Total Phospholipids in Edible Oils by In-Vial Solvent Extraction Coupled with FTIR Analysis

A simple procedure for the determination of total phospholipids (TPL) in edible oils was developed by combining a single-step, in situ methanol/acetonitrile (MeOH/ACN) extraction of the oil sample followed by Fourier transform infrared (FTIR) spectroscopic analysis of the extract. Spectral analysis of extracts in a 25 μm CaF₂ cell obtained using 1:1 MeOH/ACN added to oil in a 2:1 ratio indicated that measurements made using only the asymmetric phosphate diester PO₂^- stretching band at 1243 cm^-1 in second-derivative spectra were sufficient for the accurate measurement of TPL with minimal coextracted triglyceride interferences being encountered. FTIR calibration spectra were devised using only phosphatidylcholine (PC) as a representative phospholipid standard, covering a range of 0-50000 μg/g TPL and spiked into 1:1 MeOH/ACN, capable of tracking the added PC with an SD of <200 μg/g. The FTIR method was initially validated using model PC-spiked degummed canola oil and subsequently with commercial crude and refined soy and rapeseed oils as well as a lecithin tablet with the FTIR TPL predictions compared to those of the AOCS Ca 12-55 molybdenate method. The FTIR method tracked the AOCS results well, being somewhat more reproducible than the reference method (±3.2 vs ±4.9%), which limited its accuracy relative to the AOCS reference procedure (±2.2%). The simple in-vial solvent extraction procedure, followed by FTIR analysis of the extract, is a simple, efficient, and rapid procedure that is also amenable to automation using an autosampler-equipped FTIR if multiple samples are to be analyzed.