Abstract
Rapid and effective separation of nucleotides (NTs) and their derivatives is crucial for studying their physiological functions. In this work, we comprehensively evaluated the separation ability of a zwitterionic hydrophilic monolith, i.e., poly(N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl)ammonium betaine-co–N,N’-methylenebisacrylamide) (poly(SPP-co-MBA)) for NTs analysis, including its selectivity, chemical stability under extremely basic condition and compatibility with hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (HILIC-MS). The poly(SPP-co-MBA) monolith exhibited excellent chemical stability, as evidenced by the low relative standard deviation of retention time (0.16–1.05%) after 4000 consecutive injections over one month under strong alkaline elution condition (pH 10). After optimizing the separation conditions, including buffer pH and concentration, organic solvent content and column temperature, four nucleoside triphosphates, five nucleoside diphosphates and five nucleoside monophosphates were baseline separated within 7 min. Additionally, the mixtures containing one nucleoside and its corresponding mono-, di-, and triphosphates were baseline separated within only 3 min, respectively. It is good HILIC-MS compatibility was also confirmed by the satisfactory peak shape and high response of nine NTs. Overall, the proposed poly(SPP-co-MBA) monolith exhibited good mechanical stability and compatibility of HILIC-MS, making it a promising technique for NTs analysis.
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Funding
This research was funded by the National Natural Science Foundation of China (Grant number: 82104115, 82173781), Guangdong Basic and Applied Basic Research Foundation (Grant number: 2020A1515110867), Science and Technology Innovation Project of Guangdong Drug Administration (Grant number: 2022ZDB04), and China Postdoctoral Science Foundation (2022M721355), Key Field Projects of Guangdong Universities (Intelligent Manufacturing) (2020ZDZX2057).
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Experiment, methodology, writing—original draft preparation, LL; Experiment, methodology, partial of writing—original draft preparation, MZ; Experiment, methodology, partial of writing—original draft preparation, CL; Partial of the experiments, writing—review and editing, JQ; Writing—review and editing, project administration, DX; Conceptualization, supervision, writing—review and editing, project administration, funding acquisition, ZJ. All authors have read and agreed to the published version of the manuscript.
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Lai, L., Zhang, M., Liu, C. et al. A comprehensive evaluation of a polymeric zwitterionic hydrophilic monolith for nucleotide separation. ANAL. SCI. 40, 85–91 (2024). https://doi.org/10.1007/s44211-023-00430-5
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DOI: https://doi.org/10.1007/s44211-023-00430-5