Hongcheng Wang, Bo Xu
Daim Ntawv Thov R & D Center
Lub hauv paus rau phau ntawv
Raws li cov txheeb ze polarities ntawm chaw ruaj khov rau theem thiab mobile theem, kua chromatography (NPC) thiab thim cov chromatography (RPC). Rau RPC, kev polarity ntawm mobile theem yog muaj zog dua li ntawm theem nyob ruaj khov. RPC tau dhau los ua qhov feem ntau siv nyob rau hauv kua chromatography hom vim nws qhov kev ua tau zoo, kev daws teeb meem zoo thiab ua kom muaj kev kho kom rov zoo. Yog li RPC yog tsim nyog rau kev sib cais thiab purification ntawm ntau yam kev ua liaj ua teb, cov roj ntsha, thiab lwm yam, suav nrog rau cov pab pawg ua haujlwm, suav nrog CI18, C4, Phenyl, Cyano, Amino, thiab lwm yam ntawm cov pab pawg sib khi ua ke, feem ntau dav siv ib yog C18. Nws yog kwv yees tau tias ntau dua 80% ntawm RPC tam sim no siv C18 Bonded theem. Yog li C18 Chromatography Sab tau dhau los ua qhov yuav tsum muaj-muaj cov thoob ntiaj teb rau txhua chav kuaj.
Txawm hais tias C18 tuaj yeem siv tau rau hauv ntau cov ntawv thov, txawm li cas los xij, rau qee qhov qauv uas muaj teeb meem C18 ntau yuav muaj teeb meem thaum raug siv los ua kom cov qauv no. Nyob rau hauv RPC, feem ntau siv cov kev tshem tawm tuaj yeem raug txiav txim raws li lawv polarity: dej <Methanol <etetonol <etetonol <isopropanol. Yuav kom paub tseeb tias muaj kev cia siab zoo nyob rau hauv kem rau cov qauv no (muaj zog polar lossis hydroous system yog qhov tsim nyog yuav tsum tau siv raws li mobile theem. Txawm li cas los xij, thaum siv dej ntshiab (suav nrog cov dej ntshiab lossis cov ntsev ntshiab) raws li kev sib xyaw ua ke ntawm C18 sib xyaw ua ke, ua rau muaj peev xwm tsawg dua hauv cov ntawv khaws cia ntawm lub peev xwm lossis tsis muaj peev xwm ua tau. Qhov tshwm sim no hu ua "Hydrophobic theem vau" (raws li qhia nyob rau hauv sab laug ib feem ntawm daim duab 1). Txawm hais tias qhov xwm txheej no tau thim rov qab thaum cov ntawv yog ntxuav nrog cov kuab tshuaj organic xws li Methanol lossis Acetoniturile, nws tseem tuaj yeem ua rau kev puas tsuaj rau kem. Yog li ntawd, nws yog ib qho tsim nyog los tiv thaiv cov xwm txheej no los ntawm kev tshwm sim.
Daim duab 1. Cov schematic daim duab ntawm cov theem sib khi ntawm cov nplaim dej ntawm silica gel hauv kem niaj zaus (sab laug) thiab C18Aq (txoj cai).
Txhawm rau hais txog cov teeb meem tau hais los saum toj no, cov chratatography cov ntaub ntawv cov ntaub ntawv tau tsim kev txhim kho. Ib qho ntawm cov kev txhim kho no yog ua qee qhov kev hloov kho ntawm cov pawg Silica, xws li pom nyob hauv ib feem ntawm Hydroa Gel Ntau Hydrophilic. Yog li C18 chains ntawm Silica nto yuav ua tau tiav nyob rau hauv cov kev mob aqueous aqueous thiab hydrophobic theem kev sib tsoo tuaj yeem zam. Cov kev hloov kho C18 yog hu ua aqueous C18, uas yog tsim rau cov neeg muaj peev xwm kom tsawg dua thiab tuaj yeem tiv thaiv 100% aqueous system. C18AQ Txhua tau siv dav hauv kev sib cais thiab ua kom huv ntawm cov duabceeb polar, suav nrog cov organic acids, peptides, nucleosides thiab dej-soluble vitamins.
