The use of various hemp-derived products has been rapidly growing in the human nutrition industry and has sparked great interest in using these ingredients for companion animals as well. Thorough research is needed to determine the ingredient and safety standards required for AAFCO approval of hemp ingredients. In order to be effectively incorporated into pet food products, we must determine the nutrient content, quality, and utility of these ingredients in pet species. The objective of this study was to evaluate the nutrient composition of seeds from four different varieties of hemp, NWG 452, NWG 331, NWG 2730, X-59, and determine protein quality and true metabolizable energy using a cecectomized rooster model. The seeds were similar in macronutrient composition, with small variations in acid hydrolyzed fat, crude protein, total dietary fiber and gross energy content, as well as amino acid and long-chain fatty acid profiles. All essential amino acids were present in concentrations that exceeded the NRC (2006) recommended allowances for adult dogs and cats at maintenance with the exception of tryptophan. The long-chain fatty acid profile presented a favorable ratio of omega-6 to omega-3 fatty acids of close to 3.5:1. The results of the cecectomized rooster assay indicated no significant difference in the standardized amino acid digestibility of the indispensable amino acids among the seed varieties (P > 0.05). A significant difference in the true metabolizable energy corrected for nitrogen was observed among the seeds (P < 0.05), following the pattern of higher acid hydrolyzed fat and lower total dietary fiber content resulting in higher metabolizable energy. An adapted calculation of digestible indispensable amino acid score was made to determine protein quality of the hemp seeds using AAFCO nutrient profiles and NRC recommended allowances for adult dogs and cats at maintenance as reference points. The resulting scores determined tryptophan to be the first limiting amino acid and indicate that hemp seeds alone do not meet all the amino acid requirements for adult dogs and cats at maintenance, and would need a complimentary protein source for practical use in companion animal diets. The data from this study suggest that hemp seeds may provide a beneficial source of fat, protein, and dietary fiber, with consideration to differences in nutrient profile among seed varieties. However, further investigation in vivo is needed to determine the safety and efficacy of utilizing hemp in the diets of both canines and felines.