HPLC Separation of Oligonucleotides in Isocratic and Temperature-Programming Mode

Djordjevic, Nebojša M.; Houdiere, Fabrice; Fowler, and Patrick; Natt, François

doi:10.1021/ac9711697

Cited by 24 publications

(14 citation statements)

References 20 publications

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“…(17) and (18). We note that the applicability of the second approach discussed in Section 2.3 is very restricted since its predictions are expected to be satisfactory only if the gradients are of the same kind with those used in the fitting procedure and the conditions lie within those used for fitting.…”

Section: Gradients Of T At Constant Umentioning

confidence: 99%

Combined effect of temperature and organic modifier concentration on the retention under single mode gradient conditions in reversed‐phase HPLC

Pappa-Louisi¹,

Nikitas²,

Zisi³

et al. 2008

J of Separation Science

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The combined effect of temperature, T, and organic modifier concentration, phi, on the retention under gradient conditions in RPLC is studied by considering, both theoretically and experimentally gradients, of phi at constant T and gradients of T at constant phi. Two approaches are examined: in the first approach the prediction of the elution time of a sample solute is based on the isocratic/isothermal properties of this solute. The second approach is based on a direct fitting procedure of a proper retention model to 2-D isocratic/T-gradient or isothermal/phi-gradient retention data. These approaches were tested using alkylbenzes in eluting systems modified by ACN. We found that both approaches can give excellent predictions under certain prerequisites. However, the first approach exhibits the notable advantage that it can be used effectively to predict retention times under any kind of phi-gradients at constant T or T-gradients at constant phi. The second approach has the advantage that it is relatively simple but its applicability is very restricted since its predictions are satisfactory only if the gradients are of the same kind with those used in the fitting procedure and the conditions lie within those used for fitting.

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Section: Gradients Of T At Constant Umentioning

confidence: 99%

Combined effect of temperature and organic modifier concentration on the retention under single mode gradient conditions in reversed‐phase HPLC

Pappa-Louisi¹,

Nikitas²,

Zisi³

et al. 2008

J of Separation Science

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“…The separation and detection of these compounds is an interesting and challenging topic. The analysis of nucleosides and nucleotides has in most cases been performed by high-performance liquid chromatography (HPLC) using either reversed-phase HPLC with gradient elution or reversed-phase ion-pairing chromatography [4][5][6][7][8][9][10][11]. Electrophoresis is the major separation method in molecular biology.…”

Section: Introductionmentioning

confidence: 99%

Nucleoside monophosphates recognition using macrocyclic polyamine bonded phase in capillary electrochromatography

2002

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An open-tubular wall-coated macrocyclic polyamine capillary column (70 cm x 75 microm ID) with 50 cm effective length for the separation of nucleoside monophosphates is described. Some parameters with respect to concentration, pH, composition of the buffer, and voltage in order to optimize the separation were studied. The coated capillary showed reversed electroosmotic flow (EOF), allowing anions to be separated in the co-EOF mode. Baseline separations were achieved for the eight nucleotides in less than 26 min using a background electrolyte consisting of H(3)PO(4)-NaH(2)PO(4) (30 mM, pH 3.10), an applied voltage of -15 kV, and detection at 254 nm. The macrocyclic polyamine on the capillary wall introduced anion coordination for the interaction with the analytes, the strength of which could be moderated by the type and concentration of the competing ion used in the background electrolyte (BGE). With a low concentration of the competing ion (phosphate ion), the migration behavior followed that obtained in the electrophoretic system. Increasing the concentration of the competing ion resulted in a faster migration and more complete elution of the analyte. The method established was also employed for the analysis of nucleotides in mushrooms. Aqueous extracts of mushrooms from different species and various extraction methods were injected directly for the analysis. Uridine 5'-monophosphate, guanosine 5'-monophosphate, adenosine 5'-monophosphate, and cytidine 5'-monophosphate, were found in the sample tested.

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“…ODS-AQ stationary phase has been used for the separation of polar compounds in liquid chromatography [5]. The symmetrical peak shapes and increased solute interaction in comparison to many conventional octadecylsilica (ODS) materials, characterize liquid chromatographic separations of polar compounds on the ODS-AQ phase even in a highly aqueous mobile phase.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of ODS-AQ stationary phase for use in capillary electrochromatography

2001

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The aim of this study was to evaluate the applicability of ODS-AQ packing material as a stationary phase in capillary electrochromatography (CEC). The electroosmotic flow created on an ODS-AQ stationary phase was measured at different mobile phase compositions and at different column temperatures. It was observed that the electroosmotic flow generated in the column increased by 50% when the temperature of the system was raised from 20 degrees C to 60 degrees C, while all other conditions were kept constant. The electroosmotic flow produced by the ODS-AQ stationary phase was found to be comparable to the flow generated in a column packed with Nucleosil bare-silica material. In addition, a set of polar compounds (D-lysergic acid diethylamide derivatives) was utilized to determine the influence of temperature and mobile phase composition on their chromatographic behavior on an ODS-AQ stationary phase in a CEC mode. A linear relationship between the solute retention factor and column temperatures was seen over the temperature range studied (20 degrees C to 60 degrees C). A quadratic function was used to describe the changes in the solute retention factors with variation of acetonitrile concentration in the mobile phase.

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HPLC Separation of Oligonucleotides in Isocratic and Temperature-Programming Mode

Cited by 24 publications

References 20 publications

Combined effect of temperature and organic modifier concentration on the retention under single mode gradient conditions in reversed‐phase HPLC

Combined effect of temperature and organic modifier concentration on the retention under single mode gradient conditions in reversed‐phase HPLC

Nucleoside monophosphates recognition using macrocyclic polyamine bonded phase in capillary electrochromatography

Evaluation of ODS-AQ stationary phase for use in capillary electrochromatography

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