URui Huang, uBo Xu
Isicelo se-R&D Centre
Intshayelelo
I-Ion exchange chromatography (IEC) yindlela yechromatographic edla ngokusetyenziswa ukwahlula kunye nokucoca iikhompawundi ezivezwa ngendlela ye-ionic kwisisombululo.Ngokweemo ezahlukeneyo zentlawulo yee-ion ezitshintshisanayo, i-IEC inokohlulwa ibe ziindidi ezimbini, ichromatography yotshintshiselwano lwe-cation kunye nechromatography yotshintshiselwano lweanion.Kwikhromatografi yotshintshiselwano ngecation, amaqela aneasidi adityaniswe kumphezulu weendaba zokwahlula.Ngokomzekelo, i-sulfonic acid (-SO3H) liqela elisetyenziswa ngokuqhelekileyo kwi-cation exchange exchange (SCX), edibanisa i-H + kunye neqela elibi kakhulu -SO3-inokuthi ibhengeze ezinye ii-cations kwisisombululo.Kwi-anion exchange chromatography, amaqela e-alkaline aboshwe kumphezulu weendaba zokwahlula.Ngokomzekelo, i-amine ye-quaternary (-NR3OH, apho i-R i-hydrocarbon iqela) idla ngokusetyenziswa kwi-anion exchange eqinile (i-SAX), edibanisa i-OH- kunye neqela elihlawulwe kakuhle -N + R3 lingabhengeza ezinye ii-anion kwisisombululo, okubangelwa i-anion. utshintsho isiphumo.
Phakathi kweemveliso zendalo, i-flavonoids iye yatsala ingqalelo yabaphandi ngenxa yendima yabo ekuthinteleni nasekunyangelweni kwezifo zentliziyo.Ekubeni iimolekyuli ze-flavonoid zine-acidic ngenxa yobukho bamaqela e-phenolic hydroxyl, i-ion exchange chromatography lukhetho olulolunye ukongeza kwinqanaba eliqhelekileyo eliqhelekileyo okanye i-chromatography yesigaba esibuyiselweyo sokwahlula kunye nokucocwa kwezi khompawundi ze-acidic.Kwi-flash chromatography, imidiya yokwahlula esetyenziswa ngokuqhelekileyo kutshintshiselwano lwe-ion yi-silica gel matrix apho amaqela otshintshiselwano ion adityaniswa kumphezulu wayo.Iindlela zokutshintshiselana nge-ion ezisetyenziswa kakhulu kwi-flash chromatography yi-SCX (ngokuqhelekileyo iqela le-sulfonic acid) kunye ne-SAX (ngokuqhelekileyo iqela le-amine ye-quaternary).Kwinqaku lesicelo elipapashwe ngaphambili kunye nesihloko esithi "Ukusetyenziswa kwe-SepaFlash Strong Cation Exchange Columns Chromatography in the Purification of Alkaline Compounds" yiSanta Technologies, iikholomu ze-SCX zisetyenziselwe ukuhlanjululwa kwe-alkaline compounds.Kule post, umxube wemigangatho engathathi hlangothi kunye ne-acidic isetyenziswe njengesampuli ukuhlola ukusetyenziswa kweekholomu ze-SAX ekuhlanjululeni i-acidic compounds.
iCandelo loMfuniselo
Umzobo 1. Umzobo wesicwangciso sesigaba esimileyo esidityaniswe kumphezulu weendaba zokwahlula i-SAX.
Kule post, ikholamu ye-SAX efakwe ngaphambili kunye ne-quaternary amine bonded silica isetyenziswe (njengoko kuboniswe kuMfanekiso 1).Umxube weChromone kunye ne-2,4-dihydroxybenzoic acid isetyenziswe njengesampuli ukuba ihlanjululwe (njengoko kuboniswe kuMfanekiso 2).Umxube wachithwa kwi-methanol kwaye walayishwa kwi-flash cartridge nge-injector.Ukusetwa kovavanyo lokucocwa kweflash kudweliswe kwiThebhile yoku-1.
