Characterization of functional metabolites in broccoli by-product using green extraction with supercritical fluids

The present study aimed to valorize broccoli by-products (leaves and stems) through the extraction of bioactive compounds using supercritical fluid extraction (SFE). The influence of broccoli variety, plant part and maturation stage on total phenolic content (TPC), antioxidant activity and antimicrobial properties was evaluated. Additionally, the bioactive compounds present in the extracts were identified by HPLC-UV-electrospray ionization-tandem mass spectrometry.

Although extraction yields were lower than those obtained with conventional methods, SFE extracts exhibited high antioxidant activity, reaching values up to 1311.8 mg Trolox kg^-1 of dry extract in the CLX 3570 variety using the DPPH assay. A strong correlation between phenolic content and antioxidant activity was observed. The extracts also showed notable antimicrobial activity, with inhibition values up to 98.09% against Bacillus cereus, particularly in the Shard cultivar. This effect was associated with the presence of specific fatty acids, such as linoleic and palmitic acids, which were more abundant in stems. Furthermore, increasing maturation stage significantly enhanced extraction yield, TPC, antioxidant activity and antimicrobial capacity.

Broccoli by-products represent a promising source of functional bioactive compounds. SFE emerges as a sustainable extraction technique capable of producing high-quality extracts with significant biological activity, contributing to waste valorization and the development of functional ingredients for the food industry. © 2026 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.