The role of tetradecyltrimethylammonium bromide (TTAB) and its partition between water, chloroform, and the chloroform/water interface during the miniemulsification of a photoluminescent polymer was investigated by indirect interfacial tension/elasticity measurements. Dynamic interfacial tension and elasticity measurements showed the presence of a gas-liquid phase transition at the chloroform/water interface and the formation of a rigid interface, which was supposed to promote emulsion stability. The parameters of the adsorption isotherms and the TTAB partition coefficient were obtained from surface tension isotherms. Dynamic surface tension measurements performed after TTAB water/chloroform extraction were used to compute TTAB partition between water, chloroform, and the chloroform/water interface. Model calculations allowed identifying (for the tested conditions) the minimum size of emulsion droplets before the onset of instability and the segregation of a sizable amount of TTAB in the final polymer nanoparticles, which induced a shift in the 2,7-poly(9,9-dialkylfluorene-co-fluorenone) (PF) photoluminescence emission band. The size of the emulsion droplets of the final polymer particles and the amount of segregated TTAB were in good agreement with the corresponding experimental values.