Recent studies have raised the question whether there is a potential threat by a horizontal transfer of toxic plant constituents such as pyrrolizidine alkaloids (PAs) between donor-PA-plants and acceptor non-PA-plants. This topic raised concerns about food and feed safety in the recent years. The purpose of the study described here was to investigate and evaluate horizontal transfer of PAs between donor and acceptor-plants by conducting a series of field trials using the PA-plant Lappula squarrosa as model and realistic agricultural conditions. Additionally, the effect of PA-plant residues recycling in the form of composts or press-cakes were investigated. The PA-transfer and the PA-content of soil, plants, and plant waste products was determined in form of a single sum parameter method using high-performance liquid chromatography mass spectroscopy (HPLC-ESI-MS/MS). PA-transfer from PA-donor to acceptor-plants was frequently observed at low rates during the vegetative growing phase especially in cases of close spatial proximity. However, at the time of harvest no PAs were detected in the relevant field products (grains). For all investigated agricultural scenarios, horizontal transfer of PAs is of no concern with regard to food or feed safety. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.