l-fucose is a six-carbon sugar that has potential applications in many fields. It exerts antitumor effects and could relieve intestinal disease. It exhibits potential as an emulsifier in the food industry. It is also used as a functional food and in anti-aging skincare products. However, at present, it is not possible to prepare high-purity l-fucose on a large scale, and its preparation needs further development. This review summarizes the preparation methods of l-fucose including chemical synthesis, enzymatic synthesis, microbial fermentation, and separation and purification from algae. The detection methods of l-fucose are also introduced in detail, such as l-fucose-specific lectin, detection l-fucose dehydrogenase, cysteine-sulfuric acid method, high-performance liquid chromatography, gas chromatography, and biosensors. In this review, the properties and pharmacological effects of l-fucose; preparation methods, and the commonly used detection methods of l-fucose are reviewed to serve as a reference material.