FACILE SMALL SCALE SYNTHESIS OF NUCLEOSIDE 5 '-PHOSPHATE MIXTURES

作者：Jansen Robert S^*; Rosing Hilde; Schellens Jan H M; Beijnen Jos H

来源：Nucleosides Nucleotides & Nucleic Acids, 2010, 29(1): 14-26.

DOI：10.1080/15257770903451546

• 5 '-Triphosphates
• nucleoside analogs
• bioanalysis and mass spectrometry

We present a facile method to phosphorylate small amounts of nucleosides (0.05 mu mol) into mixtures of their 5'-mono-, di-, and triphosphates in a one-pot reaction. The nucleosides were first converted into their dichlorophosphates using a large excess (15-18 equivalents) of phosphorous oxychloride in trimethylphosphate. The large excess resulted in good dichlorophosphate yields (46-76%) for the four nucleosides tested. Upon the addition of tributylammonium-phosphate with additional tributylamine (20 equivalents both), the dichlorophosphate was converted into a mixture containing equal amounts of the mono-, di-, and triphosphate. The presented method was successfully applied to synthesize mixtures of stable isotope labeled nucleotides, which can be used as internal standards in quantitative mass spectrometric assays.

• 出版日期2010