Purification Methods
The Et3N salts are purified by dissolving 4g in 500mL of H2O (adding a drop or two of Et3N if they do not dissolve) and chromatograph by applying to a column (3 x 30cm) of DEAE-Sephadex A-25 and eluting with 1.4L of a linear gradient of Et3NH.HCO3 from 0.05 to 0.55M, pH 7.8 and 4o. The product elutes between 0.20.3M Et3N.HCO3. The pooled fractions are evaporated, and the residue is twice taken up in EtOH and evaporated to dryness to remove the last traces of Et3NH.HCO3. 31P NMR: P is a doublet at -40.81 and -40.33, and P at 7.02ppm, J, 32.96Hz. [Sheu et al. Biochemistry 18 5548 1979.]
