Ka'idoji da Hanyoyi na Ƙididdigar ƙididdigewa ta Liquid Chromatography

Tsarin rabuwa na chromatography na ruwa ya dogara ne akan bambanci a cikin kusancin abubuwan da ke cikin cakude don matakan biyu.

Dangane da matakai daban-daban na tsaye, chromatography na ruwa ya kasu kashi-kashi mai ƙarfi-chromatography, chromatography na ruwa-ruwa da chromatography lokaci.Mafi yawan amfani da su shine ruwa-m chromatography tare da silica gel a matsayin filler da bonded lokaci chromatography tare da microsilica a matsayin matrix.

Dangane da nau'i na lokaci mai tsayi, za a iya raba chromatography na ruwa zuwa chromatography na shafi, chromatography na takarda da chromatography na bakin ciki.Dangane da ƙarfin adsorption, ana iya raba shi zuwa chromatography adsorption, chromatography bangare, chromatography musayar ion da gel permeation chromatography.

A cikin 'yan shekarun nan, an ƙara tsarin ruwa mai matsananciyar matsa lamba zuwa tsarin chromatography na ruwa na ruwa don yin tafiyar lokaci ta hannu da sauri a ƙarƙashin babban matsin lamba don inganta tasirin rabuwa, don haka babban inganci (wanda aka sani da babban matsin lamba) ruwa chromatography. ya fito.

KASHI
01 Ƙa'idar Ƙididdigar Ƙididdiga na Liquid Chromatography

Don ƙididdigewa bisa tushen inganci, ana buƙatar abubuwa masu tsabta a matsayin ma'auni;

Ƙididdigar chromatography na ruwa hanya ce ta ƙididdigewa: wato, ana ƙididdige adadin adadin abin da ke cikin cakuduwar daga sanannen adadin daidaitaccen samfurin.

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02 Tushen don ƙididdigewa ta Liquid Chromatography

Adadin abin da aka auna (W) yayi daidai da ƙimar amsawa (A) (tsawon tsayi ko yanki mafi girma), W=f × A.

Ma'anar gyara ƙididdiga (f): Shi ne madaidaicin madaidaicin ma'aunin ƙididdiga, kuma ma'anarsa ta zahiri ita ce adadin abin da aka auna wanda ke wakilta ta ƙimar amsa naúrar (yankin kololuwa).

Za'a iya samun ma'aunin gyare-gyare na ƙididdigewa daga adadin da aka sani na daidaitaccen samfurin da ƙimar amsawa.

Auna ƙimar amsawar ɓangaren da ba a san shi ba, kuma ana iya samun adadin ɓangaren ta hanyar ma'aunin gyaran ƙima.

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03 Kalmomin gama gari a cikin ƙididdigewa

Samfurin (samfurin): bayani mai kunshe da nazari don nazarin chromatographic.Rarraba cikin daidaitattun samfuran da ba a sani ba.

Ma'auni: Samfuri mai tsafta tare da sanannen taro.Samfuran da ba a sani ba (wanda ba a sani ba): Cakuda wanda za a gwada ƙarfinsa.

Nauyin samfurin: Asalin awo na samfurin da za a gwada.

Dilution: Abubuwan dilution na samfurin da ba a sani ba.

Bangaren : kololuwar chromatographic da za a yi nazari akan ƙididdigewa, wato, mai nazari wanda ba a san abin da ke ciki ba.

Adadin bangaren (yawan): abun ciki (ko maida hankali) abubuwan da za a gwada.

Integerity : Tsarin lissafi na auna kololuwar yanki na kololuwar chromatographic ta kwamfuta.

Hanyar daidaitawa: Madaidaicin layi na abun ciki tare da ƙimar amsa, an kafa shi daga sanannen adadin daidaitaccen abu, ana amfani da shi don tantance abin da ba a sani ba na nazari.

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04 Ƙididdigar ƙididdiga na Chromatography na Liquid

1. Zaɓi hanyar chromatographic dacewa don ƙididdige ƙididdiga:

l Tabbatar da kololuwar abin da aka gano kuma cimma ƙuduri (R) fiye da 1.5

l Ƙayyade daidaito (tsarki) na kololuwar chromatographic na abubuwan da aka gwada

l Ƙayyade iyakar ganowa da ƙididdige ƙimar hanyar;hankali da layin layi

2. Ƙaddamar da ma'auni tare da daidaitattun samfurori na ƙididdiga daban-daban

3. Bincika daidaito da daidaitattun hanyoyin ƙididdigewa

4. Yi amfani da software na sarrafa chromatography daidai don aiwatar da tarin samfurin, sarrafa bayanai da rahoton sakamakon

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05 Gano kololuwar ƙididdigewa (mai inganci)

Ƙidaya tantance kowane kololuwar chromatographic don ƙididdige su

Na farko, yi amfani da daidaitaccen samfurin don ƙayyade lokacin riƙewa (Rt) na kololuwar ƙididdiga don ƙididdige su.Ta hanyar kwatanta lokacin riƙewa, nemo ɓangaren da ya dace da kowane kololuwar ƙira a cikin samfurin da ba a sani ba.Hanyar ƙwararrun chromatographic ita ce kwatanta lokacin riƙewa tare da daidaitaccen samfurin.Ma'aunin rashin isaƙarin tabbaci (mai inganci)

1. Daidaitaccen hanyar ƙari

2. Yi amfani da wasu hanyoyi a lokaci guda: wasu hanyoyin chromatographic (canza tsarin, kamar: amfani da ginshiƙan chromatographic daban-daban), sauran masu ganowa (PDA: kwatanta nau'i, binciken ɗakin karatu na bakan; MS: bincike mai yawa, bincike na ɗakin karatu).

