Rui Huang, Bo Xu
Ngwa R&D Center
Okwu mmalite
Ion Exchange chromatography (IEC) bụ usoro chromatographic nke a na-ejikarị kewapụ na sachaa ogige ndị e gosipụtara n'ụdị ionic na ngwọta.Dabere na steeti ụgwọ dị iche iche nke ion mgbanwe, IEC nwere ike kewaa ụzọ abụọ, chromatography mgbanwe cation na chromatography mgbanwe anion.Na cation mgbanwe chromatography, otu acidic na-ejikọta na elu nke mgbasa ozi nkewa.Dịka ọmụmaatụ, sulfonic acid (-SO3H) bụ otu a na-ejikarị eme ihe na mgbanwe mgbanwe cation siri ike (SCX), nke na-ekewa H+ na otu ebubo na-ezighị ezi -SO3- nwere ike si otú a gbasaa cations ndị ọzọ na ngwọta.Na chromatography mgbanwe anion, otu alkaline na-ejikọta na elu nke mgbasa ozi nkewa.Dịka ọmụmaatụ, amine quaternary (-NR3OH, ebe R bụ hydrocarbon Group) na-ejikarị eme ihe na mgbanwe anion siri ike (SAX), nke na-ekewa OH- na otu ebubo nke ọma -N + R3 nwere ike ịkwado anions ndị ọzọ na ngwọta, na-ebute anion. mmetụta mgbanwe.
N'ime ngwaahịa ndị sitere n'okike, flavonoids adọtala uche ndị nchọpụta n'ihi ọrụ ha na mgbochi na ọgwụgwọ ọrịa obi.Ebe ọ bụ na mkpụrụ ndụ flavonoid bụ acidic n'ihi ọnụnọ nke phenolic hydroxyl otu, ion mgbanwe chromatography bụ nhọrọ ọzọ na mgbakwunye na usoro nkịtị ma ọ bụ ntụgharị chromatography nke oge maka nkewa na ịdị ọcha nke ogige acidic ndị a.Na flash chromatography, mgbasa ozi nkewa nke a na-ejikarị maka mgbanwe ion bụ silica gel matrix ebe ndị otu mgbanwe ion na-ejikọta na elu ya.Ụdị mgbanwe ion a na-ejikarị na chromatography flash bụ SCX (na-abụkarị otu sulfonic acid) na SAX (na-abụkarị otu amine quaternary).N'ime ndetu ngwa e bipụtara na mbụ nke nwere aha "Ngwa nke SepaFlash Strong Cation Exchange Chromatography Columns in the Purification of Alkaline Compounds" nke Santai Teknụzụ, ejiri kọlụm SCX rụọ ọrụ maka ịdị ọcha nke ogige alkaline.Na post a, a na-eji ngwakọta nke ụkpụrụ na-anọpụ iche na acidic mee ihe dị ka ihe atụ iji nyochaa ngwa nke ogidi SAX na nchacha nke ogige acidic.
Ngalaba nnwale
Onyonyo 1. Eserese schematic nke usoro a na-akwụghị ọtọ jikọtara na elu mgbasa ozi nkewa SAX.
Na post a, a na-eji kọlụm SAX nke ejiri quaternary amine bonded silica mee ihe (dị ka egosiri na eserese 1).A na-eji ngwakọta nke Chromone na 2,4-dihydroxybenzoic acid mee ihe dị ka ihe nlele iji mee ka ọ dị ọcha (dị ka egosiri na foto 2).A gbazere ngwakọta a na methanol ma tinye ya na katrij flash site na injector.Edepụtara nhazi nnwale nke nchacha ọkụ na tebụl 1.
Ọgụgụ 2. Usoro kemịkalụ nke ihe abụọ dị na ngwakọta ihe atụ.
| Akụrụngwa | SepaBean™ igwe T | |||||
| Katrij | 4 g SepaFlash Standard Series flash katrij (silica oge niile, 40 - 63 μm, 60 Å, Nọmba ịtụ: S-5101-0004) | 4 g SepaFlash Bonded Series SAX flash katrij (silica oge niile, 40 - 63 μm, 60 Å, Nọmba ịtụ: SW-5001-004-IR) | ||||
| Ogologo ogologo | 254 nm (nchọpụta), 280 nm (nleba anya) | |||||
| Usoro mkpanaka | Ihe mgbaze A: N-hexane | |||||
| Ihe mgbaze B: ethyl acetate | ||||||
| Ọnụego mgbaba | 30 ml / min | 20 ml / min | ||||
| Nbudata ihe atụ | 20 mg (ngwakọta nke Component A na Component B) | |||||
| Gradient | Oge (CV) | Ihe mgbaze B (%) | Oge (CV) | Ihe mgbaze B (%) | ||
| 0 | 0 | 0 | 0 | |||
| 1.7 | 12 | 14 | 100 | |||
| 3.7 | 12 | / | / | |||
| 16 | 100 | / | / | |||
| 18 | 100 | / | / | |||
Nsonazụ na mkparịta ụka
Nke mbụ, e kewapụrụ ngwakọta ihe atụ ahụ site na katrij flash na-emekarị nke ejiri silica mgbe niile ebujuru ya.Dị ka e gosiri na foto 3, ihe abụọ ahụ dị na ihe nlele ahụ na-esi na katrij ahụ pụta n'otu n'otu.Na-esote, a na-eji katrij flash SAX maka sachaa ihe nlele ahụ.Dịka e gosiri na eserese 4, akụkụ acidic B ka edobere kpamkpam na katrij SAX.A na-eji nwayọọ nwayọọ wepụ akụkụ A na-anọpụ iche site na katrij site na elution nke usoro mkpanaka.