Umzobo 2. Ukwakhiwa kweekhemikhali zamacandelo amabini kumxube wesampuli.
| Isixhobo | Umatshini weSepaBean™ uT | |||||
| Iikhatriji | 4 g I-SepaFlash Standard Series flash cartridge (silica engaqhelekanga, 40 - 63 μm, 60 Å, inombolo ye-oda: S-5101-0004) | 4 g SepaFlash Bonded Series SAX flash cartridge (silica engaqhelekanga, 40 - 63 μm, 60 Å, Inombolo yomyalelo: SW-5001-004-IR) | ||||
| Ubude bamaza | 254 nm (ukubona), 280 nm (ukubeka iliso) | |||||
| Isigaba esiphathwayo | Isinyibilikisi A: N-hexane | |||||
| I-Solvent B: I-Ethyl acetate | ||||||
| Umgangatho wohambo | 30 mL/min | 20 mL/min | ||||
| Ukulayisha isampulu | 20 mg (umxube weCandelo A kunye neCandelo B) | |||||
| Umgangatho | Ixesha (CV) | Isinyibilikisi B (%) | Ixesha (CV) | Isinyibilikisi B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Iziphumo neengxoxo
Okokuqala, umxube wesampulu wahlulwe yi-cartridge yesigaba esiqhelekileyo esifakwe ngaphambili kunye ne-silica eqhelekileyo.Njengoko kubonisiwe kuMfanekiso 3, amacandelo amabini kwisampulu akhutshwe kwikhatriji enye emva kwenye.Emva koko, i-cartridge ye-SAX flash yasetyenziselwa ukucocwa kwesampuli.Njengoko kuboniswe kuMfanekiso we-4, iCandelo le-acidic B ligcinwe ngokupheleleyo kwi-cartridge ye-SAX.Inxalenye A engathathi hlangothi yakhutshwa kancinci kancinci kwikhatriji kunye nolution lwesigaba esihambayo.
Umzobo 3. Ikhromatogram yesampulu kwikhatriji yesigaba esiqhelekileyo.
Umzobo 4. I-chromatogram ye-flash yesampuli kwi-cartridge ye-SAX.
Ukuthelekisa i-Figure 3 kunye ne-4, i-Component A ine-peak shape engahambelaniyo kwiikhatriji ezimbini ezahlukeneyo.Ukuqinisekisa ukuba ingaba incopho ye-elution iyahambelana na necandelwana, sinokusebenzisa uphawu olupheleleyo lwe-wavelength scanning eyakhelwe kwisoftware yolawulo yomatshini weSepaBean™.Vula idatha yovavanyo yezahlulo ezimbini, tsala kumgca wesalathisi kwi-axis yexesha (CV) kwichromatogram ukuya kwindawo ephezulu kunye neyona ndawo iphakamileyo yesibini yencopho ye-lution ehambelana neCandelo A, kunye ne-spectrum yobude obupheleleyo kwezi zimbini. amanqaku aya kuboniswa ngokuzenzekelayo ngaphantsi kwechromatogram (njengoko kuboniswe kuMzobo 5 kunye noMzobo 6).Ukuthelekisa i-wavelength spectrum data epheleleyo yezi zahlulo zimbini, i-Component A ine-spectrum yokufunxa engaguqukiyo kwimifuniselo emibini.Ngenxa yesizathu seCandelo A linemilo ephezulu engahambelaniyo kwiikhatriji ezimbini ezahlukeneyo ze-flash, kucingelwa ukuba kukho ukungcola okuthe ngqo kwiCandelo A elinokugcinwa okwahlukileyo kwi-cartridge yesigaba esiqhelekileyo kunye ne-SAX cartridge.Ngoko ke, ulandelelwano eluting lwahlukile kwiCandelo A kunye nokungahlambuluki kwezi khatriji ezimbini zombane, kubangele imilo yencopho engahambelaniyo kwiichromatogram.