3. Sauran kayan aiki da hanyoyin

KASHI
06 Tabbatar da daidaiton Ƙwararrun Ƙwararru

Tabbatar da daidaito kololuwar chromatographic (tsarki)

Tabbatar cewa akwai abu guda ɗaya da aka auna a ƙarƙashin kowane kololuwar chromatographic

Bincika tsangwama daga abubuwa masu haɗaka (ƙazanta)

Hanyoyi don Tabbatar da Ƙwararrun Ƙwararrun Chromatographic (Tsarki)

Kwatanta Spectrograms tare da Masu gano Matrix Photodiode (PDA).

Kololuwar Tsabtace Identification

2996 Ka'idar Angle Tsarkake

Hanyoyin ƙididdigewa da aka saba amfani da su a PART 07

Madaidaicin hanyar lanƙwasa, an raba shi zuwa daidaitaccen hanyar waje da daidaitaccen hanyar ciki:

1. Hanyar daidaitaccen waje: mafi yawan amfani da su a cikin chromatography na ruwa

An shirya jerin samfuran ƙididdiga na ƙididdiga da aka sani ta amfani da samfurori masu tsabta na mahaɗan da za a gwada su azaman samfurori na yau da kullum.allura a cikin ginshiƙi har zuwa ƙimar amsawarsa (yanki mafi girma).
A cikin wani takamaiman kewayon, akwai kyakkyawar alaƙar layi tsakanin ƙaddamar da daidaitaccen samfurin da ƙimar amsawa, wato W = f × A , kuma an yi daidaitaccen lanƙwasa.

A ƙarƙashin ainihin yanayin gwaji iri ɗaya, allura samfurin da ba a san shi ba don samun ƙimar amsawar ɓangaren da za a auna.Bisa ga sanannun ƙididdiga f , za a iya samun maida hankali na ɓangaren da za a auna.

Fa'idodin daidaitattun hanyar waje:aiki mai sauƙi da lissafi, hanya ce ta ƙididdige yawan amfani da ita;babu buƙatar kowane sashi da za a gano kuma a ɓoye;ana buƙatar misali samfurin;yanayin ma'auni na daidaitaccen samfurin da samfurin da ba a sani ba ya kamata ya kasance daidai;girman allurar yakamata ya zama daidai.

Rashin lahani na daidaitattun hanyar waje:Ana buƙatar yanayin gwaji don zama babba, irin su ƙwarewar mai ganowa, ƙimar gudana, da abun da ke cikin tsarin wayar hannu ba za a iya canza shi ba;Ya kamata ƙarar kowace allura ta kasance mai maimaitawa mai kyau.

2. Hanyar daidaitaccen ciki: daidai, amma matsala, mafi yawan amfani da su a daidaitattun hanyoyin

An ƙara adadin da aka sani na ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ma'auni, kuma an shirya jerin ma'auni na aiki na sananniya.Matsakaicin ma'auni na molar zuwa daidaitattun ciki a cikin ma'aunin gauraye ya kasance baya canzawa.Yi allura a cikin ginshiƙi na chromatographic kuma ɗauka (misali mafi girman yanki/daidaitaccen yanki mafi girman samfurin ciki) azaman ƙimar amsawa.Dangane da alaƙar layi tsakanin ƙimar amsawa da ƙaddamar da daidaitaccen aiki, wato W = f × A , an yi daidaitaccen lanƙwasa.

Ana ƙara sanannen adadin daidaitattun ciki zuwa samfurin da ba a san shi ba kuma an allura shi cikin ginshiƙi don samun ƙimar amsawar ɓangaren da za a auna.Bisa ga sanannun ƙididdiga f , za a iya samun maida hankali na ɓangaren da za a auna.

Halayen daidaitattun hanyar ciki:A lokacin aikin, samfurin da ma'auni na ciki suna haɗuwa tare da allura a cikin ginshiƙi na chromatographic, don haka idan dai adadin adadin adadin da aka auna zuwa ma'auni na ciki a cikin cakudaccen bayani ya kasance akai-akai, canjin samfurin samfurin. ba zai shafi sakamakon ƙididdiga ba..Hanyar daidaitattun ciki tana daidaita tasirin tasirin samfurin , har ma da lokacin wayar hannu da mai ganowa, don haka ya fi daidai da daidaitattun hanyar waje.

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Abubuwa 08 Da Suke Taimakawa Sakamakon Ƙididdigar Ƙididdigar

Rashin daidaito na iya haifar da:

Haɗin yanki mara daidai, bazuwar samfurin ko ƙazanta da aka gabatar yayin shirye-shiryen samfurin, samfurin vial ba a rufe ba, samfuri ko ƙarancin ƙarfi, shirye-shiryen samfurin da ba daidai ba, matsalolin alluran samfur, shirye-shiryen daidaitattun ciki na ciki ba daidai ba.

Dalilai masu yiwuwa na rashin daidaituwa:

Haɗin kololuwa mara kyau, matsalolin allura ko injector, bazuwar samfurin ko ƙazanta da aka gabatar yayin shirye-shiryen samfurin, matsalolin chromatographic, amsawar mai ganowa mara kyau.