i-chromatography ye-liquid-liquid chromatography kanye ne-bond phase chromatography.Okusetshenziswa kakhulu i-chromatography eyi-liquid-solid enejeli ye-silica njengesigcwalisi kanye ne-bond phase chromatography ene-microsilica njenge-matrix.

Ngokohlobo lwesigaba esimile, i-chromatography ewuketshezi ingahlukaniswa ngekholomu ye-chromatography, i-chromatography yephepha kanye ne-chromatography yongqimba oluncane.Ngokomthamo we-adsorption, ingahlukaniswa nge-adsorption chromatography, partition chromatography, ion exchange chromatography kanye ne-gel permeation chromatography.

Eminyakeni yamuva nje, uhlelo lokugeleza koketshezi olunengcindezi ephezulu lwengezwe kusistimu ye-chromatography yekholomu ewuketshezi ukuze kwenziwe isigaba esihambayo sigeleze ngokushesha ngaphansi kwengcindezi ephezulu ukuze kuthuthukiswe umphumela wokuhlukanisa, ukusebenza kahle okuphezulu kakhulu (okubuye kwaziwe njenge-high-pressure) i-chromatography yoketshezi. sekuvele.

INGXENYE
01 Umgomo Wokuhlaziywa Kobuningi be-Liquid Chromatography

Ukulinganisa ngesisekelo sekhwalithi, izinto ezihlanzekile ziyadingeka njengamazinga;

I-Liquid chromatography quantification iyindlela elinganiselwe yobuningi: okungukuthi, inani le-analyte kungxube lilinganiselwa kunani elaziwayo lesampula elijwayelekile elimsulwa.

INGXENYE
02 Isisekelo Sokulinganiswa Nge-Liquid Chromatography

Inani lengxenye elinganisiwe (W) lilingana nenani lokuphendula (A) (ubude obuphakeme noma indawo ephakemeyo), W=f×A.

Isici sokulungisa sobuningi (f): Isilinganiso esingaguquguquki sefomula yokubala yobuningi, futhi incazelo yayo ebonakalayo yinani lengxenye elinganisiwe emelwe inani lokuphendula leyunithi (indawo ephakemeyo).

Isici sokulungisa sobuningi singatholwa enanini elaziwayo lesampula elijwayelekile kanye nenani layo lokuphendula.

Linganisa inani lokuphendula lengxenye engaziwa, futhi inani lengxenye lingatholwa nge-quantitative correction factor.

INGXENYE
03 Amagama ajwayelekile ekuhlaziyweni komthamo

Isampula (isampula): isisombululo esiqukethe i-analyte yokuhlaziywa kwe-chromatographic.Ihlukaniswe ngamasampula ajwayelekile nangaziwa.

Okujwayelekile: Umkhiqizo omsulwa onokugxila okwaziwayo.Isampula elingaziwa (akwaziwa): Ingxube okumele ukugxiliswa kwayo kuhlolwe.

Isisindo sesampula: Isisindo sangempela sesampula okufanele sihlolwe.

I-Dilution: I-dilution factor yesampula engaziwa.

Ingxenye : inani eliphakeme le-chromatographic okufanele lihlaziywe ngokomthamo, okungukuthi, umhlaziyi okungaziwa okuqukethwe kwakhe.

Inani lengxenye (inani): okuqukethwe (noma ukugxila) kwento ezohlolwa.

Ubuqotho : Inqubo yokubala yokulinganisa indawo ephakeme kakhulu yesiqongo se-chromatographic ngekhompuyutha.

Ijika lokulinganisa: Ijika lomugqa lokuqukethwe kwengxenye nenani lokuphendula, elisungulwe ngenani elaziwayo lento evamile, esetshenziselwa ukunquma okuqukethwe okungaziwa komhlaziyi.

INGXENYE
04 Ukuhlaziywa Kobuningi be-Liquid Chromatography

1. Khetha indlela ye-chromatographic efanele ukuhlaziya inani:

l Qinisekisa inani eliphakeme lengxenye etholiwe futhi uzuze isinqumo (R) esikhulu kuno-1.5

l Thola ukuvumelana (ukuhlanzeka) kweziqongo zechromatographic zezingxenye ezihloliwe

l Ukunquma umkhawulo wokutholwa kanye nomkhawulo wokulinganisa wendlela;ukuzwela kanye nobubanzi bomugqa

2. Sungula ijika lokulinganisa elinamasampuli ajwayelekile okugxilisana okuhlukene

3. Hlola ukunemba nokunemba kwezindlela zobuningi

4. Sebenzisa isofthiwe ehambisanayo yokuphatha i-chromatography ukuze usebenzise ukuqoqwa kwesampula, ukucutshungulwa kwedatha kanye nemiphumela yombiko

INGXENYE
05 Ukuhlonzwa kweziqongo zenani (ikhwalithi)

Khomba ngendlela efanele isiqongo ngasinye se-chromatographic okufanele silinganiswe

Okokuqala, sebenzisa isampula elijwayelekile ukuze unqume isikhathi sokugcinwa (Rt) senani eliphakeme le-chromatographic elizolinganiswa.Ngokuqhathanisa isikhathi sokugcina, thola ingxenye ehambisana nenani eliphakeme ngalinye le-chromatographic kusampuli engaziwa.Indlela yekhwalithi ye-chromatographic iwukuqhathanisa isikhathi sokugcinwa nesampula evamile.Umbandela Abaneleukuqinisekiswa okwengeziwe (qualitative)

1. Indlela yokwengeza ejwayelekile

2. Sebenzisa ezinye izindlela ngesikhathi esifanayo: ezinye izindlela ze-chromatographic (shintsha indlela yokusebenza, njengalokhu: ukusebenzisa amakholomu e-chromatographic ahlukene), ezinye izitholi (PDA: ukuqhathanisa kwe-spectrum, ukusesha komtapo we-spectrum; i-MS: ukuhlaziywa kwe-mass spectrum, ukusesha komtapo we-spectrum)

