Dietary selenium (Se) deficiency is recognized as a global problem, and exogenous Se supplementation can effectively enrich its levels in animal bodies. Offal tissues are equally important as meat in Se enrichment. Varying properties among Se species require information beyond total Se concentration to fully evaluate health risk/benefits. In the present study, the reliable inductively coupled plasma mass spectrometry (ICP‐MS) and HPLC‐ICP‐MS methods were optimized to analyze total Se content and Se speciation in the muscle and kidney of sheep, kidney and liver of pig, and liver of chicken after different Se supplementation treatments. The total Se contents in the liver and kidney were higher than in muscle. Five Se species were detected in the muscle, and selenourea was additionally detected in the liver and kidney. Sheep muscle and chicken liver mainly contained selenomethionine, and other tissues mainly contained selenocysteine. As the levels of selenomethionine or selenium‐enriched yeast increased in the feed, the proportion of selenomethionine in the sample increased, as well as the proportion of selenocysteine decreased, and almost no inorganic selenium was detected in all tissues. This study has provided insights for analyzing the Se enrichment patterns in tissues, which is significant for understanding the Se metabolism, animal health, and enriching the dietary Se supplementation for humans.