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Semaglutide and tirzepatide are now household names, but behind every dose sits an analytical problem that small-molecule thinking can’t solve. Isomers share a mass, impurities hide in the tail of the main peak, and peptides cling to tubing, vials, and column hardware. Characterizing GLP-1 Therapeutics: Analytical Challenges and Solutions explores how separation scientists are responding. An expert Peer Exchange panel weighs up orthogonal methods, 2D-LC, inert hardware, and the role of AI in peptide method development. We trace the peptide revolution from Gila monster venom to a global surge in manufacturing capacity. And a new isocratic HPLC–DAD method shows that stability testing for both blockbuster peptides can be done in just three minutes.