Berberine (BBR), a key component in Kampo medicine, is a cationic benzylisoquinoline alkaloid whose detection plays a critical role in the quality control of these traditional remedies. Traditional methods for detecting BBR often involve complex procedures, which can be time-consuming and costly. To address this challenge, our study focuses on developing a simpler, faster, and more efficient detection method for BBR in Kampo medicine formulations. We successfully developed a rapid fluorometric detection method for BBR using colloidal gold nanoparticle-based systematic evolution of ligands by exponential enrichment (GOLD-SELEX). Initially, specific single-stranded DNA (ssDNA) sequences were selected for their ability to enhance BBR's fluorescence intensity. The optimal ssDNA sequence, identified as BBR38, was further truncated to produce BBR38S, a stem-loop ssDNA that improved fluorescence upon interaction with BBR. To further enhance the fluorescence, the BBR38S aptamer underwent additional modifications, including stem truncation and nucleotide mutations, resulting in the higher fluorescence variant BBR38S-3 A10C. The final product, TetBBR38S, a tetramer version of BBR38S-3 A10C, exhibited a linear detection range of 0.780–50.0 μg mL and a limit of detection of 0.369 μg mL. The assay demonstrated sufficient selectivity and was successfully applied to analyze 128 different Kampo medicine formulations, accurately detecting BBR content with high precision. This study represents an advancement in Kampo medicine research, marking the first successful application of an aptamer-based approach for BBR detection in complex matrices. The developed method is not only simple and rapid (with a detection time of 5 min) but also cost-effective, which is crucial for widespread application.

中文翻译：

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基于适配体的荧光法快速检测汉方药物中的小檗碱


小檗碱 (BBR) 是汉方药物的关键成分，是一种阳离子苄基异喹啉生物碱，其检测在这些传统药物的质量控制中起着至关重要的作用。检测 BBR 的传统方法通常涉及复杂的程序，既耗时又昂贵。为了应对这一挑战，我们的研究重点是开发一种更简单、更快速、更有效的汉方药物制剂中 BBR 检测方法。我们使用基于胶体金纳米颗粒的指数富集配体系统进化（GOLD-SELEX）成功开发了一种 BBR 快速荧光检测方法。最初，选择特定的单链 DNA (ssDNA) 序列是因为它们能够增强 BBR 的荧光强度。最佳 ssDNA 序列被确定为 BBR38，被进一步截短以产生 BBR38S，这是一种茎环 ssDNA，可在与 BBR 相互作用时改善荧光。为了进一步增强荧光，BBR38S适体进行了额外的修饰，包括茎截断和核苷酸突变，产生了更高荧光的变体BBR38S-3 A10C。最终产品 TetBBR38S（BBR38S-3 A10C 的四聚体版本）的线性检测范围为 0.780–50.0 µg mL，检测限为 0.369 µg mL。该测定法表现出足够的选择性，并成功应用于分析 128 种不同的汉方药物配方，高精度地准确检测 BBR 含量。这项研究代表了汉方医学研究的进步，标志着基于适配体的方法首次成功应用于复杂基质中的 BBR 检测。 所开发的方法不仅简单快速（检测时间为5分钟）而且具有成本效益，这对于广泛应用至关重要。