In this study, an extraction procedure based on biosorption of Co(II), Cr(III), Mn(II), Ni(II) and Zn(II) using biocomposite of fungus (Mucor pusillis, Lindt., 1886) immobilised sepiolite powder has been developed preconcentration. For the quantitative recovery of the analytes, different parameters pH, amount of solid-phase, flow rate of the sample solution and eluent type were optimised. The interference of foreign ions on the recovery of the analytes was also investigated. Elution was carried out with 2 mL of 5% HNO3, 100 mg sorbent dosage. The recoveries of Co(II), Cr(III), Mn(II), Ni(II) and Zn(II) by Mucor pusillis (Lindt., 1886) immobilised on sepiolite powder were obtained 100 +/- 2%, 98 +/- 2% 96 +/- 2%, 99 +/- 2%, 98 +/- 3% at 95% confidence level, respectively, for spiked environmental samples. The analytical detection limits were calculated 1.2, 1.3, 1.4, 0.6 and 0.2 ng/mL for Co(II), Cr(III), Mn(II), Ni(II) and Zn(II) preconcentration, respectively. Standard reference materials of biological samples were used to validate the data. Method was applied on the real environmental samples including tap water, river water and vegetable samples to determine the concentration of the analytes. Determinations were carried out using ICP-AES.