Imigaqo kunye neendlela zoHlalutyo loMyinge ngoLiquid Chromatography

Indlela yokwahlula i-chromatography yolwelo isekelwe kumahluko wokudityaniswa kwamacandelo kumxube wezigaba ezibini.

Ngokwezigaba ezahlukeneyo zokumisa, ichromatography yolwelo yahlulwe yaba yichromatography yolwelo-oluqilima, ichromatography yolwelo kunye nesigaba se-chromatography edibeneyo.Ezona zisetyenziswa kakhulu zi-liquid-solid chromatography kunye ne-silica gel njenge-filler kunye ne-bond phase chromatography kunye ne-microsilica njenge-matrix.

Ngokohlobo lwesigaba esimileyo, ichromatography yolwelo inokohlulwa ibe yikholamu yechromatography, iphepha chromatography kunye nekhromatography ebhityileyo.Ngokomthamo we-adsorption, inokohlulwa ibe yi-adsorption chromatography, i-partition chromatography, i-ion exchange chromatography kunye ne-gel permeation chromatography.

Kwiminyaka yakutshanje, inkqubo yokuhamba kolwelo olunoxinzelelo oluphezulu yongeziweyo kwikholamu yolwelo lwechromatography inkqubo ukwenza isigaba esihambayo sihambe ngokukhawuleza phantsi koxinzelelo oluphezulu lokuphucula isiphumo sokwahlukana, ngoko ke ukusebenza kakuhle okuphezulu (okukwabizwa ngokuba yi-high-pressure) i-chromatography yolwelo. iye yavela.

INXALENYE
01 Umgaqo-siseko woVavanyo loMyinge we-Liquid Chromatography

Ukulinganisa ngesiseko somgangatho, izinto ezicocekileyo ziyafuneka njengemigangatho;

I-Liquid chromatography quantification yindlela yobungakanani: oko kukuthi, umyinge we-analyte kumxube uqikelelwa ukusuka kwisixa esaziwayo sesampulu ecocekileyo yomgangatho.

INXALENYE
02 Isiseko soBanzi ngeChromatography yoLiquid

Isixa secandelo elilinganisiweyo (W) silingana nexabiso lempendulo (A) (ubude bencopho okanye indawo encopho), W=f×A.

Umba wokulungiswa kobungakanani (f): Lulungelelwaniso olungaguqukiyo lwefomula yokubala yobungakanani, kwaye intsingiselo yayo ebonakalayo sisixa secandelo elilinganisiweyo elimelwe lixabiso lempendulo yeyunithi (indawo encochoyi).

I-quantitative correction factor inokufumaneka kwisixa esaziwayo sesampulu esemgangathweni kunye nexabiso layo lokuphendula.

Ukulinganisa ixabiso lempendulo yecandelo elingaziwayo, kwaye inani lecandelo linokufunyanwa ngumlinganiselo wokulungisa ubungakanani.

INXALENYE
03 Amagama aqhelekileyo kuhlalutyo lobungakanani

Isampuli (isampulu): isisombululo esine-analyte yohlalutyo lwe-chromatographic.Ukwahlula kwiisampuli eziqhelekileyo nezingaziwayo.

Umgangatho: Imveliso ecocekileyo ene-concentration eyaziwayo.Isampuli engaziwayo (engaziwa): Umxube oza kuvavanywa ukugxininiswa kwawo.

Ubunzima besampulu: Ubunzima bokuqala besampulu eza kuvavanywa.

Dilution: Into yokudilution yesampulu engaziwayo.

Icandelo : Incopho yechromatographic kufuneka ihlalutywe ngokobungakanani, oko kukuthi, umhlalutyi onomxholo ongaziwayo.

Isixa secandelo (isixa): Umxholo (okanye ugxininiso) lwento eza kuvavanywa.

Ingqibelelo : Inkqubo yokubala yokulinganisa ummandla wencopho yekhromatographic ngekhompyutha.

Ijika lokulinganisa: Umgca wecala lomxholo wecandelo ngokuchasene nexabiso lempendulo, esekwe kwixabiso elaziwayo lezinto eziqhelekileyo, ezisetyenziselwa ukumisela umxholo ongaziwayo womhlalutyi.

