Rui Huang, Bo Xu
Porogaramu R&D Ikigo
Intangiriro
Ion guhana chromatografiya (IEC) nuburyo bwa chromatografique bukoreshwa mugutandukanya no kweza ibice bitangwa muburyo bwa ionic mubisubizo.Ukurikije uburyo butandukanye bwishyurwa bwa ion zungurana ibitekerezo, IEC irashobora kugabanywamo ubwoko bubiri, cation yo guhana chromatografiya hamwe na chromatografi ya anion.Muri cation guhana chromatografiya, amatsinda acide ahujwe hejuru yibitangazamakuru bitandukanya.Kurugero, acide sulfonique (-SO3H) nitsinda rikunze gukoreshwa muguhana cation ikomeye (SCX), itandukanya H + hamwe nitsinda ryashizwemo nabi -SO3- rishobora rero kumenyekanisha izindi cations mugisubizo.Muri anion guhana chromatografiya, amatsinda ya alkaline ahujwe hejuru yibitangazamakuru bitandukanya.Kurugero, amine ya quaternary amine (-NR3OH, aho R ni itsinda rya hydrocarubone) isanzwe ikoreshwa muguhana anion ikomeye (SAX), itandukanya OH- kandi itsinda ryashizwemo neza -N + R3 rishobora kwamamaza izindi anion mugisubizo, bikavamo anion Ingaruka zo guhana.
Mu bicuruzwa bisanzwe, flavonoide yashimishije abashakashatsi kubera uruhare rwabo mu gukumira no kuvura indwara zifata umutima.Kubera ko molekile ya flavonoide iba acide bitewe no kuba hari amatsinda ya hydroxyl ya fenolike, ion yo guhana chromatografiya ni ubundi buryo bwiyongera ku cyiciro gisanzwe gisanzwe cyangwa icyiciro cya chromatografiya cyahinduwe cyo gutandukanya no kweza ibyo bintu bya aside.Muri flash chromatografiya, ibisanzwe bikoreshwa mubitandukanya byo guhanahana ion ni silika gel matrix aho amatsinda yo guhana ion ahujwe nubuso bwayo.Uburyo bukoreshwa cyane muburyo bwo guhanahana ion muri flash chromatografiya ni SCX (mubisanzwe itsinda rya acide sulfonique) na SAX (mubisanzwe amatsinda ya quaternary amine).Mu nyandiko isaba mbere yatangajwe ifite umutwe ugira uti "Gukoresha SepaFlash Ikomeye ya Cation Guhindura Chromatography Inkingi mu Kweza Imiti ya Alkaline" na Santai Technologies, inkingi za SCX zakoreshwaga mu kweza ibinyomoro.Muri iyi nyandiko, uruvange rwibipimo bitagira aho bibogamiye na acide byakoreshejwe nkicyitegererezo cyo gushakisha ikoreshwa ryinkingi za SAX mugusukura aside irike.
Igice cy'Ubushakashatsi
Igishushanyo 1. Igishushanyo mbonera cyicyiciro gihagaze gihujwe nubuso bwitangazamakuru rya SAX.
Muri iyi nyandiko, inkingi ya SAX yabanje gupakirwa na quaternary amine ihujwe na silika yakoreshejwe (nkuko bigaragara ku gishushanyo 1).Uruvange rwa Chromone na aside 2,4-dihydroxybenzoic yakoreshejwe nk'icyitegererezo cyo kwezwa (nkuko bigaragara ku gishushanyo cya 2).Uruvange rwashongeshejwe muri methanol hanyuma rushyirwa kuri flash cartridge nuwatewe inshinge.Ubushakashatsi bwakorewe flash yoza kurutonde murutonde 1.
Igishushanyo 2. Imiterere yimiti yibice byombi murugero rwivanze.
| Igikoresho | SepaBean ™ imashini T. | |||||
| Cartridges | 4 g SepaFlash Urutonde rusanzwe rwa flash cartridge (silika idasanzwe, 40 - 63 μ m, 60 Å, nimero yatanzwe: S-5101-0004) | 4 g SepaFlash Ihuza Urutonde SAX flash cartridge (silika idasanzwe, 40 - 63 mkm, 60 Å, nimero ya ordre ： SW-5001-004-IR) | ||||
| Uburebure | 254 nm (gutahura), 280 nm (gukurikirana) | |||||
| Icyiciro cya mobile | Umuti A: N-hexane | |||||
| Umuti B: Ethyl acetate | ||||||
| Igipimo cyo gutemba | 30 mL / min | 20 mL / min | ||||
| Icyitegererezo | 20 mg (uruvange rw'ibigize A n'ibigize B) | |||||
| Gradient | Igihe (CV) | Umuti B (%) | Igihe (CV) | Umuti B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Ibisubizo n'ibiganiro
Ubwa mbere, icyitegererezo kivanze cyatandukanijwe nicyiciro gisanzwe cya flash cartridge yabanje gupakira hamwe na silika isanzwe.Nkuko bigaragara ku gishushanyo cya 3, ibice bibiri murugero byatoranijwe kuva muri karitsiye imwe imwe.Ibikurikira, flash cartridge ya SAX yakoreshejwe mugusukura icyitegererezo.Nkuko bigaragara ku gishushanyo cya 4, Acide ya B yagumishijwe rwose kuri karitsiye ya SAX.Ibice bitagira aho bibogamiye A byavanyweho buhoro buhoro bivuye kuri karitsiye hamwe no gukuraho icyiciro cya mobile.
