Determination of the Aggregation Number of Pyrene-Labeled Gemini Surfactant Micelles by Pyrene Fluorescence Quenching Measurements

A cationic gemini surfactant referred to as Py-3-12 and composed of two alkylated diammonium bromide head groups, a propyl spacer, and dodecyl and 1-pyrenehexyl hydrophobic tails was synthesized. Its critical micellar concentration (CMC) was determined to equal 0.15 (±0.02) mM by surface tension and time-resolved fluorescence measurements. The state of the pyrene molecules, whether they were incorporated inside the Py-3-12 micelles or unassociated in the aqueous solution, was determined by applying the global model-free analysis (MFA) to the fluorescence decays acquired with Py-3-12 aqueous solutions. The unassociated Py-3-12 surfactants emitted as pyrene monomers and showed a long fluorescence lifetime. The excited pyrenyl groups located inside Py-3-12 micelles formed an excimer by a rapid encounter with a ground-state pyrene with an average rate constant equal to 0.69 (±0.06) ns–¹. After having the photophysical properties of Py-3-12 in aqueous solution characterized, the number (Nₐgg) of surfactants per micelle was determined by conducting quenching experiments with dinitrotoluene (DNT). Although DNT is fairly hydrophobic, it was found to partition itself between the Py-3-12 micelles and the aqueous phase. Fluorescence quenching experiments performed on the pyrene monomer and excimer generated by the Py-3-12 aqueous solutions yielded the concentration ([Q]b) of DNT bound to the Py-3-12 micelles and the average number ⟨n⟩d of DNT quenching an excimer by diffusive encounters. A combination of steady-state and time-resolved fluorescence quenching experiments on the excimer yielded the number (⟨n⟩ₛ) of DNT molecules that were bound to the micelles and quenched the excimer in a static manner. A plot of the sum ⟨n⟩d + ⟨n⟩ₛ as a function of [Q]b yielded an Nₐgg value of 14.0 (±0.2) Py-3-12 units per micelle. This study represents the first example in the literature where Nₐgg is determined for a micelle, where each surfactant molecule is labeled with pyrene.