In this study, the preconcentration of Cr(III) ion in Pichia kudriavzevii JD2 immobilized perlite adsorbent by solid phase extraction was investigated. The determination of Cr(III) was performed using flame atomic absorption spectrometry (AAS). The effects of pH, adsorbent amount, recovery solution volume and type, and sample solution flow rate and volume on the recovery efficiency of Cr(III) ions were investigated. Optimized preconcentration conditions for Cr(III) were established using a column technique. The optimal parameters were determined as pH 4, a recovery solution of 2 mol/L HNO