Abstract:To improve the timeliness of issuing test reports,an ultra-performance liquid chromatography-mass spectrometry/mass spectrometry （UPLC-MS/MS） method was established,in which acetonitrile was used as the extraction solvent for the pretreatment of animal-derived samples,and 15 stimulants were simultaneously determined.15 types of stimulants,included β-receptor agonists,β-blockers,protein assimilation agents,and diuretics,totaling four categories of compounds.The sample was extracted with 1% of formic acid acetonitrile,3 g of anhydrous sodium sulfate,and separated by Thermo AcclaimTM C18 column （2.2 μm,2.1 mm × 100 mm）.Gradient elution was performed using the 0.1% formic acid solution and acetonitrile as the mobile phase.It was scanned by multiple reaction monitoring mode （MRM） in electrospray positive and negative ion mode and quantified by internal standard determination.The result showed that the target substance had a good linear relationship （R>0.997 0） within different concentration ranges （0.05-300 μg/L）；the average recovery rate of the three horizontal targets ranged from 91.14% to 109.72%,and the relative standard deviations （RSDs） were less than 20%.The detection limit of adrenaline was 100 μg/kg,and the quantification limit was 200 μg/kg.The detection limit of other stimulants was 0.05-5.00 μg/kg,and the quantitation limit was 0.10-10.00 μg/kg.This method compared the results of enzymatic hydrolysis and non-enzymatic hydrolysis of some target substances through quality control samples.The non-enzymatic hydrolysis step effectively shortened the sample pretreatment time.The method can be used for the rapid detection of food borne stimulant.