Abstract
Objective: The aim of the study was to develop and validate a bioanalytical reverse-phase high-performance liquid chromatographic (HPLC) method for the estimation of nystatin in rat plasma after intranasal administration.
 Methods: The reversed-phase HPLC system was equipped with a Luna C18 column, the mobile system comprised of methanol, water, and dimethylformamide (55:30:15) and the flow rate was set at 0.9 ml/min.
 Results: The elution time for nystatin was 4.096±0.025 min. The calibration curves constructed in rat plasma were linear from 0.25 to 50 μg/ml. The lower limit of quantification (LOQ) was found to be 0.25 μg/ml. The standards for accuracy and precision of the intra- and inter-day variation studies were in the acceptable ranges as per the FDA guidelines.
 Conclusion: The LOQ value determined by the proposed method was noted to be satisfactory for inspecting the plasma pharmacokinetics of nystatin in rats’ post-administration of a nasal in situ gelling liquid crystalline precursor formulation in an in vivo study.
Highlights
The polyene class of antifungals such as nystatin, natamycin, and amphotericin B has a wide action against broad spectrum of fungi species such as Candida, Cryptococcus, Histoplasma, Blastomyces, and Aspergillus [1,2]
It is well documented that nystatin is a lyophobic drug and its solubility at room temperature in an aqueous solvent is practically insoluble while is soluble in organic solvents such as MeOH, ethanol, AcN, and DMF [21]
It is reported elsewhere that the solubility of nystatin in polar solvents can be substantially increased in the presence of 10–20% water [21,22]
Summary
The polyene class of antifungals such as nystatin, natamycin, and amphotericin B has a wide action against broad spectrum of fungi species such as Candida, Cryptococcus, Histoplasma, Blastomyces, and Aspergillus [1,2]. Initial efforts in accomplishing free drug administration have led to dose-limiting toxicity. The mucociliary clearance plays an important role in depletion of the drug; mucoadhesive formulation that enhances residence time and control drug release can be deployed to overcome these inadequacies [10]. Biopharmaceutical performance of these newer formulations has to be assessed and necessitates the development of satisfactory analytical methodologies. To estimate the intranasal in vivo performance of the in situ gelling liquid crystalline precursor system has led to the development of a suitable highperformance liquid chromatographic (HPLC) an analytical method that is accounted for here
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