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A high-throughput method for analysis of ramelteon, agomelatine, and melatonin in human plasma by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) is presented. The LC system, MS-MS system, and separation column used were Waters Acquity UPLC, Acquity TQD, and Poroshell 120 EC-C18, respectively. For extraction of the target compounds, solid-phase extraction was performed with Oasis HLB cartridges. All compounds were detected with retention times of < 3 min. The calibration curves for the compounds spiked into human plasma showed good linearities in the nanogram-per-milliliter range. The detection limit (signal-to-noise ratio = 3) was as low as 0.2-0.5 ng/ml. The method gave satisfactory recovery rates, accuracy, and precision for quality control samples spiked with these drugs. The present method should prove very useful in forensic and clinical toxicology and pharmacokinetic studies, because of its high sensitivity and rapidness. To our knowledge, this is the first trial to analyze ramelteon in a biological sample by LC-MS-MS.