Quantitative Analysis and Anti-inflammatory Activity Evaluation of the A-Type Avenanthramides in Commercial Sprouted Oat Products

Avenanthramides (AVAs) are unique phytochemicals in oat that contain two distinct groups of compounds. The first group is constituted by N-cinnamoylanthranilic acids with a single double bond (referred to as C type), and the other group is constituted by N-avenalumoylanthranilic acids with two double bonds (referred to as A type). C-type AVAs have been reported with their chemical profiles and levels in commercial oat products as well as their bioactivities. However, the accurate levels of A-type AVAs in commercial sprouted oat products and their bioactivity are still unknown. In this study, we purified seven A-type AVAs from sprouted oat bran and characterized their structures with corresponding mass spectrometry and nuclear magnetic resonance data. Among them, five compounds were isolated from oat bran for the first time. The purified A-type AVAs were used as authentic standards to establish the chemical profile of A-type AVAs in oat and to quantify the levels of all individual A-type AVAs in six commercial sprouted oat products using ultra-high-performance liquid chromatography with tandem mass spectrometry. The total A-type AVA contents in the various oat products ranged from 7.85 to 133.3 μg/g. Furthermore, the inhibition of lipopolysaccharide-induced nitric oxide production and inducible nitric oxide synthase expression by A- and C-type AVAs in macrophages were compared. The most abundant A-type AVAs (2p_d, 2c_d, and 2f_d) have similar anti-inflammatory activity to the major C-type AVAs (2p, 2c, and 2f). To the best of our knowledge, this is the first report on the bioactivity of A-type AVAs.