Rui Huang, Bo Xu
Ntchito R&D Center
Mawu Oyamba
Ion exchange chromatography (IEC) ndi njira ya chromatographic yomwe imagwiritsidwa ntchito polekanitsa ndi kuyeretsa zinthu zomwe zimaperekedwa mu mawonekedwe a ionic mu njira.Malinga ndi maiko osiyanasiyana omwe amatha kusintha ma ion, IEC ikhoza kugawidwa m'mitundu iwiri, cation exchange chromatography ndi anion exchange chromatography.Mu cation exchange chromatography, magulu acidic amamangiriridwa pamwamba pa zolekanitsa media.Mwachitsanzo, sulfonic acid (-SO3H) ndi gulu lomwe limagwiritsidwa ntchito kwambiri muzitsulo zolimba za cation (SCX), zomwe zimasiyanitsa H + ndi gulu losavomerezeka -SO3- motero likhoza kutsatsa ma cations ena mu yankho.Mu anion exchange chromatography, magulu amchere amamangiriridwa pamwamba pa zopatukana.Mwachitsanzo, quaternary amine (-NR3OH, kumene R ndi gulu la hydrocarbon) nthawi zambiri amagwiritsidwa ntchito posinthanitsa ndi anion amphamvu (SAX), yomwe imalekanitsa OH- ndi gulu lodziwika bwino -N + R3 lingathe kutulutsa anion ena mu yankho, zomwe zimapangitsa anion kusintha kwenikweni.
Pakati pa zinthu zachilengedwe, flavonoids akopa chidwi cha ofufuza chifukwa cha gawo lawo popewa komanso kuchiza matenda amtima.Popeza mamolekyu a flavonoid ndi acidic chifukwa cha kukhalapo kwa magulu a phenolic hydroxyl, ion exchange chromatography ndi njira ina kuwonjezera pa gawo lodziwika bwino kapena reversed phase chromatography pakulekanitsa ndi kuyeretsa ma acid awa.Mu flash chromatography, njira yolekanitsa yomwe imagwiritsidwa ntchito kwambiri posinthira ma ion ndi matrix a silica gel pomwe magulu osinthira ma ion amalumikizidwa pamwamba pake.Njira zosinthira ion zomwe zimagwiritsidwa ntchito kwambiri mu flash chromatography ndi SCX (nthawi zambiri gulu la sulfonic acid) ndi SAX (nthawi zambiri gulu la quaternary amine).Muzolemba zomwe zidasindikizidwa kale ndi mutu wakuti "Kugwiritsa Ntchito SepaFlash Strong Cation Exchange Chromatography Columns mu Kuyeretsedwa kwa Alkaline Compounds" ndi Santai Technologies, mizati ya SCX inagwiritsidwa ntchito pofuna kuyeretsa mankhwala a alkaline.Mu positi iyi, chisakanizo cha mfundo zosalowerera ndale ndi acidic chinagwiritsidwa ntchito ngati chitsanzo kuti tifufuze momwe mazenera a SAX akugwiritsidwa ntchito poyeretsa mankhwala a acidic.
Gawo Loyesera
Chithunzi 1. Chithunzi chojambula cha gawo loyima lolumikizidwa pamwamba pa ma TV olekanitsa a SAX.
Mu positi iyi, ndime ya SAX yodzaza ndi silica yomangika ya quaternary amine idagwiritsidwa ntchito (monga momwe tawonetsera pa Chithunzi 1).Chisakanizo cha Chromone ndi 2,4-dihydroxybenzoic acid chinagwiritsidwa ntchito ngati chitsanzo kuti chiyeretsedwe (monga momwe tawonetsera pa Chithunzi 2).Chosakanizacho chinasungunuka mu methanol ndikuyika pa cartridge ya flash ndi jekeseni.Kukonzekera koyesera kwa kuyeretsedwa kwa flash kwalembedwa mu Table 1.
Chithunzi 2. Mapangidwe a mankhwala a zigawo ziwiri muzitsulo zosakaniza.
| Chida | SepaBean™ makina T | |||||
| Makatiriji | 4 g SepaFlash Standard Series flash cartridge (silica yosakhazikika, 40 - 63 μm, 60 Å, Nambala ya Order: S-5101-0004) | 4 g SepaFlash Bonded Series SAX flash cartridge (silika yosakhazikika, 40 - 63 μm, 60 Å, Nambala ya Order: SW-5001-004-IR) | ||||
| Wavelength | 254 nm (kuzindikira), 280 nm (kuwunika) | |||||
| Gawo la mafoni | Zosungunulira A: N-hexane | |||||
| Zosungunulira B: Ethyl acetate | ||||||
| Mtengo woyenda | 30 ml / mphindi | 20 ml / mphindi | ||||
| Kutsegula kwachitsanzo | 20 mg (chisakanizo cha chigawo A ndi chigawo B) | |||||
| Gradient | Nthawi (CV) | Zosungunulira B (%) | Nthawi (CV) | Zosungunulira B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Zotsatira ndi zokambirana
Choyamba, chitsanzo osakaniza analekanitsidwa ndi yachibadwa gawo kung'anima katiriji chisanadze odzaza ndi silika wokhazikika.Monga momwe chithunzi 3 chikusonyezera, zigawo ziwiri zomwe zili mu chitsanzozo zinachotsedwa pa cartridge imodzi pambuyo pa inzake.Kenako, katiriji ya SAX flash idagwiritsidwa ntchito kuyeretsa chitsanzocho.Monga chiwonetsero cha Chithunzi 4, gawo B la acidic linasungidwa kwathunthu pa cartridge ya SAX.Gawo losalowerera ndale A linachotsedwa pang'onopang'ono kuchokera ku cartridge ndi kutulutsa kwa gawo la mafoni.
