El-Gizawy Samia M, Atia Noha N, Hosny Noha M
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut, Egypt.
Luminescence. 2018 Aug;33(5):877-884. doi: 10.1002/bio.3485. Epub 2018 Apr 24.
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 (Tb ) through fluorescence resonance energy transfer (FRET) from Tb to FBX. The formed complex was measured at λ 320 nm/λ 490 nm against a reagent blank. Fluorescence intensity of Tb 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.
开发并验证了一种快速、简单、选择性强且精确的荧光法，用于测定药物制剂和人血浆中的选择性黄嘌呤氧化酶抑制剂非布司他（FBX）。所提出的方法基于FBX通过从铽（Tb）到FBX的荧光共振能量转移（FRET）对铽（Tb）荧光强度的猝灭作用。以试剂空白为对照，在λ 320 nm/λ 490 nm处测量形成的络合物。加入FBX后，Tb的荧光强度降低。研究了形成的络合物的荧光猝灭值ΔF = FTb3 + - FFBX - Tb3 +与FBX浓度之间呈线性关系。研究了该络合物的反应条件和荧光光谱性质。所开发方法的线性范围为1.0 - 16.0 μg/ml。该方法成功应用于原料药、剂型和加标血浆样品中FBX的测定，回收率良好（96.79 - 98.89%）。此外，所开发的方法已成功应用于从健康志愿者采集的实际血浆样品中FBX的测定，回收率良好（82.06 - 85.65%）。根据国际协调会议（ICH）指南，对所开发方法获得的所有结果进行了统计分析和验证。
