Rui Huang, Bo Xu
Daim ntawv thov R&D Center
Taw qhia
Ion exchange chromatography (IEC) yog ib txoj kev chromatographic feem ntau siv los cais thiab ntxuav cov tebchaw uas tau nthuav tawm hauv daim ntawv ionic hauv kev daws.Raws li cov nqi sib txawv ntawm kev sib pauv ions, IEC tuaj yeem muab faib ua ob hom, cation exchange chromatography thiab anion exchange chromatography.Hauv cation txauv chromatography, cov pab pawg acidic tau sib koom ua ke rau saum npoo ntawm cov xov xwm sib cais.Piv txwv li, sulfonic acid (-SO3H) yog ib pab pawg siv ntau hauv kev sib pauv cation muaj zog (SCX), uas dissociates H + thiab cov pab pawg neeg tsis zoo -SO3- yog li adsorb lwm cations hauv cov tshuaj.Hauv kev sib pauv chromatography anion, alkaline pawg tau sib koom ua ke rau saum npoo ntawm cov xov xwm sib cais.Piv txwv li, quaternary amine (-NR3OH, qhov twg R yog hydrocarbon pab pawg) feem ntau yog siv nyob rau hauv muaj zog anion pauv (SAX), uas dissociates OH- thiab cov pab pawg neeg them nqi zoo -N + R3 yuav adsorb lwm anions nyob rau hauv cov tshuaj, ua rau anion. pauv nyhuv.
Ntawm cov khoom ntuj, flavonoids tau nyiam cov neeg tshawb nrhiav vim lawv lub luag haujlwm hauv kev tiv thaiv thiab kho cov kab mob plawv.Txij li thaum cov flavonoid molecules yog acidic vim muaj cov phenolic hydroxyl pawg, ion pauv chromatography yog lwm txoj kev xaiv ntxiv rau cov pa ib txwm ua los yog thim rov qab theem chromatography rau kev sib cais thiab purification ntawm cov acidic compounds.Hauv flash chromatography, feem ntau siv kev sib cais tawm rau kev sib pauv ion yog silica gel matrix qhov twg ion pauv pawg tau sib koom ua ke rau nws qhov chaw.Feem ntau siv ion pauv hom hauv flash chromatography yog SCX (feem ntau sulfonic acid pawg) thiab SAX (feem ntau quaternary amine pawg).Nyob rau hauv daim ntawv tshaj tawm yav dhau los daim ntawv thov nrog lub npe "Kev Siv SepaFlash Strong Cation Exchange Chromatography Kem hauv Kev Ua Kom Huv Ntawm Alkaline Cheeb Tsam" los ntawm Santai Technologies, SCX kab tau ua haujlwm rau kev ua kom huv ntawm alkaline tebchaw.Hauv cov ntawv tshaj tawm no, kev sib xyaw ntawm cov qauv nruab nrab thiab cov kua qaub tau siv los ua cov qauv los tshawb txog daim ntawv thov ntawm SAX kab hauv kev ua kom huv ntawm cov kua qaub.
Ntu kev sim
Daim duab 1. Daim duab schematic ntawm cov theem nyob ruaj ruaj txuas rau saum npoo ntawm SAX sib cais tawm.
Hauv cov ntawv tshaj tawm no, SAX kem ua ntej ntim nrog quaternary amine bonded silica tau siv (raws li qhia hauv daim duab 1).Kev sib xyaw ntawm Chromone thiab 2,4-dihydroxybenzoic acid tau siv los ua tus qauv kom huv (raws li qhia hauv daim duab 2).Cov sib tov yog yaj nyob rau hauv methanol thiab loaded mus rau lub flash cartridge los ntawm ib tug injector.Kev sim teeb tsa ntawm lub flash purification yog teev nyob rau hauv Table 1.
