Capillary zone electrophoresis of oligonucleotides

Dolnı́k, Vladislav; Liu, Jinping; Banks, J. Fred; Novotný, Miloš V.; Boček, Peter

doi:10.1016/s0021-9673(01)84301-6

Cited by 70 publications

(15 citation statements)

References 15 publications

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“…Other workers [23] observed no change in selectivity when investigating the effect of BGE concentration on polycytidine migration but this is probably attributable to 20 mM being the lowest BGE concentration compared with 10 mM in our experiments. From these experiments, it appears that BGE concentration has little influence on the selectivity of nucleotide separations.…”

Section: Influence Of Bge Concentration On Nucleotide Migration and Econtrasting

confidence: 84%

Electrophoretic behaviour of oligonucleotides and mono-, di- and triphosphate nucleotides by capillary zone electrophoresis

McKeown

Shaw

2001

Electrophoresis

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A systematic investigation has been made into the mechanisms of the capillary zone electrophoresis (CZE) separation of 12 common nucleotides (mono-, di- and triphosphorylated) and polydeoxythymidylic acid oligonucleotides (pd(T)5-18) using electrophoretic mobility values calculated from migration time data. Relationships between electrophoretic mobility and the physicochemical characteristics of the analytes (charge, dissociation constants, charge-to-mass ratio) and the background electrolyte conditions (buffer strength, percentage organic modifier and buffer pH) were characterised. Nucleotide migration was dominated by the negatively charged phosphate groups. Additionally, there were important contributions to migration behaviour from the ionised amide groups of the nucleobases guanine and uracil at higher buffer pH values or with the presence of methanol in the electrolyte. Calculated electrophoretic mobility values for the nucleotides showed a substantially improved (5-fold) inter-run repeatability compared with migration time data. These studies show the value of representing nucleotide migration data as electrophoretic mobility in CZE for obtaining a more thorough analysis of separation mechanisms and to compensate for variation in migration time data caused by small changes in electrosmotic flow. Oligonucleotides pd(T)5-11 could be adequately resolved from their nearest neighbour, but the limit of single-base separation was pd(T)10 from pd(T)11 under the conditions used. It was calculated that a difference in charge-to-mass ratio of 2.64 x 10(-5) was required for resolution under the CZE conditions used.

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Section: Influence Of Bge Concentration On Nucleotide Migration and Econtrasting

confidence: 84%

Electrophoretic behaviour of oligonucleotides and mono-, di- and triphosphate nucleotides by capillary zone electrophoresis

McKeown

Shaw

2001

Electrophoresis

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“…A similar effect was observed with 0.5 M glucose [31]. Under these neutral conditions no interaction of analytes with spermine [32] was observed either. Substitution of NaOH with borates (BGE "C", Section 2.3) improved selectivity of C and AraC with an optimum resolution for a 40 mM SDS concentration ( R = 2.2 for methanol and AraC, R = 1.5 for AraC and C).…”

Section: Separation In Neutral Mediasupporting

confidence: 81%

Separation of aracytidine and cytidine by capillary electrophoretic techniques

et al. 1996

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Aracytidine (cytarabine, 1-beta-D-arabinofuranosylcytosine) is a synthetic analog of cytidine in which ribose is substituted by arabinose; it is used as a drug for the treatment of leukemia. A fast and reliable capillary electrophoretic method for the analysis of cytarabine and cytidine is described. The procedure utilizes the interactions with sodium dodecyl sulfate (SDS) micelles and borate, present in the background electrolyte, for the mobilization and selective separation of the analytes. The detection is carried out by UV absorbance at 275 nm. The method was applied both to pharmaceutical preparations and human serum. Analysis of an untreated serum requires 15 min; the detection limit is 0.8 microgram/mL and the relative standard deviation (RSD) is 5.3%.

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“…In this trend, freesolution mobility data are available from the work by Dolník et al [39] for polycytidine (3 £ N £ 10) in a 30 mM ionic strength histidine-glutamic acid buffer, pH 6. The two pK a values for the cytidine-5©-monophosphoric acid are 4.35 and 6.35 [40].…”

Section: Polycytidinesmentioning

confidence: 99%