Evaluating the "wrong-way-round" electrospray ionization of antiretroviral drugs for improved detection sensitivity

The presence of antiretroviral drugs (ARVDs) in the aquatic environment poses a significant health risk to the ecosystem. The dilution of these compounds during wastewater treatment processes, followed by discharge into the environment, results in extremely low concentrations in the range of ng/L. Therefore, to enable detection of these low concentrations, it is important to determine the most efficient electrospray ionization (ESI) mode using the right mobile phase modifier and to establish a selective extraction procedure. In this study, we compared the ESI intensity in the positive and negative mode using both formic acid (FA) and ammonium hydroxide (NH₄OH) as mobile phase modifiers. The results revealed a phenomenon known as the "wrong-way-round" (WWR) ESI in which high intensity [M + H]⁺ ions were detected under basic conditions using NH₄OH as modifier and, similarly, high intensity [M-H]^- ions were detected under acidic conditions using FA as modifier. Furthermore, mixed-mode strong cation (MCX) and mixed-mode strong anion (MAX) exchange sorbents were evaluated for extraction recoveries, which yielded extraction recoveries between 60 and 100%. Finally, the recoveries obtained using mixed-mode ion exchange sorbents compared to ion production during the ESI process provide evidence that ions produced in solution do not necessarily reflect the ions that are produced during the ESI process. Based on the results of this study, it is recommended to evaluate the optimal ionization mode under basic and acidic conditions, instead of defaulting to the use of acidic modifiers with positive ion detection.