1. Development of a highly sensitive method for the quantitative analysis of modified nucleosides using UHPLC-UniSpray-MS/MS
- Author
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Aya Harada Takeda, Toshiya Morie, Kentaro Jingushi, Masatatsu Yamamoto, Souta Iyama, Ikumi Ohshio, Hiroaki Hase, Shintarou Fujii, Masami Sato, Zenzaburo Tozuka, Yuko Ueda, Kazuhiro Ueda, Kazutake Tsujikawa, Shohei Sato, Kentaro Minami, Takahiro Kogaki, Daisuke Saigusa, Tatsuhiko Furukawa, Toshiyuki Nagata, Masaya Aoki, Kaori Kitae, Kohichi Kawahara, and Hasumi Ura more...
- Subjects
Accuracy and precision ,Spectrometry, Mass, Electrospray Ionization ,Lung Neoplasms ,Clinical Biochemistry ,Pharmaceutical Science ,Mass spectrometry ,01 natural sciences ,Uhplc ms ms ,Modified nucleosides ,Analytical Chemistry ,Mice ,Tandem Mass Spectrometry ,Carcinoma, Non-Small-Cell Lung ,Drug Discovery ,Animals ,Spectroscopy ,Chromatography, High Pressure Liquid ,Chromatography ,010405 organic chemistry ,Chemistry ,010401 analytical chemistry ,Nucleosides ,0104 chemical sciences ,Highly sensitive ,Non small cell ,Quantitative analysis (chemistry) - Abstract
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 more...
- Published
- 2020