Abstract
Sea cucumbers are valuablemarine invertebrates known for their biologically active compoundswith health-promoting properties.
However, research has largely focused on select high-value species, overlooking others with promising bioactive potential. This
study presents the first untargeted metabolomic analysis of Holothuria cinerascens from KwaZulu-Natal, South Africa, using 1Hnuclear
magnetic resonance (NMR) and ultra performance liquid chromatography quadruple time-of-flight mass spectrometry
(UPLC–QTOF–MS) to assess metabolic and seasonal variability across three tissues: body wall, gonad and gut/mesentery. The
body wall exhibited the highest metabolite diversity, with elevated levels of amino acids and potential triterpene glycosides, likely
linked to stress or defence responses, whereas the gut/mesentery showed higher levels of sugars (galactose, xylose) and glycerol,
possibly reflecting energy metabolism, diet or microbial activity. The gonad showed the lowest overall metabolite abundance but
the highest levels of betaine and pyruvate. Seasonal differences were most pronounced in the gonad and gut/mesentery, likely
related to reproductive activity and increased food availability. Compound identification was limited by structural isomerism and
gaps in holothurian literature and databases, leaving several metabolites unidentified. These findings highlight the untapped
potential of H. cinerascens and underscore the need to expand holothurian research. Future studies should prioritise compound
characterisation and examine environmental influences on metabolite profiles to inform sustainable cultivation practices and
natural product discovery.