Lub hauv paus ntsiab lus thiab cov txheej txheem ntawm Kev Ntsuas Quantitative los ntawm Liquid Chromatography

Cov txheej txheem sib cais ntawm cov kua chromatography yog nyob ntawm qhov sib txawv ntawm cov affinity ntawm cov khoom hauv qhov sib tov rau ob theem.

Raws li cov theem sib txawv nyob ruaj ruaj, kua chromatography muab faib ua kua-khoom chromatography, kua-kua chromatography thiab bonded theem chromatography.Feem ntau siv yog kua-khoom chromatography nrog silica gel raws li muab tub lim thiab bonded theem chromatography nrog microsilica li matrix.

Raws li daim ntawv ntawm cov theem nyob ruaj khov, kua chromatography tuaj yeem muab faib ua kem chromatography, ntawv chromatography thiab nyias txheej chromatography.Raws li lub peev xwm adsorption, nws tuaj yeem muab faib ua adsorption chromatography, muab faib chromatography, ion pauv chromatography thiab gel permeation chromatography.

Nyob rau hauv xyoo tas los no, lub tshuab ua kua dej siab tau ntxiv rau hauv cov kua kem chromatography system ua rau cov theem ntawm lub xov tooj ntawm tes ntws sai hauv kev kub siab los txhim kho cov nyhuv sib cais, yog li kev ua haujlwm siab (tseem hu ua high-pressure) kua chromatography. tau tshwm sim.

PART
01 Lub hauv paus ntsiab lus ntawm Quantitative Analysis of Liquid Chromatography

Txhawm rau kom muaj nuj nqis raws li qhov muaj txiaj ntsig, cov tshuaj ntshiab yog xav tau raws li cov qauv;

Cov kua dej chromatography quantification yog ib txoj hauv kev kom muaj nuj nqis: uas yog, tus nqi ntawm cov tshuaj ntsuam xyuas hauv qhov sib xyaw yog kwv yees los ntawm tus lej paub ntawm tus qauv ntshiab.

PART
02 Lub hauv paus rau Quantification los ntawm Liquid Chromatography

Tus nqi ntawm cov ntsuas ntsuas (W) yog proportional rau tus nqi teb (A) (nruab qhov siab lossis qhov siab tshaj), W = f × A.

Quantitative correction factor (f): Nws yog qhov proportionality tas li ntawm tus lej suav, thiab nws lub ntsiab lus ntawm lub cev yog tus nqi ntawm cov ntsuas ntsuas uas sawv cev los ntawm chav tsev teb tus nqi (peak cheeb tsam).

Qhov kev kho kom muaj nuj nqis tuaj yeem tau txais los ntawm qhov paub tus nqi ntawm tus qauv qauv thiab nws cov nqi teb.

Ntsuas cov lus teb tus nqi ntawm cov khoom tsis paub, thiab tus nqi ntawm cov khoom tuaj yeem tau txais los ntawm kev kho qhov ntau.

PART
03 Cov ntsiab lus dav dav hauv kev txheeb xyuas qhov ntau

Qauv (piv txwv): ib qho kev daws teeb meem uas muaj cov tshuaj ntsuam xyuas chromatographic.Muab faib ua cov qauv thiab tsis paub cov qauv.

Standard: Ib yam khoom ntshiab nrog ib tug paub concentration.Tsis paub tus qauv (unknow): Qhov sib tov uas nws concentration yuav tsum tau kuaj.

Qauv hnyav: Thawj qhov hnyav ntawm tus qauv yuav tsum tau kuaj.

Dilution: Lub dilution yam ntawm tus qauv tsis paub.

Cheebtsam: lub ncov chromatographic yuav tsum tau soj ntsuam quantitatively, uas yog, tus analyte uas nws cov ntsiab lus tsis paub.

Tus nqi ntawm cov khoom (ntau npaum li cas): cov ntsiab lus (lossis concentration) ntawm cov khoom yuav tsum tau kuaj.

Integerity : Cov txheej txheem suav ntawm kev ntsuas thaj tsam siab tshaj ntawm chromatographic ncov los ntawm lub computer.

