A new bio-MSPE sorbent based on the use of C. micaceus and γ-Fe2O3 magnetic nanoparticle was prepared for the preconcentrations of Co(II) and Hg(II). Critical parameters including pH, flow rate, quantity of C. micaceus, quantity of γ-Fe2O3 magnetic nanoparticle, eluent (type, concentration and volume), sample volume, and foreign ions were examined. Surface structure and variations after interaction with Co(II) and Hg(II) of bio-MSPE sorbent were investigated by FT-IR, SEM, and EDX. The impact of bio-MSPE column reusage was also tested. The biosorption capacities were determined as 24.7 mg g-1 and 26.2 mg g-1, respectively for Co(II) and Hg(II). Certified reference materials were utilized to find out the accuracy of the prepared bio-MSPE method. This novel bio-MSPE method was accomplished by being applied to real food and water samples. In particular, it will be possible to make use of C. micaceus as new alternatives, in environmental biotechnology applications.