Kev txiav txim siab yog ib qho ntawm cov ntawv thov c18Aq hauv kev ua kom muaj cov ntsev lossis cov qauv ntawm cov qauv hauv cov qauv ntawm kev tshawb fawb tom ntej. Hauv cov ncej no, xiav xiav fcf nrog muaj zog polarity tau siv ua tus qauv thiab ua kom huv ntawm C18Aq. Cov qauv sam thiaj tau hloov los ntawm cov organic huab cua los ntawm kev daws teeb meem, yog li ua raws li cov kev daws teeb meem qeeb qeeb qeeb qeeb qeeb qeeb thiab ua haujlwm kom txuag tau. Tsis tas li ntawd, txoj kev huv ntawm cov hnoos qeev tau txhim kho los ntawm kev tshem qee qhov impurities hauv cov qauv.
Kev sim ntu
Daim duab 2. Cov qauv tshuaj ntawm cov qauv.
FCF xiav xiav FCF tau siv los ua cov qauv hauv cov ntawv no. Lub purity ntawm cov qauv raw yog 86% thiab cov qauv tshuaj muaj nyob hauv daim duab 2. Los npaj cov qauv ua piv txwv hauv 1 m nah2po4 buffer daws thiab shook zoo los ua ib qho kev daws teeb meem meej. Cov qauv kev daws teeb meem yog tom qab ntawd txhaj rau hauv flash rau los ntawm kev muaj kev tiv thaiv. Kev sim teeb tsa ntawm Flash Purification yog teev nyob rau hauv lub rooj 1.
| Lub cuab tam | Tshuab Ntxhua Tshuab Sepababean2 | |||
| Tus saib xyuas | 12 g Sepaflash C18 RP Flash Carica, Spherical Silica, 20 - 45 μm, 100 Å, tus lej xaj: Sw-5222-012-sp) | 12 g Sepaflash C18aq RP Flash Carica, 20 - 45 μm, 100 Å, tus lej xaj: Sw-5222-0- sp (aq)) | ||
| Lub paj | 254 nm | |||
| Txawb theem | Hnyav A: Dej Cov kuab tshuaj B: Methanol | |||
| Txaus tus nqi | 30 ml / min | |||
| Qauv Chaw thau khoom | 300 mg (xiav xiav fcf nrog lub purity ntawm 86%) | |||
| Grongent | Lub sijhawm (CV) | Cov kuab tshuaj B (%) | Lub sijhawm (CV) | Cov kuab tshuaj B (%) |
| 0 | 10 | 0 | 0 | |
| 10 | 10 | 10 | 0 | |
| 10.1 | 100 | 10.1 | 100 | |
| 17.5 | 100 | 17.5 | 100 | |
| 17.6 | 10 | 17.6 | 0 | |
| 22.6 | 10 | 22.6 | 0 | |
Cov txiaj ntsig thiab kev sib tham
A Sepaflash C18Aq RP Flash Cartridge tau siv rau cov qauv desalting thiab purification. Cov kauj ruam Gradient tau siv nyob rau hauv cov dej ntshiab tau siv raws li cov theem mobile thaum pib kev xaiv tsa thiab khiav rau 10 daim ntawv tshaj tawm (CV). Raws li muaj nyob hauv daim duab 3, thaum siv dej ntshiab li cov mobile theem, cov qauv tau khaws tag nrho ntawm lub nyem cartridge. Tom ntej no, methanol hauv cov theem mobile tau nce ncaj qha rau 100% thiab cov gradient tau tswj hwm rau 7.5 CV. Cov qauv tau raug kaw los ntawm 11.5 txog 13,5 CV. Hauv kev sau cov zauv feem, cov qauv daws tau hloov los ntawm NAH2POPO4 Buffer daws rau methanol. Muab piv nrog cov tshuaj aqueous, Methanol tau yooj yim dua kom raug tshem tawm ntawm cov kauj ruam qeeb qeeb, uas ua rau cov kev tshawb fawb tom qab.