3. Amanye amathuluzi nezindlela

INGXENYE
06 Ukuqinisekiswa Kokungaguquguquki Kwenani Eliphakeme Eliphakeme

Qinisekisa ukuvumelana kwe-chromatographic peak (ubumsulwa)

Qinisekisa ukuthi kunengxenye eyodwa kuphela elinganisiwe ngaphansi kokuphakama ngakunye kwe-chromatographic

Bheka ukugxambukela ezintweni ezi-co-eluting (ukungcola)

Izindlela Zokuqinisekisa Ukuvumelana Kwe-Chromatographic Peak (Ubumsulwa)

Ukuqhathanisa ama-Spectrograms namaDetectors e-Photodiode Matrix (PDA).

I-Peak Purity Identification

2996 Purity Angle Theory

Izindlela zokulinganisa ezivame ukusetshenziswa INGXENYE 07

Indlela yejika evamile, ihlukaniswe yaba indlela yangaphandle evamile kanye nendlela evamile yangaphakathi:

1. Indlela ejwayelekile yangaphandle: esetshenziswa kakhulu ku-chromatography ewuketshezi

Uchungechunge lwamasampuli ajwayelekile okugxiliswa kwawo okwaziwayo alungiswa kusetshenziswa amasampula amsulwa ezinhlanganisela azohlolwa njengamasampula ajwayelekile.ifakwe kukholamu kuze kufike kunani layo lokuphendula (indawo ephakeme kakhulu).
Phakathi kobubanzi obuthile, kunobudlelwano obuhle bomugqa phakathi kokugxilwa kwesampula evamile kanye nenani lokuphendula, okungukuthi W= f×A , futhi kwenziwa ijika elijwayelekile.

Ngaphansi kwezimo zokuhlola ezifanayo ncamashi, jova isampula engaziwa ukuze uthole inani lokuphendula lengxenye ezolinganiswa .Ngokusho kwe-coefficient f eyaziwayo, ukugxila kwengxenye ezolinganiswa kungatholakala.

Izinzuzo zendlela yangaphandle evamile:ukusebenza okulula nokubala, kuyindlela evame ukusetshenziswa yobuningi;asikho isidingo sokuthi ingxenye ngayinye itholwe futhi ichazwe;isampula ejwayelekile iyadingeka;izimo zokulinganisa zesampula evamile kanye nesampula engaziwa kufanele zihambisane;umthamo womjovo kufanele ucace.

Ububi bendlela yangaphandle evamile:Izimo zokuhlola zidingeka ukuthi zibe phezulu, njengokuzwela komtshina, izinga lokugeleza, nokwakheka kwesigaba esihambayo akukwazi ukushintshwa;umthamo womjovo ngamunye kufanele ube nokuphindaphinda okuhle.

2. Indlela ejwayelekile yangaphakathi: enembile, kodwa enzima, esetshenziswa kakhulu ezindleleni ezijwayelekile

Inani elaziwayo lezinga langaphakathi liyengezwa esilinganisweni ukuze kwenziwe izinga elixubile, futhi uchungechunge lwamazinga okusebenza okugxilisa ingqondo okwaziwayo luyalungiswa.Isilinganiso se-molar sendinganiso nesilinganiso sangaphakathi esilinganisweni esixubile sihlala singashintshile.Jova kukholamu ye-chromatographic futhi uthathe (indawo ephakeme yesampula evamile/indawo ephakeme yesampula evamile) njengenani lokuphendula.Ngokobudlelwano bomugqa phakathi kwevelu yokuphendula nokugxilisana kwezinga lokusebenza, okungukuthi W= f×A, kwenziwa ijika elijwayelekile.

Inani elaziwayo lezinga langaphakathi liyengezwa kusampula engaziwa futhi lifakwe kukholomu ukuze kutholwe inani lokuphendula lengxenye ezolinganiswa .Ngokusho kwe-coefficient f eyaziwayo, ukugxila kwengxenye ezolinganiswa kungatholakala.

Izici zendlela yangaphakathi evamile:Ngesikhathi sokusebenza, isampula nezinga langaphakathi kuxutshwa ndawonye futhi kujovelwe kukholamu ye-chromatographic, inqobo nje uma isilinganiso senani lengxenye elinganisiwe nezinga langaphakathi kusixazululo esixubile singaguquki, ukuguqulwa kwevolumu yesampula. ngeke kuthinte imiphumela yobuningi..Indlela esezingeni yangaphakathi isusa umthelela wevolumu yesampula , ngisho nesigaba seselula nomtshina, ngakho inembe kakhulu kunendlela evamile yangaphandle.

INGXENYE
08 Izinto Ezithinta Imiphumela Yokuhlaziya Okuningi

Ukunemba okungalungile kungabangelwa:

Ukuhlanganiswa kwendawo ephakeme engafanele, ukubola kwesampula noma ukungcola okwethulwe ngesikhathi sokulungiswa kwesampula, isampula yebhodlela elingavalwanga, isampula noma ukuvuvuka kwe-solvent, ukulungiswa kwesampula okungalungile, izinkinga zomjovo wesampula, ukulungiswa okungalungile kwangaphakathi.

Izizathu ezingaba khona zokunemba okungalungile:

Ukuhlanganiswa okuphezulu okungalungile, izinkinga zomjovo noma umjovo, ukubola kwesampula noma ukungcola okwethulwa ngesikhathi sokulungiswa kwesampula, izinkinga ze-chromatographic, impendulo yomtshina eyonakele.