Amperometric salicylate microbiosensors were constructed by salicylate hydroxylase (SH) immobilization on different carbon fibers electrodes. The characteristics of the microbiosensor prepared with different carbon fibers were investigated. Five different carbon fibers were investigated. Toray M40 was very unstable, although reasonable performance compared with others was observed. The carbon fiber Toray‐T800 was found to be the best amperometric microbiosensor for salicylate, presenting two linear response ranges between 0.1–1.93 µmol L−1 (sensitivity of 56.0±0.4 mA mol −1 L) and 1.96–12.0 µmol L−1 (sensitivity of 20.8±0.4 mA mol−1 L), with a detection limit of 6.4±0.9×10−8 m L−1 and response time of 3.5 seconds. The relative standard deviation for 10 determinations was 4.1 %. The high sensitivity of the microbiosensor was assigned to the chemical amplification process. Salicylate determination can be carried out with applied potentials higher than 150 mV (vs. SCE), with good precision. The useful lifetime of the biosensor was 100 continuous determinations without significant change.