Griffey R H, Greig M J, Gaus H J, Liu K, Monteith D, Winniman M, Cummins L L
Isis Pharmaceuticals, Carlsbad, CA 92008, USA.
J Mass Spectrom. 1997 Mar;32(3):305-13. doi: 10.1002/(SICI)1096-9888(199703)32:3<305::AID-JMS482>3.0.CO;2-R.
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.
使用液相色谱/电喷雾质谱（LC/ESI-MS）和配备四极杆离子阱质谱仪的LC/ESI-MS/MS，确定了在猪肾中观察到的反义硫代磷酸酯寡核苷酸的核酸酶降解模式。代谢物通过使用含六氟异丙醇-三乙胺水溶液和甲醇梯度的反相高效液相色谱按长度进行分离。通过去卷积确定每个LC峰中代谢物的个体质量，并将其转化为潜在的核苷酸组成。核苷酸组成用于在已知的寡聚物序列中定位代谢物。使用在线LC/MS/MS生成适合序列验证的碎片离子，从而确认代谢物的身份。观察到数量有限的较短寡核苷酸片段，表明体内代谢可能依赖于序列。
