Donkuru, McDonaldMichel, DeborahAwad, HananKatselis, GeorgeEl-Aneed, Anas2016-12-192016-12-192016-05-13Donkuru, M., Michel, D., Awad, H. et al. J Chromatogr A. 2016 1446: 114-24.http://hdl.handle.net/10388/7636Diquaternary gemini surfactants have successfully been used to form lipid-based nanoparticles that are able to compact, protect, and deliver genetic materials into cells. However, what happens to the gemini surfactants after they have released their therapeutic cargo is unknown. Such knowledge is critical to assess the quality, safety, and efficacy of gemini surfactant nanoparticles. We have developed a simple and rapid liquid chromatography electrospray ionization-tandem mass spectrometry (LC-ESI–MS/MS) method for the quantitative determination of various structures of gemini surfactants in cells. Hydrophilic interaction liquid chromatography (HILIC) was employed allowing for a short simple isocratic run of only 4 minutes. The lower limit of detection (LLOD) was 3 ng/mL. The method was valid to 18 structures of gemini surfactants belonging to two different structural families. A full method validation was performed for two lead compounds according to USFDA guidelines. The HILIC-MS/MS was compatible with the physicochemical properties of gemini surfactants that bear a permanent positive charge with both hydrophilic and hydrophobic elements within their molecular structure. In addition, an effective liquid-liquid extraction method (98% recovery) was employed surpassing previously used extraction methods. The analysis of nanoparticle-treated cells showed an initial rise in the analyte intracellular concentration followed by a maximum and a somewhat more gradual decrease of the intracellular concentration. The observed intracellular depletion of the gemini surfactants may be attributable to the bio­transformation into metabolites and exocytosis from the host cells. Obtained cellular data showed a pattern that grants additional investigations , evaluating metabolite formation and assessing the subcellular distribution of tested compounds.enAttribution-NonCommercial-NoDerivs 2.5 CanadaDrug delivery nanoparticles; Gemini surfactants; HILIC chromatography; LC–MS/MSPostprint10.1016/j.chroma.2016.04.01327086283