Rui Huang, Bo Xu
Isicelo se-R&D Center
Isingeniso
I-Ion exchange chromatography (IEC) iyindlela yechromatographic evame ukusetshenziswa ukuhlukanisa nokuhlanza izinhlanganisela ezethulwa ngendlela ye-ionic njengesixazululo.Ngokwezimo zokushaja ezihlukene zama-ion ashintshanayo, i-IEC ingahlukaniswa ngezinhlobo ezimbili, i-cation exchange chromatography kanye ne-anion exchange chromatography.Ku-chromatography yokushintshisana nge-cation, amaqembu ane-asidi ahlanganiswe phezulu kwemidiya yokuhlukanisa.Isibonelo, i-sulfonic acid (-SO3H) iyiqembu elivame ukusetshenziswa ekuhwebeni okuqinile kwe-cation (SCX), ehlukanisa i-H + kanye neqembu elikhokhiswa kabi -SO3- ngaleyo ndlela lingakwazi ukukhangisa amanye ama-cations kusixazululo.Ku-anion exchange chromatography, amaqembu ane-alkaline ahlanganiswe phezulu kwemidiya yokuhlukanisa.Isibonelo, i-quaternary amine (-NR3OH, lapho i-R iyiqembu le-hydrocarbon) ivame ukusetshenziswa ekushintsheni okuqinile kwe-anion (SAX), ehlukanisa i-OH- kanye neqembu elikhokhiswa kahle -N+R3 lingakwazi ukukhangisa amanye ama-anion esixazululweni, okuholela ku-anion. ukushintshana umphumela.
Phakathi kwemikhiqizo yemvelo, ama-flavonoid adonse ukunaka kwabacwaningi ngenxa yendima yabo ekuvimbeleni nasekwelapheni izifo zenhliziyo.Njengoba ama-molecule e-flavonoid ene-acidic ngenxa yokuba khona kwamaqembu e-phenolic hydroxyl, i-ion exchange chromatography ingenye inketho ngaphezu kwesigaba esivamile esivamile noma i-chromatography yesigaba ehlehlisiwe yokuhlukaniswa nokuhlanzwa kwalezi zinhlanganisela ezine-asidi.Ku-flash chromatography, imidiya ehlukanisayo evame ukusetshenziswa ekushintshisaneni kwe-ion i-silica gel matrix lapho amaqembu okushintshanisa ama-ion ehlanganiswa khona phezulu.Izindlela zokushintshaniswa kwe-ion ezisetshenziswa kakhulu ku-flash chromatography yi-SCX (imvamisa iqembu le-sulfonic acid) kanye ne-SAX (imvamisa iqembu le-quaternary amine).Encwadini yesicelo eshicilelwe ngaphambilini enesihloko esithi "Ukusetshenziswa Kwe-SepaFlash Strong Cation Exchange Chromatography Columns in the Purification of Alkaline Compounds" yi-Santai Technologies, amakholomu e-SCX asetshenziselwa ukuhlanzwa kwezinhlanganisela ze-alkaline.Kulokhu okuthunyelwe, inhlanganisela yamazinga angathathi hlangothi kanye ne-acidic yasetshenziswa njengesampula ukuhlola ukusetshenziswa kwamakholomu e-SAX ekuhlanzweni kwama-acidic compounds.
Isigaba Sokuhlola
Umfanekiso 1. Umdwebo wohlelo lwesigaba esimile esiboshelwe ebusweni bemidiya yokuhlukanisa i-SAX.
Kulokhu okuthunyelwe, ikholomu ye-SAX efakwe ngaphambili nge-quaternary amine bonded silica isetshenzisiwe (njengoba kuboniswe kuMfanekiso 1).Ingxube ye-Chromone ne-2,4-dihydroxybenzoic acid isetshenziswe njengesampula okufanele ihlanzwe (njengoba kuboniswe kumfanekiso 2).Ingxube yachithwa ku-methanol futhi yalayishwa ku-flash cartridge ngomjovo.Ukusethwa kokuhlolwa kokuhlanzwa kwe-flash kufakwe kuhlu kuThebula 1.
