InstrumentationHPLC Instrument: The HPLC system used was Hitachi L-2000 series (Japan), equipped with a Model L-2130 pump, an L injection loop and a UV-Vis detector L2420. The separation was carried out in a Abstract This paper describes an enhanced High-performance liquid chromatography (HPLC) method for the analysis of miconazole in water samples. In this study, determination of miconazole has been carried out according to standard method for water and wastewater analysis. Samples of collected water were agriculture stream water, River Nile (Surface water samples) water and Hospital wastewater samples from El-gharbia governorate in Egypt. Miconazole was extracted by liquid-liquid extraction and analyzed by HPLC. The chromatographic separation was performed using a Phenomenex C8 column, flow rate of 0.8mL/min, and UV detection at 220nm. The optimized HPLC system was achieved using mobile phase composition containing methanol: water (85:15v/v). The intra-day and interday precisions were lower than 0.58 while the accuracy ranged from 99.06% to 101.53%. Finally, liquid-liquid phase extraction in combination with HPLC is a sensitive and effective method for the determination of Miconazole Nitrate in water samples. Miconazole was observed in some agricultural streams and waste water samples of El-gharbia governorate hospitals.