Pantothenic acid (vitamin B5) is an essential component of food and a precursor of important biologically active agents in the body. It is involved in energy metabolism and quite common in foods. Vitamin B5 deficiency occurs in certain categories of the population. The production of vitamin B5-enriched dietary supplements, foods, premixes, and feeds is practiced to overcome this problem. Since pantothenic acid is quite labile to the action of high temperatures, for the enrichment of food and feed is usually used its more stable form—calcium pantothenate. The most common and easy to implement method of its identification is high-performance liquid chromatography, which is described in detail in this chapter.