title: Facile small scale synthesis of nucleoside 5'-phosphate mixtures.
abstract: 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.
authors.affiliation: Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Amsterdam, The Netherlands.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Jansen
authors.forename: Hilde
authors.initials: H
authors.lastname: Rosing
authors.forename: Jan H M
authors.initials: JH
authors.lastname: Schellens
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
mesh_terms: 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
chemical_list: D002021:Buffers; D003597:Cytosine Nucleotides; D007202:Indicators and Reagents; D009711:Nucleotides; D017553:Phosphorus Compounds; D000249:Adenosine Monophosphate; C112125:cladribine monophosphate; C013196:phosphoryl chloride