A surface-enhanced Raman scattering (SERS) method for measuring nitrate nitrogen in aquaculture water was developed using a substrate of β-cyclodextrin-modified gold nanoparticles (SH-β-CD@AuNPs). Addressing the issues of low sensitivity, narrow linear range, and relatively poor selectivity of single metal nanoparticles in the SERS detection of nitrate nitrogen, we combined metal nanoparticles with cyclodextrin supramolecular compounds to prepare a AuNPs substrate enveloped by cyclodextrin, which exhibits ultra-high selectivity and Raman activity. Subsequently, vanadium(III) chloride was used to convert nitrate ions into nitrite ions. The adsorption mechanism between the reaction product benzotriazole (BTAH) of o-phenylenediamine (OPD) and nitrite ions on the SH-β-CD@AuNPs substrate was studied through SERS, achieving the simultaneous detection of nitrate nitrogen and nitrite nitrogen. The experimental results show that BTAH exhibits distinct SERS characteristic peaks at 1168, 1240, 1375, and 1600 cm−1, with the lowest detection limits of 3.33 × 10−2, 5.84 × 10−2, 2.40 × 10−2, and 1.05 × 10−2 μmol/L, respectively, and a linear range of 0.1–30.0 μmol/L. The proposed method provides an effective tool for the selective and accurate online detection of nitrite and nitrate nitrogen in aquaculture water.