Nheyo uye Nzira dzeKuongorora Kwehuwandu neLiquid Chromatography

Iyo yekuparadzanisa nzira yemvura chromatography yakavakirwa pamusiyano mukubatanidzwa kwezvikamu mumusanganiswa wezvikamu zviviri izvi.

Zvinoenderana nematanho akasiyana-siyana akamira, liquid chromatography yakakamurwa kuita liquid-solid chromatography, liquid-liquid chromatography uye bonded phase chromatography.Iyo inonyanya kushandiswa ndeye liquid-solid chromatography ine silica gel sekuzadza uye bonded phase chromatography ine microsilica sematrix.

Zvinoenderana nechimiro cheyakamira chikamu, liquid chromatography inogona kukamurwa kuita column chromatography, bepa chromatography uye nhete layer chromatography.Zvinoenderana neiyo adsorption kugona, inogona kukamurwa kuita adsorption chromatography, partition chromatography, ion exchange chromatography uye gel permeation chromatography.

Mumakore achangopfuura, yakakwira-kudzvanywa kwemvura kuyerera sisitimu yakawedzerwa kune yemvura column chromatography system kuita kuti nharembozha ifambe nekukasira pasi pekumanikidzwa kwepamusoro kuvandudza mhedzisiro yekuparadzanisa, saka yakanyanya-kushanda (inozivikanwawo seyepamusoro-kudzvanywa) liquid chromatography. yabuda.

CHIKAMU
01 Nheyo yeKuongorora Kwehuwandu hweLiquid Chromatography

Kuverengera pahwaro hwemhando, zvinhu zvakachena zvinodiwa seyero;

Liquid chromatography quantification inzira ine mwero: kureva kuti, huwandu hweanalyte mumusanganiswa unofungidzirwa kubva pachiyero chinozivikanwa cheiyo pure standard sample.

CHIKAMU
02 Nheyo yeQuantification neLiquid Chromatography

Huwandu hwechikamu chakayerwa (W) chinoenzana nehukoshi hwemhinduro (A) (hurefu hwepamusoro kana nzvimbo yepamusoro), W=f×A.

Quantitative correction factor (f): Ndiko kuenzana kwehuwandu hweformula yehuwandu hwekuverenga, uye chirevo chayo chemuviri chiyero chechikamu chakayerwa chinomiririrwa neiyo unit mhinduro kukosha (nzvimbo yepamusoro).

The quantitative correction factor inogona kuwanikwa kubva kuhuwandu hunozivikanwa hwemuenzaniso wemuenzaniso uye kukosha kwayo kwekupindura.

Edza kukosha kwekupindura kwechikamu chisingazivikanwi, uye chiyero chechikamu chinogona kuwanikwa nehuwandu hwekugadzirisa chinhu.

CHIKAMU
03 Mazwi akajairika mukuongorora kwehuwandu

Sample (sample): mhinduro ine analyte yechromatographic analysis.Yakakamurwa kuita zviyero uye zvisingazivikanwe sampuli.

Chiyero: Chigadzirwa chakachena chine hunhu hunozivikanwa.Sampuli isingazivikanwe (isingazivikanwe): Musanganiswa uyo kutarisisa kunofanirwa kuongororwa.

Sample uremu: Yekutanga kuyerwa kwesample kuti iongororwe.

Dilution: The dilution factor yemuenzaniso usingazivikanwi.

Chikamu : iyo chromatographic peak kuti iongororwe zvakanyanya, kureva, muongorori ane izvo zvisingazivikanwe.

Huwandu hwechikamu (huwandu): zviri mukati (kana kusungirirwa) kwechinhu chinoedzwa.

Kutendeseka : Iyo computational maitiro ekuyera nzvimbo yepamusoro yechromatographic peak nekombuta.

Calibration curve: Mutsara wemutsetse wechikamu chemukati maringe nekupindura kukosha, yakagadzwa kubva kune inozivikanwa huwandu hwechinhu chakajairwa, inoshandiswa kuona izvo zvisingazivikanwe zviri muanalyte.

CHIKAMU
04 Quantitative Analysis yeLiquid Chromatography

1. Sarudza nzira yechromatographic yakakodzera kuongororwa kwehuwandu:

l Simbisa iyo yepamusoro yechikamu chakaonekwa uye uwane sarudzo (R) yakakura kupfuura 1.5

l Sarudza kuwirirana (kuchena) kweiyo chromatographic peaks yezvakaedzwa zvikamu

l Sarudza iyo yekuona muganho uye quantification muganho weiyo nzira;sensitivity uye linear range

2. Gadzira calibration curve nemasampuli akajairwa ezvikamu zvakasiyana

3. Tarisa kururamisa uye kurongeka kwehuwandu nzira

4. Shandisa inoenderana chromatography manejimendi software kuti uise sampuli yekuunganidza, kugadzirisa data uye mishumo yemhedzisiro

CHIKAMU
05 Kuzivikanwa kwehuwandu hwepamusoro (hunhu)

Nehuno ratidza imwe neimwe chromatographic peak kuti iverengerwe

Chekutanga, shandisa yakajairwa sampuli kuona nguva yekuchengeta (Rt) yechromatographic peak kuti iverengerwe.Nekuenzanisa nguva yekuchengeta, tsvaga chikamu chinoenderana neimwe chromatographic peak mumuenzaniso usingazivikanwe.Iyo chromatographic qualitative nzira ndeyekuenzanisa nguva yekuchengeta neyakajairwa sampuli.The criterion Insufficientkumwe kusimbisa (unhu)

