Lepidium meyenii Walpers, a well-known crop with a long history in Peru, has been utilized in the Andean region as foodstuff and medical supply. In our study, ethanol extract was obtained from the dried root of this plant and then chemically investigated. Two new imidazole alkaloids, namely, lepidiline C (1) and D (2), along with two known imidazole alkaloids (lepidiline A and B), were isolated through semi-preparative high-performance liquid chromatography (HPLC). Their structures were elucidated through HPLC-mass spectrometry, 1D and 2D proton nuclear magnetic resonance, Fourier transform infrared spectroscopy, and ultraviolet spectrophotometry. (C) 2016 Phytochemical Society of Europe. Published by Elsevier Ltd. All rights reserved.