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Characterization of Oligonucleotide Metabolism In Vivo via Liquid Chromatography/Electrospray Tandem Mass Spectrometry with a Quadrupole Ion Trap Mass Spectrometer
Author(s) -
Griffey Richard H.,
Greig Michael J.,
Gaus Hans J.,
Liu Kenneth,
Monteith David,
Winniman Michael,
Cummins Lendell L.
Publication year - 1997
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/(sici)1096-9888(199703)32:3<305::aid-jms482>3.0.co;2-r
Subject(s) - chemistry , mass spectrometry , chromatography , electrospray , top down proteomics , ion trap , selected reaction monitoring , hybrid mass spectrometer , tandem mass spectrometry , sample preparation in mass spectrometry , quadrupole mass analyzer , protein mass spectrometry , triple quadrupole mass spectrometer , quadrupole ion trap , liquid chromatography–mass spectrometry , analytical chemistry (journal) , electrospray ionization
The pattern of nuclease degradation observed for an antisense phosphorothioate oligonucleotide in pig kidney was determined using liquid chromatography/electrospray mass spectrometry (LC/ESI‐MS) and LC/ESI‐MS/MS with a quadrupole ion trap mass spectrometer. Metabolites were separated by length using reversed‐phase high‐performance liquid chromatography with aqueous hexafluoropropan‐2‐ol–triethylamine and a methanol gradient. The individual masses of metabolites in each LC peak were determined via deconvolution and converted into potential nucleotide compositions. The nucleotide composition was used to locate metabolites within the known oligomer sequence. The identity of metabolites was confirmed using on‐line LC/MS/MS to generate fragment ions suitable for sequence verification. A limited number of shorter oligonucleotide fragments were observed, suggesting that metabolism in vivo may be sequence dependent. © 1997 by John Wiley & Sons, Ltd.