This review describes a comprehensive investigation in our laboratory focusing on development of sensitive analytical methods by utilizing luminescence detections, and leading to practical applications for quantification of biologically related compounds, especially drugs of abuse. The developed methods mainly used a high-performance liquid chromatographic separation technique with fluorescence and chemiluminescence detections. The main targets of our investigation are ultra-small amounts of biologically active compounds, especially centered on abused drugs such as stimulants, narcotics and anorectics in various matrices. Furthermore, drug-drug interaction of these compounds was also examined. The results obtained in our study might contribute to the prediction of and the protection of human health from the risks of abused drugs. The developed methods might be useful for pharmaceutical, clinical and forensic studies on drugs of abuse as well as other important biologically active compounds.