INXALENYE
04 Uhlalutyo lobungakanani be-Liquid Chromatography

1. Khetha indlela yechromatographic elungele uhlalutyo lobungakanani:

l Qinisekisa incopho yecandelo elichongiweyo kwaye uzuze isisombululo (R) esingaphezulu kwe-1.5

l Qinisekisa ukuhambelana (ukucoceka) kweencopho zechromatographic zamacandelo avavanyiweyo

l Ukumisela umda wokufumanisa kunye nomda wobungakanani bendlela;ubuntununtunu kunye noluhlu lomgca

2. Ukuseka i-curve calibration kunye neesampuli ezisemgangathweni zogxininiso oluhlukeneyo

3. Jonga ukuchaneka kunye nokuchaneka kweendlela zobuninzi

4. Sebenzisa isoftware yolawulo lwechromatography ehambelanayo ukuphumeza ukuqokelela iisampulu, ukusetyenzwa kwedatha kunye neziphumo zengxelo

INXALENYE
05 Ukuchongwa kweencopho zobungakanani (ubunjani)

Chonga ngokufanelekileyo incopho nganye yechromatographic eza kubalwa

Okokuqala, sebenzisa isampulu eqhelekileyo ukumisela ixesha lokugcinwa (Rt) lencopho yechromatographic ukuba ilinganiswe.Ngokuthelekisa ixesha lokugcinwa, fumana icandelo elihambelana nencopho nganye yechromatographic kwisampulu engaziwayo.Indlela yomgangatho wechromatographic kukuthelekisa ixesha lokugcinwa kunye nesampulu eqhelekileyo.Iinqobo zokugweba azanelangauqinisekiso olongezelelweyo (umgangatho)

1. Indlela yokudibanisa esemgangathweni

2. Sebenzisa ezinye iindlela ngaxeshanye: ezinye iindlela zechromatographic (tshintsha indlela yokusebenza, efana: usebenzisa iikholamu ezahlukeneyo zechromatographic), ezinye ii-detectors (PDA: uthelekiso lwe-spectrum, ukukhangela kwethala leencwadi; i-MS: uhlalutyo lwe-massspectrum, ukukhangela kwethala leencwadi le-spectrum)

3. Ezinye izixhobo kunye neendlela

INXALENYE
06 Ukuqinisekiswa kokuNxibelelana kweNcopho yoMyinge

Qinisekisa ukuhambelana kwencopho yechromatographic (ubunyulu)

Qinisekisa ukuba kukho icandelo elinye elilinganisiweyo phantsi kwencopho nganye yechromatographic

Jonga uphazamiseko kwizinto ezixutywayo (ubumdaka)

Iindlela zoQinisekiso lweChromatographic Peak Consistency (Ubunyulu)

Ukuthelekisa iiSpectrograms kunye nePhotodiode Matrix (PDA) Detectors

Incopho yokuchongwa kobunyulu

2996 Ithiyori yeAngle enyulu

Iindlela zobungakanani eziqhele ukusetyenziswa kwiCANDELO 07

Indlela yegophe esemgangathweni, yahlulwe yaba yindlela yomgangatho wangaphandle kunye nendlela yomgangatho wangaphakathi:

1. Indlela yomgangatho wangaphandle: isetyenziswa kakhulu kwichromatography yolwelo

Uluhlu lweesampulu ezisemgangathweni zogxininiso olwaziwayo lwalungiswa kusetyenziswa iisampuli ezicocekileyo zekhompawundi eziza kuvavanywa njengeesampuli eziqhelekileyo.ifakwe kwikholamu ukuya kwixabiso layo lokuphendula (indawo ephezulu).
Ngaphakathi koluhlu oluthile, kukho ubudlelwane obuhle bomgca phakathi koxinaniso lwesampulu esemgangathweni kunye nexabiso lempendulo, oko kukuthi W= f×A, kwaye ijika eliqhelekileyo lenziwe.

