{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:48:07Z","timestamp":1787028487137,"version":"build-2736575974"},"reference-count":30,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2022,9,5]],"date-time":"2022-09-05T00:00:00Z","timestamp":1662336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The application of oligonucleotides as drugs for different genetic diseases is increasing rapidly. Since 2016 they are used during spinal muscular atrophy treatment with the use of nusinersen oligonucleotide. The purpose of this study was to improve methods for the analysis of serum samples of patients treated with nusinersen. The results showed that liquid-liquid extraction (with phenol\/chloroform) is insufficient and an additional purification step using solid-phase extraction is necessary. The best results were obtained for microextraction by packed sorbents. Important parameters in the optimization of the method were mainly the type of amine in the mobile phase and the stationary phase. Both influenced the selectivity of metabolite separation and thus their correct identification; while amine type impacted also the intensity of signals. Finally, the highest resolution of separation and the highest peak areas were obtained for N,N-dimethylbutylamine or N,N-diisopropylthylamine with an octadecyl column with a terminal aryl group. Over a dozen of metabolites were successfully identified with the use of methods developed during the study. The 3\u2032 exonucleases and 5\u2032 exonucleases were mainly responsible for nusinersen metabolism, consequently, 3\u2032end shortmers, and 5\u2032end shortmers were observed, as well as metabolites with simultaneous loss of bases at both ends of the sequence. However, some depurination and depyrimidination products were also identified. To the best of our knowledge, this is the first report on nusinersen and its metabolite identification in serum samples by liquid chromatography and mass spectrometry.<\/jats:p>","DOI":"10.3390\/ijms231710166","type":"journal-article","created":{"date-parts":[[2022,9,6]],"date-time":"2022-09-06T05:17:23Z","timestamp":1662441443000},"page":"10166","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Development of the Method for Nusinersen and Its Metabolites Identification in the Serum Samples of Children Treated with Spinraza for Spinal Muscular Atrophy"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4658-7092","authenticated-orcid":false,"given":"Sylwia","family":"Studzi\u0144ska","sequence":"first","affiliation":[{"name":"Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toru\u0144, 7 Gagarin Str., PL-87-100 Toru\u0144, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9405-5066","authenticated-orcid":false,"given":"Maria","family":"Mazurkiewicz-Be\u0142dzi\u0144ska","sequence":"additional","affiliation":[{"name":"Department of Developmental Neurology, Medical University of Gdansk, 7 D\u0119binki Str., PL-80-952 Gda\u0144sk, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bogus\u0142aw","family":"Buszewski","sequence":"additional","affiliation":[{"name":"Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University in Toru\u0144, 7 Gagarin Str., PL-87-100 Toru\u0144, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"100416","DOI":"10.1016\/j.jbc.2021.100416","article-title":"Antisense technology: A review","volume":"296","author":"Crooke","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_2","unstructured":"Institute for Clinical and Economic Review (ICER) (2022, July 25). Spinraza\u00ae and Zolgensma\u00ae for Spinal Muscular Atrophy: Effectiveness and Value. Final Evidence Report. Available online: https:\/\/icerreview.org\/wp-content\/uploads\/2018\/07\/ICER_SMA_Final_Evidence_Report_040319.pdf."},{"key":"ref_3","unstructured":"US Food and Drug Administration (FDA) (2022, July 25). SPINRAZA. 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