基于智能手机的高通量荧光法测定人尿液中组氨酸含量的 96 孔板法
Smartphone-Based High-Throughput Fluorimetric Assay for Histidine Quantification in Human Urine Using 96-Well Plates.
机构信息
Laboratory of Analytical Chemistry, School of Chemistry, Faculty of Sciences, Aristotle University of Thessaloniki, GR-54124 Thessaloniki, Greece.
Laboratory of Pharmaceutical Analysis, School of Pharmacy, Aristotle University of Thessaloniki, GR-54124 Thessaloniki, Greece.
出版信息
Molecules. 2023 Aug 23;28(17):6205. doi: 10.3390/molecules28176205.
A high-throughput fluorimetric assay for histidine was developed, using a 96-well plates platform. The analyte reacts selectively with -phthalaldehyde under mild alkaline conditions to form a stable derivative. Instrumental-free detection was carried out using a smartphone after illumination under UV light (365 nm). The method was proved to be linear up to 100 μM histidine, with an LLOQ (lower limit of quantification) of 10 μM. The assay was only prone to interference from glutathione and histamine that exist in the urine samples at levels that are orders of magnitude lower compared to histidine. Human urine samples were analyzed following minimum treatment and were found to contain histidine in the range of 280 to 1540 μM. The results were in good agreement with an HPLC corroborative method.
开发了一种基于 96 孔板平台的用于组氨酸的高通量荧光分析方法。在温和的碱性条件下，分析物与邻苯二甲醛选择性反应，形成稳定的衍生物。在紫外光（365nm）照射下，使用智能手机进行无仪器检测。该方法在 100μM 组氨酸范围内呈线性，LLOQ（定量下限）为 10μM。该测定法仅易受存在于尿液样品中的谷胱甘肽和组氨酸的干扰，其浓度比组氨酸低几个数量级。对经过最低限度处理的人尿样进行分析，结果显示组氨酸的浓度范围为 280 至 1540μM。结果与 HPLC 确证方法吻合良好。
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