Polyglycerol esters (PGEs), produced by esterification of fatty acids on polyglycerols, were analysed by High Resolution Mass Spectrometry (HRMS), HPLC-MS and U-HPLC-MS. A structural study of PGEs in 4 samples synthesised by the Gattefossé company was carried out using an elemental analysis of HRMS spectra and modelling of all probable isomers and cyclic structures. The results were used to construct a structural database of all species present in the 4 samples. After an assessment of the selectivity of 5 reversed phase columns: Aeris Widepore XB-C8, 3.6 μm, 2.1 × 150 mm (Phenomenex), Acquity CSH C18 1.7 μm 2.1 × 50 mm, Acquity CSH Phenyl-Hexyl 1.7 μm 2.1 × 50 mm, Acquity CSH Fluoro-Phenyl 1.7 μm 2.1 × 50 mm (Waters Co.) and Kinetex F5 1.7 μm 2.1 × 100 mm (Phenomenex), HPLC-MS and U-HPLC-MS analyses were performed on an Aeris Widepore XB-C8 (Phenomenex) column (HPLC) and Acquity CSH Fluoro-Phenyl (Waters) column (U-HPLC) with aqueous formic acid /acetonitrile in gradient mode. The separation was optimised with 10 min (HPLC) and 5 min (U-HPLC) of gradient. The detection, performed on a QDA detector (Waters), produced extracted ion chromatograms (XICs) based on all adducts identified in the HRMS analysis. HPLC and U-HPLC analyses showed the different mono-and di-ester species and provided relative quantification of all identified constituents. The combined analyses of the HRMS, HPLC-MS and U-HPLC-MS results were used to compare the different PGE batches and quantify the molecular constituents according to their relative abundance, for these complex mixtures. With HPLC and U-HPLC analyses, using 2 different gradient times and 2 different selectivity columns, and comparing the retention factors and log P of the different species, it was possible to link structural identification and relative quantification of all PGEs identified in the samples.