Daim duab 3. Lub Flash Chromoram ntawm cov qauv ntawm C18AQ Cartridge.
Txhawm rau sib piv cov kev ua tsis tau ntawm C18aq Cartridge thiab cov cartridge rau cov qauv ntawm muaj zog polarity, kev sib piv sib piv tau ua. A Sepaflash C18 RP Flash Cartridge tau siv thiab Flash ChromoRam rau cov qauv c18 cartridges, lub siab tshaj plaws zam aqueous theem yog li 90%. Vim li no pib pib gradient tau teem rau 10% methanol hauv 90% dej. Raws li muaj nyob hauv Daim Duab 4, vim yog cov saw hlau dollapse ntawm C18 cov qauv tshwm sim los ntawm cov kab mob C18 tsis tu ncua thiab raug kaw ncaj qha los ntawm cov theem mobile. Raws li qhov tshwm sim, kev ua haujlwm ntawm cov qauv uas ntxim nyiam lossis ua kom huv tsis tuaj yeem ua tiav.
Daim duab 4. Lub Flash ChromoRamram ntawm cov qauv ntawm cov cartridge li niaj zaus c18 cartridge.
Piv nrog linear gradient, kev siv kauj ruam gradient muaj cov txiaj ntsig hauv qab no:
1. Kev siv cov kev siv thiab khiav lub sijhawm rau cov piv txwv purification yog txo.
2. Lub hom phiaj khoom tau txais txiaj ntsig hauv lub ntsej muag ntse, uas txo qhov ntim ntawm cov leb feem thiab yog li ua rau cov neeg sib sau ua ke rau kev txuag lub sijhawm.
3. Cov khoom khaws tau yog nyob hauv Methanol uas yooj yim yuav tsum tau evaporated, yog li ntawd kom qhuav lub sijhawm yog txo.
Hauv kev xaus, rau kev ua kom huv ntawm cov qauv uas muaj zog polar lossis hydrooghilic, Sepaflash C18aq RP flash carratogrigges sepabograph
Txog Sepaflash Bonded Series C18 RP Flash Cartridges
Muaj cov koob c18aq RP flash cartridges nrog cov sib txawv los ntawm cov thev naus laus zis sib txawv (raws li qhia hauv Table 2).
| Yam khoom | Kem li | Txaus tus nqi (ml / min) | Max.pressure (psi / bar) |
| SW-5222-004-SP (AQ) | 5.4 g | 5-15 | 400 / 27.5 |
| SW-5222-012-AQ) | 20 g | 10-25 | 400 / 27.5 |
| SW-5222-025-SP (AQ) | 33 g | 10-25 | 400 / 27.5 |
| SW-5222-040-SP (AQ) | 48 g | 15-30 | 400 / 27.5 |
| SW-5222-080-SP (AQ) | 105 g | 25-50 | 350 / 24.0 |
| SW-522-120-SP (AQ) | 150 g | 30-60 | 300 / 20.7 |
| SW-5222-220-SP (AQ) | 300 g | 40-80 | 300 / 20.7 |
| SW-522-330-SP (AQ) | 420 g | 40-80 | 250 / 17.2 |
Rooj 2. Sepaflash C18aq RP Flash cartridges.
Cov ntaub ntawv ntim: High-Efficiticquicical spherical c18 (aq) -bonded silica, 20 - 45 μ5 μ5 μm, 100 Å.
Logy (raws li qhia hauv Table 2).
Lub sij hawm Post: Lub Yim Hli 27-2018