<p>High-sensitivity and multiplex detection of urinary metabolites remain challenging in clinical diagnosis. In this work, a novel nanocomposite consisting of gold nanoparticles (AuNPs) deposited on the metal–organic framework ZIF-8 (denoted as ZIF-8@AuNPs) was designed and employed as a surface-enhanced Raman scattering (SERS) substrate for the simultaneous quantitative detection of adenosine triphosphate (ATP), uric acid (UA), adenine (Ade), and creatinine (Cr). The SERS substrate synergistically integrates the electromagnetic enhancement of AuNPs with the molecular enrichment capability of ZIF-8. The ZIF-8@AuNPs composite was comprehensively characterized using scanning electron microscopy, transmission electron microscopy, N₂ adsorption-desorption isotherms and atomic force microscopy. SERS spectra were performed to investigate the detection performance of SERS substrate toward four urinary metabolites. This substrate exhibits outstanding SERS performance with a enhancement factor (EF) of up to 1.81 × 10⁵. Taking ATP detection as an example, the linear range of the SERS sensor prepared is 10^− 10 to 10^− 2 M with a detection limit of 7.07 × 10^− 11 M. The fabricated SERS sensor was then successfully employed to detect the metabolites in urine, and yielding recoveries ranging from 87.10% to 111.00%. This study highlights the significant potential of ZIF-8@AuNPs in the development of next-generation SERS sensors for point-of-care diagnostics and metabolomics analysis.</p> Graphical Abstract <p></p>