Chithunzi 3. Kung'anima kwa chromatogram ya chitsanzo pa cartridge yokhazikika yokhazikika.
Chithunzi 4. Chromatogram yowala ya chitsanzo pa cartridge ya SAX.
Poyerekeza Chithunzi 3 ndi Chithunzi 4, Chigawo A chili ndi mawonekedwe apamwamba osagwirizana pa makatiriji awiri osiyana.Kuti titsimikize ngati chiwongola dzanjacho chikugwirizana ndi gawo, titha kugwiritsa ntchito mawonekedwe onse a kutalika kwa mawonekedwe omwe ali mu pulogalamu yowongolera ya makina a SepaBean™.Tsegulani deta yoyesera ya kulekanitsa kuwiriko, kokerani ku mzere wosonyeza nthawi (CV) mu chromatogram kufika pamtunda wapamwamba kwambiri ndi nsonga yachiwiri yapamwamba kwambiri ya elution yogwirizana ndi Chigawo A, ndi mawonekedwe a kutalika kwa mafunde awiriwa. mfundo adzakhala basi anasonyeza pansipa chromatogram (monga momwe chithunzi 5 ndi Chithunzi 6).Poyerekeza deta yonse ya kutalika kwa mawonekedwe a magawo awiriwa, Gawo A limakhala ndi mayamwidwe osasinthasintha muzoyesera ziwiri.Pazifukwa za Chigawo A chili ndi mawonekedwe osagwirizana pachimake pamakatiriji awiri osiyana, zimaganiziridwa kuti pali zodetsa zenizeni mu Chigawo A chomwe chimakhala ndi kusungidwa kosiyana pa cartridge yanthawi zonse ndi cartridge ya SAX.Choncho, ndondomeko ya eluting ndi yosiyana ndi Chigawo A ndi kusayera kwa makatiriji awiriwa, zomwe zimapangitsa kuti mawonekedwe apamwamba asamagwirizane pa ma chromatogram.
Chithunzi 5. Chiwonetsero chonse cha kutalika kwa mawonekedwe a Chigawo A ndi zonyansa zolekanitsidwa ndi cartridge yanthawi zonse.
Chithunzi 6. Chiwonetsero chonse cha kutalika kwa gawo A ndi zonyansa zolekanitsidwa ndi cartridge ya SAX.
Ngati chinthu chomwe chikuyenera kusonkhanitsidwa ndi gawo losalowerera ndale A, ntchito yoyeretsa imatha kutha mosavuta pogwiritsa ntchito cartridge ya SAX kuti iwonongeke pambuyo pa kutsitsa kwachitsanzo.Kumbali ina, ngati chinthu chomwe chikuyenera kusonkhanitsidwa ndi gawo la acidic B, njira yotulutsira kulanda ikhoza kutengedwa ndikungosintha pang'ono pamasitepe oyesera: pomwe chitsanzocho chidayikidwa pa cartridge ya SAX ndi gawo losalowerera ndale A. anali kwathunthu eluted ndi yachibadwa gawo organic zosungunulira, kusinthana gawo mafoni kuti methanol njira munali 5% asidi asidi.Ma ion a acetate mu gawo la mafoni adzapikisana ndi Gawo B kuti agwirizane ndi magulu a quaternary amine ion pagawo lokhazikika la cartridge ya SAX, potero amachotsa Gawo B kuchokera pa katiriji kuti apeze zomwe akufuna.Chromatogram yachitsanzo chopatulidwa munjira yosinthira ion chikuwonetsedwa Chithunzi 7.
Chithunzi 7. Chromatogram ya kung'anima ya Chigawo B idasinthidwa munjira yosinthira ion pa cartridge ya SAX.
Pomaliza, zitsanzo za acidic kapena ndale zitha kuyeretsedwa mwachangu ndi katiriji ya SAX kuphatikiza katiriji wanthawi zonse pogwiritsa ntchito njira zosiyanasiyana zoyeretsera.Kuphatikiza apo, mothandizidwa ndi mawonekedwe athunthu owunikira mafunde omwe amapangidwa mu pulogalamu yowongolera ya makina a SepaBean™, mawonekedwe a mayamwidwe a tizigawo tating'onoting'ono amatha kufananizidwa ndikutsimikiziridwa mosavuta, kuthandiza ofufuza kudziwa mwachangu kapangidwe ndi kuyera kwa tizigawo tating'onoting'ono ndikuwongolera. ntchito bwino.
| Nambala Yachinthu | Kukula kwa Mzere | Mtengo Woyenda (mL/mphindi) | Max.Pressure (psi/bar) |
| SW-5001-004-IR | 5.9g pa | 10-20 | 400/27.5 |
| SW-5001-012-IR | 23 g pa | 15-30 | 400/27.5 |
| SW-5001-025-IR | 38g pa | 15-30 | 400/27.5 |
| SW-5001-040-IR | 55g pa | 20-40 | 400/27.5 |
| SW-5001-080-IR | 122 g pa | 30-60 | 350/24.0 |
| SW-5001-120-IR | 180 g pa | 40-80 | 300/20.7 |
| SW-5001-220-IR | 340 g pa | 50-100 | 300/20.7 |
| SW-5001-330-IR | 475g pa | 50-100 | 250/17.2
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Table 2. SepaFlash Bonded Series SAX flash cartridges.Zida zonyamula: Silika yopangidwa ndi ultra-pure yosakhazikika ya SAX, 40 - 63 μm, 60 Å.
Kuti mumve zambiri zatsatanetsatane wa SepaBean™makina, kapena kuyitanitsa zambiri pa SepaFlash mndandanda kung'anima makatiriji, chonde pitani patsamba lathu.
Nthawi yotumiza: Nov-09-2018