Ngaphantsi kweemeko ezifanayo zovavanyo, faka isampuli engaziwayo ukufumana ixabiso lempendulo yecandelo eliza kulinganiswa.Ngokutsho kwe-coefficient f eyaziwayo, ukuxinwa kwecandelo eliza kulinganiswa kunokufumaneka.

Uncedo lwendlela yomgangatho wangaphandle:ukusebenza okulula kunye nokubala, yindlela esetyenziswa ngokuqhelekileyo yobungakanani;akukho mfuneko yokuba icandelo ngalinye lichongwe kwaye licocwe;isampuli eqhelekileyo iyafuneka;iimeko zokulinganisa isampula eqhelekileyo kunye nesampuli engaziwayo kufuneka ihambelane;umthamo wokutofa kufuneka uchaneke.

Ukungalungi kwendlela yomgangatho wangaphandle:Iimeko zovavanyo zifunekayo ukuba zibe phezulu, ezifana novakalelo lwe-detector, izinga lokuhamba, kunye nokubunjwa kwesigaba seselula asinakuguqulwa;Umthamo wenaliti nganye kufuneka ube nokuphindaphinda okulungileyo.

2. Indlela yomgangatho wangaphakathi: ichanekile, kodwa iyinkathazo, isetyenziswa kakhulu kwiindlela eziqhelekileyo

Umlinganiselo owaziwayo womgangatho wangaphakathi wongezwa kumgangatho wokwenza umgangatho odibeneyo, kwaye uchungechunge lwemigangatho yokusebenza yoxinaniso olwaziwayo lulungiswa.Umlinganiselo we-molar womgangatho ukuya kumgangatho wangaphakathi kumgangatho odibeneyo uhlala ungatshintshi.Tofa kwikholamu yechromatographic kwaye uthathe (indawo yencopho yesampuli eqhelekileyo/indawo yesampulu esemgangathweni yangaphakathi) njengexabiso lokuphendula.Ngokobudlelwane bomgca phakathi kwexabiso lempendulo kunye nokugxininiswa komgangatho wokusebenza, oko kukuthi W= f× A, ijika eliqhelekileyo lenziwa.

Isixa esaziwayo somgangatho wangaphakathi wongezwa kwisampuli engaziwayo kwaye ifakwe kwikholomu ukufumana ixabiso lempendulo yecandelo eliza kulinganiswa.Ngokutsho kwe-coefficient f eyaziwayo, ukuxinwa kwecandelo eliza kulinganiswa kunokufumaneka.

Iimpawu zendlela yomgangatho wangaphakathi:Ngexesha lokusebenza, isampuli kunye nomgangatho wangaphakathi zixutywe kunye kwaye zifakwe kwikholamu yechromatographic, nje ukuba umlinganiselo wecandelo elilinganisiweyo ukuya kumgangatho wangaphakathi kwisisombululo esixubileyo sihlala sitshintshile, ukuguqulwa komthamo wesampuli. ayiyi kuchaphazela iziphumo zobungakanani..Indlela yomgangatho wangaphakathi isusa impembelelo yevolumu yesampuli , kunye nesigaba seselula kunye ne-detector, ngoko ichaneke ngakumbi kunendlela yomgangatho wangaphandle.

INXALENYE
08 Imiba echaphazela iziphumo zoHlalutyo lobungakanani

Ukuchaneka kakubi kunokubangelwa:

Ukudityaniswa kwendawo eyincopho engafanelekanga, ukubola kwesampulu okanye ukungcola okwaziswe ngexesha lolungiselelo lwesampulu, isampuli yevial ​​ayitywinwanga, iisampulu okanye ukunyibilika kwenyibiliko, ukulungiswa kwesampulu engalunganga, iingxaki zesampulu yesitofu, ukulungiswa kwemigangatho engachanekanga.

Izizathu ezinokwenzeka zokungachaneki kakuhle:

Ukudityaniswa kwencopho engafanelekanga, inaliti okanye iingxaki zesitofu, ukubola kwesampulu okanye ukungcola okungeniswa ngexesha lolungiselelo lwesampulu, iingxaki zechromatographic, ukuthotywa kwempendulo yomtshini