Umzobo 5. I-spectrum yobude obupheleleyo beCandelo A kunye nokungcola okwahlulwe ngekhatriji yesigaba sesiqhelo.
Umzobo 6. Umzobo opheleleyo we-wavelength yeCandelo A kunye nokungcola okuhlukaniswe yi-cartridge ye-SAX.
Ukuba imveliso ekujoliswe kuyo ukuba iqokelelwe yiNgxelo ye-A engathathi hlangothi, umsebenzi wokucoca unokugqitywa ngokulula ngokusebenzisa i-cartridge ye-SAX ye-lution emva kokulayishwa kwesampuli.Ngakolunye uhlangothi, ukuba imveliso ekujoliswe kuyo ukuba iqokelelwe i-Acid Component B, indlela yokukhupha i-capture-release ingamkelwa kuphela ngohlengahlengiso oluncinci kumanyathelo okulinga: xa isampuli ilayishwe kwi-cartridge ye-SAX kunye neCandelo elingathathi hlangothi A. yakhutshwa ngokupheleleyo kunye nesigaba esiqhelekileyo sokunyibilikisa kwe-organic, tshintshela isigaba esihambayo kwisisombululo se-methanol esine-5% ye-acetic acid.Iiyoni zeacetate kwisigaba esihambayo ziya kukhuphisana neCandelo B lokubophelela kumaqela e-amine ion e-quaternary kwinqanaba elimileyo lekhatriji ye-SAX, ngaloo ndlela ikhupha iCandelo B ukusuka kwikhatriji ukufumana imveliso ekujoliswe kuyo.I-chromatogram yesampulu ehlukaniswe kwimodi yokutshintshiselana kwe-ion iboniswe kwi-Figure 7.
Umzobo 7. Ikhromatogram ekhanyayo yeCandelo B icaciswe kwimowudi yokutshintshiselana nge-ion kwikhatriji ye-SAX.
Ukuqukumbela, isampulu eneasidi okanye engathathi hlangothi inokucocwa ngokukhawuleza ngekhatriji ye-SAX idityaniswe nekhatriji yesigaba esiqhelekileyo kusetyenziswa iindlela ezahlukeneyo zokucoca.Ngaphaya koko, ngoncedo lwesici esipheleleyo sokuskena se-wavelength eyakhelwe kwisoftware yolawulo yomatshini we-SepaBean™, uphawu lokufunxa uphawu lwamaqhezu akhutshiweyo lunothelekiseka lula kwaye luqinisekiswe, ukunceda abaphandi baqonde ngokukhawuleza ukubunjwa kunye nokusulungeka kwamaqhezu akhutshiweyo kwaye ngaloo ndlela kuphuculwe. ukusebenza kakuhle.
| Inombolo yento | Ubungakanani beKholam | Umgangatho wohambo (mL/min) | Uxinzelelo olukhulu (psi/bar) |
| SW-5001-004-IR | 5.9 g | 10-20 | 400/27.5 |
| SW-5001-012-IR | 23 g | 15-30 | 400/27.5 |
| SW-5001-025-IR | 38 g | 15-30 | 400/27.5 |
| SW-5001-040-IR | 55 g | 20-40 | 400/27.5 |
| SW-5001-080-IR | 122 g | 30-60 | 350/24.0 |
| SW-5001-120-IR | 180 g | 40-80 | 300/20.7 |
| SW-5001-220-IR | 340 g | 50-100 | 300/20.7 |
| SW-5001-330-IR | 475 g | 50-100 | 250/17.2
|
Itheyibhile 2. SepaFlash Bonded Series SAX iikhatriji flash.Izinto zokupakisha: I-silica ye-Ultra-pure engaqhelekanga ye-SAX, i-40 - 63 μm, 60 Å.
Ngolwazi oluthe vetshe ngeenkcukacha zeSepaBean™umatshini, okanye ulwazi lokuodola SepaFlash series flash cartridges, nceda undwendwele website yethu.
Ixesha lokuposa: Nov-09-2018