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Semaglutide is a glucagon-like peptide 1 (GLP-1) agonist that has found widespread popularity as a weight loss product. Initially developed as a treatment for diabetes, the drug acts to stimulate the secretion of insulin, as well as suppress appetite by delaying gastric emptying and promoting satiety. By allowing users to moderate their weight, the drug may provide a competitive advantage to those individuals participating in sports that are regulated by weight classes, such as wrestling and boxing. Most current testing regimes for semaglutide rely on the use of blood samples. Here, a simple alternative LC-MS/MS test is presented that relies on urine, a much simpler matrix to obtain. The method is based on the detection of two unique semaglutide metabolites: U6 and U7. It was demonstrated to have an LOD of 50 pg/mL based on recovery from 2 mL of urine.