In this study, d‐penicillamine‐functionalized graphene quantum dots (DPA‐GQD) has been synthesized, which significantly increases the fluorescence intensity of GQD. We used this simple fluorescent probe for metal ions detection in human plasma samples. Designed DPA‐GQD respond to Hg2+, Cu2+, Au2+, Ag+, Co2+, Zn2+, and Pb2+ with high sensitivity. The fluorescence intensity of this probe decreased significantly in the presence of metal ions such as, Hg2+, Cu2+, Au2+, Ag+, Co2+, Zn2+, and Pb2+. In this work, a promising probe for ions monitoring was introduced. Moreover, DPA‐GQD probe has been tested in plasma samples. The functionalized DPA‐GQDs exhibits great promise as an alternative to previous fluorescent probes for bio‐labeling, sensing, and other biomedical applications in aqueous solution.