Development and Validation of an LC–MS/MS Assay for Quantitative Analysis of Nusinersen in Human CSF and Plasma

IF 1.8 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Biomedical Chromatography Pub Date : 2025-06-13 DOI:10.1002/bmc.70138

Tingting Xu, Wei Zuo, Zhuo Sun, Simin Zhang, Zhengqing Qiu, Bo Zhang, Yi Dai

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引用次数: 0

Abstract

Nusinersen is the first antisense oligonucleotide (ASO) drug approved for the treatment of spinal muscular atrophy (SMA) in China; however, its pharmacokinetics (PK) in Chinese SMA patients remains unknown. The objective of this study was to develop and validate a liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) methodology for quantifying nusinersen in human plasma and cerebrospinal fluid (CSF) samples. The samples were prepared by protein precipitation, and then the gradient was eluted on a column of Acquity UPLC Xbridge C18 by using acetonitrile with 0.5% triethylamine (TEA) and 0.5% hexafluoroisopropanol (HFIP) and water with 0.5% TEA and 0.5% HFIP as the mobile phase. Detection was performed on a QTRAP6500⁺ tandem mass spectrometer in the negative ion multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The optimized method was successfully qualified for the nusinersen in human plasma and CSF samples over the range of 5.00 to 1000 ng/mL and 2.00 to 400 ng/mL, respectively. Importantly, our work is the first report of quantification of nusinersen in human plasma and CSF by LC–MS/MS methodology. The developed methodology is reliable and will be applied to PK study of nusinersen in Chinese SMA patients.

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人脑脊液和血浆中Nusinersen的LC-MS /MS定量分析方法的建立和验证

Nusinersen是国内首个获批用于治疗脊髓性肌萎缩症（SMA）的反义寡核苷酸（ASO）药物；然而，其在中国SMA患者中的药代动力学（PK）尚不清楚。本研究的目的是建立和验证液相色谱-串联质谱（LC-MS /MS）方法，用于定量人血浆和脑脊液（CSF）样品中的nusinersen。采用蛋白质沉淀法制备样品，以含0.5%三乙胺（TEA）和0.5%六氟异丙醇（HFIP）的乙腈和含0.5% TEA和0.5% HFIP的水为流动相，在Acquity UPLC Xbridge C18色谱柱上洗脱梯度。在QTRAP6500+串联质谱仪上进行检测，采用电喷雾电离（ESI），负离子多重反应监测（MRM）模式。优化后的方法能在5.00 ~ 1000 ng/mL和2.00 ~ 400 ng/mL范围内成功检测人血浆和脑脊液样品中的nusinsen。重要的是，我们的工作是第一个用LC-MS /MS方法定量人血浆和脑脊液中的nusinersen的报告。所建立的方法是可靠的，可应用于我国SMA患者nusinsen的PK研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomedical Chromatography 生物-分析化学

CiteScore

3.60

自引率

5.60%

发文量

268

审稿时长

2.3 months

期刊介绍： Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.