Daim duab 2. Cov qauv tshuaj ntawm ob yam khoom hauv cov qauv sib xyaw.
| Ntsuas | SepaBean™ tshuab T | |||||
| Cartridges | 4 g SepaFlash Standard Series flash cartridge (silica tsis xwm yeem, 40 - 63 μm, 60 Å, Order naj npawb: S-5101-0004) | 4 g SepaFlash Bonded Series SAX flash cartridge (tsis xwm yeem silica, 40 - 63 μm, 60 Å, Order naj npawb: SW-5001-004-IR) | ||||
| Lub wavelength | 254 nm (nrhiav), 280 nm (saib xyuas) | |||||
| Mobile theem | Solvent A: N-hexane | |||||
| Cov kuab tshuaj B: Ethyl acetate | ||||||
| Tus nqi ntws | 30 ml / min | 20 ml / min | ||||
| Sample loading | 20 mg (ib qho sib xyaw ntawm Cov Khoom Siv A thiab Cov Khoom B) | |||||
| Gradient | Lub sijhawm (CV) | Cov kuab tshuaj B (%) | Lub sijhawm (CV) | Cov kuab tshuaj B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Cov txiaj ntsig thiab kev sib tham
Ua ntej, cov qauv sib xyaw yog sib cais los ntawm ib txwm theem flash cartridge pre-packed nrog silica li niaj zaus.Raws li pom nyob rau hauv daim duab 3, ob qho tib si nyob rau hauv cov qauv raug eluted ntawm lub cartridge ib tug tom qab lwm.Tom ntej no, SAX flash cartridge tau siv rau kev ua kom huv ntawm cov qauv.Raws li pom nyob rau hauv daim duab 4, cov acidic Cheebtsam B tau khaws cia tag nrho ntawm SAX cartridge.Qhov nruab nrab cov ntsiab lus A tau maj mam eluted los ntawm lub cartridge nrog lub elution ntawm lub mobile theem.
Daim duab 3. Lub teeb nyem chromatogram ntawm cov qauv ntawm cov cartridge ib txwm muaj.
Daim duab 4. Lub teeb chromatogram ntawm cov qauv ntawm SAX cartridge.
Muab piv rau daim duab 3 thiab daim duab 4, Cov Khoom Siv A muaj qhov tsis sib haum xeeb ncov ntawm ob daim cartridges sib txawv.Txhawm rau kom paub meej tias qhov elution ncov yog sib haum rau cov khoom siv, peb tuaj yeem siv tag nrho cov wavelength scanning feature uas tau tsim rau hauv kev tswj software ntawm SepaBean ™ tshuab.Qhib cov ntaub ntawv sim ntawm ob qhov kev sib cais, rub mus rau qhov taw qhia kab ntawm lub sijhawm axis (CV) hauv chromatogram mus rau qhov siab tshaj plaws thiab qhov thib ob siab tshaj plaws ntawm qhov siab tshaj plaws ntawm elution ncov sib xws rau Cov Khoom A, thiab tag nrho cov wavelength spectrum ntawm ob. cov ntsiab lus yuav cia li pom hauv qab ntawm chromatogram (raws li qhia hauv daim duab 5 thiab daim duab 6).Sib piv cov ntaub ntawv wavelength spectrum tag nrho ntawm ob qhov kev sib cais no, Cov Khoom Siv A muaj qhov nqus tau zoo ib yam hauv ob qhov kev sim.Rau qhov laj thawj ntawm Cov Khoom Siv A muaj qhov tsis sib haum xeeb ntawm cov duab sib txawv ntawm ob daim cartridges sib txawv, nws tau kwv yees tias muaj cov impurity tshwj xeeb hauv Cov Khoom Siv A uas muaj qhov sib txawv ntawm qhov sib txawv ntawm cov theem ib txwm cartridge thiab SAX cartridge.Yog li, qhov sib txawv ntawm qhov sib txawv ntawm cov khoom A thiab cov impurity ntawm ob lub flash cartridges, uas ua rau cov duab tsis sib xws ntawm chromatograms.
