Facile small scale synthesis of nucleoside 5'-phosphate mixtures.
We present a facile method to phosphorylate small amounts of nucleosides (0.05 mumol) 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.
Jansen
Robert S
RS
Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Rosing
Hilde
H
Schellens
Jan H M
JH
Beijnen
Jos H
JH
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D002021:Buffers; D002625:Chemistry, Organic / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003597:Cytosine Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D006868:Hydrolysis; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D009711:Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D017553:Phosphorus Compounds / Q000737:chemistry
D002021:Buffers; D003597:Cytosine Nucleotides; D007202:Indicators and Reagents; D009711:Nucleotides; D017553:Phosphorus Compounds; D000249:Adenosine Monophosphate; C112125:cladribine monophosphate; C013196:phosphoryl chloride