Calibration nkhaus: Ib txoj kab nkhaus ntawm cov ntsiab lus ntawm cov ntsiab lus piv rau cov lus teb tus nqi, tsim los ntawm tus lej paub ntawm cov khoom siv, siv los txiav txim siab cov ntsiab lus tsis paub ntawm qhov ntsuas.

PART
04 Quantitative Analysis of Liquid Chromatography

1. Xaiv ib txoj kev chromatographic haum rau kev soj ntsuam ntau:

l Paub meej qhov ncov ntawm qhov ntsuas pom thiab ua tiav qhov kev daws teeb meem (R) ntau dua 1.5

l Txiav txim siab qhov sib xws (purity) ntawm chromatographic peaks ntawm cov khoom kuaj

l Txiav txim xyuas qhov txwv tsis pub thiab kev txwv ntau ntawm txoj kev;rhiab heev thiab linear ntau yam

2. Tsim kom muaj qhov nkhaus calibration nrog cov qauv qauv ntawm ntau qhov sib txawv

3. Tshawb xyuas qhov tseeb thiab qhov tseeb ntawm ntau txoj hauv kev

4. Siv cov software tswj chromatography sib thooj los siv cov qauv sau, ua cov ntaub ntawv thiab tshaj tawm cov txiaj ntsig

PART
05 Kev txheeb xyuas qhov muaj nuj nqis siab tshaj (zoo)

Ua kom zoo txheeb xyuas txhua qhov chromatographic ncov kom muaj nuj nqis

Ua ntej, siv tus qauv qauv los txiav txim lub sijhawm khaws cia (Rt) ntawm chromatographic ncov kom muaj nuj nqis.Los ntawm kev sib piv lub sijhawm khaws cia, nrhiav cov khoom sib txuam rau txhua qhov chromatographic ncov hauv cov qauv tsis paub.Cov txheej txheem chromatographic zoo yog los sib piv lub sijhawm khaws cia nrog tus qauv qauv.Qhov ntsuas tsis txausntxiv kev pom zoo (zoo)

1. Cov txheej txheem ntxiv

2. Siv lwm txoj hauv kev tib lub sijhawm: lwm txoj hauv kev chromatographic (hloov cov txheej txheem, xws li: siv cov kab chromatographic sib txawv), lwm yam khoom siv (PDA: spectrum sib piv, spectrum library search; MS: mass spectrum analysis, spectrum library search)

3. Lwm yam cuab yeej thiab txoj kev

PART
06 Kev lees paub ntawm Quantitative Peak Consistency

Paub meej tias chromatographic ncov siab sib xws (purity)

Xyuas kom meej tias tsuas muaj ib qho kev ntsuas nyob rau hauv txhua qhov chromatographic ncov

Tshawb xyuas kev cuam tshuam los ntawm co-eluting tshuaj (impurities)

Cov txheej txheem rau kev lees paub ntawm Chromatographic Peak Consistency (Purity)

Sib piv Spectrograms nrog Photodiode Matrix (PDA) Detectors

Peak Purity Identification

2996 Purity Angle Theory

Cov txheej txheem ntau feem ntau siv hauv PART 07

Standard nkhaus txoj kev, muab faib mus rau hauv cov txheej txheem sab nraud thiab sab hauv txheej txheem:

1. Cov txheej txheem sab nraud: feem ntau siv hauv cov kua chromatography

Ib cov qauv qauv ntawm cov kev paub paub ntau tau npaj siv cov qauv ntshiab ntawm cov tebchaw los kuaj ua cov qauv qauv.txhaj rau hauv kem mus txog nws cov lus teb tus nqi (peak area).
Nyob rau hauv ib tug tej yam ntau yam, muaj ib tug zoo linear kev sib raug zoo ntawm cov concentration ntawm tus qauv qauv thiab cov lus teb tus nqi, uas yog W = f × A, thiab ib tug qauv nkhaus yog ua.

