High-performance liquid chromatographic determination of liposomal nystatin in plasma and tissues for pharmacokinetic and tissue distribution studies

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 735; no. 1; pp. 51 - 62
• Main Authors: Groll, Andreas H, Mickiene, Diana, Werner, Kathy, Piscitelli, Stephen C, Walsh, Thomas J
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 26.11.1999; Elsevier Science
• Subjects: Acetonitriles; Animals; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antifungal agents; Antifungal Agents - analysis; Antifungal Agents - blood; Antifungal Agents - pharmacokinetics; Biological and medical sciences; Centrifugation; Chromatography, High Pressure Liquid - methods; Drug Stability; Edetic Acid; Hydrogen-Ion Concentration; Liposomes; Medical sciences; Methanol; Nystatin; Nystatin - analysis; Nystatin - blood; Nystatin - pharmacokinetics; Pharmacology. Drug treatments; Phosphates; Rabbits; Reproducibility of Results; Sensitivity and Specificity; Tissue Distribution; Nystatin; Delivery system; Animal model; HPLC chromatography; Rabbit; Liposome; Lagomorpha; Blood plasma; Tissue; Antibiotic; Antifungal agent; Vertebrata; Mammalia; Analytical method; Pharmacokinetics; Quantitative analysis
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(99)00396-5

A reliable reversed-phase high-performance liquid chromatographic method was developed for the determination of liposomal nystatin in plasma. Nystatin is extracted by 1:2 (v/v) liquid–liquid extraction with methanol. Separation is achieved by HPLC after direct injection on a μBondapak™ C 18 analytical column with a mobile phase composed of 10 m M sodium phosphate, 1 m M EDTA, 30% methanol and 30% acetonitrile adjusted to pH 6. Detection is by ultraviolet absorbance at 305 nm. Quantitation is based on the sum of the peak area concentration of the two major isomers of nystatin, which elute at 7.5–8.5 and 9.5–10.5 min. The assay was linear over the concentration range of 0.05 to 50 μg/ml. The lower limit of quantitation was 0.05 μg/ml, sufficient for investigating the plasma pharmacokinetics of liposomal nystatin in preclinical studies. Accuracies and intra- and inter-day precision showed good reproducibility. With minor modifications, this method also was used for assaying nystatin in various non-plasma body fluids and tissues.