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Investigating fluorogenic labelling of amine coupled with first-order derivative spectrofluorimetry as a versatile analytical methodology: Application to estimation of benoxinate in its eye drops
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy ( IF 4.3 ) Pub Date : 2023-09-26 , DOI: 10.1016/j.saa.2023.123410 Rasha Aboshabana 1 , Heba Samir Elama 1 , Heba Elmansi 1 , Samah Abo El Abass 2
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy ( IF 4.3 ) Pub Date : 2023-09-26 , DOI: 10.1016/j.saa.2023.123410 Rasha Aboshabana 1 , Heba Samir Elama 1 , Heba Elmansi 1 , Samah Abo El Abass 2
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In this research, fluorogenic labelling followed by applying first-order derivative spectrofluorimetry for the developed fluorophore is discussed as an alternative, sensitive and selective analytical approach. Benoxinate, containing a primary amine and fluorescamine reagent were selected for the study. Then the proposed methodology relies on the reaction between the primary amine in benoxinate with fluorescamine that selectively reacts with primary amines to develop highly fluorescent products. The fluorescamine-benoxinate developed fluorophore is identified by its sharp first-order derivative peak at 465 nm following excitation at 386 nm in borate buffer, pH 8. The optimum reaction conditions were ascertained. Following ICH validation guidelines, the first order derivative of the relative fluorescence intensity for the developed fluorophore was linearly related to benoxinate concentration and ranged from 20.0 to 200.0 ngmL with a detection limit of 3.36 ng/mL and a quantitation limit of 10.19 ng/mL, moreover, satisfying accuracy and precision values were obtained upon statistical analysis of results. The offered analytical method was successfully applied to quantify benoxinate in raw material and Benox® eye drops as a direct application to a commercial formulation.
中文翻译：
研究胺的荧光标记与一阶导数荧光光度法作为通用分析方法：应用于滴眼液中苯氧酯的估计
在本研究中，讨论了荧光标记，然后对开发的荧光团应用一阶导数荧光光谱法，作为一种替代的、灵敏的和选择性的分析方法。本研究选择含有伯胺和荧光胺试剂的 Benoxinate。然后，所提出的方法依赖于苯氧酯中的伯胺与荧光胺之间的反应，荧光胺选择性地与伯胺反应以开发高荧光产物。在 pH 8 的硼酸盐缓冲液中，在 386 nm 处激发后，荧光胺-苯肟酸开发的荧光团可通过其在 465 nm 处尖锐的一阶导数峰来识别。确定了最佳反应条件。根据 ICH 验证指南，所开发的荧光团的相对荧光强度的一阶导数与苯氧酯浓度呈线性相关，范围为 20.0 至 200.0 ngmL，检测限为 3.36 ng/mL，定量限为 10.19 ng/mL，此外，通过对结果进行统计分析，获得了令人满意的准确度和精密度值。所提供的分析方法已成功应用于量化原材料和 Benox® 滴眼剂中的苯氧酯，并直接应用于商业配方。
更新日期：2023-09-26
中文翻译：
研究胺的荧光标记与一阶导数荧光光度法作为通用分析方法：应用于滴眼液中苯氧酯的估计
在本研究中，讨论了荧光标记，然后对开发的荧光团应用一阶导数荧光光谱法，作为一种替代的、灵敏的和选择性的分析方法。本研究选择含有伯胺和荧光胺试剂的 Benoxinate。然后，所提出的方法依赖于苯氧酯中的伯胺与荧光胺之间的反应，荧光胺选择性地与伯胺反应以开发高荧光产物。在 pH 8 的硼酸盐缓冲液中，在 386 nm 处激发后，荧光胺-苯肟酸开发的荧光团可通过其在 465 nm 处尖锐的一阶导数峰来识别。确定了最佳反应条件。根据 ICH 验证指南，所开发的荧光团的相对荧光强度的一阶导数与苯氧酯浓度呈线性相关，范围为 20.0 至 200.0 ngmL，检测限为 3.36 ng/mL，定量限为 10.19 ng/mL，此外，通过对结果进行统计分析，获得了令人满意的准确度和精密度值。所提供的分析方法已成功应用于量化原材料和 Benox® 滴眼剂中的苯氧酯，并直接应用于商业配方。
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