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Background and Objectives: Amitraz is an insecticide and pesticide that, due to exceptionally strong parasiticide activities, is widely used for biological pollution monitoring. The purpose of this research was to develop and validate a liquid phase extraction technique with hollow fibers (HF-LPME) in order to determine amitraz residue in honey samples. Materials and Methods: Method of Hollow fiber liquid phase microextraction (HF-LPME) was used as simple, fast, low cost and sensitive method for determination of the amitraz from honey samples by gas chromatography flame ionization detection. Effects of five experimental factors: type of the extracting solvent, pH of the sample, stirring rate, extraction time and percent of NaCl were studied and optimized. Results: Analysis of variance showed that percent of salt and stirring rate have major effect on the extraction of amitraz (p = 0.05). Method in honey matrix was linear in the range from 20 to 1000 ng/mL with RSD lower than 14% and r2> 0.9901. Enrichment factor was 75 also the limit of detection (LOD) based on S/N = 3 was 10 ng/mL. Conclusion: According to obtained high recovery (>90%) of Amitraz by applying HF-LPME method in honey samples, this method is suggested as simple, rapid and sensitive technique. Keywords: Amitraz, Hollow fiber liquid phase microextraction, Orthogonal array design, Honey sample