Development of an improved online comprehensive hydrophilic interaction chromatography × reversed‐phase ultra‐high‐pressure liquid chromatography platform for complex multiclass polyphenolic sample analysis

Sommella, Eduardo; Ismail, Omar H.; Pagano, Francesco; Pepe, Giacomo; Ostacolo, Carmine; Mazzoccanti, Giulia; Russo, Mariateresa; Novellino, Ettore; Gasparrini, Francesco; Campiglia, Pietro

Development of an improved online comprehensive hydrophilic interaction chromatography × reversed‐phase ultra‐high‐pressure liquid chromatography platform for complex multiclass polyphenolic sample analysis

2017

In this study, an improved online comprehensive two‐dimensional liquid chromatography platform coupled to tandem mass spectrometry was developed for the analysis of complex polyphenolic samples. A narrowbore hydrophilic interaction chromatography column (150 × 2.0 mm, 3.0 μm, cross‐linked diol) was employed in the first dimension, while a reversed‐phase column based on monodisperse sub‐2 μm fully porous particles (50 × 3.0 mm, 1.9 μm d.p.) with high surface area (410 m²/g) was employed in the second dimension. The combination of a trapping column modulation interface with the high retentive fully porous monodisperse reversed‐phase column in the second dimension resulted in higher peak capacity values (1146 versus 867), increased sensitivity, sharper and more symmetrical peaks in comparison with a conventional loop‐based method, with the same analysis time (70 min). The system was challenged against a complex polyphenolic extract of a typical Italian apple cultivar, enabling the simultaneous separation of multiple polyphenolic classes, including oligomeric procyanidins, up to degree of polymerization of 10. Hyphenation with an ion trap time‐of‐flight mass spectrometer led to the tentative identification of 121 analytes, showing how this platform could be a powerful analytical tool for the accurate profiling of complex polyphenolic samples.

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