Identification and quantification of triacylglycerols in human milk fat using ultra-performance convergence chromatography and quadrupole time-of-flight mass spectrometery with supercritical carbon dioxide as a mobile phase

An ultra-performance convergence chromatography (UPC2) method coupled with quadrupole time-of-flight mass spectrometry (Q-TOF-MS), was developed for the determination of complicated triacylglycerols (TAGs) in human milk fat. The use of supercritical carbon dioxide as the mobile phase improved the chromatographic separation of the TAGs significantly. By optimizing the scan modes of Q-TOF-MS and selecting parent ions for MS/MS ionization, the fragment ions of each TAG including TAG isomers with overlapping retention time were adequately resolved for identification and quantification. A total of 95 different TAGs were identified in the human milk samples from Chinese mother volunteers, with O-P-L representing the main TAG, followed by O-P-O and O-L-L. In addition, the compositions and contents of TAGs in different fats and oils were successfully measured. The developed method can serve as an advanced and reliable analytical tool for the determination of complicated TAGs in various biological samples.