A high-performance liquid chromatographic procedure was developed for the analysis of calcium pantothenate in nutritional supplements. The method involves a simple extraction using phosphate buffer and sonication. Chromatographic separation is obtained using an aminopropyl-loaded silica gel column in the reverse-phase mode. A UV detector set at 210 nm was used to monitor the effluent. Quantitative recoveries were obtained, and precision of the method is discussed. The method is applicable to multivitamin tablets, calcium pantothenate raw material, and yeast grown in the presence of high levels of calcium pantothenate. The results of the method are compared with results obtained from the USP microbiological method of analysis. It was concluded that the procedure is rapid, accurate, easily automated, and practical for routine quality control use.