Surface-enhanced Raman scattering (SERS) Spectroscopy and atomic force microscopy (AFM) were employed to characterize the Langmuir-Blodgett (LB) monolayers of an amphiphilic nitroaniline derivatives, n-docosyl-2-methyl-4-nitroaniline (DCMNA) on both Ag surface and mica. DCMNA formed a stable, organized and homogeneous monolayer at the air-water interface only under the specific conditions. Structural changes at the interface induced by surface pressure, temperature and pH of the subphase, and the mole ratio of mixed molecules have been studied. The stable monolayers of DCMNA were obtained on a neutral subphase maintained lower than 10 degrees C. The compressibility of the monolayer was improved by addition of arachidic acid or spreading the monolayer on an alkaline subphase. Micro-Raman and AFM studies indicated that pure DCMNA on a neutral subphase spontaneously formed a well ordered crystalline phase as well as an organized monolayer even under a lower surface pressure. AFM and lateral force microscopy (LFM) confirmed the arachidic acid minimizes self-aggregation of DCMNA and induces formation of monodispersed nano sized domain in the mixed monolayer.