1. Standard kuwedzera nzira

2. Shandisa dzimwe nzira panguva imwe chete: dzimwe nzira dzechromatographic (shandura nzira, dzakadai se: kushandisa chromatographic columns dzakasiyana), mamwe madhigirii (PDA: kuenzanisa kwemavara, kutsvaga kweraibhurari yemagetsi; MS: kuongororwa kwehuwandu, kutsvaga kweraibhurari ye spectrum)

3. Zvimwe zviridzwa uye nzira

CHIKAMU
06 Kusimbisa Kwehuwandu Peak Consistency

Simbisa chromatographic peak kuenderana (kuchena)

Ita shuwa kuti pane chikamu chimwe chete chakayerwa pasi peyega yega chromatographic peak

Tarisa kukanganiswa kubva ku-co-eluting zvinhu (kusvibiswa)

Nzira dzeSimbiso yeChromatographic Peak Consistency (Kuchena)

Kuenzanisa Spectrograms ne Photodiode Matrix (PDA) Detectors

Peak Purity Identification

2996 Purity Angle Theory

Quantitative nzira dzinowanzoshandiswa muCHIKAMU 07

Yakajairika curve nzira, yakakamurwa kuva yekunze chiyero nzira uye yemukati chiyero nzira:

1. External mwero nzira: inonyanya kushandiswa mune yemvura chromatography

Mutsara wemasampuli akajairwa ezviyero zvinozivikanwa zvakagadzirirwa uchishandisa yakachena masampuli emakomponi kuti aedzwe seyakajairwa sampuli.jekiseni mukoramu kusvika kune yayo mhinduro kukosha (peak nzvimbo).
Mukati mehumwe mutsara, pane hukama hwakanaka hwemutsara pakati pekumisikidzwa kweyakajairwa sampuli uye kukosha kwekupindura, zvinoti W= f×A, uye mwero wecurve unogadzirwa.

Pasi pemamiriro ezvinhu chaiwo ekuedza, jekiseni muenzaniso usingazivikanwi kuti uwane kukosha kwekupindura kwechikamu chinofanirwa kuyerwa.Maererano neiyo inozivikanwa coefficient f , kusungirirwa kwechikamu chinofanirwa kuyerwa chinogona kuwanikwa.

Zvakanakira zvekunze chiyero nzira:kushanda nyore uye kuverenga, inowanzoshandiswa nzira yehuwandu;hapana chikonzero chekuti chikamu chega chega chionekwe uye kuburitswa;chiyero chemuenzaniso chinodiwa;kuyerwa kwemamiriro emuenzaniso wakajairika uye sampuli isingazivikanwe inofanira kuenderana;vhoriyamu yejekiseni inofanira kuva yakarurama.

Zvakaipa zvekunze chiyero nzira:Mamiriro ekuedza anodiwa kuti ave akakwirira, akadai sekunzwa kwe detector, chiyero chekuyerera, uye kuumbwa kwechikamu chekufambisa hakugoni kuchinjwa;huwandu hwejekiseni rega rega hunofanirwa kuve nekudzokororwa kwakanaka.

2. Internal mwero nzira: chakarurama, asi chinonetsa, chinoshandiswa zvakanyanya mumaitiro akajairwa

Chiyero chinozivikanwa chechiyero chemukati chinowedzerwa kune chiyero kuita chiyero chakavhenganiswa, uye nhevedzano yekushanda kwemaitiro ekuzivikanwa kunozivikanwa anogadzirirwa.Iyo molar ratio yechiyero kune yemukati mwero mune yakasanganiswa mwero inoramba isina kuchinjika.Jekiseni mukoramu yechromatographic uye tora (yakajairwa sampuli peak nzvimbo/yemukati mwero wepamusoro sampuli nzvimbo) sekukosha kwekupindura.Zvinoenderana nehukama hwemutsara pakati pekukosha kwemhinduro uye kusungirirwa kwechiyero chekushanda, chinoti W= f×A, curve yakajairwa inogadzirwa.

Nhamba inozivikanwa yechiyero chemukati inowedzerwa kune isingazivikanwe sampuli uye inopinzwa muchikamu kuti uwane kukosha kwekupindura kwechikamu chinoyerwa.Maererano neiyo inozivikanwa coefficient f , kusungirirwa kwechikamu chinofanirwa kuyerwa chinogona kuwanikwa.

Hunhu hwemukati mwero nzira:Panguva yekushanda, sampu uye chiyero chemukati zvinosanganiswa pamwe chete uye zvinoiswa mukromatographic column, chero bedzi chiyero chehuwandu hwechikamu chakayerwa kusvika kuchiyero chemukati mune yakasanganiswa mhinduro inoramba iripo, shanduko yevhoriyamu yemuenzaniso. hazvizokanganisa huwandu hwemhedzisiro..Iyo yemukati chiyero nzira inobvisa pesvedzero yevhoriyamu yemuenzaniso, uye kunyange nharembozha uye detector, saka yakanyatsojeka kupfuura yekunze chiyero nzira.

CHIKAMU
08 Zvinhu Zvinokanganisa Quantitative Analysis Results

Kusakwana kwechokwadi kunogona kukonzerwa ne:

Kubatanidzwa kwakashata kwepamusoro penzvimbo, kuparara kwemuenzaniso kana kusvibiswa kwakaunzwa panguva yekugadzirira sampuli, sampuli vial isina kuvharwa, sampuli kana solvent volatilization, gadziriro isiriyo yemuenzaniso, matambudziko ejekiseni, kusarongeka kwemukati gadziriro.

Zvikonzero zvinogoneka zvekushomeka kwakashata:

Kusanganisa peak kusanganisa, jekiseni kana jekiseni matambudziko, kuparara kwemuenzaniso kana tsvina yakaunzwa panguva yekugadzirira sampuli, matambudziko echromatographic, yakaderedzwa detector mhinduro.