Federal University of Rio de Janeiro, Institute of Chemistry, Metabolomics Laboratory, Rio de Janeiro 21941-598, RJ, Brazil.
Fluminense Federal University, Institute of Chemistry, Department of Analytical Chemistry, Laboratory of Fundamental and Applied Analytical Chemistry, Niterói 24020-141, RJ, Brazil.
Anal Methods. 2023 Aug 17;15(32):3932-3939. doi: 10.1039/d3ay00587a.
Pompe disease (PD) is an inborn error of metabolism caused by α-glucosidase acid enzyme deficiency. It significantly impacts patients' health and life quality and may lead to death in the first few years of life. Among the well-established diagnostic methods, urinary glucose tetrasaccharide (Glc) screening by high performance-liquid chromatography has been helpful in monitoring Glc levels in patients on enzyme replacement therapy, demonstrating therapy efficacy. However, the specimen shipping process from a sample collecting location to a specialized laboratory for monitoring the Glc is costly and presents preanalytical challenges. In this work, we developed a filter paper based-urine collection kit to facilitate specimen shipment, and liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) analysis to determine Glc and creatinine in dried urine on filter paper. The LC-HRMS was based on a combination of targeted and untargeted screening on the same specimen injection and was successfully developed and validated. Bland-Altman statistics revealed a good relationship between dried and liquid urine samples and Glc and creatinine. Glc and other metabolites in dried urine showed stability for at least 7 days at 4 and 22 °C, and 3 days at 50 °C. The stability of the analytes and the efficiency of the kit were tested simulating real conditions by sending it by post. After two days in transit without refrigeration, the stability of compounds was maintained, showing the reliability of the urine collection kit and analysis method to determine the PD biomarker Glc.
庞贝病（PD）是一种由α-葡萄糖苷酸酶缺乏引起的先天性代谢错误。它严重影响患者的健康和生活质量，并可能导致患者在生命的最初几年内死亡。在已确立的诊断方法中，通过高效液相色谱法对尿葡萄糖四糖（Glc）进行筛查有助于监测接受酶替代治疗的患者的 Glc 水平，从而证明治疗效果。然而，将样本从采集地点运送到专门的实验室监测 Glc 的过程成本高昂，并存在分析前挑战。在这项工作中，我们开发了一种基于滤纸的尿液收集试剂盒，以方便标本运输，并使用液相色谱-高分辨率质谱（LC-HRMS）分析来确定滤纸上干燥尿液中的 Glc 和肌酐。LC-HRMS 基于在同一标本进样上进行靶向和非靶向筛选的组合，并成功开发和验证。Bland-Altman 统计显示，干燥尿液和液体尿液样本以及 Glc 和肌酐之间存在良好的关系。干燥尿液中的 Glc 和其他代谢物在 4°C 和 22°C 下至少稳定 7 天，在 50°C 下稳定 3 天。通过邮寄模拟实际情况来测试分析物的稳定性和试剂盒的效率。在没有冷藏的情况下运输两天后，化合物的稳定性得以维持，这表明尿液收集试剂盒和分析方法用于确定 PD 生物标志物 Glc 是可靠的。
