A Practical Method for Analysis of Triacylglycerol Isomers Using Supercritical Fluid Chromatography

Triacylglycerol (TAG) isomers have been reported to have differing physical and nutritional properties. The analysis of TAG isomers is therefore important for understanding the physical properties of lipids as well as their digestion and absorption. However, methods for the quantitative analysis of TAG regioisomers and enantiomers in vegetable oils and biological samples are still under development. Recently, methods using recycle high-performance liquid chromatography (HPLC) and silver ion column-HPLC have been reported. However the recycle HPLC method requires more than 1 hour, in general, for each sample that is analyzed. Furthermore, existing methods are unable to quantify regioisomers and enantiomers simultaneously. Thus, we aimed to develop a practical method to simultaneously quantify regioisomers and enantiomers of TAG. Three isomers of sn-POO, OPO, and OOP were separated by supercritical fluid chromatography coupled with triple quadrupole mass spectrometer (SFC/MS/MS) using a CHIRALPAK^® IG-U column with acetonitrile and methanol as mobile phase. The separation was completed in 40 min, which is a shorter run time than the conventional techniques published to date. Linear calibration curves with standards were obtained and used to quantify sn-OPO, sn-POO, and sn-OOP in extra virgin olive oil, refined olive oil, palm oil, palm olein, and interesterified palm olein.