New rapid detection method of total chlorogenic acids in plants using SERS based on reusable Cu₂O-Ag substrate

A new method for rapidly detecting of total chlorogenic acids (CGAs) in plants by surface-enhanced Raman spectroscopy (SERS) based on reusable Cu₂O-Ag substrate was developed in this study. The Cu₂O-Ag substrate prepared by the in-situ growth method had high uniformity with peak intensity relative standard deviation (RSD) of 5.27%, repeatability with peak intensity RSD of 3.58%, and sensitivity with an analytical enhancement factor of 1.27 × 10⁵ for detecting CGAs. Furthermore, the substrate had excellent reusability, after it was reused for seven cycles, the signal strength of CGAs was still above 80% of initial. Compared with the standard method of high-performance liquid chromatography (HPLC), the SERS method can successfully analyze the contents of total CGAs in plants, such as Stevia rebaudiana leaves, coffee beans, Lonicera japonica leaves, and Eucommia ulmoides flowers, with recovery rate from 93.26% to 112.65%, and the limit of detection was 0.13 μg/mL. The total CGAs content of Stevia rebaudiana leaves samples detected by HPLC and SERS have good consistency with R = 0.9760 and RMSE = 3286 mg/kg. Furthermore, the SERS method only needed less than 1 min, one standard and reusable substrate in this study to analyze, which can further reduce the cost of method analysis. Therefore, the SERS method with the appropriate substrate can provide a rapid, accurate, and economic way to detect the total CGAs in plants.