Omeprazole, a gastric acid pump inhibitor, dose-dependently controls gastric acid secretion; the drug has greater antisecretory activity than histamine H(2)-receptor antagonists. Omeprazole has been determined in formulations and biological fluids by a variety of methods such as spectrophotometry, high-performance liquid chromatography with ultraviolet detection and liquid chromatography coupled with tandem mass spectrometry. The overview includes the most relevant analytical methodologies used in its determination since the origin still today.