Fucoxanthin, a bioactive carotenoid, is well known for its antioxidant, anti-inflammatory, anti-obesity, and anticancer effects, making it a vital compound in nutraceuticals, pharmaceuticals, and cosmetics. Predominantly derived from diatoms such as Phaeodactylum tricornutum, fucoxanthin plays a pivotal role in photosynthesis and photoprotection. However, its commercial use is often hampered by extraction and purification issues owing to its limited natural abundance and sensitivity to deterioration under light and heat. This chapter presents a procedure for the extraction and purification of fucoxanthin from Phaeodactylum biomass using silica gel column chromatography. The methodology involves using solvent-based extraction to obtain fucoxanthin from microalgal biomass and silica gel column chromatography to purify fucoxanthin. This protocol provides a scalable and efficient method for producing high-quality fucoxanthin, making it suitable for use in a wide range of industrial and research applications.