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HCC is usually diagnosed by abdominal sonography and serum alpha\u2010fetoprotein (AFP) detection. Modified nucleosides, regarded as indicators for the whole\u2010body turnover of RNAs, are excreted in abnormal amounts in the urine of patients with malignancies and can serve as tumor markers. We analyzed the excretion patterns of urinary nucleosides from 25 HCC patients and 20 healthy volunteers by high\u2010performance liquid chromatography\/electrospray ionization tandem mass spectrometry (HPLC\/ESI\u2010MS\/MS) under optimized conditions. The HPLC\/ESI\u2010MS\/MS approach with selective reaction monitoring (SRM) allowed for the sensitive determination of nucleosides in human urine samples. The mean levels of the urinary nucleosides adenosine, cytidine, and inosine were significantly higher in HCC patients than healthy volunteers (average of 1.78\u2010, 2.26\u2010, and 1.47\u2010fold, respectively). However, the mean levels of urinary 1\u2010methyladenosine, 3\u2010methylcytidine, uridine, and 2\u2032\u2010deoxyguanosine were not significantly different. Combined with the determination of serum AFP levels, the higher levels of urinary adenosine, cytidine, and inosine may be additional diagnosis markers for HCC in Taiwanese patients. 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