Technical Abstract: Fruits of 63 accessions of Capsicum chinense Jacq. from the USDA/ARS Capsicum germplasm collection were analyzed for two major capsaicinoids, capsaicin and dihydrocapsaicin, content using gas chromatography with nitrogen phosphorus detectin (GC/NPD). The objectives of the present investigation were: 1) to quantify the major capsaicinoids (capsaicin and dihydrocapsaicin) in fruits of Capsicum chinense accessions and, 2) to identify accessions containing great concentrations of capsaicinoids among countries of hot pepper origin. Seeds of C. chinense accessions received from Belize, Brazil, Colombia, Ecuador, Mexico, Peru, Puerto Rico, and United States were field grown in a silty-loam soil. Mature fruits were analyzed for major capsaicinoids content. Capsaicin concentrations were generally greater than dihydrocapsaicin. Fruits of C. chinense accession PI640900 (USA) contained the greatest concentration of capsaicin (1.52 mg/g fruit) and dihydrocapsaicin (1.16 mg/g fruit), while total major capsaicinoids in the fruits of PI438648 (Mexico) averaged 2 mg/g fruit. These two accessions were identified as potential candidates for mass production of major capsaicinoids that have health promoting properties and for use as a source of pest control agents in the agricultural fields.