31P NMR Quantification of Phospholipids and Lysophospholipids in Food Emulsions

For food emulsions containing enzymatically modified egg yolk, the conventional Folch extraction does not fully recover the polar lysophospholipids. This can be overcome by repeated methanol extractions. After solvent evaporation, the extracted (lyso)phospholipids are solubilized into mixed micelles with cholate as a detergent. The solubilized (lyso)phospholipids can be accurately quantified by 31P NMR with recoveries ranging between 96% and 108%. Detection at a high (16.4 T) relative to a mainstream (9.4 T) magnetic field strength did not offer a significant advantage since the slow molecular tumbling of the mixed micelles increased line widths. This was due to field-strength-dependent chemical shift anisotropy relaxation. Method precision is similar at 9.4 and 16.4 T, with within-laboratory reproducibilities of 7-22% and 12-25%, respectively. The method can be implemented as a routine analytical procedure at 9.4 T (400 MHz NMR spectrometer), and the limits of detection and quantification are adequate for the verification of the standard of identity of a mayonnaise prepared with enzymatically modified egg yolk.