Umfanekiso 2. Isakhiwo samakhemikhali sezingxenye ezimbili zengxube yesampula.
| Insimbi | SepaBean™ umshini T | |||||
| Amakhatriji | 4 g SepaFlash Standard Series flash cartridge (silica engajwayelekile, 40 - 63 μm, 60 Å, Inombolo ye-oda: S-5101-0004) | 4 g I-SepaFlash Bonded Series SAX flash cartridge (i-silica engajwayelekile, 40 - 63 μm, 60 Å, inombolo ye-oda:SW-5001-004-IR) | ||||
| Ubude begagasi | 254 nm (ukutholwa), 280 nm (ukuqapha) | |||||
| Isigaba seselula | I-solvent A: I-N-hexane | |||||
| I-solvent B: I-Ethyl acetate | ||||||
| Izinga lokugeleza | 30 mL/min | 20 mL/min | ||||
| Isampula iyalayisha | 20 mg (inhlanganisela Yesakhi A kanye Nesakhi B) | |||||
| I-Gradient | Isikhathi (CV) | I-solvent B (%) | Isikhathi (CV) | I-solvent B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Imiphumela nengxoxo
Okokuqala, ingxube yesampula yahlukaniswa yi-cartridge yesigaba esivamile efakwe ngaphambili nge-silica evamile.Njengoba kubonisiwe kuMfanekiso 3, izingxenye ezimbili zesampula zakhishwa ku-cartridge ngokulandelana.Okulandelayo, kwasetshenziswa i-flash cartridge ye-SAX ukuze kuhlanzwe isampula.Njengombukiso kuMfanekiso 4, i-acidic Component B igcinwe ngokuphelele kukhatriji ye-SAX.Ingxenye A engathathi hlangothi yakhishwa kancane kancane kukhathriji ngokuchazwa kwesigaba seselula.
Umfanekiso 3. I-flash chromatogram yesampula kukhatriji yesigaba esivamile.
Umfanekiso 4. I-flash chromatogram yesampula kukhatriji ye-SAX.
Uma kuqhathaniswa Umfanekiso 3 kanye noMfanekiso 4, Ingxenye A inomumo ophakeme ongaguquki kuma-flash cartridge amabili ahlukene.Ukuqinisekisa ukuthi ukuphakama kwe-elution kuyahambisana yini nengxenye, singasebenzisa isici esigcwele sokuskena se-wavelength esakhelwe kusofthiwe yokulawula yomshini we-SepaBean™.Vula idatha yokuhlola yokuhlukaniswa okubili, hudula uye kumugqa wenkomba ku-eksisi yesikhathi (CV) ku-chromatogram uye endaweni ephakeme kakhulu kanye nephoyinti lesibili eliphakeme kakhulu le-elution peak elihambisana ne-Component A, kanye nobubanzi obugcwele be-wavelength balezi zinto ezimbili. amaphuzu azovezwa ngokuzenzakalelayo ngaphansi kwechromatogram (njengoba kukhonjisiwe kuMfanekiso 5 kanye noMfanekiso 6).Uma kuqhathaniswa idatha ye-spectrum ye-wavelength egcwele yalokhu kuhlukaniswa okubili, Ingxenye A ine-spectrum yokumunca engashintshi ekuhlolweni okubili.Ngenxa yesizathu seNgxenye A inokwakheka okuphezulu okungahambisani kuma-flash cartridges amabili ahlukene, kucatshangwa ukuthi kukhona ukungcola okuthile Engxenyeni A enokugcinwa okuhlukile kukhatriji yesigaba esivamile kanye nekhatriji ye-SAX.Ngakho-ke, ukulandelana kwe-luting kuhlukile Kungxenye A kanye nokungcola kulawa ma-flash cartridges amabili, okuholela ekumeni kwesiqongo okungahambisani kuma-chromatogram.
