Column wall modifications using polyethylene oxide for capillary electrophoresis of ribonucleotides

Abstract

SummaryIn this study, a variety of fused silica capillaries with different combinations and sequences of treatments with HMDS and polyethylene oxide were prepared in order to develop an optimized column modification method for analysis of ribonucleotides. The 12 most common ribonucleotides (UTP, CTP, ATP, GTP, UDP, CDP, ADP, GDP, UMP, CMP, AMP, and GMP) in human cells were used as test solutes. Column performance measurements, including electroosmotic flow '(EOF), solute migration speed and retention, column efficiency, peak shape, and resolution were investigated. By analyzing solute migration speed and retention of various hydrophilic/hydrophobic solutes, the column wall effects (EOF and adsorption) can be distinguished. This analysis method can give guidance in optimizing polymer coating properties (hydrophilicity/hydrophobicity) for CE columns. By studying the performance of these columns after various surface treatments, we were able to improve the separation of ribonucleotides from real samples to within 16 minutes with high efficiency and stability (over 300 analyses) using columns first deactivated with hexamethyldisilazane, and then coated with polyethylene oxide.

Critical factors for high-performance physically adsorbed (dynamic) polymeric wall coatings for capillary electrophoresis of DNA

“…However, LPA coatings have limited stability and lifetime. To address these limitations, other covalently bound coatings have been developed, including those based on poly(vinyl alcohol) (PVA) [9,10], poly(acryloylaminoethoxy-ethanol) (poly(AAEE)) [11], and others [12][13][14][15][16][17]. Some covalently linked coatings, including PVA and poly(AAEE), can be stable for over 100 runs [10].…”

Nucleoside monophosphates recognition using macrocyclic polyamine bonded phase in capillary electrochromatography

“…CE has been proven to have several advantages over HPLC, particularly in terms of its high efficiency and small volume requirements. Here, we only studied those for the separation of nucleoside monophosphates [12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31]. Among them, a number of studies involved the addition of quaternary ammonium salt as EOF modifier [12,25,27,29,30], cyclodextrin [15,23], acetonitrile [20,22], mixture of cyclodextrin and MgCl 2 [23], or metal ions [23,25,27,30] in the background electrolyte (BGE).…”

“…Among them, a number of studies involved the addition of quaternary ammonium salt as EOF modifier [12,25,27,29,30], cyclodextrin [15,23], acetonitrile [20,22], mixture of cyclodextrin and MgCl 2 [23], or metal ions [23,25,27,30] in the background electrolyte (BGE). Examples also included micellar electrokinetic chromatography (MEKC) [23,24], the use of cross-linked polyacrylamide [13], polyethylene oxide [26], diol-bonded [28], and Ucon-coated columns [14,16,18,22], or medium at pH above 10 [19,21,31] to achieve a better separation.…”