Comparative urinary globotriaosylceramide analysis by thin-layer chromatography-immunostaining and liquid chromatography-tandem mass spectrometry in patients with Fabry disease

• Published in: Molecular genetics and metabolism reports 2021-12, Vol.29, p.100804-100804, Article 100804
• Main Authors: Shiga, Tomoko, Tsukimura, Takahiro, Namai, Yurie, Togawa, Tadayasu, Sakuraba, Hitoshi
• Format: Article
• Language: English
• Subjects: Fabry disease; Globotriaosylceramide; Liquid chromatography-tandem mass spectrometry; Research Paper; Thin-layer chromatography-immunostaining
• ISSN: 2214-4269; 2214-4269
• DOI: 10.1016/j.ymgmr.2021.100804

In Fabry disease, accumulation of glycolipids, predominantly globotriaosylceramide (Gb3), affects the kidneys, and nephropathy is one of the important disorders that influence the disease severity and prognosis of patients. Urinary Gb3 has been analyzed for diagnosis and monitoring of Fabry disease. In this study, we analyzed urinary Gb3 by thin-layer chromatography (TLC)-immunostaining and liquid chromatography (LC)-tandem mass spectrometry (MS/MS). An improved qualitative method, TLC-immunostaining, revealed excessive urinary Gb3 excretion in 100 (8/8), 88 (14/16), and 74% (45/61) of the classic Fabry males, later-onset Fabry males, and Fabry females examined, respectively. This authentic method is robust, easy, economic, and hardly affected by abundant urinary sediment, and this is useful for diagnosing individual Fabry patients. LC-MS/MS can determine the level of Gb3 in urine with high sensitivity, and it revealed that the Gb3 excretion level was higher in the order of classic Fabry males, later-onset Fabry males, Fabry females, and controls, respectively, and this is expected to be a useful quantitative method not only for diagnosis but also for predicting the progression of Fabry nephropathy. As to the relation of the urinary Gb3 level and renal events, our study revealed that the urinary Gb3 level in Fabry patients experiencing renal events tended to be higher than that in ones who did not have any renal events in each phenotypic group of the disease. •Urinary Gb3 excretion was analyzed by means of TLC-immunostaining and LC-MS/MS.•Excessive urinary Gb3 excretion was detected in most of the Fabry patients on TLC-immunostaining.•The urinary Gb3 levelwas higher in the order of classic Fabry males, later-onset Fabry males, Fabry females, and controls.•The urinary Gb3 level in the Fabry patients who exhibited renal events tended to be higher than that in those who did not.