Sea cucumber tentacles (aquapharyngeal bulb) are a rich source of biologically active compounds, including phenolics, however they are mainly discarded as processing waste. This study evaluated free, esterified, and insoluble-bound phenolics of Atlantic sea cucumber (Cucumaria frondosa) tentacles and their antioxidant activity for the first time. Biological properties such as inhibitory activities against α-glucosidase, tyrosinase, and the formation of AGEs as well as LDL-cholesterol and DNA oxidation were investigated. The antioxidant activity of the phenolic extracts was also evaluated in a fish model system. In the UHPLC-QTOF-MS/MS analysis, 31 phenolic compounds, mainly phenolic acids and flavonoids, were identified and quantified. Among them, eight compounds were detected for the first time in any species of sea cucumber. The free phenolic fraction was the major form of phenolics, mainly protocatechuic acid, p-coumaric acid, gallic acid, ellagic acid, catechin, and quercetin, exhibiting strong antioxidant and biological activities. Fresh Atlantic salmon treated with sea cucumber phenolics delayed lipid oxidation as measured by the thiobarbituric acid reactive substances (TBARS) assay. Therefore, Atlantic sea cucumber tentacles may serve as a viable source of functional food ingredients with protective antioxidant properties.