Fluorimetric determination of febuxostat in dosage forms and in real human plasma via Förster resonance energy transfer

• Published in: Luminescence (Chichester, England) England), 2018-08, Vol.33 (5), p.877-884
• Main Authors: El‐Gizawy, Samia M., Atia, Noha N., Hosny, Noha M.
• Format: Article
• Language: English
• Subjects: Blood plasma; Calibration; Dosage; dosage forms; Energy transfer; Febuxostat; Febuxostat - analysis; Febuxostat - blood; Fluorescence; Fluorescence resonance energy transfer; Fluorescence Resonance Energy Transfer - methods; Formulations; human plasma; Humans; Limit of Detection; Linearity; Plasma; Powder; Powders - analysis; Quenching; Reagents; Reproducibility of Results; Resonance; Solvents - chemistry; Spectrometry, Fluorescence - methods; Spectrophotometry, Ultraviolet; Statistical analysis; Statistical methods; Temperature; Terbium; Terbium - chemistry; Xanthine oxidase
• ISSN: 1522-7235; 1522-7243
• DOI: 10.1002/bio.3485

A rapid, simple, selective and precise fluorimetric method was developed and validated for determination of a selective xanthine oxidase inhibitor; febuxostat (FBX) in pharmaceutical formulations and in human plasma. The proposed method is based on quenching effect of FBX on the fluorescence intensity of terbium (Tb3+) through fluorescence resonance energy transfer (FRET) from Tb3+ to FBX. The formed complex was measured at λex. 320 nm/λem. 490 nm against a reagent blank. Fluorescence intensity of Tb3+ was diminished when FBX was added. A linear relationship between the fluorescence quenching value of the formed complex ΔF=FTb3+−FFBX−Tb3+ and the concentration of FBX was investigated. The reaction conditions and the fluorescence spectral properties of the complex have been studied. The linearity range of the developed method was 1.0–16.0 μg/ml. The suggested method was applied successfully for the estimation of FBX in bulk powder, dosage forms and spiked plasma samples with excellent recoveries (96.79–98.89%). In addition, the developed method has been successfully applied for determination of FBX in real plasma samples collected from healthy volunteers with good recoveries (82.06–85.65%). All obtained results of the developed method were statistically analyzed and validated according to ICH (International Conference on Harmonization) guidelines.