The structure of sea spray aerosols (SSAs) has been described as a saline core coated by organic surfactants. The presence of surface-active compounds at the air-water interface can have a large impact on physical, chemical and optical properties of SSAs. The surfactant molecules chosen for this study, palmitic acid (PA), stearic acid (SA), arachidic acid (AA), methyl palmitate (MP), methyl stearate (MS) and methyl arachidate (MA), were used to investigate the effect of alkyl chain-length, head-groups and sea salts on the surface properties of these monolayers. A Langmuir trough was used for measuring surface pressure−area (π−A) isotherms to reveal macroscopic phase behavior of the surface films at the air-water interface. Infrared reflection absorption spectroscopy (IRRAS) was employed to have a molecular-level understanding of the interfacial molecular organization. The π−A isotherms indicated that sea salts, present in the subphase, exert a strong condensing effect on fatty acid monolayers, while exerting expanding effect on fatty acid methyl ester monolayers, which was confirmed by results from IRRAS experiments. IRRAS further revealed that the alkyl chains were in an all-trans conformation, which can be evidenced by the relatively low νa(CH2) and νs(CH2) stretching frequencies. The conformational order changes in the alkyl chains of different film-forming species (C16 < C18 < C20) were directly revealed by analyzing the relative intensity of the νa(CH2) and νs(CH2) peaks in the C-H stretching region. Thus, all the three factors alter the phase behavior and molecular packing of the monolayers at the air-aqueous interface.