Development of a highly sensitive method for the quantitative analysis of modified nucleosides using UHPLC-UniSpray-MS/MS
Development of a highly sensitive method for the quantitative analysis of modified nucleosides using UHPLC-UniSpray-MS/MS
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开发一种使用 UHPLC-UniSpray-MS/MS 定量分析修饰核苷的高灵敏度方法
DOI：
10.1016/j.jpba.2021.113943
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发表时间：
2021-02-15
影响因子：
3.4
通讯作者：
Tsujikawa, Kazutake
中科院分区：
文献类型：
--
作者：
Kogaki, Takahiro;Ohshio, Ikumi;Tsujikawa, Kazutake
有超过150种自然发生的修饰核苷，它们被认为参与了各种生物过程。近年来，建立了一种高效液相色谱-电喷雾电离-串联质谱（UHPLC-ESI-MS/MS）技术来测定低水平的修饰核苷。对修饰核苷的全面分析将有助于更好地理解细胞内核糖核酸修饰，但这种分析需要高灵敏度的测量。为此，我们利用新开发的离子源UniSpray建立了一种高灵敏度和定量的方法。质谱仪与UniSpray源在正离子模式下使用。采用uhplc - unisprayy -MS/MS方法，在15 min内分离检测4种主要核苷、42种修饰核苷和dG15N5（内标）。UniSpray方法为所有分析的核苷提供了良好的相关系数（>0.99），并且对46种核苷中的35种具有较宽的线性范围。准确度和精密度值均满足
There are more than 150 types of naturally occurring modified nucleosides, which are believed to be involved in various biological processes. Recently, an ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) technique has been developed to measure low levels of modified nucleosides. A comprehensive analysis of modified nucleosides will lead to a better understanding of intracellular ribonucleic acid modification, but this analysis requires high-sensitivity measurements. In this perspective, we established a highly sensitive and quantitative method using the newly developed ion source, UniSpray. A mass spectrometer was used with a UniSpray source in positive ion mode. Our UHPLC-UniSpray-MS/MS methodology separated and detected the four major nucleosides, 42 modified nucleosides, and dG15N5 (internal standard) in 15 min. The UniSpray method provided good correlation coefficients (>0.99) for all analyzed nucleosides, and a wide range of linearity for 35 of the 46 nucleosides. Additionally, the accuracy and precision values satisfied the criteria of