Quantitation of Ketoconazole in Biological Fluids Using High-Performance Liquid Chromatography

• Published in: Journal of pharmaceutical sciences 1983-12, Vol.72 (12), p.1467-1469
• Main Authors: Pascucci, V.L., Bennett, J., Narang, P.K., Chatterji, D.C.
• Format: Article
• Language: English
• Subjects: Analysis; Biological and medical sciences; Biological Assay; Chromatography, High Pressure Liquid - methods; General pharmacology; high-performance liquid chromatography; High-performance liquid chromatography-quantitation of ketoconazole in biological fluids; High-performance liquid chromatography—quantitation of ketoconazole in biological fluids, humans; Humans; Ketoconazole - analysis; Ketoconazole - blood; Ketoconazole-quantitation in biological fluids; Ketoconazole—quantitation in biological fluids, highperformance liquid chromatography, humans; Ketoconazole—quantitation in biological fluids, high‐performance liquid chromatography, humans; Medical sciences; Pharmacology. Drug treatments; Spectrometry, Fluorescence - methods; Spectrophotometry, Ultraviolet - methods
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.2600721225

□ A rapid, specific procedure is described for the quantitation of ketoconazole in biological fluids using high-performance liquid chromatography (HPLC). The procedure involves sample preparation using a reverse-phase C-18 cartridge prior to chromatography and quantitation using peak height ratios (UV absorbance detection, 231 nm) of ketoconazole to the internal standard, phenothiazine. A sensitivity of 0.2 μg/ml was achieved using a 0.5-ml sample. The mean recovery was 86.2%, and overall coefficient of variation of the procedure was 7.1%. This procedure has been used to determine ketoconazole levels in human serum, plasma, CSF, and synovial fluid. A comparison with a microbiological assay is presented, and adaptability of this procedure to quantitation by fluorescence to increase the sensitivity fivefold is discussed.