Identification of Novel Bioactive Molecules in Black Chiloe's Giant Garlic (Allium ampeloprasum L.) by Green Microwave-Assisted Extraction and Effect-Directed Analysis Using High-Performance Thin Layer Chromatography-Bioassay and Mass Spectrometry.

Black Chiloe's giant garlic is a functional food produced by a mild Maillard reaction that contains relevant bioactive molecules like organosulfur compounds (OSCs) and (poly)phenols (PPs). Compared with raw garlic, black garlic has a higher content of PPs and S-allyl cysteine (SAC), a key OSC due to its bioactivities. The objective of the present work was to optimize by chemometric tools a green microwave-assisted extraction (MAE) of SAC and PPs present in black Chiloe's giant garlic to detect and identify novel bioactive molecules with antioxidant and/or inhibitory activities over cyclooxygenase, α-glucosidase, and acetylcholinesterase enzymes. The MAE factors were optimized using a central composite design, establishing optimal PP and SAC yields at 67 °C, 0% ethanol, 12 min and 30 °C, 40% ethanol, 3 min, respectively. PP and SAC values were 9.19 ± 0.18 mg GAE/g DW and 2.55 ± 0.10 mg SAC/g DW. Applying effect-directed analysis using high-performance thin layer chromatography-bioassay and mass spectrometry, the bioactive molecules present in the MAE extract with antioxidant and inhibitory activities over cyclooxygenase, α-glucosidase, and acetylcholinesterase enzymes were identified as N-fructosyl-glutamyl-S-(1-propenyl)cysteine, N-fructosyl-glutamylphenylalanine, and Harmane.