Abstract
A method for the synthesis of phosphocitrate is described using 2-cyanoethyl phosphate to phosphorylate triethyl citrate. Following alkaline hydrolysis of the coupled intermediate, phosphocitrate was purified by ion-exchange chromatography on an AG 1-X8 (HCO 3 −) column. The method was also used to prepare [ 32P]phosphocitrate. Phosphocitrate was characterized by 1H NMR, 31P NMR, and 13C NMR spectroscopy. In addition methods for thin-layer chromatography and enzyme assay are detailed for the detection of phosphocitrate.
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