Clinical Pharmacology Research Center, Peking Union Medical College Hospital, State Key Laboratory of Complex Severe and Rare Diseases, NMPA Key Laboratory for Clinical Research and Evaluation of Drug, Beijing Key Laboratory of Clinical PK & PD Investigation for Innovative Drugs, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
Department of Pediatrics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Sep 1;1229:123900. doi: 10.1016/j.jchromb.2023.123900. Epub 2023 Oct 1.
Urinary 1,5-anhydroglucitol (1, 5-AG), 6-α-D-glucopyranosyl-maltotriose (Glc) and maltotetraose (M) are important biomarkers for glycogen storage disease (types Ib and Ⅱ). This study aimed to develop and validate an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to detect these three urinary saccharide metabolites. Urine samples were diluted and then analyzed. Chromatographic separation was performed on an Acquity™ UPLC Amide column (2.1 × 100 mm, 1.7 μm) with gradient elution. The quantitation of analytes was achieved on a 5500 Qtrap mass spectrometer using negative multiple reaction monitoring (MRM) mode. The calibration curves for all analytes were linear over the range of 0.500 to 100 μg/mL with a correlation coefficient, R ≥ 0.999. The percent relative standard deviations (RSD%) were ≤12.8%, and the percent relative errors (RE%) were in the range of -11.7%-11.0%. The relative matrix effects of all analytes were between 87.2% and 104% with RSD% < 3.10% across three concentrations. The developed analytical method was simple, accurate, and reliable for rapid and simultaneous analysis of these three urinary saccharide metabolites. It was applied to healthy volunteers and patients. To our knowledge, it was the first validated assay for urinary maltotetraose quantification. This work provides support for exploring the potential of maltotetraose as a biomarker for Pompe disease.
尿 1,5-脱水山梨醇（1,5-AG）、6-α-D-葡萄糖基麦芽糖三糖（Glc）和麦芽四糖（M）是糖原贮积病（Ib 型和 II 型）的重要生物标志物。本研究旨在开发和验证一种超高效液相色谱-串联质谱（UPLC-MS/MS）法，用于检测这三种尿糖代谢物。尿液样本经稀释后进行分析。采用 Acquity™ UPLC Amide 柱（2.1×100mm，1.7μm）进行色谱分离，采用梯度洗脱。采用 5500 Qtrap 质谱仪以负多重反应监测（MRM）模式进行分析物定量。所有分析物的校准曲线在 0.500 至 100μg/mL 范围内呈线性，相关系数 R≥0.999。相对标准偏差（RSD%）≤12.8%，相对误差（RE%）在-11.7%-11.0%范围内。所有分析物的相对基质效应均在 87.2%-104%之间，三个浓度的 RSD%<3.10%。所建立的分析方法简单、准确、可靠，可用于快速同时分析这三种尿糖代谢物。该方法已应用于健康志愿者和患者。据我们所知，这是首个用于定量尿麦芽四糖的验证方法。这项工作为探索麦芽四糖作为庞贝病生物标志物的潜力提供了支持。
