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In this report, we have studied the formation of iron-oxide nanoparticle at biologically relevant phospholipids, DPPC Langmuir monolayer at air/water interface. Water subphase contains FeCl₃. Adsorption and agglomeration of Fe³⁺ ions at DPPC head group have being monitored by Langmuir and Langmuir Blodgett (LB) technique. Adsorption kinetics (π–t) as well as the surface pressure area (π–A) isotherms measurement demonstrate the incorporation of Fe³⁺ ion at DPPC monolayer. The amount of incorporation of Fe³⁺ to the DPPC monolayer is FeCl₃ concentration and time dependent. This reaction kinetics is well fitted by single exponential association equation. The composite monolayers transferred to different substrates are characterized by UV–vis absorption spectroscopy and electron microscopy (FE-SEM and HR-TEM). Study shows the formation of monodisperse Fe₃O₄ nanoparticle having size …