A novel approach on fluorescence quenching of tyrosine and l-tryptophan is presented for spectrofluorimetric determination of aniracetam in drug substances and products. The quenching mechanism was investigated using Stern-Volmer plots and ultraviolet spectra figures of quencher-fluorophore mixtures. Binding constant and stoichiometry were calculated using double-log plots. The spectrofluorimetric method was optimized for the experimental conditions affecting fluorescence quenching including fluorophore concentration, diluent, and reaction time. Moreover, the pH-rate profile of aniracetam was studied using simple kinetics and found to be stable within the pH range 5-8. Fluorescence quenching of tyrosine and l-tryptophan were observed on addition of aniracetam in aqueous medium at pH 5.5-6.5. Aniracetam quenched the fluorescence of tyrosine and l-tryptophan in the concentration range 1-20 μg/ml and 0.3-20 μg/ml, respectively, with binomial relationships between quenching values (ΔF) and aniracetam concentration. Limits of detection were found to be 0.10 μg/ml for tyrosine-aniracetam and 0.14 μg/ml for l-tryptophan-aniracetam. Method validation was performed as per ICH guidelines and demonstrated that the developed spectrofluorimetric method was accurate, precise, specific, and suitable for analysis of aniracetam in routine quality control laboratories. All experimental materials and solvents used are eco-friendly, indicating that the cited spectrofluorimetric procedure is an excellent green method.

译文

提出了一种用于酪氨酸和l-色氨酸荧光猝灭的新方法,用于分光光度法测定药物和产品中的阿尼西坦。使用Stern-Volmer图和猝灭剂-荧光团混合物的紫外光谱图研究了猝灭机理。使用双对数图计算结合常数和化学计量。针对影响荧光猝灭的实验条件 (包括荧光团浓度,稀释剂和反应时间) 进行了优化。此外,使用简单的动力学研究了阿尼西坦的pH速率分布,发现在5-8的pH范围内稳定。在pH 5.5-6.5的水介质中加入茴拉西坦后,观察到酪氨酸和l-色氨酸的荧光猝灭。茴拉西坦分别在1-20μg/ml和0.3-20 μ g/ml的浓度范围内猝灭酪氨酸和l-色氨酸的荧光,猝灭值 (Δ f) 与茴拉西坦浓度之间具有二项式关系。发现酪氨酸-茴拉西坦的检出限为0.10 μ g/ml,l-色氨酸-茴拉西坦的检出限为0.14 μ g/ml。方法验证根据ICH指南进行,并证明所开发的荧光光谱法是准确,精确,特异性的,并且适用于常规质量控制实验室中的阿尼西坦分析。使用的所有实验材料和溶剂都是环保的,这表明所引用的荧光光谱法是一种出色的绿色方法。

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