Igishushanyo 3. Flash chromatogramu yicyitegererezo kuri karitsiye isanzwe isanzwe.
Igicapo 4. Flash chromatogramu yicyitegererezo kuri karitsiye ya SAX.
Ugereranije Igishushanyo cya 3 nishusho ya 4, Igice A gifite imiterere yimpinga idahuye kuri flash cartrid ebyiri zitandukanye.Kugirango twemeze niba impinga ya elution ihuye nibigize, turashobora gukoresha uburyo bwuzuye bwo gusikana umurongo wubatswe muri software igenzura imashini ya SepaBean ™.Fungura amakuru yubushakashatsi bwibice bibiri bitandukanijwe, kurura kumurongo werekana kumurongo wigihe (CV) muri chromatogramu kugeza kumurongo wo hejuru hamwe na kabiri murwego rwo hejuru rwo hejuru ya elution ihuye nibigize A, hamwe nuburebure bwuzuye bwumurambararo wibi byombi ingingo zizahita zerekanwa munsi ya chromatogramu (nkuko bigaragara ku gishushanyo cya 5 na 6).Ugereranije umurongo wuzuye wumurongo wamakuru yibi gutandukana byombi, Ibigize A bifite uburyo bwo kwinjiza ibintu muburyo bubiri.Kubwimpamvu yibigize A ifite imiterere yimpinga idahuye kumashanyarazi abiri atandukanye ya flash, haravugwa ko hariho umwanda wihariye mubice A bifite uburyo butandukanye kuri karitsiye yicyiciro gisanzwe na karitsiye ya SAX.Kubwibyo, gutondekanya amajwi biratandukanye kubigize A hamwe nubuhumane kuri aya makaritsiye abiri ya flash, bivamo imiterere yimpinga idahuye kuri chromatogramu.
Igicapo 5. Uburebure bwuzuye bwumurongo wibigize A hamwe numwanda utandukanijwe na karitsiye isanzwe.
Igicapo 6. Uburebure bwuzuye bwumurongo wibigize A hamwe numwanda utandukanijwe na SAX cartridge.
Niba ibicuruzwa bigomba gukusanyirizwa hamwe bitagira aho bibogamiye A, umurimo wo kweza urashobora kurangizwa byoroshye ukoresheje ikarita ya SAX kugirango ikurweho nyuma yo gupakira icyitegererezo.Ku rundi ruhande, niba ibicuruzwa bigomba gukusanyirizwa hamwe bigize aside B, uburyo bwo gufata-kurekura bushobora gukurikizwa hahinduwe bike gusa muntambwe yubushakashatsi: mugihe icyitegererezo cyapakiwe kuri karitsiye ya SAX hamwe nibice bitagira aho bibogamiye A yasibwe burundu hamwe nicyiciro gisanzwe cyumuti, hindura icyiciro kigendanwa kumuti wa methanol urimo aside 5% ya acide.Ion ya acetate mugice kigendanwa izahatana nigice B kugirango ihuze amatsinda ya kane ya amine ion kumurongo uhagaze ya karitsiye ya SAX, bityo ikuremo B B kuva muri karitsiye kugirango ibone ibicuruzwa bigenewe.Chromatogramu yicyitegererezo yatandukanijwe muburyo bwo guhanahana ion yerekanwe mubishusho 7.
Igicapo 7. Flash chromatogramu yibigize B yavuzwe muburyo bwo guhana ion kuri karitsiye ya SAX.
Mu gusoza, icyitegererezo cya acide cyangwa kidafite aho kibogamiye gishobora kwezwa byihuse na SAX ya karitsiye hamwe na karitsiye isanzwe ikoresha ingamba zitandukanye zo kweza.Byongeye kandi, hifashishijwe uburyo bwuzuye bwo gusikana umurongo wubatswe muri software igenzura imashini ya SepaBean ™ imashini, ibintu biranga kwinjiza ibice byavunitse bishobora kugereranywa byoroshye kandi bikemezwa, bifasha abashakashatsi kumenya byihuse imiterere nubuziranenge bwibice byavuzwe bityo bigatera imbere gukora neza.
| Umubare w'ingingo | Ingano yinkingi | Igipimo cy'Uruzi (mL / min) | Icyiciro (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
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Imbonerahamwe 2. SepaFlash Bonded Series SAX flash cartridges.Ibikoresho byo gupakira: Ultra-yera idasanzwe SAX ihujwe na silika, 40 - 63 mm, 60 Å.
Kubindi bisobanuro birambuye kuri SepaBean ™imashini, cyangwa amakuru atumiza kuri SepaFlash yuruhererekane rwa flash cartridges, nyamuneka sura urubuga.
Igihe cyo kohereza: Ugushyingo-09-2018