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Spectrofluorimetric methods for the determination of mirabegron by quenching tyrosine and L-tryptophan fluorophores: Recognition of quenching mechanism by stern volmer relationship, evaluation of binding constants and binding sites
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy ( IF 4.3 ) Pub Date : 2023-02-09 , DOI: 10.1016/j.saa.2023.122473
Naira A Farid 1 , Nadia F Youssef 1 , Hisham E Abdellatef 2 , Yasmine A Sharaf 2
Affiliation  

Green spectrofluorimetric methods have been adopted for the determination of Mirabegron (MG) in pure drug and pharmaceutical dosage form. The developed methods based on fluorescence quenching of tyrosine and L-tryptophan amino acids fluorophores by the effect of Mirabegron as a quencher. Experimental conditions of the reaction were studied and optimized. The Fluorescence quenching (ΔF) values were proportional to the concentration range of MG 2–20 μg/ml for the tyrosine-MG system in buffered media pH 2 and 1–30 μg/ml for L-tryptophan-MG system pH 6. Good correlation coefficients with low detection limits of 0.163 and 0.234 μg/ml for the two systems respectively. Method validation was applied according to ICH guidelines. The cited methods were successively applied for MG determination in tablet formulation. No statistically significant difference between the results of the cited and the reference methods regarding t and F tests. The proposed spectrofluorimetric methods are simple, rapid, eco-friendly and can contribute to MG’s methodologies in quality control labs. Stern-Volmer relationship, the effect of temperature, quenching constant (Kq), and UV spectra were studied to identify the mechanism by which the quenching might occur. The results demonstrated that fluorescence quenching of tyrosine was a dynamic quenching process and L-tryptophan was static. The double log plots were constructed to determine the binding constants and binding sites. The greenness profile of the developed methods has been assessed by Green Analytical procedure index (GAPI) and Analytical Greenness Metric Approach (AGREE).



中文翻译:

通过猝灭酪氨酸和 L-色氨酸荧光团测定米拉贝隆的荧光光谱法:通过斯特恩沃尔默关系识别猝灭机制,结合常数和结合位点的评估

已采用绿色荧光分光光度法测定纯药物和药物剂型中的米拉贝隆 (MG)。开发的方法基于通过米拉贝隆作为淬灭剂的作用对酪氨酸和 L-色氨酸氨基酸荧光团进行荧光淬灭。对反应的实验条件进行了研究和优化。荧光猝灭 (ΔF) 值与缓冲介质 pH 2 中酪氨酸-MG 系统的 MG 浓度范围 2-20 μg/ml 和 L-色氨酸-MG 系统 pH 6 中的 1-30 μg/ml 的浓度范围成正比。良好两个系统的相关系数分别为 0.163 和 0.234 μg/ml 的低检测限。根据 ICH 指南应用方法验证。所引用的方法相继应用于片剂中MG的测定。关于 t 和 F 检验,引用方法和参考方法的结果在统计学上没有显着差异。拟议的荧光光谱法简单、快速、环保,可以为 MG 在质量控制实验室中的方法做出贡献。Stern-Volmer关系,温度的影响,淬火常数(Kq ), 和 UV 光谱被研究以确定猝灭可能发生的机制。结果表明酪氨酸的荧光猝灭是一个动态猝灭过程,而L-色氨酸是静态的。构建双对数图以确定结合常数和结合位点。所开发方法的绿色度概况已通过绿色分析程序指数 (GAPI) 和分析绿色度度量方法 (AGREE) 进行了评估。

更新日期:2023-02-09
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