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In this work, a facile and environmentally friendly solid-phase microextraction assay based on on-fiber derivatization coupled with gas chromatography and mass spectrometry was developed for determining four nonvolatile index biogenic amines (putrescine, cadaverine, histamine, and tyramine) in fish samples. In the assay, the fiber was firstly dipped into a solution with isobutyl chloroformate as derivatization reagent and isooctane as extraction solvent. Thus, a thin organic liquid membrane coating was developed. Then the modified fiber was immersed into sample solution to extract four important bioamines. Afterwards, the fiber was directly inserted into gas chromatography injection port for thermal desorption. 1,7-Diaminoheptane was employed as internal standard reagent for quantification of the targets. The limits of detection of the method were 2.98-45.3 μg/kg. The proposed method was successfully applied to the detection of bioamines in several fish samples with recoveries ranging 78.9-110%. The organic reagent used for extraction was as few as microliter that can greatly reduce the harm to manipulator and environment. Moreover, the extraction procedures were very simple without concentration and elution procedures, which can greatly simplify the pretreatment process. The assay can be extended to the in situ screening of other pollutant in food safety by changing the derivatization reagent. [ABSTRACT FROM AUTHOR]