The genus Capsicum is a member of the Solanaceae or nightshade family. It contains five commonly cultivated species (C. annuum L., C. frutescens L., C. chinense, C. baccatum L. and C. pubescens). Varieties of these, and other Capsicum spp., exhibit varying degrees of pungency that reflect relative concentrations of capsaicin, dihydrocapsaicin, nordihydrocapsaicin. Environmentally friendly pest-control agents for use on vegetable crops are needed to replace pesticides that are ineffective, that have been withdrawn for regulatory reasons or whose costs are prohibitive. The need for new control agents is also a result of the increasing difficulty of managing pesticide resistance. Botanical pesticides offer potential as substitutes for or supplements to be used with, synthetic pesticides. The USDA Capsicum germplasm collection contains many thousands of accessions of Capsicum spp., although only limited information is currently available on the capsaicinoid content of the fruit of these accessions. The objectives of the present investigation were 1) to quantify the major capsaicinoids in fruits of Capsicum chinense, C. frutescens, C. baccatum, C. annuum and C. pubescens accessions obtained from the USDA Capsicum germplasm collection, and 2) to screen and select candidate accessions from among those for mass production of capsaicinoids for future use as alternative insecticides. Materials and Methods Fruits from accessions of Capsicum chinense, accessions of C. frutescens, accessions of C. baccatum, accessions of C. annuum, and one accession of C. pubescens were harvested at random from field-grown plants for capsaicinoids analysis. Total capsaicinoids were extracted by blending ten fresh fruit of comparable size in methanol for one minute. The solvent extracts were concentrated to 10 mL of methanol. Each extract was subsequently passed through a 0.45 μm GD/X disposable syringe filter. One μL of this filtrate was injected into a gas chromatograph (GC) equipped with a nitrogen-phosphorus detector. GC separations were accomplished using a 25 m × 0.20 mm ID capillary column with 0.33 μm film thickness (HP-1). Quantifications were based on average peak areas of 1 μL injections obtained from external standard solutions of capsaicinoids prepared in methanol. Under these conditions, retention times (R_t) were