The determination of aflatoxin B-1 (AFB1) in pistachio has been accomplished by normal and synchronous fluorimetry in combination with some multivariate calibration methods and derivative techniques. Extending the two-dimensional synchronous fluorescence scan to a three-dimensional total synchronous fluorescence scan was used to obtain the optimized Delta lambda for AFB1 in pistachio sample. The methods are based on the enhanced fluorescence of AFB1 by beta-cyclodextrine in 10% (v/v) methanol-water solution. Twenty-six pistachio samples containing AFB1 in the range 0-15 ppb were used as calibration set. Eighteen combinational methods were tested to make best model for prediction of AFB1 and finally best results obtained using a method based on synchronous fluorimetry in combination with multiple linear regressions (MLR). For concentrations ranging from 0 to 15 ppb of AFB1 in 22 pistachio samples as prediction set, the values of root mean square difference (RMSD) and relative error of prediction (REP) using MLR were 0.328 and 4.453%, respectively were observed. Two naturally contaminated pistachio samples were analyzed by synchronous fluorimetry using MLR and compared with HPLC results. (c) 2006 Elsevier B.V. All rights reserved.