A new simple analytical method for monitoring propyl propane thiosulfonate (PTSO) in animal feed is presented. PTSO is an active ingredient from Allium spp. (like onion and shallot), proposed as a natural additive for feed being an efficient alternative to antibiotics used as growth promoter due to its efficiency improving animal health. Reversed-phase liquid chromatography with UV detection has been used and a previous sample treatment based on solid-liquid extraction has been developed and optimized in order to extract PTSO from a feed for laying hens using acetone as extraction solvent. The method has been characterized obtaining limits of detection and quantification of 11.2 and 37.3 mg kg−1, respectively, which are lower than the concentrations expected in samples containing this additive. The intra- and interday relative standard deviations (RSDs) were lower than 8.3 % in all the cases, and recoveries varied from 90.2 to 94.6 %. Finally, in order to check the unequivocal identification of PTSO, mass spectrometry detection was applied. The proposed method is a simple procedure for monitoring PTSO in commercial feed, being possible to implement it in routine laboratories for quantification purposes and stability studies of the distributed products.