Raws li qhov kev sim tib yam nkaus, txhaj cov qauv uas tsis paub meej kom tau txais cov lus teb tus nqi ntawm cov khoom siv ntsuas.Raws li paub coefficient f, qhov concentration ntawm cov khoom yuav tsum tau ntsuas tau.

Qhov zoo ntawm cov txheej txheem sab nraud:Kev ua haujlwm yooj yim thiab kev suav, nws yog ib txoj hauv kev siv ntau ntau;tsis tas yuav tsum tau rau txhua yam khoom kom raug kuaj thiab eluted;tus qauv yuav tsum muaj;cov kev ntsuas ntawm tus qauv qauv thiab cov qauv tsis paub yuav tsum ua kom zoo ib yam;lub ntim ntim yuav tsum yog qhov tseeb.

Disadvantages ntawm tus txheej txheem sab nraud:Cov kev sim yuav tsum tau ua kom siab, xws li rhiab heev ntawm lub ntes, tus nqi ntws, thiab cov muaj pes tsawg leeg ntawm lub xov tooj ntawm tes yuav hloov tsis tau;qhov ntim ntawm txhua qhov txhaj tshuaj yuav tsum muaj qhov rov ua tau zoo.

2. Cov txheej txheem sab hauv: muaj tseeb, tab sis teeb meem, feem ntau siv hauv cov txheej txheem txheem

Ib tug paub tus nqi ntawm cov txheej txheem sab hauv yog ntxiv rau tus qauv los ua tus qauv sib xyaw, thiab cov txheej txheem kev ua haujlwm ntawm kev paub txog concentration yog npaj.Cov molar piv ntawm tus qauv rau tus qauv sab hauv hauv cov qauv sib xyaw tseem tsis hloov pauv.Txhaj rau hauv chromatographic kem thiab coj (tus qauv piv txwv thaj chaw siab tshaj / sab hauv tus qauv piv txwv thaj tsam) raws li tus nqi teb.Raws li kev sib raug zoo ntawm tus nqi teb thiab qhov concentration ntawm cov qauv ua haujlwm, xws li W = f × A, tus qauv nkhaus yog tsim.

Ib tug paub tus nqi ntawm tus qauv sab hauv yog ntxiv rau cov qauv tsis paub thiab txhaj rau hauv kem kom tau txais cov lus teb tus nqi ntawm cov khoom yuav ntsuas.Raws li paub coefficient f, qhov concentration ntawm cov khoom yuav tsum tau ntsuas tau.

Cov yam ntxwv ntawm tus txheej txheem sab hauv:Thaum lub sijhawm ua haujlwm, cov qauv thiab cov qauv sab hauv yog sib xyaw ua ke thiab txhaj rau hauv kab chromatographic, yog li ntev npaum li qhov piv ntawm tus nqi ntawm cov ntsuas ntsuas mus rau tus qauv sab hauv hauv cov tshuaj sib tov yog tas li, kev hloov pauv ntawm cov qauv ntim. yuav tsis cuam tshuam cov txiaj ntsig ntau..Cov txheej txheem sab hauv offsets tus cwj pwm ntawm cov qauv ntim, thiab txawm tias lub xov tooj ntawm tes thiab ntes, yog li nws yog qhov tseeb dua li cov qauv txheej txheem sab nraud.

PART
08 Yam Uas Muaj Feem Cuam Tshuam Txog Kev Tshawb Fawb Txog Kev Tshawb Fawb

Qhov tseeb tsis zoo tuaj yeem tshwm sim los ntawm:

Kev sib koom ua ke ntawm thaj chaw ncov tsis raug, qauv decomposition lossis impurities qhia thaum lub sijhawm ua qauv, qauv vial tsis kaw, qauv lossis hnyav volatilization, tsis raug coj mus kuaj, teeb meem kev txhaj tshuaj, kev npaj tsis raug

Yog vim li cas rau qhov tsis zoo precision:

Kev sib xyaw ncov tsis raug, kev txhaj tshuaj lossis cov teeb meem injector, qauv decomposition lossis impurities qhia thaum lub sijhawm ua qauv, teeb meem chromatographic, degraded detector teb