A new method for the determination of Cu(II) and Pb(II) by flame atomic absorption spectrometry (FAAS) after preconcentrating on a column containing Anoxybacillus sp. SO B1-immobilized Amberlite XAD-16 was developed. The functional groups of Anoxybacillus sp. SO B1 immobilized on Amberlite XAD-16 were characterized in KBr tablets by Fourier transform infrared (FT-IR) spectrometry. Various parameters such as pH, amount of the adsorbent, eluent type and volume, and flow rate of the sample solution were studied. The optimum pH values of quantitative sorption for Cu(II) and Pb(II) were found to be pH 7.0 and 5.0 and Cu(II) and Pb(II) ions could be quantitatively eluted with 5.0 ml of 1.0 mol L ¡1 HCI and 10.0 ml of 0.25 mol L ¡1 HNO 3 , respectively. Recoveries of Cu(II) and Pb(II) were found to be 100.9 § 1.57% and 100.3 § 0.49% (N D 5), the limits of detection of Cu(II) and Pb(II) in the determination by FAAS (3 s, N D 10) were found to be 0.8 and 1.6 mg L ¡1 , respectively. The proposed enrichment method was applied for metal ion determination from water samples such as two parts of Tigris River water in Diyarbakır and Elazı g, Lake of Hazar in Elazı g, and tap water in Diyarbakır. Furthermore, the accuracy of the proposed method was verified by studying the analytical recovery and by analyzing certified reference material (NCS-DC 73350 leaves of poplar).