Hongcheng Wang, Bo Xu
Applicationem R & D Center
Introductio
Secundum ad relativum polareties ex stationarius tempus et mobile tempus, liquid chromatography potest dividitur in normalis phase Chromatography (NPC) et reversed Phase Chromatography (RPC). Nam RPC, ad vectigalis mobile tempus fortior est quam in stationibus Phase. RPC est facti maxime late usus est in liquid chromatography separationis modi ex eius princeps efficientiam, bonum resolutio et patet retention mechanism. Igitur RPC est idoneam ad separationem et purificationem varii Suspendisse vel non-Sulpicius componit, comprehendo alkaloids, carbohydrates, pingue acida, steroids, nucleic acida, in RPC, peptides, nucleic, etc. in RPC matrix, C8, C8, C4, C18, C8, C4: Cyano, Amino, etc. Inter haec bonded eget coetus, maxime late usus est c18. Est aestimari quod plus quam LXXX% of RPC nunc per C18 religata tempus. Igitur C18 Chromatography columna facta est esse, ut universalis columnae pro omnibus laboratorium.
Licet C18 columnae potest esse in late range of applications tamen aliqua exempla quae est Suspendisse aut altus hydrophilic, iusto C18 columnas potest habere problems cum solebat purificare huiusmodi exempla. In RPC, the commonly used elution solvents can be ordered according to their polarity: water < methanol < acetonitrile < ethanol < tetrahydrofuran < isopropanol. Certe bonum retention in columna horum exempla (fortis Suspendisse aut altus hydrophilic), summus ratio aqueum ratio necesse est utendum mobile tempus. However, when using pure water system (including pure water or pure salt solution) as the mobile phase, the long carbon chain on the stationary phase of C18 column tends to avoid the water and mix with each other, resulting in an instantaneous decrease in the retention capacity of the column or even no retention. Hoc phaenomenon dicitur "Hydrophobic phase ruina" (ut ostensum est in sinistra parte figure I). Etsi hoc situ est reversible cum columna lavabitur cum organicum solvents ut methanolum aut acetonitrile, quod adhuc potest damnum ad columnam. Igitur necessarium ne hoc situ ex fieri.
Figura I. Et schematic diagram in ponderibus in superficie Silica gel in iusto C18 columna (left) et C18aq columna (ius).
Ad oratio in supradictis problems, chromatographic sarcina materiae manufacturers fecerunt technica improvements. Unum ex his melioramentis est facere quaedam modifications super superficiem de silica matrix, ut introductio hydrophilic cyano coetibus (ut ostensum est in dextro figure I), ut superficies Silica gel magis hydrophilic. Ita c18 vincula in silica superficies potuit plene extenditur in altus aqueum condiciones et hydrophobic tempus ruina vitanda. Haec mutatio C18 columnas dicuntur Aqueum C18 columnas, nimirum c18aq columnas, quae sunt disposito altus aqueous Helke condiciones et potest tolerare C% aqueous ratio. C18AQ columnas sunt late applicantur in separationem et purificationem fortis Suspendisse componit, inter organicum acida, peptides, nucleosides et aqua-solutum vitamins.
Desalting est unus de typical applications de c18aq columnas in Flash purificacionem pro samples, quod removet sal et quiddam components in sample solvente ad faciliorem applicationem in sample in sequentibus studiis. Hic post, in clarissimum hyacintho FCF cum fortis verticitas usus est ut sample et purificatum in C18aq columnae. Suus samptu solvendo reponi a SOLUTIO SOLUTIO solvendo, ita facilius sequenti gyratorius evaporatio tum salvo solvents et operating est. Ceterum puritas sample est emendatam ab removendo aliquid immunditia in sample.
Sectionalis
Figura II. In eget structuram de sample.
Et egregie hyacintho FCF cum usus est in sample hoc post. Et puritas est rudis sample erat LXXXVI% et chemical structuram de sample ostensum est in Figura II. Ut parare sample solutio, CCC mg pulsu in I m nah2po4 quiddam solution et excussit in I m nah2po4 solution. Et sample solution est tunc infusum in mico columna per injector. Et experimentum setup de mico purificatione est enumerantur in mensa I.
