Steadfast efforts have been made to develop novel materials and incorporate them into functional devices for practical applications, pushing the research on electroactive materials to the forefront of nano electronics. Liquid/liquid interface-assisted polymerization offers a scalable methodology to fabricate hybrid materials with multifunctional applications, in contrast to the conventional and ubiquitous routes. Here, we explored this efficient and versatile approach toward the in situ tailoring of Au-Ag alloy nanostructures with a conducting polymer, poly(3,4-ethylene-dioxythiophene) (PEDOT). With the appropriate choice of organic and inorganic phases for the distribution of monomer and oxidant, the miscibility restraints of the reactants in a single phase were alleviated. Effective nanostructure tuning of highly crystalline and electroactive PEDOT/Au-Ag alloy has been achieved by varying the molar ratio of Au3+/Ag+ in the reaction mixture. The as-synthesized composite is further explored to detect neuromodulator histamine (HA), which displays high sensitivity with a limit of detection (LOD) of 1.5 nM, and selectivity even in the presence of various interfering analogs of 10-fold concentration. Subsequently, density functional theory (DFT) simulations are employed to assess the mode of interaction between HA and the electroactive surfaces. The competency to detect HA in preserved food entails its potential in food spoilage monitoring. Furthermore, the detection of histamine generated by sub-cultured human neuronal cells SH-SY5Y proves its practical viability in health monitoring devices.