Hongcheng wang, bo xu
Porogaramu R & D Centre
Intangiriro
Dukurikije imiyoboro ijyanye na provie na phrate ya mobile, chromatografiya ishobora kugabanywamo icyiciro gisanzwe cya chromatografiya (NPC) na PRASE CHROMATOGRAPHY (RPC). Kuri RPC, umuyoboro wicyiciro cya mobile urakomeye kuruta uw'icyiciro gihagaze. RPC yakoreshejwe cyane muburyo bwa chromatografiya yo gutandukanya imiterere kubera imikorere yayo yose, imyanzuro myiza no kugenzura uburyo bwo kugenzura. Kubwibyo GPC ibereye gutandukana no kwezwa amakipe atandukanye cyangwa adapamba, harimo na karubone, acide, acide, muri poptine, muri poptine, muri PROPOIQUES, C8, C8, Phenyl, Cyano, Adino, nibindi muriyi matsinda yimikorere, yakoreshejwe cyane ni C18. Bigereranijwe ko 80% bya RPC ari ubu bakoresheje icyiciro cya C18. Niyo mpamvu c18 chromatography inkingi yabaye igikwiye-kugira inkingi yose kuri buri laboratoire.
Nubwo C18 inkingi ishobora gukoreshwa muburyo butandukanye cyane kubisabwa, ariko, kubishusho bimwe na bimwe bya polar cyangwa hydrophilic nyinshi cyangwa hydrophilic, inkingi zisanzwe za c18 zishobora kugira ibibazo mugihe zikoreshwa mugusukura ingero nkizo. Muri RPC, bikunze gukoreshwa hejuru yimyenda irashobora gutegekwa ukurikije polarity yawe: amazi <methanol <acetonitrile <acetonitrile <ethanol <tetrahydrofuran <isopropanol. Kwemeza kugumana neza ku nkingi (polar ikomeye cyangwa hydrophilic)), igipimo kinini cya sisitemu irakenewe kugirango ikoreshwe nkicyiciro cya mobile. Ariko, mugihe ukoresheje sisitemu y'amazi meza (harimo n'amazi meza cyangwa igisubizo cyiza cyumunyu) nkigikoresho kirekire cya karubone, bikavanga mu cyiciro cyagenwe cya C18 gihari, kikaba cyagabanijwe. Iki kintu cyitwa "Icyiciro cya Hydrophobike" (nkuko bigaragara mugice cyibumoso cyishusho 1). Nubwo iki kibazo gihinduka mugihe inkingi yogejwe hamwe na methanol cyangwa acetitol, iracyashobora kwangiza inkingi. Kubwibyo, birakenewe kubuza iki kibazo kubaho.
Igishushanyo 1. Igishushanyo cya Schematic cyicyiciro cyahujwe hejuru ya Silica Gel muri C18: Ibumoso) na C18AQ inkingi (iburyo).
Kugirango ukemure ibibazo byavuzwe haruguru, ibikoresho byo gupakira chromatografiabakora bakoze iterambere rya tekiniki. Kimwe muri ibyo byiyongereyeho ni bimwe bihindura hejuru ya silica matrix, nko gutangiza amatsinda ya hydrophilic ya cyano (nkuko bigaragara mugice cyiza cyishusho 1), kugirango ugaragaze igice cyishusho 1), kugirango ugaragaze gel ya silika. Gutyo, iminyururu ya C18 ku buso bwa silica bushobora kwagurwa burundu mu bihe by'amazi menshi kandi icyiciro cya hydrophobike gishobora kwirindwa. Iyi nkingi zahinduwe zitwa C18 zitwa C18, aribyo inkingi za C18AQ, zagenewe kwishyurwa cyane kandi zirashobora kwihanganira sisitemu 100%. Inkingi za C18AQ zakoreshejwe cyane mugutandukana no kweza ibigo bikomeye, birimo aside organide, buringaniye, nocleoside hamwe na vitamine zifatika.
Kugereranya ni kimwe mubisabwa bisanzwe byinkingi ya C18AQ muri flash yo kweza ingero, zikuraho uruziga ruke murwego rworoshya kugirango borohereze icyitegererezo mugihe cyakurikiyeho. Muri iyi nyandiko, fcf nziza yubururu ifite polarity yakoreshejwe nkicyitegererezo kandi isukurwa kuri c18AQ. Icyitegererezo Cyicyitegererezo cyasimbuwe na kama mubisubizo bya buffer, bityo uroborora guhumeka ibinyuramo kimwe no kuzigama intege nke nigihe cyo gukora. Byongeye kandi, ubuziranenge bwicyitegererezo bwaratejwe imbere mugukuraho umwanda murugero.
Icyiciro cyageragejwe
Igishushanyo cya 2. Imiterere yimiti yicyitegererezo.
