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In the present investigation, a novel approach towards a complete separation of all 15 protease and reverse transcriptase inhibitors which are currently approved for use in highly active antiretroviral therapy in a single analytical run is presented. The developed method employs an acidic background electrolyte with sodium polyanethol sulfonate (SPAS) as polyanionic electroosmotic flow (EOF) modifier to establish a strong cathodic EOF, sodium dodecyl sulfate (SDS) as pseudostationary selector, and acetonitrile and ethanol as organic modifiers. Separation of the analytes is based on two different mechanisms. The more basic analytes are protonated at the prevailing pH conditions and thus migrate in front of the cathodic EOF, whereas the less basic and neutral analytes interact with the SDS and are retained after the EOF. By optimizing electrolyte pH, the amount of solvents and SDS concentrations in the background electrolyte it is possible to completely separate all compounds of interest.

In the present investigation, a co-electroosmotic capillary zone electrophoretic method is shown for the simultaneous separation of protease inhibitors and reverse transcriptase inhibitors, which are used as antiretroviral therapy drags against the human immunodeficiency virus (HIV). The investigated drugs carry basic amino groups, thus the electrophoretic system takes advantage of an acidic buffer electrolyte. In order to establish a strong cathodic electroosmotic flow (EOF), a poly-anionic surfactant is added to the background electrolyte. Thus, fast migration times due to a co-directional migration of analytes and EOF (co-electroosmotic CE) are obtained. The developed separation system exhibits good selectivities for the investigated compounds and sufficient sensitivity to monitor drug levels in the low ppm range in HIV positive patients who are treated by highly active anitiretroviral therapy.