Quantitative analysis of H2O2-related biomarkers using surface-enhanced Raman spectroscopy technology based on TMB-Au@PB nanoparticles

Background

Hydrogen peroxide (H₂O₂), a pivotal component of reactive oxygen species, is intricately intertwined with numerous pathological and physiological processes within organisms. In the disease diagnosis domain, the precise detection of H₂O₂ and its associated biomarkers, which play crucial roles in discerning disease states, is of paramount importance. However, current methods often suffer from limited sensitivity and reliability. There is an urgent need for a reliable, highly accurate method to detect these substances, aiming to enhance diagnostic precision and enable early disease intervention.

Results

A novel Surface Enhanced Raman Scattering (SERS) substrate was developed by coating Prussian blue (PB) on gold nanoparticles (Au@PB NPs) for the detection of H₂O₂ and biomarkers such as alanine aminotransferase (ALT), cholesterol, and uric acid. The PB Raman signal serves as an ideal and non-interfering internal standard to correct the Raman intensity fluctuations of analytes for accurate quantification. The Au@PB NPs acted as a peroxidase-mimicking enzyme, catalyzing the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H₂O₂. The method achieved a detection limit of 0.19 × 10⁻¹² M for H₂O_2, with a linear range of 10⁻¹² to 10⁻² M. Linear detection ranges for ALT, cholesterol and uric acid were 0.5–100 U/L, 10–500 μM, and 5–600 μM, respectively. The specificity was assessed through interference and cross-reactivity experiments. The results showed negligible interference, stable Raman signals, and a relative error of less than 3 %. Clinical serum samples validation exhibited a high degree of consistency, ranging from 92.49 % to 105.92 %, with an average discrepancy of less than 5 %, demonstrating the method's precision and reliability.

Significance

This study presents a highly sensitive and accurate method for detecting H₂O₂ and key biomarkers, offering significant potential for early diagnosis of liver and kidney damage. The approach provides a robust tool for clinical applications, enabling precise monitoring of disease-related biochemical alterations.