Ubururu bwa FCF bwakoreshejwe nkicyitegererezo muriyi nyandiko. Isuku yicyitegererezo mbisi yari 86% hamwe nuburyo bwa shimi ya sample bwerekanwe ku gishushanyo cya 2. Gutegura igisubizo cyicyitegererezo cya FCF cyashonze muri 1 M NAH2PO4 Icyitegererezo cyicyitegererezo cyatewe mu nkingi ya flash by injino. Gukora igerageza rya flash ryerekanwe ku mbonerahamwe ya 1.
| Igikoresho | Sepabean ™ imashini2 | |||
| Amakarito | 12 G DissFlash C18 Rp Flash Cartridge (SPERICACT SILICA, 20 - 45 μM, 100 å, Inomero ya 100 å-5222-012-SP) | 12 G DissFlash C18AQ Rp Flash Cartridge (SPERICACH SILICA, 20 - 45 μM, 100 å, SW-5222-012-SP (AQ)) | ||
| Uburebure | 254 nm | |||
| Icyiciro cya mobile | Gukemura a: amazi Yamazaki B: methanol | |||
| Igipimo | 30 ml / min | |||
| Icyitegererezo | 300 mg (fcf nziza yubururu hamwe nisuku ya 86%) | |||
| Gradient | Igihe (CV) | Yamazaki B (%) | Igihe (CV) | Yamazaki B (%) |
| 0 | 10 | 0 | 0 | |
| 10 | 10 | 10 | 0 | |
| 10.1 | 100 | 10.1 | 100 | |
| 17.5 | 100 | 17.5 | 100 | |
| 17.6 | 10 | 17.6 | 0 | |
| 22.6 | 10 | 22.6 | 0 | |
Ibisubizo n'ibiganiro
Gutandukanya C18AQ rp flash cartridge yakoreshejwe kugirango icyitegererezo gitere no kwezwa. Intambwe Nziza yakoreshejwe mu mazi meza yakoreshejwe nk'icyiciro cya mobile mu ntangiriro yo kwishura no kwiruka ku mubumbe 10 (CV). Nkuko bigaragara ku gishushanyo cya 3, mugihe ukoresheje amazi meza nkicyiciro cya mobile, icyitegererezo cyagumishijwe rwose kuri karitsiye ya flash. Ubukurikira, methanol muri icyiciro cya mobile yariyongereye ku 100% ndetse na Gradient yakomezaga kuri 7.5 CV. Icyitegererezo cyasohotse kuva 11.5 kugeza 13.5 cv. Mubice byakusanyirijwe hamwe, igisubizo cyicyitegererezo cyasimbuwe na nah2po 4 Igisubizo cya Buffer kuri methanol. Kugereranya igisubizo kiratangaje cyane, methanol yari yoroshye cyane gukurwaho na rotary guhumeka mu ntambwe ikurikira, yorohereza ubushakashatsi bukurikira.
Igishushanyo cya 3. Flash chromatogram yumutego kuri cartridge ya C18AQ.
Kugereranya imyitwarire yo kugumana C18AQ Cartridge na Carridge isanzwe ya C18 kuri portularity, ikizamini kibangikanye cyakozwe. DiSTFLASH C18 RP Flash Cartridge yakoreshejwe na flaromatogram kuri sample yerekanwe ku gishushanyo cya 4. Kuri Carridges isanzwe ya C18, igipimo kinini cyicyiciro cya 90%. Kubwibyo rero Gradient yashyizwe kuri Methanol 10% mumazi 90%. Nkuko bigaragara ku gishushanyo cya 4, kubera icyiciro cya hydrophobike gisenyuka ku munyururu wa C18 cyatewe n'ikigereranyo kinini cya Cartrid kandi cyavanywe mu cyiciro cya mobile. Nkigisubizo, imikorere yicyitegererezo cyangwa isuku ntishobora kurangira.
Igishushanyo cya 4. Flash chromatogram yumutego kuri cartridge isanzwe ya c18.
Kugereranya na lishar, ikoreshwa ryintambwe Gradient ifite ibyiza bikurikira:
1. Umuyoboro wo gukoresha no gukora igihe kugirango usukurwe neza.
2. Ibicuruzwa bigamije ellites igororotse, bigabanya ingano ya coction yakusanyirijwe bityo byorohereze umwuka ukurikira ndetse nigihe cyo kuzigama.
3. Ibicuruzwa byakusanyirijwe biri muri methanol biroroshye guhumeka, bityo igihe cyumisha kiragabanuka.
Mu gusoza, kugirango usukure icyitegererezo cya polar cyangwa hydrophilic nini cyangwa hydrophilic, stafflash c18AQ rp flash carridges sisitemu ya flaromaatography spabean ™ imashini irashobora gutanga igisubizo cyihuse kandi cyiza.
Kubijyanye na restaflash kumurongo c18 rp flash cartridges
Hano hari urukurikirane rwa Startflash C18AQ rp flash cartridges hamwe nibisobanuro bitandukanye kuva tekinoroji ya santai (nkuko bigaragara mu mbonerahamwe ya 2).
| Umubare w'ikintu | Ingano y'inkingi | Igipimo (ML / Min) | Max.kwishimira (PSI / BAR) |
| SW-5222-004-SP (AQ) | 5.4 g | 5-15 | 400/1 27.5 |
| SW-5222-012-SP (AQ) | 20 g | 10-25 | 400/1 27.5 |
| SW-5222-025-SP (AQ) | 33 g | 10-25 | 400/1 27.5 |
| Sw-5222-040-SP (AQ) | 48 g | 15-30 | 400/1 27.5 |
| Sw-5222-080-sp (aq) | 105 g | 25-50 | 350 / 24.0 |
| Sw-5222-120-SP (AQ) | 155 g | 30-60 | 300 / 20.7 |
| SW-5222-220-SP (AQ) | 300 g | 40-80 | 300 / 20.7 |
| SW-5222-330-SP (AQ) | 420 G. | 40-80 | 250 / 17.2 |
Imbonerahamwe ya 2. Gutandukanya C18AQ RP FL Flash Cartridges.
Ibikoresho byo gupakira: Gukora neza-Imikorere ya Spherical C18 (AQ) -Yambere Silica, 20 - 45 μm, 100 å.
logy (nkuko bigaragara mu mbonerahamwe ya 2).
Igihe cya nyuma: Aug-27-2018