LC-MS/MS quantification of nusinersen in rat cerebrospinal fluid and preclinical pharmacokinetics study application

  • Bioanalysis. 2025 Jul;17(13):839-846. doi: 10.1080/17576180.2025.2535949.
Yujie Li  1 ,  Shu Zhang  2 ,  Xiayi Wang  3 ,  Xiaochuan Li  4 ,  Lizhong Guo  1
Affiliations
  • 1. School of Life Sciences, Qingdao Agricultural University, Qingdao, Shandong, China.
  • 2. School of Pharmacy, Faculty of Medicine, Macau University of Science and Technology, Macau SAR, China.
  • 3. School of Pharmacy, Yantai University, Yantai, PR China.
  • 4. College of Biomedical Industry, Guilin Medical University, Guilin, Guangxi, China.
PMID: 40689874 DOI: 10.1080/17576180.2025.2535949
Abstract

Background: Background: An oligonucleotide drug named nusinersen sodium is used to treat Spinal Muscular Atrophy (SMA), requires accurate detection for therapeutic research. There are no published reports on liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for detecting nusinersen in rat cerebrospinal fluid (CSF).

Methods: An LC-MS/MS method has been created and verified to detect nusinersen in Sprague-Dawley (SD) rat CSF. The method employed solid-phase extraction for post-extraction analysis and used dT20 as an internal standard. Negative ion multiple reaction monitoring (MRM) mode scanning and the electrospray ionization (ESI) source were used. The method was validated over a concentration range of 5-2000 ng/mL with a Lower Limit of Quantification (LLOQ) of for nusinersen at 5 ng/mL.

Results and conclusions: The method achieves extremely high accuracy and precision, good linearity, high extraction recovery, and provides a useful approach for evaluating the pharmacokinetics of nusinersen in rats.

Keywords
Antisense oligonucleotide drugs; HPLC-MS/MS; cerebrospinal fluid; nusinersen; pharmacokinetics; solid phase extraction.
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