Cereus hildmannianus (Cactaceae) is a medicinal plant with various pharmacological and industrial applications. Plant biotechnology is an attractive approach for specialized metabolites production under controlled conditions, with the elicitation process being considered a highly effective strategy for increasing the production of bioactive compounds. In this regard, the present study investigated the effects of elicitors, sucrose (S, 1-8 g L − 1 ), UV-C light (UV-C, 10-80 min), salicylic acid (SA, 50-200 µmol), and jasmonic acid (JA,, on the pro le of esterase isozymes, total phenolic and total avonoid contents, antioxidant activity (FRAP, DPPH, and ABTS), and the dereplication method by Ultra-High-Performance Liquid Chromatography coupled with Mass Spectrometry (UHPLC-ESI-Q-TOF-MS/MS).The isozyme pro le was not signi cantly changed. The greatest accumulations of total phenolics were observed in callus tissues induced with 100 µmol JA (390.8), 200 µmol SA (355.5), 20 min UV-C (182.9), and 6 g L − 1 S (122.1 µg mg − 1 DW); the greatest concentrations of total avonoids were observed following culture with 200 µmol SA (89.8) and 2 g L − 1 S (28.4 µg mg − 1 DW). Furthermore, the dereplication study allowed putative identi cation of thirty-six compounds (thirty phenolic and organic acids, and six avonoids). The greatest antioxidant activities were obtained with callus tissues induced with 100 µmol JA, 200 µmol SA, 80 min UV-C, and 1 g L − 1 S. The elicitors were able to alter the production of phenolic compounds in callus cultures of C. hildmannianus. These results are promising for the clean and sustainable bioproduction of bioactive molecules for pharmaceutical and cosmeceutical purposes, prioritizing the conservation of the species.