We describe experimental and theoretical studies of the third-order nonlinear optical coefficients of methyl orange solutions under different pH conditions. A combination of semiempirical and ab initio methods was adopted to investigate the most stable geometrical structures possible for this molecule. The experimental data obtained using the Z-scan technique for the third-order nonlinear optical properties of this compound has allowed the determination of the nonlinear refractive index and nonlinear absorption coefficient under picosecond excitation in the visible (532 nm) spectral region. From those experimental results, the second hyperpolarizability of methyl orange was inferred both for acidic and alkaline solutions. Comparison of these values to the results predicted by semiempirical methods suggests that even at low pH, when the probability of cis-trans isomerization is increased, the trans conformation of the methyl orange molecule should dominate the nonlinear spectra of this compound. The theoretical results were used as an auxiliary tool to identify possible trends on the nonlinear properties changes as a function of the distinct molecular conformations adopted by the methyl orange molecule under different pH conditions.