In this paper, the critical micelle concentration (cmc) values of sodium dodecyl benzene sulfonate (SDBS) in aqueous solution at different concentrations of hydroxypropyl-β-cyclodextrin (HP-β-CD) are determined by synchronous fluorescence spectrometry. The results indicated that, when the scanning wavelength difference of the synchronous fluorescence spectrum is 25 nm, both the critical micelle concentration value and the fluorescence intensity of SDBS will greatly increase in the presence of HP-β-CD. The standard molar Gibbs free energy for SDBS from aqueous HP-β-CD solution to the micelle, ΔγGΘm, increases with increasing HP-β-CD concentration in aqueous solution, which showed that SDBS molecules are more likely to form inclusion complexes with HP-β-CD rather than micelles. The results of Job’s plot for inclusion complexation of SDBS with HP-β-CD indicated that “β-CD/SDBS” inclusion should be formed with the molar ratio of 1∶1 in aqueous solution. The effect of the formation of SDBS micelle on the quantitative determination of SDBS could be greatly reduced by adding HP-β-CD with the molar ratio of 1∶1. Thus, whether higher or lower than the critical micelle concentration value, the concentration of SDBS in water samples from T5-X15 and T9-X4 sites in LinPan oilfield could be calculated by establishing the quantitative standard curve of SDBS in aqueous HP-β-CD solution, and the recovery rate of SDBS was 100.5%~101.2%. The results of 1H-NMR and FT-IR showed that the phenyl group of SDBS molecule is likely located within the broad mouth of HP-β-CD molecule.