| Instrumentum | Sepabean ™ Machina2 | |||
| Cartridges | XII G Sepeflashsh C18 RP Flash Cartridge (Sphaerical Silica, XX - XLV μm, C å, ut Number: Sw-5222-012, SP) | XII G Sepeflashsh c18aq RP Flash Cartridge (Sphaerical Silica, XX - XLV μm, C å, ut Number: Sw-5222-012-SP (AQ)) | ||
| RECONCINNUS | CCLIV NM | |||
| Mobile Phase | Solvent A: Aqua Solvent B: Methanol | |||
| Rate fluunt | XXX ml / min | |||
| Sample loading | CCC mg (egregie hyacintho FCF cum puritate LXXXVI%) | |||
| Clivus | Tempus (CV) | Solvent B (%) | Tempus (CV) | Solvent B (%) |
| 0 | 10 | 0 | 0 | |
| 10 | 10 | 10 | 0 | |
| 10,1 | C | 10,1 | C | |
| 17.5 | C | 17.5 | C | |
| 17,6 | 10 | 17,6 | 0 | |
| 22.6 | 10 | 22.6 | 0 | |
Results et disputationem
A separabis c18aq RP mico cartridge est usus ad sample desalting et purificatio. Step gradientis usus est in quo pura aqua erat usus est ut mobile tempus ad initium eludendi et currere ad X columna voluminibus (cv). Ut ostensum est in Figura III, cum per aquam aquam ut mobile tempus, in sample erat omnino retinuit in mico cartridge. Deinde, in Methanol in mobile tempus erat directe auctus ad C% et gradiente esse servetur 7.5 c. De sample erat eluted ex 11,5 ad 13.5 CV. In collectis partibus, in sample solutio reponitur a Nah2po4 quiddam solution ad methanolum. Comparet cum altus aqueum solution, Methanol erat multo facilius remota per gyranni evaporatio in subsequentem gradum, quae faciliorem sequentibus investigationis.
Figura III. Et Flash chromatogram de sample in c18aq cartridge.
Compare retention moribus C18AQ cartridge et iusto C18 cartridge ad exempla fortis verticitatem parallelarum test fetus. A separabis c18 RP Flash Cartridge erat usus et Flash chromatogram pro sample ostensum est in Figura IV. Nam iusto C18 cartridges, summa tolerata aqueous phase de XC%. Igitur initium gradiente erat posuit ad X% Methanol in XC% aqua. Ut ostensum est in Figura IV, debitum ad Hydrophobic Phase ruina C18 vincula per altum aqueum Ratio, in sample vix retinuit in regularis C18 cartridge et directe elutu ex mobili tempus. Ex causa, quod operatio specimen desalting et purificatio non perficitur.
Figura IV. In Flash chromatogram de sample in iusto C18 cartridge.
Comparet cum linearibus gradiente, in usum gradiente habet sequenti commoda:
I. solvendo usus et currere tempus pro specimen purificationis reducitur.
II. In scopum productum elutis in acuto apicem, quod reducit volumine collectis partium ita faciliorem sequenti gyratorius evaporatio tum salvo tempore.
III. In collected uber est in Methanol quod est facile evaporationem, sic siccatio tempus reducitur.
In conclusione, ad purificationem sample quod est valde Suspendisse vel altus hydrophilic, Sepaflash c18aq RP mico cartridge combining cum praeparative mico chromatography system Sepabean ™ apparatus poterat offerre ieiunium et efficientem solutio.
De Series Bonded Series C18 RP Flash Cartridges
Sunt series in Sepaflash c18aq RP mico cartridges cum diversis specifications a Santai technology (ut ostensum est in Table II).
| Item numerus | Columna | Rate fluunt (M / min) | Max.pressure (Psi / Bar) |
| Sw-5222-004-SP (AQ) | 5.4 g | 5-15 | CD / 27.5 |
| Sw-5222-012, SP (AQ) | XX g | 10-25 | CD / 27.5 |
| Sw-5222-25-SP (AQ) | XXXIII g | 10-25 | CD / 27.5 |
| Sw-5222-040-SP (AQ) | XLVIII G | 15-30 | CD / 27.5 |
| Sw-5222-080-SP (AQ) | CV g | 25-50 | CCCL / 24.0 |
| Sw-5222-120, SP (AQ) | CLV g | 30-60 | CCC / 20.7 |
| Sw-5222-220-SP (AQ) | CCC g | 40-80 | CCC / 20.7 |
| Sw-5222-330-SP (AQ) | CDXX G | 40-80 | CCL / 17.2 |
Table II. Sepaflash c18aq RP mico cartridges.
Stipare materiae, summus efficientia sphaerici C18 (AQ) -Bonded Silica, XX - μm XLV, C a.
logy (ut ostensum est in Tabula II).
Post tempus: Aug-27-2018