Methods Mol Biol 2026 Jan 01;3054:133-142. doi: 10.1007/978-1-0716-5384-5_10.

Characterization of Bioactive Saponins from Sea Cucumbers.

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Sea cucumbers are highly valued for their bioactive constituents, particularly triterpenoid saponins, which exhibit diverse pharmacological properties. The complex amphiphilic nature of saponins poses significant challenges in their extraction and purification. Their structural diversity and amphiphilicity demand carefully optimized protocols to maximize yield and purity. Moreover, the variability of saponin profiles between different sea cucumber species and tissues necessitates reproducible methods that can be scaled from small laboratory samples to larger quantities suitable for industrial applications. This chapter describes comprehensive, reproducible protocols for both small- and large-scale extraction of saponins from sea cucumber tissues, followed by chromatographic separation and bioactivity-guided fractionation. Subsequently, analytical techniques such as thin-layer chromatography (TLC), yeast growth inhibition assays, and liquid chromatography-mass spectrometry (LC-MS) are detailed. These detailed extraction, purification, and analytical protocols facilitate downstream applications such as structural characterization, quantification, and functional studies of saponins.

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