Sarcosine is an important marker for the diagnosis of prostate cancer in urine. A chemiluminescence (CL) sensor based upon silver doped iron-based metal-organic gels (Ag/Fe-MOGs) was constructed for sarcosine determination. The Ag/Fe-MOGs with peroxidase-like activities were synthesized by a simple room temperature method to catalyze hydrogen peroxide to generate reactive oxygen species. During detection, sarcosine was oxidized by sarcosine oxidase (SOx) to produce H2O2 which was catalyzed by Ag/Fe-MOGs and accompanied by luminol chemiluminescence. Due to the peroxidase-like activity and high stability of Ag/Fe-MOGs, the luminol chemiluminescence was applied to measure sarcosine from 1 to 50 mM with a detection limit of 0.23 mM. Practical applicability was validated by determining sarcosine in human urine. This work not only offers a simple approach to a prepare catalyst with enzyme-like activity, but also provides an efficient and sensitive strategy for other substances in a rapid and cost-effective approach by replacing enzymes.