Mfundo ndi Njira Zowunika Kuchuluka kwa Liquid Chromatography

Njira yolekanitsa ya chromatography yamadzimadzi imatengera kusiyana kwa kuyanjana kwa zigawo zomwe zili mu osakaniza a magawo awiriwa.

Malinga ndi magawo osiyanasiyana oyima, chromatography yamadzimadzi imagawidwa mu liquid-solid chromatography, liquid-liquid chromatography ndi bonded phase chromatography.Zomwe zimagwiritsidwa ntchito kwambiri ndi chromatography yamadzi-solid yokhala ndi silika gel ngati filler ndi bonded phase chromatography yokhala ndi microsilica ngati matrix.

Malinga ndi mawonekedwe a gawo loyima, chromatography yamadzi imatha kugawidwa mu column chromatography, paper chromatography ndi thin layer chromatography.Malinga ndi mphamvu yotsatsa, imatha kugawidwa mu adsorption chromatography, partition chromatography, ion exchange chromatography ndi gel permeation chromatography.

M'zaka zaposachedwa, makina othamanga kwambiri amadzimadzi adawonjezedwa pamakina amadzimadzi amtundu wa chromatography kuti gawo loyenda liziyenda mwachangu pansi pamavuto akulu kuti apititse patsogolo kulekanitsa, kuchita bwino kwambiri (kotchedwanso high-pressure) liquid chromatography. chatulukira.

GAWO
01 Mfundo Yowunikira Kuchuluka kwa Liquid Chromatography

Kuwerengera pamaziko a zinthu zabwino, zoyera zimafunikira ngati miyezo;

Liquid chromatography quantification ndi njira yochulukirachulukira: ndiye kuti, kuchuluka kwa analyte mu osakaniza akuyerekezedwa kuchokera ku kuchuluka kodziwika kwa zitsanzo zoyera.

GAWO
02 Maziko a Quantification ndi Liquid Chromatography

Kuchuluka kwa chigawo choyezedwa (W) n'chofanana ndi mtengo woyankhira (A) (utali wamtali kapena malo okwera), W=f×A.

Quantitative correction factor (f): Ndiko kufanana kosalekeza kwa chiŵerengero cha kachulukidwe, ndipo tanthauzo lake lenileni ndi kuchuluka kwa chigawo choyezedwa choimiridwa ndi mtengo wakuyankhidwa kwa unit (malo apamwamba).

Kuwongolera kwachulukidwe kachulukidwe kumatha kupezeka kuchokera ku kuchuluka kodziwika kwa zitsanzo zokhazikika komanso kuyankha kwake.

Yezerani mtengo woyankhira wa gawo losadziwika, ndipo kuchuluka kwa gawoli kungapezeke ndi kuchuluka kwa kuwongolera.

GAWO
03 Mawu wamba pakuwunika kuchuluka

Zitsanzo (zitsanzo): yankho lomwe lili ndi analyte pakuwunika kwa chromatographic.Amagawidwa kukhala zitsanzo zodziwika bwino komanso zosadziwika.

Standard: Chopangidwa choyera chokhala ndi ndende yodziwika bwino.Zitsanzo zosadziwika (zosadziwika): Kusakaniza komwe kuyenera kuyesedwa.

Kulemera kwachitsanzo: Kuyeza koyambirira kwa chitsanzo kuti chiyesedwe.

Dilution: The dilution factor of the sample osadziwika.

Chigawo : nsonga ya chromatographic kuti iwunikidwe mochulukira, ndiye kuti, wowunika yemwe zomwe zili zake sizikudziwika.

Kuchuluka kwa gawo (kuchuluka): zomwe zili (kapena ndende) zazinthu zomwe ziyenera kuyesedwa.

Kukhulupirika : Njira yowerengera yoyezera nsonga ya nsonga ya chromatographic ndi kompyuta.