Umfanekiso 5. Ububanzi obugcwele be-wavelength beSakhiwo A kanye nokungcola okuhlukaniswa yikhatriji yesigaba esivamile.
Umfanekiso 6. Ububanzi obugcwele be-wavelength beSakhiwo A kanye nokungcola okuhlukaniswa yikhatriji ye-SAX.
Uma umkhiqizo ohlosiwe ozoqoqwa uyiNgxenye A engathathi hlangothi, umsebenzi wokuhlanza ungaqedwa kalula ngokusebenzisa i-cartridge ye-SAX ngokuqondile ngemva kokulayisha isampula.Ngakolunye uhlangothi, uma umkhiqizo oqondiwe ozoqoqwa Uyingxenye B ene-acidic, indlela yokukhishwa kokuthwebula ingase yamukelwe ngokulungiswa okuncane kuphela ezinyathelweni zokuhlola: lapho isampuli ilayishwa kukhatriji ye-SAX kanye Nengxenye Ephakathi nendawo A. yakhishwa ngokuphelele ngezincibilikisi ze-organic zesigaba esivamile, shintsha isigaba seselula sibe isixazululo se-methanol esiqukethe i-acetic acid engu-5%.Ama-acetate esigabeni esihambayo azoqhudelana neNgxenye B ngokubophezela kumaqembu e-amine e-quaternary esigabeni esimile se-cartridge ye-SAX, ngaleyo ndlela akhiphe Ingxenye B kukhathriji ukuze kutholwe umkhiqizo oqondiwe.I-chromatogram yesampula ehlukaniswe ngemodi yokushintshanisa i-ion iboniswe kuMfanekiso 7.
Umfanekiso 7. I-flash chromatogram Yengxenye B ikhishwe kumodi yokushintshanisa i-ion kukhatriji ye-SAX.
Sengiphetha, isampula eline-asidi noma elingathathi hlangothi lingahlanzwa ngokushesha ngekhatriji ye-SAX ehlanganiswe nekhatriji yesigaba esivamile kusetshenziswa amasu ahlukene okuhlanza.Ngaphezu kwalokho, ngosizo lwesici esigcwele sokuskena se-wavelength esakhelwe kusofthiwe yokulawula yomshini we-SepaBean™, i-spectrum yokumuncwa kwesici sezingxenyana ezikhishiwe ingase iqhathaniswe kalula futhi iqinisekiswe, kusize abacwaningi basheshe banqume ukwakheka nobumsulwa bezingxenyana ezihlutshiwe futhi ngaleyo ndlela kuthuthukiswe. ukusebenza kahle.
| Inombolo Yento | Usayizi Wekholomu | Isilinganiso sokugeleza (mL/min) | Ubukhulu.Ingcindezi (i-psi/ibha) |
| I-SW-5001-004-IR | 5.9g | 10-20 | 400/27.5 |
| I-SW-5001-012-IR | 23 g | 15-30 | 400/27.5 |
| I-SW-5001-025-IR | awu 38g | 15-30 | 400/27.5 |
| I-SW-5001-040-IR | awu 55g | 20-40 | 400/27.5 |
| I-SW-5001-080-IR | awu 122g | 30-60 | 350/24.0 |
| I-SW-5001-120-IR | 180 g | 40-80 | 300/20.7 |
| I-SW-5001-220-IR | awu 340g | 50-100 | 300/20.7 |
| I-SW-5001-330-IR | awu 475g | 50-100 | 250/17.2
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Ithebula 2. I-SepaFlash Bonded Series SAX cartridges flash.Izinto zokupakisha: I-silica eboshwe nge-SAX ehlanzekile kakhulu engajwayelekile, 40 - 63 μm, 60 Å.
Ukuze uthole ukwaziswa okwengeziwe mayelana nokucaciswa okuningiliziwe kwe-SepaBean™umshini, noma imininingwane yoku-oda ku-SepaFlash series flash cartridges, sicela uvakashele iwebhusayithi yethu.
Isikhathi sokuthumela: Nov-09-2018