Hydrogen peroxide (H₂O₂) is an important chemical substance produced in the metabolic process of organisms. Excess or less production could lead to serious effects on the body. Therefore, the development of advanced technology to accurately detect the content of H₂O₂ is of great significance. Herein, we developed a new ratiometric SERS nanoprobe based on shell-isolated nanoparticles (SHINs) for rapidly quantitative detection of H₂O₂. Because of the small Raman cross-section of H₂O₂, the ratiometric nanoprobe is an effective method for indirect detection of H₂O₂, which is designed based on the reaction of p-mercaptophenylboric acid (MPB) with H₂O₂ to form p-mercaptophenol (MP). Meanwhile, the nanoprobe was used to achieve quantitative detection of H₂O₂ and applied in quantitative detection of actual sample¾glucose, whose linear correlation coefficient could reach 0.9947 and 0.9812, respectively. This method expands the application of SERS technology, especially provides a reference for the detection of molecules with small Raman cross-section.