Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toruń, 7 Gagarin St., 87-100 Toruń, Poland.
Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus University in Toruń, 4 Wilenska St., 87-100 Toruń, Poland.
Analyst. 2024 Jul 8;149(14):3739-3746. doi: 10.1039/d4an00436a.
The study aimed to analyze nusinersen metabolites in the cerebrospinal fluid samples using ion-pair reversed-phase ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Three different sample preparation methods were tested for extraction and purification, but solid phase extraction appeared to be the most suitable, allowing a significant sample enrichment (40-fold). This step was necessary to detect and identify metabolites of nusinersen in the cerebrospinal fluid. The developed and applied analytical procedure enabled the identification of nusinersen metabolites: sequences shorter by several nucleotides from the 3' end; shorter by several nucleotides from both the 3' and 5' ends; and some depurination products. To the best of our knowledge, this is the first report on the analysis and identification of nusinersen metabolites in cerebrospinal fluid samples taken from children with spinal muscular atrophy treated with Spinraza.
本研究旨在使用离子对反相超高效液相色谱-四极杆飞行时间质谱法分析脑脊液样本中的nusinersen 代谢物。测试了三种不同的样品前处理方法进行提取和纯化，但固相萃取似乎是最合适的方法，可显著富集样品（40 倍）。这一步骤对于检测和鉴定 nusinersen 在脑脊液中的代谢物是必要的。所开发和应用的分析程序能够识别 nusinersen 的代谢物：从 3'端短几个核苷酸的序列；从 3'和 5'端短几个核苷酸的序列；以及一些脱嘌呤产物。据我们所知，这是关于在接受 Spinraza 治疗的脊髓性肌萎缩症儿童的脑脊液样本中分析和鉴定 nusinersen 代谢物的首次报告。
