This paper deals with the synthesis of a series of hexadecyl o-xylene sulfonate isomers (with the o-xylene ring located at different positions along the n-hexadecyl chain) by a Friedel-Crafts reaction, and the Grignard reaction followed by a hydrogenation. The structure was confirmed by 1 H NMR. All analytical methods indicated high levels of purities of the isomers with the orthoxylene ring located at the first, third, fifth and seventh carbon atom on the n-hexadecane chain. The critical micelle concentration (CMC), surface tension and maximum surface excess concentration at CMC and area per molecule at the interface were determined. As the o-xylene sulfonate group is shifted toward the center of the hexadecyl chain, the branching degree is enhanced and the surfactant molecule tends to produce a much looser packing at the gas-liquid interface. Accordingly, at CMC, the adsorption density decreases, the CMC increases and the tension reduction is weakened.