Duan Zheng, Wang Chen, Wang Yinsheng
Department of Chemistry, University of California Riverside, Riverside, California 92521, United States.
J Am Soc Mass Spectrom. 2025 Oct 1;36(10):2017-2021. doi: 10.1021/jasms.5c00245. Epub 2025 Aug 27.
Liquid chromatography-tandem mass spectrometry has been widely used to quantify modified nucleosides enzymatically released from RNA and DNA. Compared to MS/MS, MS offers higher specificity for analytes in complex sample matrices because coeluting chemical interferences can be more effectively filtered out. To date, the detection of uracil- and thymine-containing nucleosides and their modified derivatives still suffers from poor sensitivity, owing, in part, to their relatively low proton affinities. In this study, we explored various settings of values in the MS analysis of unmodified and chemically modified uridine and thymidine nucleosides on a linear ion trap mass spectrometer. Our results showed that increasing the value led to markedly improved sensitivities for detecting some of these nucleosides using MS. Our work suggests that optimizing the value constitutes a useful approach for the highly sensitive detection of modified nucleosides and other types of analytes by MS.
液相色谱-串联质谱法已被广泛用于定量从RNA和DNA中酶促释放的修饰核苷。与串联质谱（MS/MS）相比，质谱（MS）对复杂样品基质中的分析物具有更高的特异性，因为共洗脱的化学干扰物可以更有效地被滤除。迄今为止，含尿嘧啶和胸腺嘧啶的核苷及其修饰衍生物的检测灵敏度仍然较低，部分原因是它们相对较低的质子亲和力。在本研究中，我们在线性离子阱质谱仪上探索了未修饰和化学修饰的尿苷和胸苷核苷的质谱分析中各种碰撞能量（CE）值设置。我们的结果表明，增加CE值会显著提高使用质谱检测其中一些核苷的灵敏度。我们的工作表明，优化CE值是通过质谱对修饰核苷和其他类型分析物进行高灵敏度检测的一种有用方法。