Mzere wokhotakhota: Mzere wokhotakhota wa zinthu zomwe zili ndi gawo motsutsana ndi mtengo wamayankhidwe, wokhazikitsidwa kuchokera ku kuchuluka kodziwika kwa chinthu chokhazikika, chomwe chimagwiritsidwa ntchito kudziwa zomwe sizikudziwika mu analyte.

GAWO
04 Kuwunika Kwambiri kwa Liquid Chromatography

1. Sankhani njira ya chromatographic yoyenera kusanthula kuchuluka kwake:

l Tsimikizirani kuchuluka kwa gawo lomwe lapezeka ndikukwaniritsa chigamulo (R) choposa 1.5

l Dziwani kusasinthasintha (kuyera) kwa nsonga za chromatographic za zigawo zoyesedwa

l Dziwani malire ozindikira ndi kuchuluka kwa njira;sensitivity ndi liniya osiyanasiyana

2. Khazikitsani mayendedwe opendekera ndi zitsanzo zokhazikika zamagulu osiyanasiyana

3. Onani kulondola ndi kulondola kwa njira zochulukira

4. Gwiritsani ntchito pulogalamu yoyang'anira chromatography kuti mugwiritse ntchito kusonkhanitsa zitsanzo, kukonza deta ndi zotsatira za malipoti

GAWO
05 Kuzindikiritsa nsonga zochulukira (zabwino)

Dziwani bwino nsonga iliyonse ya chromatographic kuti iwerengedwe

Choyamba, gwiritsani ntchito chitsanzo chodziwika kuti mudziwe nthawi yosungira (Rt) ya nsonga ya chromatographic kuti iwerengedwe.Poyerekeza nthawi yosungira, pezani gawo lomwe likugwirizana ndi nsonga ya chromatographic iliyonse pachitsanzo chosadziwika.Njira yopangira chromatographic ndikufanizira nthawi yosungira ndi chitsanzo chokhazikika.Mulingo Wosakwanirachitsimikizo china (chabwino)

1. Njira yowonjezera yowonjezera

2. Gwiritsani ntchito njira zina panthawi imodzimodzi: njira zina za chromatographic (kusintha makina, monga: kugwiritsa ntchito ma chromatographic columns), zodziwira zina (PDA: kufananitsa kwa sipekitiramu, kufufuza kwa laibulale ya sipekitiramu; MS: kusanthula kwa masipekitiramu ambiri, kufufuza mu library ya sipekitiramu)

3. Zida zina ndi njira

GAWO
06 Chitsimikizo Chakuchulukira Pamwamba Kwambiri

Tsimikizirani kusasinthika kwapachromatographic (kuyera)

Onetsetsani kuti pali gawo limodzi lokha loyezedwa pansi pa nsonga iliyonse ya chromatographic

Yang'anani kusokonezedwa ndi co-eluting substances (zonyansa)

Njira Zotsimikizira Kuti Chromatographic Peak Consistency (Kuyera)

Kuyerekeza Ma Spectrograms ndi Photodiode Matrix (PDA) Detectors

Peak Purity Identification

2996 Purity Angle Theory

Njira zochulukira zomwe zimagwiritsidwa ntchito mu GAWO 07

Njira yokhotakhota yokhazikika, yogawidwa munjira yakunja ndi njira yamkati:

1. Njira yakunja yokhazikika: amagwiritsidwa ntchito kwambiri mu chromatography yamadzi

Mndandanda wa zitsanzo zodziwika bwino zodziwika zidakonzedwa pogwiritsa ntchito zitsanzo zoyera zamagulu kuti ayesedwe ngati zitsanzo zokhazikika.jekeseni pamzati mpaka pamtengo wake (malo apamwamba).
Mumtundu wina, pali ubale wabwino wa mzere pakati pa kuchuluka kwa zitsanzo zokhazikika ndi mtengo wamayankhidwe, womwe ndi W= f×A, ndipo piritsi lokhazikika limapangidwa.

