Electrochemical impedance immunosensor, with its high sensitivity from electrochemical impedance analysis and ideal specificity from the immunoassay, is increasingly used in the detection of a kind of phenoxy acid herbicides which is 2,4-Dichlorophenoxybutyric acid (2,4-DB). In this experiment, synthetic 2,4-DB antibodies were immobilized on the electrode by the crosslinking of L-Cysteine/glutaraldehyde, and 2,4-DB were measured by the increase of electron-transfer resistance when the immune reaction occurred, with Fe(CN)(6)(3-)/Fe(CN)(6)(4-) as the probe. Under optimal conditions, the change of resistance is in a linear relationship with the logarithm of the concentration in the range of 1.0 x 10(-7)-1.0 x 10(-3) g/L (R=0.994) with the detection limit of 1.0 x 10(-7) g/L (0.1 ppb). This method bears such merits as simplicity in operation, high sensitivity, wide linear range, specificity, reproducibility and good stability. The actual soybean samples were analyzed with the recovery of 82.8%-1023%. (C) 2012 Elsevier B.V. All rights reserved.