Analysis of nucleic acid derivatives by micellar electrokinetic capillary chromatography

Abstract

Summary 2 Materials and MethodsThe use of micellar electrokinetic capillary chromatography (MECC) for the analysis of the major nucleobases, nucleosides, and nucleotides, and their chemically modified derivatives, has been developed and refined.The dimensions of the separating capillaries, the compos~t~on of the buffering systems, and the conditions used for electrophoresis were investigated in order to obtain the best performance. ~~~~~~bphosphodlesterase, micrococcal nuclease, bases, nucleosides, and nuc!eotldes standards were Purchased from Sigma Sodium dodecyl sulfate (SDS) and other chemicals (of HPLC grade) were purchased from Aldrich thymus DNA, Particular emphasis was placed on the identification of the physiological constituents of nucleic acids and their chemically modified analogs: in vitm studies on calf thymus DNA exposed to genotoxic agents have demonstrated that adducted bases and nucleolides can be identified by MECC.

Separation of nucleosides and nucleotide‐3′‐monophosphates by micellar electrokinetic capillary chromatography

“…Different buffers have been previously evaluated [7] for reaching the conditions for simultaneous separation of nucleosides and nucleotide-3 'mP. Among them were monohydrogen phosphate, dihydrogen phosphate, borate, carbonate, glycine, phosphate-borate (pH 6 to 10 and ionic strength 10 to 25 mM), organic modifier (ACN 5 to 1096, isopropyl alcohol l o % , urea 1 M), and SDS concentration (50, 75, 100, 200 &).…”

“…Near neutral pH, nucleosides are neutral molecules, whereas nucleotides have an ionized acidic phosphate group that imparts a double negative charge to the molecule. In a previous paper [7], we reported MECC separations of normal and some modified nucleic acid bases, including deoxy-and ribonucleosides, and deoxynucleotide 5'and 3'-monophosphates (the form in which nucleotides are phosphorylated in the postlabeling technique). An interesting characteristic was that, because of their different behavior in micellar conditions, deoxyribonucleosides were well separated from the ribonucleosides and from the nucleotides.…”

Development and validation of a high-performance liquid chromatography assay and a capillary electrophoresis assay for the analysis of adenosine and the degradation product adenine in infusions

“…CE has also been applied to study the stability of drugs. Several reports describe the separation of adenine and adenosine along with other purine and pyrimidine bases as well as nucleosides and nucleotides by CE [5][6][7][8] including their analysis in biological samples, such as urine [9] or human cord plasma [10], in Xenopus oocytes [11], or in beer samples [12]. However, the analysis of adenosine and adenine in pharmaceuticals, has not been investigated by CE, to date.…”

Enhanced analysis of purine and pyrimidine bases by the use of double-strand polyaniline coatings in micellar electrokinetic capillary chromatography

“…Most purine and pyrimidine bases have a pK a above 8 [6], however, CZE analysis of such bases is mostly performed at high pH values [7][8][9][10][11][12][13][14][15][16]. Alternatively, separations of these bases in acidic media are mostly performed using MEKC [17][18][19][20][21][22][23][24]. The pK a values of the bases used in this study are displayed in Table 1 [25].…”