Daim duab 5. Tag nrho cov wavelength spectrum ntawm lub Cheebtsam A thiab cov impurity sib cais los ntawm ib txwm theem cartridge.
Daim duab 6. Tag nrho cov wavelength spectrum ntawm cov khoom A thiab cov impurity sib cais los ntawm SAX cartridge.
Yog hais tias lub hom phiaj cov khoom yuav tsum tau sau yog nruab nrab Cheebtsam A, purification txoj hauj lwm yuav yooj yim ua tiav los ntawm ncaj qha siv lub SAX cartridge rau elution tom qab coj mus kuaj.Ntawm qhov tod tes, yog tias lub hom phiaj cov khoom yuav raug sau yog acidic Cheebtsam B, qhov kev ntes-tso tawm tuaj yeem tau txais los ntawm kev hloov kho me ntsis hauv cov kauj ruam kev sim: thaum tus qauv raug thauj khoom ntawm SAX cartridge thiab cov khoom nruab nrab A. tau tag nrho eluted tawm nrog cov theem ib txwm organic solvents, hloov lub mobile theem mus rau methanol tov uas muaj 5% acetic acid.Cov acetate ions nyob rau hauv lub xov tooj ntawm tes yuav sib tw nrog cov khoom B rau kev khi rau quaternary amine ion pawg nyob rau theem nyob ruaj khov ntawm SAX cartridge, yog li eluting Cheebtsam B ntawm lub cartridge kom tau txais cov khoom lag luam.Cov chromatogram ntawm cov qauv sib cais hauv ion pauv hom tau pom hauv daim duab 7.
Daim duab 7. Lub teeb chromatogram ntawm lub Cheebtsam B eluted nyob rau hauv ion pauv hom ntawm SAX cartridge.
Hauv kev xaus, cov qauv acidic lossis nruab nrab tuaj yeem ua kom huv sai sai los ntawm SAX cartridge ua ke nrog cov txheej txheem ib txwm siv cov tswv yim sib txawv.Tsis tas li ntawd, nrog kev pab los ntawm tag nrho cov wavelength scanning feature ua rau hauv kev tswj software ntawm SepaBean ™ tshuab, cov yam ntxwv nqus spectrum ntawm cov eluted feem yuav muab tau yooj yim piv thiab paub tseeb, pab soj ntsuam sai sai txiav txim seb muaj pes tsawg leeg thiab purity ntawm cov eluted feem thiab yog li txhim kho. ua hauj lwm efficiency.
| Yam khoom naj npawb | Kem Loj | Tus nqi ntws (ml / min) | Max. Siab (psi/bar) |
| SW-5001-004-IR | 5,9g ua | 10-20 | 400/27.5 |
| SW-5001-012-IR | 23g ua | 15-30 | 400/27.5 |
| SW-5001-025-IR | 38g ua | 15-30 | 400/27.5 |
| SW-5001-040-IR | 55g ua | 20-40 | 400/27.5 |
| SW-5001-080-IR | 12 2g | 30-60 | 350/24.0 Nws |
| SW-5001-120-IR | 180 g | 40-80 : kuv | 300/20.7 |
| SW-5001-220-IR | 340g ua | 50-100 | 300/20.7 |
| SW-5001-330-IR | 475g ua | 50-100 | 250/17.2
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Table 2. SepaFlash Bonded Series SAX flash cartridges.Cov ntaub ntawv ntim: Ultra-ntshiab irregular SAX-bonded silica, 40 - 63 μm, 60 Å.
Yog xav paub ntxiv txog cov ncauj lus kom ntxaws ntawm SepaBean ™tshuab, lossis cov ntaub ntawv xaj ntawm SepaFlash series flash cartridges, thov mus saib peb lub vev xaib.
Post lub sij hawm: Nov-09-2018