Isolation and identification of urinary nucleosides. Applications of high-performance liquid chromatographic methods to the synthesis of 5'-deoxyxanthosine and the simultaneous determination of 5,6-dihydrouridine and pseudouridine

• Published in: Journal of chromatography Vol. 515; p. 537
• Main Authors: Nakano, K, Yasaka, T, Schram, K H, Reimer, M L, McClure, T D, Nakao, T, Yamamoto, H
• Format: Journal Article
• Language: English
• Published: Netherlands 31.08.1990
• Subjects: Chromatography, High Pressure Liquid - methods; Creatine - urine; Deoxyribonucleosides; Female; Gas Chromatography-Mass Spectrometry; Gastrointestinal Neoplasms - urine; Humans; Male; Nucleosides - isolation & purification; Nucleosides - urine; Pseudouridine - urine; Ribonucleosides - biosynthesis; Uridine - analogs & derivatives; Uridine - urine
• DOI: 10.1016/S0021-9673(01)89352-3

Modified nucleosides from pooled normal human urine were extracted using a boronate affinity gel column and fractionated by reversed-phase high-performance liquid chromatography (RP-HPLC). The major constituents in each of the 30 RP-HPLC fractions were determined by gas chromatography-mass spectrometry of the trimethylsilyl derivatives of the fractions. The same RP-HPLC method was used in the synthesis of 5'-deoxyxanthosine from authentic 5'-deoxyadenosine. In addition, the simultaneous determination of urinary 5,6-dihydrouridine (D) and pseudouridine (psi) was carried out by RP-HPLC using two ODS columns in series. The level of D in pooled normal urine was 4.87 nmol/mumols creatinine. The RP-HPLC method was applied to the measurement of D and psi levels in urines collected before and after surgery from four patients with gastrointestinal cancer. A large decline in both nucleoside levels in urines after surgery was observed in three of the four cancer patients.