Concurrent analysis of nucleoside reverse transcriptase inhibitors in a pool of endogenous nucleosides by short-end injection-capillary electrochromatography on a β-cyclodextrin-bonded stationary phase

Abstract

As part of our on-going study of the analysis of anti-human immunodeficiency virus (HIV) nucleosides, a capillary electrochromatography (CEC) method has been developed for the concurrent analysis of nucleoside HIV reverse transcriptase inhibitors (NRTIs) in a pool of endogenous nucleosides. Up to now, beta-cyclodextrin-bonded silica stationary phases have mainly been dedicated to the separation of enantiomers; however, these polysaccharides can be also be used in achiral way. This work aims at showing how CEC performed on a beta-cyclodextrin-bonded silica stationary phase can be used to concurrently resolve zidovudine (AZT), lamivudine (3TC), didanosine (ddA) and its administrated form (ddI), stavudine (d4T) and hivid (ddC) in a mixture of adenosine (A), cytidine (C), guanosine (G), thymidine (T) and uridine (U). The influence of several parameters (pH buffer, ionic strength, acetonitrile content, temperature and voltage) on both the retention times and the retention factors has been investigated using the short-end injection technique to achieve baseline separation in a short-time analysis before quantitation. Moreover, the retention factors of the charged solutes in short-end injection-CEC were calculated using theoretically derived equations, allowing for the actual voltage drop in the packed section of the semipacked CEC capillary.

Nucleoside reverse transcriptase inhibitors and their phosphorylated metabolites in human immunodeficiency virus-infected human matrices

“…In this method, a beta-cyclodextrin-bonded silica packed column was applied to separate the NRTIs, taking advantage of the internal hydrophobicity of the polysaccharide. The influence of several parameters such as buffer pH, ionic strength, ACN content, temperature and voltage has been investigated using the short-end injection technique to achieve baseline separation in a short-time analysis [61,62].…”

CEC of phytochemical bioactive compounds

“…More than ten nucleosides and their related compounds, including adenine, adenosine, uracil, uridine, guanine, guanosine, hypoxanthine, inosine, thymine, thymidine, deoxyuridine and cordycepin, have been isolated and/or identified 105. For the analysis of nucleosides and their analogues, granular packed CEC 29, 33, 106–111, monolithic CEC 74, 112, 113 and OT‐CEC 114, 115 have been used, but few was employed for the analysis of real samples 33, 74 (Table 2). Therefore, it is worthy to employ CEC especially monolithic and OT‐CEC for the analysis of nucleosides and their analogues in different matrices.…”

The separation of biomolecules using capillary electrochromatography

“…Mesplet et al [43,44] separated and quanti®ed the nucleoside HIV reverse transcriptase inhibitors by short-end injection capillary electrochromatography on a b-cyclodextrin-bonded silica stationary phase, proving that CEC would be a suitable tool for both the quantitative determination and the assay of a mixture of nucleosides. …”