Pansi pa zoyeserera zomwezo, bayani zitsanzo zosadziwika kuti mupeze yankho la gawo lomwe liyenera kuyezedwa.Malingana ndi coefficient yodziwika f, chiwerengero cha chigawo chomwe chiyenera kuyezedwa chikhoza kupezeka.

Ubwino wa njira yakunja yokhazikika:ntchito yosavuta komanso yowerengera, ndi njira yomwe imagwiritsidwa ntchito mochulukira;palibe chifukwa choti chigawo chilichonse chidziwike ndikusinthidwa;muyezo muyezo chofunika;miyeso yachitsanzo chodziwika bwino ndi zitsanzo zosadziwika ziyenera kukhala zofanana;kuchuluka kwa jakisoni kuyenera kukhala kolondola.

Kuipa kwa njira yakunja yokhazikika:Mikhalidwe yoyesera imayenera kukhala yapamwamba, monga kukhudzidwa kwa detector, kuthamanga kwa kuthamanga, ndi mapangidwe a gawo la mafoni sangathe kusinthidwa;voliyumu ya jekeseni iliyonse iyenera kukhala yobwerezabwereza bwino.

2. Internal muyezo njira: yolondola, koma yovuta, yomwe imagwiritsidwa ntchito kwambiri munjira zokhazikika

Kuchuluka kodziwika kwa ndondomeko yamkati kumawonjezeredwa ku muyezo kuti apange muyezo wosakanikirana, ndipo mndandanda wa ntchito zogwirira ntchito zodziwika bwino zimakonzedwa.Chiŵerengero cha molar cha muyezo kwa muyezo wamkati muyeso wosakanikirana sichinasinthe.Lowetsani mugawo la chromatographic ndikutenga (gawo lokhazikika lachitsanzo / gawo lachitsanzo lamkati) ngati mtengo woyankhira.Malingana ndi mgwirizano wa mzere pakati pa mtengo woyankhira ndi kuyika kwa mulingo wogwirira ntchito, womwe ndi W= f×A, piritsi lokhazikika limapangidwa.

Kuchuluka kodziwika kwa muyezo wamkati kumawonjezedwa ku zitsanzo zosadziwika ndikulowetsedwa muzanja kuti mupeze yankho la gawo lomwe liyenera kuyezedwa .Malingana ndi coefficient yodziwika f, chiwerengero cha chigawo chomwe chiyenera kuyezedwa chikhoza kupezeka.

Makhalidwe a Internal Standard method:Pa opareshoni, chitsanzo ndi muyezo wamkati zimasakanizidwa pamodzi ndi jekeseni mu chromatographic ndime, bola ngati chiŵerengero cha kuchuluka kwa chigawo choyezera ndi muyezo wamkati mu njira osakaniza ndi mosalekeza, kusintha kwa voliyumu chitsanzo. sichidzakhudza zotsatira za kuchuluka..Njira yokhazikika yamkati imachotsa chikoka cha voliyumu yachitsanzo, komanso gawo la mafoni ndi chowunikira, chifukwa chake ndi cholondola kuposa njira yakunja.

GAWO
08 Zinthu Zomwe Zimakhudza Zotsatira Zakuwunika Kwambiri

Kusalondola bwino kungayambitsidwe ndi:

Kuphatikizika kolakwika kwa nsonga ya nsonga, kuwonongeka kwa zitsanzo kapena zonyansa zomwe zidayambika pokonzekera zitsanzo, vial yachitsanzo yomwe sinasindikizidwe, kuyesa kwachitsanzo kapena kusungunulira kwa zosungunulira, kukonzekera kwachitsanzo molakwika, zovuta za jekeseni, kukonzekera kolakwika kwamkati.

Zifukwa zomwe zingakhale zosalondola bwino:

Kuphatikizika kwapamwamba kolakwika, zovuta za jakisoni kapena jekeseni, kuwonongeka kwa zitsanzo kapena zonyansa zomwe zidayambika pokonzekera zitsanzo, zovuta za chromatographic, kuyankha kolakwika kwa chowunikira.