Development of a bioanalytical method for an antisense therapeutic using high-resolution mass spectrometry.
Yuchen SunShin-Ichiro NittaKosuke SaitoRyuta HosogaiKeiko NakaiRyoya GodaMasaaki KakehiKazuyuki MurataTakeru YamaguchiTakeshi OkuzonoShinichi YamaneYoshimasa EnokiMitsuhiko KawabataKentaro TakaharaShimpei SatoTokuyuki YoshidaTakao InoueYoshiro SaitoPublished in: Bioanalysis (2020)
Background: Ion-pairing reverse-phase LC coupled with high-resolution mass spectrometry (IP-LC/HRMS) has gained attention in oligonucleotide therapeutic bioanalyses owing to its high sensitivity and selectivity. However, optimization and validation of IP-LC/HRMS-based methods are rare. The objective of this study is the development of a sensitive and reproducible IP-LC/HRMS-based bioanalytical method using clinically approved mipomersen as a model for antisense oligonucleotides. Materials & methods/results: Mipomersen was extracted from rat plasma using Clarity OTX SPE and quantified by IP-LC/HRMS. The calibration range was 0.5-250.0 ng/ml. The developed method met the general regulatory criteria for accuracy, precision, carry-over, selectivity, matrix effect and dilution integrity. Conclusion: A highly sensitive and reliable method for mipomersen measurement with potential antisense oligonucleotide bioanalysis applications has been developed.
Keyphrases
- high resolution mass spectrometry
- liquid chromatography
- mass spectrometry
- ultra high performance liquid chromatography
- gas chromatography
- simultaneous determination
- tandem mass spectrometry
- solid phase extraction
- liquid chromatography tandem mass spectrometry
- nucleic acid
- working memory
- transcription factor
- tyrosine kinase