Concurrent analysis of nucleoside reverse transcriptase inhibitors in a pool of endogenous nucleosides by short-end injection-capillary electrochromatography on a beta-cyclodextrin-bonded stationary phase.
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.
Mesplet
Nathalie
N
Institut de Chimie Organique et Analytique, Université d'Orléans, Orléans, France. nathalie.mesplet@univ-orleans.fr
Morin
Philippe
P
Agrofoglio
Luigi A
LA
D000097:Acetonitriles / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000302:isolation & purification; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D003505:Cyclodextrins / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008962:Models, Theoretical; D009705:Nucleosides / Q000302:isolation & purification*; D009994:Osmolar Concentration; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D047392:beta-Cyclodextrins
D000097:Acetonitriles; D019380:Anti-HIV Agents; D002021:Buffers; D003505:Cyclodextrins; D009705:Nucleosides; D018894:Reverse Transcriptase Inhibitors; D047392:beta-Cyclodextrins; C031215:betadex; C032159:acetonitrile