High-performance liquid chromatography (HPLC) and laser-desorption Fourier-transform mass spectrometry (LD FTMS) have been applied for direct measurements of radiation-induced products of nucleic acid constituents containing thymidine. Laser desorption FTMS could be used for the direct detection (neither hydrolyzed nor derivatized) of X ray-induced decomposition products of aqueous thymidine monophosphate. After these initial experiments, a variety of hydrogenated and hydroxylated thymine standards were acquired and examined by FTMS to assist in the identification of unknown radiation-induced decomposition products of thymine-containing nucleotides and dinucleotides. To extend these studies to dinucleotides, the radiation-induced products generated by the gamma radiolysis of thymidylyl (3′→5′) thymidine (TpT) were isolated by reverse-phase HPLC and identified by LD FTMS. Thymine and thymidine 3′-monophosphate were observed as the major products in this case. Several of the minor products of the HPLC profile were pooled in a single fraction and characterized simultaneously by LD FTMS. The resulting mass spectra indicated the presence of hydroxy-5,6-dihydrothymidine monophosphate, 5,6-dihydrothymidine monophosphate and thymidine monophosphate, thymine glycol, hydroxy-5,6-dihydrothymine, 5-hydroxy-methyluracil and 5,6-dihydrothymine. The combination of HPLC purification and LD FTMS structural characterization provides a useful tool for the direct measurement of radiation-induced products of nucleotides and dinucleotides.
Characterization of Radiation-Induced Products of Thymidine 3′-Monophosphate and Thymidylyl (3′→5′) Thymidine by High-Performance Liquid Chromatography and Laser-Desorption Fourier-Transform Mass Spectrometry
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H. Yoshida, R. L. Hettich; Characterization of Radiation-Induced Products of Thymidine 3′-Monophosphate and Thymidylyl (3′→5′) Thymidine by High-Performance Liquid Chromatography and Laser-Desorption Fourier-Transform Mass Spectrometry. Radiat Res 1 September 1994; 139 (3): 271–279. doi: https://doi.org/10.2307/3578824
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