Ọgụgụ 3. chromatogram flash nke ihe nlele na katrij oge nkịtị.
Ọgụgụ 4. chromatogram flash nke ihe nlele na katrij SAX.
N'iji atụnyere eserese 3 na eserese 4, akụrụngwa A nwere ọdịdị elu na-ekwekọghị ekwekọ na katrij ọkụ abụọ dị iche iche.Iji gosi ma elution elu ọ dabara na akụrụngwa, anyị nwere ike iji njirimara nyocha ogologo ogologo nke arụnyere n'ime ngwanrọ njikwa nke igwe SepaBean™.Mepee data nnwale nke nkewa abụọ ahụ, dọrọ gaa na akara ngosi na axis oge (CV) na chromatogram ruo ebe kachasị elu na nke abụọ kachasị elu nke elution elu kwekọrọ na Component A, yana ụdị nhụsianya zuru oke nke abụọ a. A ga-egosipụta isi ihe na-akpaghị aka n'okpuru chromatogram (dị ka egosiri na foto 5 na eserese 6).N'ịtụnyere data ụdịdị dị iche iche nke ogologo ogologo ogologo nke nkewa abụọ a, akụrụngwa A nwere ụdịdị nnabata na-agbanwe agbanwe na nnwale abụọ.N'ihi na akụrụngwa A nwere ọdịdị elu na-ekwekọghị ekwekọ na katrij flash abụọ dị iche iche, a na-eche na enwere adịghị ọcha akọwapụtara na akụrụngwa A nke nwere njigide dị iche na katrij nke nkịtị na katrij SAX.Ya mere, usoro eluting dị iche iche maka Component A na adịghị ọcha na katrij ọkụ abụọ a, na-ebute ọdịdị elu na-ekwekọghị ekwekọ na chromatograms.
Onyonyo 5. Ụdị dị iche iche nke akụkụ A na adịghị ọcha kewapụrụ site na katrij oge nkịtị.
Ọnụọgụ 6. Ụdị dị iche iche nke akụkụ A na adịghị ọcha nke SAX cartridge kewapụrụ.
Ọ bụrụ na ngwaahịa a ga-anakọta bụ ihe na-anọpụ iche A, ọrụ nchacha ahụ nwere ike ịrụcha ya ngwa ngwa site na iji SAX katrij maka elution mgbe nbudata sample.N'aka nke ọzọ, ọ bụrụ na ngwaahịa a ga-anakọta bụ acidic Component B, a ga-anakwere ụzọ ntọhapụ ahụ site na iji obere mgbanwe na usoro nnwale: mgbe a na-ebunye ihe nlele ahụ na katrij SAX na akụkụ A na-anọpụ iche. Ejiri ihe mgbaze na-edozi ahụ na-eme ka ọ bụrụ ihe na-edozi ahụ kpamkpam, gbanwee usoro mkpanaka gaa na ngwọta methanol nwere 5% acetic acid.Ions acetate dị na usoro mkpanaka ga-asọmpi na Component B maka ijikọ otu amine ion dị n'akụkụ ọdụ nke SAX cartridge, si otú ahụ na-ebupụ akụkụ B site na katrij iji nweta ngwaahịa ezubere iche.E gosipụtara chromatogram nke nlele ahụ kewapụrụ na ọnọdụ mgbanwe ion na foto 7.
Onyonyo 7. Kromatogram flash nke akụrụngwa B gbalitere na ọnọdụ mgbanwe ion na katrij SAX.
Na mmechi, ihe nlele acidic ma ọ bụ nnọpụiche nwere ike sachaa ngwa ngwa site na SAX cartridge jikọtara ya na katrij oge nkịtị na-eji atụmatụ dị ọcha dị iche iche.Ọzọkwa, site n'enyemaka nke njiri nyocha ogologo ogologo nke arụnyere n'ime ngwanrọ njikwa nke igwe SepaBean ™, enwere ike iji ya tụnyere njirimara nke irighiri irighiri ahụ n'ụzọ dị mfe, na-enyere ndị nyocha aka ịchọpụta ngwa ngwa na ịdị ọcha nke irighiri irighiri ahụ wee si otú a na-emeziwanye. arụ ọrụ arụmọrụ.
| Nọmba ihe | Nha kọlụm | Ọnụego mgbaba (mL/min) | Oke. Nrụgide (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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Tebụl 2. SepaFlash Bonded Series SAX flash cartridges.Ihe nkwakọ ngwaahịa: Ultra-dị ọcha silica na-ejikọtaghị SAX, 40 - 63 μm, 60 Å.
Maka ozi ndị ọzọ gbasara nkọwa zuru ezu nke SepaBean™igwe, ma ọ bụ ozi ịtụ na sepaFlash usoro flash katrij, biko gaa na webụsaịtị anyị.
Oge nzipu: Nov-09-2018