The synthesis of unlabeled and 32P-labeled phosphocitrate and analytical systems for its identification
- Resource Type
- Authors
- John D. Sallis; Gethin Williams
- Source
- Analytical biochemistry. 102(2)
- Subject
- Phosphocitrate
Chromatography
Magnetic Resonance Spectroscopy
biology
Chemistry
Biophysics
Cell Biology
Carbon-13 NMR
Alkaline hydrolysis (body disposal)
Phosphate
Chromatography, Ion Exchange
Biochemistry
Enzyme assay
chemistry.chemical_compound
Triethyl citrate
Isotope Labeling
Proton NMR
biology.protein
Chromatography, Thin Layer
Citrates
Molecular Biology
Phosphorus Radioisotopes
- Language
- ISSN
- 0003-2697
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 [ 32 P]phosphocitrate. Phosphocitrate was characterized by 1 H NMR, 31 P NMR, and 13 C NMR spectroscopy. In addition methods for thin-layer chromatography and enzyme assay are detailed for the detection of phosphocitrate.