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- HPLC analysis: Elution solvent 1:9 methanol/water, flow 2 mL/min, UV detection at 263 nm, column: Waters Resolve C18, 10 um, 8mm × 100mm, Retention times: 1 6.5 min, 8-bromoguanine 8.4 min, 2 28.0 min. 1H NMR analysis: Spectra of the product were obtained in D2O. The presence of 1 in the sample could be determined by appearance of resonances at 7.904 and 6.410 ppm corresponding to the C-H8 and H-l′ protons, respectively, of 1