Abstract The purpose of the study was the quantitative determination of fluconazole by IR spectrophotometry which was realized by the application of the KBr disc technique. In this study dehydrocholic acid was used as internal standard. The absorption bands at 960 and 675 cm−1 were chosen for fluconazole and 1705 cm−1 for dehydrocholic acid. The concentration range between 0.4–1.6% w/w in KBr disc showed compliance with Beer's law. Besides IR spectroscopy, UV spectroscopy and HPLC methods were also used In the quantitative determinations. In the UV spectroscopy method the absorbance value at 261 nm in MeOH was used for fluconazole. In quantitative determinations which were performed by using HPLC, ketoconazole was the internal standard. Different regression equations were applied for each method in order to complete the quantitative determination. In UV spectroscopy and HPLC methods, relative standards were found as 0.74, 1.04% and in IR spectroscopy (675, 960 cm−1) relative standards were found as 1.1 an...