Martineau Tristan, Maranda Bruno, Auray-Blais Christiane
Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Centre de Recherche-CHUS, 3001, 12th Avenue North, Sherbrooke, QC J1H 5N4, Canada.
Biomolecules. 2024 Dec 17;14(12):1612. doi: 10.3390/biom14121612.
Early detection of sphingolipidoses is crucial to prevent irreversible complications and improve patient outcomes. The use of urine samples dried on filter paper (DUS) is a non-invasive strategy that simplifies the collection, storage, and shipping of samples compared to using liquid urine specimens.
(1) Develop and validate a multiplex ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methodology using DUS to quantify twenty-one lysosphingolipids normalized to creatinine for eight different sphingolipidoses. (2) Establish normal reference values to evaluate the clinical utility of the methodology.
Samples were eluted from a 5 cm filter paper disk (~1 mL of urine) and extracted on Oasis MCX solid-phase extraction cartridges prior to injection in the UPLC-MS/MS system.
Urinary lysosphingolipids were stable on DUS at -80 °C and -30 °C for 117 days, at 21.5 °C and 4 °C for at least 26 days, and at 35 °C for 3 days. Globotriaosylsphingosine, glucosylsphingosine, and their analogs were elevated in patients with Fabry disease and Gaucher disease, respectively, compared to controls (-value < 0.0001). The analysis of related analog profiles suggests a better overall reliability in detecting patients early, especially for Fabry patients.
This approach is feasible and might be useful for the early detection, monitoring, and follow-up of patients with sphingolipidoses.
早期发现鞘脂贮积症对于预防不可逆并发症和改善患者预后至关重要。与使用液态尿液样本相比，使用滤纸上干燥的尿液样本（DUS）是一种非侵入性策略，可简化样本的采集、储存和运输。
（1）开发并验证一种使用DUS的多重超高效液相色谱 - 串联质谱（UPLC-MS/MS）方法，以定量针对八种不同鞘脂贮积症的二十一种溶血鞘脂，并以肌酐进行标准化。（2）建立正常参考值以评估该方法的临床实用性。
从5厘米滤纸片（约1毫升尿液）上洗脱样本，并在注入UPLC-MS/MS系统之前在Oasis MCX固相萃取柱上进行萃取。
尿液溶血鞘脂在DUS上于-80°C和-30°C下可稳定保存117天，在21.5°C和4°C下至少可稳定保存26天，在35°C下可稳定保存3天。与对照组相比，法布里病和戈谢病患者的球三糖基鞘氨醇、葡萄糖基鞘氨醇及其类似物分别升高（P值<0.0001）。相关类似物谱分析表明，在早期检测患者方面，尤其是法布里病患者，整体可靠性更高。
这种方法是可行的，可能有助于鞘脂贮积症患者的早期检测、监测和随访。
